A previous study of 26 Japanese, VHL families was extended to 41 additional families. Germline mutations were detected in 55 of 77 Japanese families using SSCP, direct sequencing, and Southern Blot analysis of peripheral blood samples. The L198Gfs*57 (c.592_593del) germline mutation was found in a VHL type 1 patient presenting with CNS hemangioblastoma, retinal angioma, pancreatic cyst or tumor (unspecified) and renal cell carcinoma (family ID 67).

A previous study of 26 Japanese, VHL families was extended to 41 additional families. Germline mutations were detected in 55 of 77 Japanese families using SSCP, direct sequencing, and Southern Blot analysis of peripheral blood samples. The L178P (c.533T>C) germline mutation was found in a VHL type 1 patient presenting with CNS hemangioblastoma, retinal angioma and renal cell carcinoma (family ID 34). This family was previously reported in PMID 8634692.

A previous study of 26 Japanese, VHL families was extended to 41 additional families. Germline mutations were detected in 55 of 77 Japanese families using SSCP, direct sequencing, and Southern Blot analysis of peripheral blood samples. The E173Rfs*29 (c.516del) germline mutation was found in a VHL type 1 patient presenting with CNS hemangioblastoma, pancreatic cyst or tumor (unspecified) and renal cell carcinoma (family ID 105).

Thirty-five unrelated patients suspected of having von Hippel-Lindau disease were analyzed and 32 VHL gene variants were found in 35 patients using PCR and SSCP by peripheral blood leukocytes. An association of clear cell renal carcinoma development with a relatively high loss of structural stability in VHL protein missense mutations was discovered. A mutation, F76del(c.227_229del) (noted as g.941_943delTCT in the paper originally) was found in a patient with cerebellar hemangioblastomas, renal cyst, and pancreatic cyst, the pateints age on onset was 31 years old. No family history was mentioned, other than a note of no familial antecedents with the disease.

Thirty-five unrelated patients suspected of having von Hippel-Lindau disease were analyzed and 32 VHL gene variants were found in 35 patients using PCR and SSCP by peripheral blood leukocytes. An association of clear cell renal carcinoma development with a relatively high loss of structural stability in VHL protein missense mutations was discovered. A mutation, P154= (c.462A>G) (noted as a splice-site mutation: g.8665A>G, in the paper originally) was found in a patient with retinal hemangioblastomas, cerebellar hemangioblastomas, and pancreatic cyst, the pateints age on onset was 13 years old. The family history also included incidences of clear cell renal cell carcinoma and renal cysts.

The authors identify a new VHL cryptic exon (termed E1') deep in intron 1 that is naturally expressed in many tissues. Mutations in E1' are identified in 7 families with erythrocytosis and in 1 large family with typical VHL disease but without any alteration in the other VHL exons. Whole blood was isolated from patients and sequenced using sanger sequencing and whole-genome sequencing performed at the clinically accredited Molecular Diagnostics Laboratory at the John Radcliffe Hospital using the Hi-Sequation 4000 platform (Illumina Inc., San Diego, CA) in high-throughput mode. As well, paired-end sequencing was carried out in a single lane on the HiSeq 2500 system (Illumina).In this study, the authors show that the mutations in E1' induced dysregulation of VHL splicing with excessive retention of E1' and were associated with a downregulation of VHL protein expression. Three patients were found with both the c.598C>T germline VHL mutation and the c.340+770T>C mutation resulting in p.Ser179Pro mutation on potential X1 protein encoded by E1' exon. In family 2, II.1 was the only person who had both of these mutations. In family 3, brothers II.1 and II.2 had these two mutations. Biological material from the parents of these three children were not available but chuvash core haplotype analysis demonstrated that all patients were compound heterozygous with one mutation being inherited from each parent. All three cases described above had erythrocytosis.

The authors identify a new VHL cryptic exon (termed E1') deep in intron 1 that is naturally expressed in many tissues. Mutations in E1' are identified in 7 families with erythrocytosis and in 1 large family with typical VHL disease but without any alteration in the other VHL exons. In this study, the authors show that the mutations in E1' induced a dysregulation of VHL splicing with excessive retention of E1' and were associated with a downregulation of VHL protein expression. Patient II.1 of family F4, was found to have two germline mutations in the VHL gene: R200W (c.598C>T) and c.340+694_340+711dup. This patient was diagnosed with erythrocytosis, deep vein thrombosis, intracerebral hemorrhage at age 7 (DOB: 1990). This patient's asymptomatic mother was also genetically tested and found to have the germline c.340+694_340+711dup variant of the VHL gene. Her father was not tested but is assumed to carry the R200W (c.598C>T) germline variant in VHL.

The authors identify a new VHL cryptic exon (termed E1') deep in intron 1 that is naturally expressed in many tissues. Mutations in E1' are identified in 7 families with erythrocytosis and in 1 large family with typical VHL disease but without any alteration in the other VHL exons. In this study, the authors show that the mutations in E1' induced a dysregulation of VHL splicing with excessive retention of E1' and were associated with a downregulation of VHL protein expression. Patient II.1 (family F9) was found to have the germline variant D143= (c.429C>T). The patient was diagnosed with erythrocytosis at 5 months old (DOB: 1997). This variant is homozygous in the patient. This patient was also found to have the HB Sitia germline variant. Both asymptomatic parents were genetically sequenced and found to be carriers of the variant. Patient II.1 (family F10) was also found to have this variant. This patient was diagnosed with erythrocytosis (Polycythemia) and splenomegaly at 4.5 years old (DOB: 2002). This patient was also found to have the Beta Thalassemia germline variant. This patient’s asymptomatic patients were both found to be carriers of the same variant in VHL. Their other child (II.2) also carried the gene. Patient II.1 was the only one who was homozygous for the variant.

Germline mutations in 45 unrelated Japanese VHL patients were determined from blood samples using PCR-SSCP or Southern blot analyses. The authors reported a nucleotide deletion (i.e. 452delG) which coincides with legacy variant nomenclature (AF010238.1) and was mapped to c.239del (NM_000551.3). PCR-SSCP or Southern blot analyses of the DNA from family members showed that mutations were segregated with disease phenotype in VHL families, although this data was not shown. Data on the presence of retinal angioma, CNS hemangioblastoma (Hb) renal cell carcinoma (RCC), and pheochromocytoma was available. Case 22 had p.Ser80Ilelfs*79 (c.239del) mutation. One member was affected with retinal capillary hemangioma and hemangioblastoma. Other demographics of the members were not described.

Germline mutations in 45 unrelated Japanese VHL patients were determined from blood samples using PCR-SSCP or Southern blot analyses. The authors reported a nucleotide deletion (i.e. 445A>C) which coincides with legacy variant nomenclature (AF010238.1) and was mapped to c.232A>C (NM_000551.3). PCR-SSCP or Southern blot analyses of the DNA from family members showed that mutations were segregated with disease phenotype in VHL families, although this data was not shown. Data on the presence of retinal angioma, CNS Hb, RCC, and Pheo was available. Case 1 had p.Asn78His mutation and one affected member with retinal angioma and hemangioblastoma.

The data of 18 patients with solid renal tumours associated with VHL disease were analyzed. Seventeen patients had a VHL family history, the only patient that had no family history of VHL was patient 12. The patients came from 12 Chinese families and 5 were from the same family. Extrarenal manifestations of VHL disease were detected in 17 patients. Of 18 patients, 15 (including 6 asymptomatic) volunteered for DNA testing as previously described. Patient 16 had genetic testing and was found to have a germline mutation c.286C>T (Gln96*). This missense mutation is noted as c.499C>T in the paper due to a difference in the VHL reference sequence notation used. Patient 16 was a 50-year-old female with a medical history of solid renal tumour and CNS hemangioblastoma.

The data of 18 patients with solid renal tumours associated with VHL disease were analyzed. All patients except for Patient 12 came from families with a definite VHL family history (the paper does not provide further details on this.) The patients came from 12 Chinese families and 5 were from the same family. Extrarenal manifestations of VHL disease were detected in 17 patients. Of 18 patients, 15 (including 6 asymptomatic) volunteered for DNA testing as previously described. Patients 6 and 7 had genetic testing and were found to have the same germline frameshift mutation c.447del (p.Asn150Ilefs*)(noted as c.661delA in the paper due to a difference in the VHL reference sequence notation used.). It is unclear if these patients are related. Patient 7 had family study DNA test as their reason for treatment. Patient 6 was a 50-year-old female with a medical history of synchronous bilateral solid renal tumors some which were further characterized as clear cell RCC, multiple renal cysts, pancreatic cyst. Patient 7 was a 48-year-old male with a medical history of solid renal tumours some which were further characterized as clear cell RCC, multiple renal cysts, CNS hemangioblastoma , pancreatic cyts. AMCG evidence codes: ’PP4’ because the patients phenotype is highly specific for a mutation in the VHL gene.

Germline mutational data on VHL was gathered from various countries to assess variation in germline mutations that produce disease within different populations. Families were grouped according to three possible outcomes of disease: renal cell carcinoma without pheochromocytoma, renal cell carcinoma with pheochromocytoma, or pheochromocytoma alone. SSCP and denaturing gradient gel electrophoresis identified mutations in 300/469 (63%) of families affected with VHL. This germline familial VHL mutation (C162Y (c.485G>A)) was taken from the Department of Uroglogy, Yokohama database, with an unknown number of members belonging to Family ID 11YO. It was reported in the paper as c.698G>A due to a difference in transcript reference. As reported in Table 3, this mutation is found in VHL families without pheochromocytoma, and with renal cell carcinoma.

Germline mutational data on VHL was gathered from various countries to assess variation in germline mutations that produce disease within different populations. Families were grouped according to three possible outcomes of disease: renal cell carcinoma without pheochromocytoma, renal cell carcinoma with pheochromocytoma, or pheochromocytoma alone. SSCP and denaturing gradient gel electrophoresis identified mutations in 300/469 (63%) of families affected with VHL. This germline familial VHL mutation (L89P (c.266T>C)) was taken from multiple databases including: the Cambridge, Department of Pathology database and the Munich, university of Freiburg and Institute of Pathology database, with an unknown number of members belonging to Family IDs 8CE, 59CA, 21CE. It was reported in the paper as c.479T>C due to a difference in transcript reference. As reported in Table 3, this mutation is found in VHL families without pheochromocytoma, and with renal cell carcinoma.

Germline mutational data on VHL was gathered from various countries to assess variation in germline mutations that produce disease within different populations. Families were grouped according to three possible outcomes of disease: renal cell carcinoma without pheochromocytoma, renal cell carcinoma with pheochromocytoma, or pheochromocytoma alone. SSCP and denaturing gradient gel electrophoresis identified mutations in 300/469 (63%) of families affected with VHL. This germline familial VHL mutation (G93D (c.278G>A)) was taken from the National Cancer Institute database, with an unknown number of members belonging to Family ID 2547NCI. It was reported in the paper as c.491G>A due to a difference in transcript reference. As reported in Table 4, this mutation is found in VHL families with pheochromocytoma. Other phenotypes are likely, including renal cell carcinoma, however not detailed here. Later publications reference this cohort as being reported in PMID: 7728151.

Germline mutational data on VHL was gathered from various countries to assess variation in germline mutations that produce disease within different populations. Families were grouped according to three possible outcomes of disease: renal cell carcinoma without pheochromocytoma, renal cell carcinoma with pheochromocytoma, or pheochromocytoma alone. SSCP and denaturing gradient gel electrophoresis identified mutations in 300/469 (63%) of families affected with VHL. This germline familial VHL mutation (G57fs (c.167_168insA)) was taken from the Cambridge, Department of Pathology database, with an unknown number of members belonging to Family ID 110CA. It was reported in the paper as c.380insA due to a difference in transcript reference. As reported in Table 3, this mutation is found in VHL family without pheochromocytoma, and with renal cell carcinoma.

Germline mutational data on VHL was gathered from various countries to assess variation in germline mutations that produce disease within different populations. Families were grouped according to three possible outcomes of disease: renal cell carcinoma without pheochromocytoma, renal cell carcinoma with pheochromocytoma, or pheochromocytoma alone. SSCP and denaturing gradient gel electrophoresis identified mutations in 300/469 (63%) of families affected with VHL. This germline familial VHL mutation (S111fs (c.331delA)) was taken from the National Cancer Institute database, with an unknown number of members belonging to Family ID 3669NCI. It was reported in the paper as c.544delA due to a difference in transcript reference. As reported in Table 3, this mutation is found in VHL families without pheochromocytoma, and with renal cell carcinoma.

Germline mutational data on VHL was gathered from various countries to assess variation in germline mutations that produce disease within different populations. Families were grouped according to three possible outcomes of disease: renal cell carcinoma without pheochromocytoma, renal cell carcinoma with pheochromocytoma, or pheochromocytoma alone. SSCP and denaturing gradient gel electrophoresis identified mutations in 300/469 (63%) of families affected with VHL. This germline familial VHL mutation (S80N (c.239G>A)) was taken from the Munich, university of Freiburg and Institute of Pathology database, with an unknown number of members belonging to Family ID 39CE. It was reported in the paper as c.452G>A due to a difference in transcript reference. As reported in Table 3, this mutation is found in VHL families without pheochromocytoma, and with renal cell carcinoma.

GroupID/ KindredID: 102CA

DiseaseAssertion: VHL

GroupSize: N/A

GroupNumberMales: N/A

GroupNumberFemales: N/A

GroupEthnicity: N/A

GroupAgeRange: N/A

GroupInfo: This germline familial VHL mutation was taken from the Cambridge, Department of Pathology database, with an unknown number of members belonging to Family ID 102CA. Table 3 lists all ID and mutations associated with individuals possessing renal carcinoma, without pheochromocytoma.

GroupPresentingHPOs: HP:0005548 (renal cell carcinoma)

GroupHPOFreeText: Described as a VHL family with RCC, without pheochromocytoma. Other phenotypes are likely, however not detailed in this publication

GroupNotHPOs: HP:0002666 (pheochromocytoma)

GroupNotHPOFreeText: N/A

GroupPreviousTesting: N/A

PreviouslyPublished: N/A

SupplementalData: N/A

Variant: p.D126Rfs*6 (c.375_376insC)

LegacyVariant: FS (c.588insC)

CaseProblemVariantFreeText: c.375_376insC automaps to NM_000551.3:c.375dupC

ClinVarID: N/A

CAID: CA16621931 https://reg.clinicalgenome.org/redmine/projects/registry/genboree_registry/allele?hgvsOrDescriptor=NM_000551.3%3Ac.375dupC

gnomAD: N/A

VariantEvidence: This paper declares this variant is predicted to compromise the Elongin-binding site either by truncating the protein.

MutationType: duplication; plus_1_frameshift_variant

CivicName: D126fs (c.375dupC)

MultipleGeneVariants: N/A

Germline mutational data on VHL was gathered from various countries to assess variation in germline mutations that produce disease within different populations. Families were grouped according to three possible outcomes of disease: renal cell carcinoma without pheochromocytoma, renal cell carcinoma with pheochromocytoma, or pheochromocytoma alone. SSCP and denaturing gradient gel electrophoresis identified mutations in 300/469 (63%) of families affected with VHL. This germline familial VHL mutation (G104Hfs*27 (c.309_310delTG)) was taken from the Cambridge, Department of Pathology database, with an unknown number of members belonging to Family ID 53CA. It was reported in the paper as c.522delTG due to a difference in transcript reference. As reported in Table 3, this mutation is found in VHL families without pheochromocytoma, and with renal cell carcinoma.

Germline mutational data on VHL was gathered from various countries to assess variation in germline mutations that produce disease within different populations. Families were grouped according to three possible outcomes of disease: renal cell carcinoma without pheochromocytoma, renal cell carcinoma with pheochromocytoma, or pheochromocytoma alone. SSCP and denaturing gradient gel electrophoresis identified mutations in 300/469 (63%) of families affected with VHL. This germline familial VHL mutation (L158V (c.472C>G)) was taken from the Cambridge, Department of Pathology database, with an unknown number of members belonging to Family ID 91CA. It was reported in the paper as c.685C>G due to a difference in transcript reference. As reported in Table 3, this mutation is found in VHL families without pheochromocytoma, and with renal cell carcinoma.

Germline mutational data on VHL was gathered from various countries to assess variation in germline mutations that produce disease within different populations. Families were grouped according to three possible outcomes of disease: renal cell carcinoma without pheochromocytoma, renal cell carcinoma with pheochromocytoma, or pheochromocytoma alone. SSCP and denaturing gradient gel electrophoresis identified mutations in 300/469 (63%) of families affected with VHL. This germline familial VHL mutation (V170F (c.508G>T)) was taken from the Cambridge, Department of Pathology database, with an unknown number of members belonging to Family ID 26CA. It was reported in the paper as c.721G>T due to a difference in transcript reference. As reported in Table 4, this mutation is found in VHL families with pheochromocytoma. Other phenotypes are likely, including renal cell carcinoma, however not detailed here.

Germline mutational data on VHL was gathered from various countries to assess variation in germline mutations that produce disease within different populations. Families were grouped according to three possible outcomes of disease: renal cell carcinoma without pheochromocytoma, renal cell carcinoma with pheochromocytoma, or pheochromocytoma alone. SSCP and denaturing gradient gel electrophoresis identified mutations in 300/469 (63%) of families affected with VHL. This germline familial VHL mutation (W88S (c.263G>C)) was taken from the Department of Uroglogy, Yokohama database, with an unknown number of members belonging to Family ID 8YO. It was reported in the paper as c.476G>C due to a difference in transcript reference. As reported in Table 3, this mutation is found in VHL families without pheochromocytoma, and with renal cell carcinoma.

An Israeli specialized centre identified 32 VHL patients belonging to 16 different kindreds. This retrospective collection and analysis of clinical, laboratory and imaging data of VHL patients involved 3 individuals belonging to kindred V. Genetic analysis via direct VHL gene sequencing and quantitative Southern blot identified the L188del (c.562_564delCTG) mutation. This kindred includes 3 patients: a father and his two children. The father, 55Y, had bilateral adrenal pheochromocytoma, along with his two children (13Y, 17Y).

Between 1994 and 2001, the Dutch VHL working group conducted DNA analysis on 146 probands with VHL-associated manifestations. DNA of probands was extracted from peripheral blood samples. Exons 1, 2 and 3 of the VHL gene and their flanking untranslated and intronic coding sequences were amplified by PCR and subjected to semi-automated sequence analysis. Screening for structural rearrangements, including deletions, involving the VHL gene was performed by Southern blot analysis and the multiplex ligation-dependent probe amplification (MLPA) assay. Deletions encompassing the entire VHL gene were confirmed by FISH analysis on lymphocytes from affected individuals. VHL mutations were identified in 43 families. This delins (c.393_394delinsA) mutation was found in two VHL type 1 family members (family no. 26). Both had retinal hemangioblastomas, one had renal cysts and one had pancreatic cysts.

Between 1994 and 2001, the Dutch VHL working group conducted DNA analysis on 146 probands with VHL-associated manifestations. DNA of probands was extracted from peripheral blood samples. Exons 1, 2 and 3 of the VHL gene and their flanking untranslated and intronic coding sequences were amplified by PCR and subjected to semi-automated sequence analysis. Screening for structural rearrangements, including deletions, involving the VHL gene was performed by Southern blot analysis and the multiplex ligation-dependent probe amplification (MLPA) assay. Deletions encompassing the entire VHL gene were confirmed by FISH analysis on lymphocytes from affected individuals. VHL mutations were identified in 43 families. Family 20 has one affected member. Genetic analysis reveals mutation R161*(c.481C>T) in the VHL gene. The patient has a family history of VHL disease and presented with retinal capillary hemangioma, renal cyst and pancreatic cysts. ACMG: 'PP4' because the patient's phenotype and family history are highly specific for a disease with single genetic etiology.

Between 1994 and 2001, the Dutch VHL working group conducted DNA analysis on 146 probands with VHL-associated manifestations. DNA of probands was extracted from peripheral blood samples. Exons 1, 2 and 3 of the VHL gene and their flanking untranslated and intronic coding sequences were amplified by PCR and subjected to semi-automated sequence analysis. Screening for structural rearrangements, including deletions, involving the VHL gene was performed by Southern blot analysis and the multiplex ligation-dependent probe amplification (MLPA) assay. Deletions encompassing the entire VHL gene were confirmed by FISH analysis on lymphocytes from affected individuals. VHL mutations were identified in 43 families. Family 3 has one affected member. Genetic analysis reveals mutation S65W(c.194C>G)in the VHL gene. The patient presented with renal cell carcinoma, CNS hemangioblastoma, retinal capillary hemangioma, renal cyst and pancreatic cysts. ACMG: 'PP4' because the patient's phenotypes are highly specific for a disease with single genetic etiology.

Between 1994 and 2001, the Dutch VHL working group conducted DNA analysis on 146 probands with VHL-associated manifestations. DNA of probands was extracted from peripheral blood samples. Exons 1, 2 and 3 of the VHL gene and their flanking untranslated and intronic coding sequences were amplified by PCR and subjected to semi-automated sequence analysis. Screening for structural rearrangements, including deletions, involving the VHL gene was performed by Southern blot analysis and the multiplex ligation-dependent probe amplification (MLPA) assay. Deletions encompassing the entire VHL gene were confirmed by FISH analysis on lymphocytes from affected individuals. VHL mutations were identified in 43 families. This in-frame deletion variant (c.227_229delTCT) was detected in 2 VHL families: 24 and 25. Family no. 24 was a type 2 family with 1 patient with central nervous system hemangioblastoma, pheochromocytoma, renal cell carcinoma, and epididymal cysts. They were identified as sporadic with their parents being excluded from having a germline mutation, although they were not available for DNA analysis. Family no.25 had 2 members with retinal hemangioblastomas and renal cell carcinoma, as well as 1 member with CNS hemangioblastoma. This was published in a future publication: PMID: 32179488.

A novel heterozygous mutation (c.349 T > A, p.W117R) was detected in affected family members of a chinese family. No mutation was detected in unaffected family members or in the 150 normal controls. The mutation segregated with the disease phenotype throughout three generations. Case II:1 (the proband) was a 32 year old female who complained of blurred vision for the past 2 years in her left eye. Fundus examinations showed several retinal hemangioblastomas in the temporal area of the left eye. The patient had previously undergone the operation for cerebellar hemangioblastoma 3 years prior. Case III:1 (the proband’s son) was an asymptomatic 6-year-old male, who was found to have a small hemangioblastoma in the distant peripheral retina of his right eye. Case III:2 (the proband’s daughter) was a 12 year old female who complained of a curtain coming across the vision in her left eye during the screening examination. Fundus examinations revealed a small hemangioblastoma in the infratemporal region of the peripheral retina in her right eye. A sizeable hemangioblastoma with 2.5 papilla diameters and prominant feeder vessels were detected in the inferior peripheral retina of her left eye. Case I:2 (the proband’s mother) had blindness in one eye and died of renal carcinoma, genetic testing was not preformed on this patient. Genomic DNA was extracted from peripheral blood using a DNA extraction kit (TIANGEN). Genomic DNA from all available family members were obtained for PCR and next-generation sequencing (NGS). Sites of variation were identified through a comparison of DNA sequences with the corresponding GenBank reference sequences.

This paper presented 2 cases of VHL syndrome presented simultaneously in twins. Molecular studies were performed for all subjects of the family. DNA was extracted from peripheral blood. Mutation was detected though southern blotting hybridization to radiolabelled VHLcDNA (g7 probe). DNA changes in VHL were only detected in the twins, suggesting a de novo missense mutation in the exon 1B of VHL gene N78S (c.233A>G) (This mutation was labelled as a/G transition at codon 446 in the paper.) Case 1 (III.2) was a 16Y M who presented with right cerebellar hemangioblastoma, medullary and spinal cord solid tumors. Case 2 (III.3) was a 16Y M who presented with right cerebellar hemagioblastoma and nasal pharygeal bleeding fibroangioma.

Twelve patients with hereditary non-SDHx head and neck paragangliomas were analyzed. Eleven of the 12 patients were found to carry germline mutations in the VHL gene. Blood was used for molecular genetic analyses and the NM_000551.2 reference sequence was used. The paper includes 3 additional VHL patients: case 22, 23, 27. Case 24 is a 24Y female who was found to harbour the p.T152I (c.455C>T) mutation of the VHL gene. The variant was written in paper under p.Tyr152Ile. This is incorrect however is paired to a paper where this variant it described (PMID: 18031321) referenced Thr152Ile, so this mutation was deduced. She presented with a a carotid body (paraganglioma), with no other details. A negative family history was described.

Twelve patients with hereditary non-SDHx head and neck paragangliomas were analyzed. Eleven of the twelve patients were found to carry germline mutations in the VHL gene. Blood was used for molecular genetic analyses and the NM_000551.2 reference sequence was used. A total of five patients in this cohort were found to harbour the p.Tyr98His germline mutation of the VHL gene. As well as one patient from literature (previously published in 9880225). In family 1, the proband (case 1) was a 7 year old male diagnosed with malignant extra-adrenal pheochromocytoma of the left retroperitoneum with regional lymph node metastases and treated with resection of the primary tumor, lymphadenectomy, and chemotherapy. At age 9, he presented with a local reocurrance which was treated with resection and external radiotherapy of 20Gy. At age 13, he presented a right adrenal pheochromocytoma which was resected. At age 15 a left carotid body tumor was clinically detected and removed, but recurred at age 16 and again was resected. At age 17, a pheochromocytoma at the renal pelvis was diagnosed by MRI and resected. At age 18, he was disease free except for a lesion in the L1 vertebral body suggestive of hemangioma or pheochromocytoma. His mother was asymptomatic but had the mutation. His cousin had the mutation and presented with a pheochromocytoma. His aunt also had the mutation and a history of retinal hemangioblastomas. The fifth patient with this mutation (case 10), an unrelated 34 year old female, presented with no personal or family history of VHL and persistent pharyngeal discomfort. MRI demonstrated a 4cm tumor in the parapharyngeal space. The tumor was removed, and histology revealed a paraganglioma. Postoperatively, she had Horner’s syndrome and a transitory hypoglossal paresis. At age 46, bilateral adrenal pheochromocytoma and a paraaortic paraganglioma were removed. Later, her niece was diagnosed with a pheochromocytoma, and her brother suffered from pheochromocytoma, renal cell carcinoma, and retinal hemangioblastomas. The proband’s mother was then found to have pheochromocytoma. Case 23 was from the literature and was a female presenting with pheochromocytoma, paraganglioma of head and neck, retinal capillary hemangioma and pancreatic cysts. The patient had a positive family history.

A cohort of 141 patients with hemangioblastomas of the central nervous system was analyzed. 81 patients had germline mutations in the VHL gene, analyzed through peripheral blood by Southern blotting and SSCP analysis. This missense mutation (c.533T>C) was recorded in the paper as c.746T>C due to a difference in transcript sequence. It was found in one patient with a hemangioblastoma of the central nervous system. Any family information is unknown. No other phenotypes described.

A cohort of 141 patients with hemangioblastomas of the central nervous system was analyzed. 81 patients had germline mutations in the VHL gene, analyzed through peripheral blood by Southern blotting and SSCP analysis. This inframe mutation (c.224_226delTCT) was recorded in the paper as c.437delTCT due to a difference in transcript sequence. It was found in 2 patients with hemangioblastomas of the central nervous system. Any relation between patients is unknown. No other phenotypes described.

A cohort of 141 patients with hemangioblastomas of the central nervous system was analyzed. 81 patients had germline mutations in the VHL gene, analyzed through peripheral blood by Southern blotting and SSCP analysis. This missense mutation (c.194C>G) was recorded in the paper as c.407C>G due to a difference in transcript sequence. It was found in 2 patients with hemangioblastomas of the central nervous system. Any relation is unknown. No other phenotype described.

A cohort of 141 patients with hemangioblastomas of the central nervous system was analyzed. 81 patients had germline mutations in the VHL gene, analyzed through peripheral blood by Southern blotting and SSCP analysis. This inframe mutation (c.230_231insTCT) was recorded in the paper as c.443insTCT due to a difference in transcript sequence. It was found in 1 patient with a hemangioblastoma of the central nervous system. Any family information is unknown. No other phenotypes described.

In a study of 426 unrelated VHL patients, 111 were discovered to have alterations in the VHL gene. 18 novel variants were identified in VHL patients, but none were present in 200 unaffected control individuals. DNA samples from peripheral blood leukocytes underwent polymerase chain reaction (PCR) amplification, denaturing high-performance liquid chromatography (DHPLC), and direct sequence analysis of the entire coding sequence and intron-exon boundaries. The P61= (c.183C>G) synonymous germline mutation of the VHL gene was found in a 58-year-old VHL patient with hemangioblastomas of the central nervous system (index case 52).

In a study of 426 unrelated VHL patients, 111 were discovered to have alterations in the VHL gene. 18 novel variants were identified in VHL patients, but none were present in 200 unaffected control individuals. DNA samples from peripheral blood leukocytes underwent polymerase chain reaction (PCR) amplification, denaturing high-performance liquid chromatography (DHPLC), and direct sequence analysis of the entire coding sequence and intron-exon boundaries. The D126Rfs*6 (c.375dup) germline variant of the VHL gene was found in a 24 year old patient (ID 107) presenting with CNS hemangioblastoma and retinal hemangioblastoma. Parents and siblings were negative for the mutation confirming it as de novo in the proband. No phenotype details are noted for these family members. The variant was reported as c.375_376insC and was mapped to c.375dup. This variant also potentially alters the beta domain of the VHL protein.

In a study of 426 unrelated VHL patients, 111 were discovered to have alterations in the VHL gene. 18 novel variants were identified in VHL patients, but none were present in 200 unaffected control individuals. DNA samples from peripheral blood leukocytes underwent polymerase chain reaction (PCR) amplification, denaturing high-performance liquid chromatography (DHPLC), and direct sequence analysis of the entire coding sequence and intron-exon boundaries. The P138T (c.412C>A) missense mutation was found in a 21 yr old VHL patient with retinal hemangioblastomas and pheochromocytoma (index case 115). Proband and father were found to carry the P138T (c.412C>A) germline mutation of the VHL gene. Father, siblings and cousins were analyzed but no phenotype information was provided for these family members. Genotype information for siblings and cousins was not provided.

In a study of 426 unrelated VHL patients, 111 were discovered to have alterations in the VHL gene. 18 novel variants were identified in VHL patients, but none were present in 200 unaffected control individuals. DNA samples from peripheral blood leukocytes underwent polymerase chain reaction (PCR) amplification, denaturing high-performance liquid chromatography (DHPLC), and direct sequence analysis of the entire coding sequence and intron-exon boundaries. Germline mutation c.-65_-55dup11 (mapped to c.-61_-51dup) was identified in prodand 221, 230 and 231. Proband 221 (12Y) presented with retinal capillary hemangioma. Proband 230 (40Y) presented with CNS hemangioblastoma. Proband 231 (9Y) presented with CNS hemangioblastoma. All three patients were previously published in PMID 17006605.

In a study of 426 unrelated VHL patients, 111 were discovered to have alterations in the VHL gene. 18 novel variants were identified in VHL patients, but none were present in 200 unaffected control individuals. DNA samples from peripheral blood leukocytes underwent polymerase chain reaction (PCR) amplification, denaturing high-performance liquid chromatography (DHPLC), and direct sequence analysis of the entire coding sequence and intron-exon boundaries. This novel, missense mutation was found in a 16 yr old VHL patient with pheochromocytoma (index case 55). Variant was transmitted from a parent (both parents were analyzed) but it is unclear which parent was positive for the variant.

In a study of 426 unrelated VHL patients, 111 were discovered to have alterations in the VHL gene. 18 novel variants were identified in VHL patients, but none were present in 200 unaffected control individuals. DNA samples from peripheral blood leukocytes underwent polymerase chain reaction (PCR) amplification, denaturing high-performance liquid chromatography (DHPLC), and direct sequence analysis of the entire coding sequence and intron-exon boundaries. This de novo, missense mutation, c.233A>G, was found in 2 VHL patients. A 24-year-old patient had retinal capillary hemangioma, pancreatic lesions, and renal cysts (index case 152). The other patient, a 33-year-old, presented with hemangioblastomas of the central nervous system and pancreatic lesions (index case 45). This variant also potentially alters the stability of the beta domain of the VHL protein. There is strong evidence for pathogenicity because this variant is de novo in a patient with the disease and no family history.

In a study of 426 unrelated VHL patients, 111 were discovered to have alterations in the VHL gene. 18 novel variants were identified in VHL patients, but none were present in 200 unaffected control individuals. DNA samples from peripheral blood leukocytes underwent polymerase chain reaction (PCR) amplification, denaturing high-performance liquid chromatography (DHPLC), and direct sequence analysis of the entire coding sequence and intron-exon boundaries. The R107Pfs*25 (c.314dup) germline variant of the VHL gene was found in a 16 year old patient (ID 225) presenting with CNS hemangioblastoma. The variant is noted as Familial (i.e. “transmission of the variant from a parent”). Sibling was analyzed but details are not provided. The variant was reported as c.314_315insC and was mapped to c.314dup. There is co-segregation of disease with mutation.

Eleven RCCs and 16 renal cysts from eight patients with VHL were analyzed. All patients had known germline VHL mutations. One male patient had a germline missense mutation (c.233A>C; p.N78T) in the VHL gene and presented with bilateral RCC and cysts, CNS hemangioblastoma, and pancreatic cysts. Family history was not described for the patient.

Eleven RCCs and 16 renal cysts from eight patients with VHL were analyzed. All patients had known germline VHL mutations. One male patient had a missense germline mutation (c.351G>T; p.W117C) in VHL gene and presented with bilateral RCC and cysts, CNS and retinal hemangioblastoma, and pancreatic cysts. Family history was not described for the patient.

All patients included in the study were VHL mutation carriers of 18 years or older examined between 1972 and 2012 in the VHL expertise centers University Medical Center Groningen (UMCG) and University Medical Center Utrecht (UMCU). Clinical data of 75 VHL mutation carriers were retrospectively retrieved from electronic and written patient record. Of these patients, one was found to harbour the c.358A>G germline mutation of the VHL gene. They presented with a spinal hemangioblastoma. Their age at the last follow-up was 15. Previously published in: PMID 24132471.

All patients included in the study were VHL mutation carriers of 18 years or older examined between 1972 and 2012 in the VHL expertise centers University Medical Center Groningen (UMCG) and University Medical Center Utrecht (UMCU). Clinical data of 75 VHL mutation carriers were retrospectively retrieved from electronic and written patient record. Of these patients, one was found to harbour the c.341-59_341-14del germline mutation of the VHL gene. They presented with a retinal hemangioblastoma, cerebellar hemangioblastoma, renal cell carcinoma, renal cyst, pheochromocytoma and pancreatic cysts. Their age at the last follow-up was 67. Previously published in: PMID 24132471.

The authors described the growth kinetics of 96 renal tumours in 64 VHL patients (32 males, 32 females). All patients had VHL disease with renal involvement. 54/64 had undergone renal surgery or a radiofrequency thermal ablation procedure for RCC. In 10/64 patients, at least one or more RCC was diagnosed. However, at the time of diagnosis, an indication for treatment was not yet present. For all patients, several CT/MRI scans existed for retrospective description of tumour growth kinetics. 10-ml EDTA anticoagulated blood samples were available from each patient for mutation screening of all exons of the VHL gene. Genomic DNA from blood samples was extracted using standard methods. Small deletions/insertions or point mutations were identified by DNA sequencing followed by complete mutation screening using PCR/single-strand conformation polymorphism and denaturing high-performance liquid chromatography. A germline nonsense variant p.S65*(c.194C>A) was identified in patient #5. This mutation was noted as c.407C>A in the paper due to a difference in reference transcript used (see Table S2).

The authors described the growth kinetics of 96 renal tumours in 64 VHL patients (32 males, 32 females). All patients had VHL disease with renal involvement. 54/64 had undergone renal surgery or a radiofrequency thermal ablation procedure for RCC. In 10/64 patients, at least one or more RCC was diagnosed. However, at the time of diagnosis, an indication for treatment was not yet present. For all patients, several CT/MRI scans existed for retrospective description of tumour growth kinetics. 10-ml EDTA anticoagulated blood samples were available from each patient for mutation screening of all exons of the VHL gene. Genomic DNA from blood samples was extracted using standard methods. Small deletions/insertions or point mutations were identified by DNA sequencing followed by complete mutation screening using PCR/single-strand conformation polymorphism and denaturing high-performance liquid chromatography. A germline missense variant p.Q164L(c.491A>T) was identified in patient #48. This mutation was noted as c.704A>T in the paper due to a difference in reference transcript used (see Table S1). No family information is available for this patient.

The authors described the growth kinetics of 96 renal tumours in 64 VHL patients (32 males, 32 females). All patients had VHL disease with renal involvement. 54/64 had undergone renal surgery or a radiofrequency thermal ablation procedure for RCC. In 10/64 patients, at least one or more RCC was diagnosed. However, at the time of diagnosis, an indication for treatment was not yet present. For all patients, several CT/MRI scans existed for retrospective description of tumour growth kinetics. 10-ml EDTA anticoagulated blood samples were available from each patient for mutation screening of all exons of the VHL gene. Genomic DNA from blood samples was extracted using standard methods. Small deletions/insertions or point mutations were identified by DNA sequencing followed by complete mutation screening using PCR/single-strand conformation polymorphism and denaturing high-performance liquid chromatography. A germline missense variant p.H115R(c.344A>G) was identified in patient #12. This mutation was noted as c.557A>G in the paper due to a difference in reference transcript used (see Table S1). No family information is available for this patient.

The authors described the growth kinetics of 96 renal tumours in 64 VHL patients (32 males, 32 females). All patients had VHL disease with renal involvement. 54/64 had undergone renal surgery or a radiofrequency thermal ablation procedure for RCC. In 10/64 patients, at least one or more RCC was diagnosed. However, at the time of diagnosis, an indication for treatment was not yet present. For all patients, several CT/MRI scans existed for retrospective description of tumour growth kinetics. 10-ml EDTA anticoagulated blood samples were available from each patient for mutation screening of all exons of the VHL gene. Genomic DNA from blood samples was extracted using standard methods. Small deletions/insertions or point mutations were identified by DNA sequencing followed by complete mutation screening using PCR/single-strand conformation polymorphism and denaturing high-performance liquid chromatography. A germline missense variant p.P154L(c.461C>T) was identified in patient #14. This mutation was noted as c.674C>T in the paper due to a difference in reference transcript used (see Table S1). No family information is available for this patient.

A study of 103 patients with VHL retinal manifestations and 108 patients without VHL retinal manifestations extracted from the French VHL database revealed that the number of hemangioblastomas appeared higher in patients who had a substitution when compared with patients with a truncated protein. Detected by PCR and SSCP, the missense mutation (c.337C>T (p.Arg113Ter)) was found in 2 patients stated to have retinal HBs. Other manifestations are possible but not clear. This is possibly previously published in PMID: 15300849

A study of 103 patients with VHL retinal manifestations and 108 patients without VHL retinal manifestations extracted from the French VHL database revealed that the number of hemangioblastomas appeared higher in patients who had a substitution when compared with patients with a truncated protein. Detected by PCR and SSCP, this mutation p.Y156C (c.467A>G) was found in 3 VHL patients. 1 member was said to have retinal hemangioblastomas, while the other 2 were said not to have this phenotype. Other manifestations are not clearly stated. No family information or relation was provided.

A study of 103 patients with VHL retinal manifestations and 108 patients without VHL retinal manifestations extracted from the French VHL database revealed that the number of hemangioblastomas appeared higher in patients who had a substitution when compared with patients with a truncated protein. Detected by PCR and SSCP, this mutation p.R82P (c.245G>C) was found in 3 VHL patients. 1 member was said to have retinal hemangioblastomas, while the other 2 were said not to have this phenotype. Other manifestations are not clearly stated. No family information or relation was provided. This is possibly previously published in PMID: 15300849

A study of 103 patients with VHL retinal manifestations and 108 patients without VHL retinal manifestations extracted from the French VHL database revealed that the number of hemangioblastomas appeared higher in patients who had a substitution when compared with patients with a truncated protein. Detected by PCR and SSCP, this mutation p.Y156* (c.468T>G) was found in 3 VHL patients. All 3 were said not to have retinal hemangioblastomas, while other manifestations are not clearly stated. No family information or relation was provided. This is possibly previously published in PMID: 15300849

A study of 103 patients with VHL retinal manifestations and 108 patients without VHL retinal manifestations extracted from the French VHL database revealed that the number of hemangioblastomas appeared higher in patients who had a substitution when compared with patients with a truncated protein. Detected by PCR and SSCP, this mutation (p.Q73* (c.217C>T)) was found in 5 VHL patients. 4 members were said to have retinal hemangioblastomas, while the other was said not to have this phenotype. Other manifestations are not clearly stated. No family information or relation was provided. This is possibly previously published in PMID: 15300849

A study of 103 patients with VHL retinal manifestations and 108 patients without VHL retinal manifestations extracted from the French VHL database revealed that the number of hemangioblastomas appeared higher in patients who had a substitution when compared with patients with a truncated protein. Detected by PCR and SSCP, this mutation p.W117C (c.351G>T) was found in 5 VHL patients. 4 members were said to have retinal hemangioblastomas, while the other member was said not to have this phenotype. Other manifestations are not clearly stated. No family information or relation was provided. This is possibly previously published in PMID: 15300849

A study of 103 patients with VHL retinal manifestations and 108 patients without VHL retinal manifestations extracted from the French VHL database revealed that the number of hemangioblastomas appeared higher in patients who had a substitution when compared with patients with a truncated protein. Detected by PCR and SSCP, this stop mutation (p.E46* (c.136G>T)) was found in a single patient with VHL retinal manifestation, which coincides with mutations confirmed to be associated with VHL type 1 in this study. No family information was provided. This is possibly previously published in PMID: 15300849

The patient cohort consisted of 57 patients (26 male and 31 female) from Southern Italy, most of whom (55 of 57) presented with solitary HGB of the CNS: 6 were syndromic and 51 were apparently sporadic. 8 patients had germline VHL mutations. Both peripheral blood and tumor tissue samples were sequenced using Big Dye Terminator Ready Reaction mix v.1.1 (Life Technologies, Foster City, CA). Sequencing reactions were loaded on an ABI 3100 capillary sequencer (Life Technologies) and analyzed using the Sequencing Analysis software v.3.7 (PE Applied Biosystems, Grand Island, NY). A 61 year old Italian male patient (Case ID HGB-25) was found to harbor a germline missense mutation (c.74C>T; p.Pro25Leu) in the VHL gene. The patient was also found to have two somatic mutations, a c.375insA was found in tumor sample 2 and a c.163delG was found in sample 1a. He was diagnosed at age 53 with cerebellar hemangioblastoma. The patient has positive family history for VHL disease.

A retrospective analysis of 164 patients waiting for surgery for hemangioblastomas at Freiburg University Medical Center, Freiburg, Germany was conducted. Bidirectional direct sequencing by use of an ABI PRISM 3130 DNA Genetic Analyzer (Applied Biosystems) was used to identify mutations in abnormal samples. Patient #3 (male, 47Y) was found to carry the germline VHL nonsense mutation c.481C>T, p.Arg161* (stated as c.694C>T in the paper which used the Latif sequence as reference) and presented with brainstem hemangioblastoma. Family history was not described. ACMG: PP4, PM1

Genotype-phenotype correlations of 573 VHL patients from 200 kindreds were analyzed. Higher risk of pheochromocytoma was associated with missense mutations that result in substitution of a surface amino acid (p<=0.004) than for patients with missense mutations that disrupt the structural integrity of the VHL gene product (pVHL) or other loss of function mutations (either germline deletions or truncating mutations). This missense mutation (c.499C>T) was found in 12 families with 58 affected members: 36 had retinal angioma, 18 had cerebellar hemangioblastoma, 15 had renal cell carcinoma, 31 had pheochromocytoma. ACMG PP4

Genotype-phenotype correlations of 573 VHL patients from 200 kindreds were analyzed and confirmed that higher risk of pheochromocytoma is associated with missense mutations that result in substitution of a surface amino acid or that disrupt the structural integrity of the VHL gene product (pVHL). Age-related risks of retinal hemangoblastomas and renal cell carcinoma were higher in patients with nonsense or frameshift mutations. Southern blot and/or MLPA analysis by whole blood detected this mutation. R113FS (c.337del) was found in a patient with retinal hemangioblastomas diagnosed at age 28Y.

Genotype-phenotype correlations of 573 VHL patients were analyzed and confirmed that higher risk of pheochromocytoma is associated with missense mutations that result in substitution of a surface amino acid or that disrupt the structural integrity of the VHL gene product (pVHL). A missense mutation (c.346C>G), affecting residue with pVHL protein core, was detected in a VHL type 1 family with 3 affected individuals. All 3 patients had retinal angioma.

Genotype-phenotype correlations of 573 VHL patients from 200 kindreds were analyzed and confirmed that higher risk of pheochromocytoma is associated with missense mutations that result in substitution of a surface amino acid or that disrupt the structural integrity of the VHL gene product (pVHL). Age-related risks of retinal hemangoblastomas and renal cell carcinoma were higher in patients with nonsense or frameshift mutations. Southern blot and/or MLPA analysis by whole blood detected this nonsense mutation (Q73*) was found in 3 VHL families with 9 patients altogether. Seven had retinal hemangioblastomas, 3 had cerebellar hemangioblastomas, and 5 had renal cell carcinoma.

Genotype-phenotype correlations of 573 VHL patients from 200 kindreds were analyzed. Higher risk of pheochromocytoma was associated with missense mutations that result in substitution of a surface amino acid (p<=0.004) than for patients with missense mutations that disrupt the structural integrity of the VHL gene product (pVHL) or other loss of function mutations (either germline deletions or truncating mutations). This missense mutation (c.395A>C), affecting a residue on surface of pVHL, was found in a VHL family of 2, 1 with retinal angiomas diagnosed at 18Y, 1 with cerebellar hemangioblastomas at 37Y, and 1 with renal cell carcinoma at 49Y.

Genotype-phenotype correlations of 573 VHL patients from 200 kindreds were analyzed and confirmed that higher risk of pheochromocytoma is associated with missense mutations that result in substitution of a surface amino acid or that disrupt the structural integrity of the VHL gene product (pVHL). Age-related risks of retinal hemangoblastomas and renal cell carcinoma were higher in patients with nonsense or frameshift mutations. Southern blot and/or MLPA analysis by whole blood detected this nonsense mutation. c.529A>T was found in 3 VHL families total. 2 families were grouped and detailed as affected with a total of 10 patients, 8 with retinal angioma, 4 with cerebellar hemangioblastoma, 1 with renal cell carcinoma (diagnosed at 50Y), and 1 with pheochromocytoma (diagnosed at 27Y). Another individual belonging to a separate family was written with the same nonsense mutation and retinal angiomas diagnosed at 28Y, cerebellar hemangioblastomas diagnosed at 22Y, and renal cell carcinoma diagnosed at 37Y.

Genotype-phenotype correlations of 573 VHL patients were analyzed and confirmed that higher risk of pheochromocytoma is associated with missense mutations that result in substitution of a surface amino acid or that disrupt the structural integrity of the VHL gene product (pVHL). Age-related risks of retinal hemangoblastomas and renal cell carcinoma were higher in patients with nonsense or frameshift mutations. Southern blot and/or MLPA analysis by whole blood detected this nonsense mutation (c.375insT: p.Glu55*) in a VHL family with 2 affected members. They both had hemangioblatomas. No other info was given.

The mutation was written as follows: AFO10238:375_376insT; NM_000551.2:c.162_163insT; p.54MetX. Author Maher was contacted and confirmed "A single nucleotide insertion (T) was detected at position 375 (375insT)” from 2000 lab report - June 1, 2021 - therefore assume p. transcribed incorrectly.

112 unrelated patients with pheochromocytoma or paraganglioma were screened for mutations in the VHL gene, as well as the SDHx related genes (SDHD, SDHB, SDHC). Mutation analysis was performed on patient blood samples: DNA was extracted using a DNA purification kit (Gentra), exon-intron boundaries of the VHL gene were amplified by PCR, mutation screening was performed by single strand conformation polymorphism (SSCP) and heteroduplex analysis, or by denaturing high performance liquid chromatography (dHPLC) analysis (WAVE 3500 HT system (Transgenomic, Inc., Omaha, NE, USA)) with confirmatory sequencing on CEQ2000 capillary sequencer (Beckman Coulter). Two patients without features suggestive of VHL syndrome were found to have germline VHL mutations. The second patient was associated with the presence of a right pneumogastric nerve paraganglioma diagnosed at 49 years, and was found to have a germline p.His125Tyr mutation. Only the amino acid change was given in the paper, the nucleotide change was not specified in the paper, inferred change is c.373G>A (H125Y).

VHL germline mutation carriers who had an ophthalmologic examination at the Erasmus Medical Centre or the Rotterdam Eye Hospital between 1984 and 2017, and were diagnosed with retinal hemangioblastoma (RH) were included in this study. A diagnosis of RH was based on the clinical appearance of a vasoproliferative retinal tumour. Peripheral blood samples from every member that had clinically suspected VHL disease of each family were collected for VHL mutation analysis. A total of 21 patients from 10 families were included on the basis of both a VHL germline mutation and ocular manifestations. Copy number analyses were carried out by multiplex ligation dependent probe amplification (MLPA) and fluorescence in situ hybridization (FISH). Mutations were described as one of the following two groups: truncating (complete deletion, partial deletion, nonsense mutation, splice site mutation, duplication and insertion) or missense. Ocular manifestations were not reported on an individual level, but rather were broken down based on the two groups. From Pedigree (family) 14, two individuals were suspected of VHL disease and mutational analysis revealed a R64P(c.191G>C) missense variant. Both individuals from this family did not develop RH.

In a study of 138 unrelated VHL families, 109 germline mutations were found. Missense mutations were more frequently found in families with pheochromocytoma (PC). Deletions, nonsense, and frameshift mutations were more frequently found in families without PC. Large deletions and mutations predicted to cause truncated protein were associated with lower risk of PC. The mutation of N78S (c.233A>G) was referred to in the paper by c.446A>G due to a difference in transcript sequence. This missense mutation was found in a VHL family (family ID 160). No phenotype provided.

Germline mutations were found in twenty VHL probands and their families, of whom 17 presented with a family history (15 from Brazil, one from Portugal, and one from Ecuador) . An intronic variant, c.340+7G>A, (reported as IVS1+7G>A in the paper) was found in 2 affected family members that both presented with with hemangioblastomas of the CNS. They were able to analyze the germline mutation linkage to disease in this family. The pathogenic nature of the variant was not confirmed at the mRNA level; however, the fact that this alteration was not detected in one unaffected first degree relative (after clinical and radiological screening) suggests that it may be associated with the disease. Additionally, the reported novel mutation were not represented in 120 alleles from normal subjects in the Brazilian population suggesting that these base changes are not common polymorphisms.

Germline mutations were found in 20 Brazilian, VHL probands and their families. The R107P (c.320G>C) germline missense mutation was found in a VHL type 1 family of 5 individuals, 4 with hemangioblastomas of the central nervous system, 3 with retinal angiomas, 2 with renal carcinoma, and a proband that presented with a non-functional paraganglioma (VHL family 7).

Among patients prospectively followed by the members of the French VHL Study Group, 35 consecutive patients with well-documented endocrine pancreatic tumour (20 women and 15 men; median age,37 [21Y-57] years) were included. The data from 7 patients with EPT have been partially reported previously in Hammel et al., 1995;2000. All 35 patients had VHL germline mutations and were assessed for the presence of pheochromocytoma and renal cancer in addition to pancreatic tumours. The presence of other phenotypes was not described. Patient No. 1 from Family 81 was found with a c.509T>G germline missense mutation and a pancreatic endocrine tumour. A second patient, was also found with this mutation: Patient No. 9 from family 81, and presented with pancreatic endocrine tumour and pheochromcytoma. Both the patient's phenotype is highly specific to VHL disease (PP4) and disease-causing missense variants are common in VHL disease (PP2).

Among patients prospectively followed by the members of the French VHL Study Group, 35 consecutive patients with well-documented endocrine pancreatic tumour (20 women and 15 men; median age,37 [21Y-57] years) were included. All 35 patients had VHL germline mutations and were assessed for the presence of pheochromocytoma and renal carcinoma in addition to pancreatic tumours. The presence of other phenotypes was not described. DNA was isolated from peripheral blood lymphocytes and mutations were detected using direct sequencing of the 3 exons of VHL and/or southern blot analysis. A germline missense mutation (c.256C>T) was found in 3 patients, 2 of which were related. Patient No 7 from family 503 presented with pancreatic endocrine tumour, pheochromocytoma, and renal carcinoma and patient No 15 from family 503 presented with pancreatic endocrine tumour. Their familial relation was not described. Patient No. 27 from family 11 also had the same mutation and presented with pancreatic endocrine tumour, pheochromocytoma, and renal carcinoma. The patients' phenotypes are highly specific to VHL disease (ACMG PP4).

PatientID: 27

KindredID: 11

Case: Sex unknown, Age unknown, French

DiseaseAssertion: VHL

FamilyInfo: None

CasePresentingHPOs: HP:0030405; HP:0002666; HP:0005584 (pancreatic endocrine tumour, pheochromocytoma, renal cell carcinoma)

CaseHPOFreeText: N/A

CaseNotHPOs: N/A

CaseNotHPOFreeText: N/A

CasePreviousTesting: N/A

PreviouslyPublished: N/A

SupplementalData: N/A

Variant: p.P86S (c.256C>T)

LegacyVariant: N/A

CaseProblemVariantFreeText: N/A

ClinVarID: 178692 https://www.ncbi.nlm.nih.gov/clinvar/variation/178692/

CAID: N/A

gnomAD: N/A

VariantEvidence:N/A

MutationType: missense_variant;transition

CivicName: P86S (c.256C>T)

MultipleGeneVariants: N/A

Among patients prospectively followed by the members of the French VHL Study Group, 35 consecutive patients with well-documented endocrine pancreatic tumour (20 women and 15 men; median age,37 [21Y-57] years) were included. The data from 7 patients with EPT have been partially reported previously in Hammel et al., 1995;2000. All 35 patients had VHL germline mutations and were assessed for the presence of pheochromocytoma and renal cancer in addition to pancreatic tumours. The presence of other phenotypes was not described. The c.500G>A mutation was found in 2 unrelated patients. Patient no 13 from family 354 presented with pancreatic endocrine tumour and pheochromocytoma. Patient no. 29 from family 372 presented with pancreatic endocrine tumour and pheochromocytoma. No family history was mentioned. The patients' phenotypes are highly specific to VHL disease (PP4) and disease-causing missense variants are common in VHL disease (PP2). Further, the mutation is occuring at a VHL hotspot (PM1).

Thirty-two surgically resected HB specimens from 28 patients treated at the University of Tokyo Hospital between 1995 and 2012 were analyzed retrospectively. Of the 32 specimens, 11 were VHL-related HBs from 7 patients. Two brothers identified as hemangioblastoma patients were confirmed with a germline c.216delC deletion in the VHL gene. Identical single base-pair deletions were found in 3 hemangioblastomas from these 2 patients with VHL disease from the same family, indicating that this small frameshift deletion was a germline mutation. One brother had hemangioblastoma in the cerebellum at 22Y and then again at 24Y, and the 30Y brother had a hemangioblastoma in the brainstem. No other phenotypic information was available.

A retrospective cohort study included all the VHL patients diagnosed at Peking University First Hospital (Beijing, China) prior to June 1 2016. A total of 291 patients from 115 different families were included for analysis. This missense mutation was confirmed in 2 families (#18 and #19). Family #18: Male proband was diagnosed with CNS hemangioblastoma at 31Y, renal cell carcinoma at 32Y, and pancreatic cysts or pancreatic tumours at 32Y (mutation confirmed); daughter diagnosed with retinal angioma at 11Y (mutation confirmed); father diagnosed with renal cell carcinoma at 52Y (passed away before diagnosis/ genetic confirmation). Family #19: female proband diagosed with retinal angioma at 12Y, renal cell carcinoma at 33Y (mutation confirmed), and multiple pancreatic cysts or pancreatic tumours at 33Y; her daughter has no clinical manifestations and her age is unknown (mutation confirmed). This cohort was also published in a paper by Wang et al., 2018 (PMID: 29330336).

A retrospective cohort study included all the VHL patients diagnosed at Peking University First Hospital (Beijing, China) prior to June 1 2016. A total of 291 patients from 115 different families were included for analysis. This nonsense mutation was found in two families (#30 and #31). Family #30: a male proband (patient 93) was diagnosed with CNS hemangioblastoma at 31Y (mutation confirmed). No other family information was provided. Family #31: a male proband (patient 94) was diagnosed with CNS hemangioblastoma at 56Y, renal cell carcinoma at 53Y, multiple pancreatic cysts or tumours at 53Y (mutation confirmed); his son (patient 95) was diagnosed with retinal angioma at 29Y, renal cell carcinoma at 29Y, multiple pancreatic cysts or tumours at 29Y, and pheochromocytoma at 29Y. This cohort was also published in a paper by Wang et al., 2018 (PMID: 29330336).

A retrospective cohort study included all the VHL patients diagnosed at Peking University First Hospital (Beijing, China) prior to June 1 2016. A total of 291 patients from 115 different families were included for analysis. This familial mutation was confirmed in family #42. A male proband (patient 114) was diagnosed with CNS hemangioblastoma at 36Y, retinal angioma at 44Y, RCC at 44Y, and multiple pancreatic lesions at 44Y. His son (patient 115) was diagnosed with CNS hemangioblastoma at 12Y and retinal angioma at 11Y. His first sister (patient 116) was diagnosed with multiple pancreatic lesions at 53Y. His second sister (patient 117), who was not genetically confirmed for the mutation, was diagnosed with multiple pancreatic lesions at 46Y. His niece (daughter of second sister) (patient 118) was asymptomatic. His first brother (patient 119), who was not genetically confirmed for the mutation, was diagnosed with CNS hemangioblastoma at 47Y, RCC at 59Y, and multiple pancreatic lesions at 59Y. His second brother (patient 120), who was not genetically confirmed for the mutation, was diagnosed with CNS hemangioblastoma at 44Y, retinal angioma at 43Y, RCC at 44Y, and multiple pancreatic lesions at 44Y. His nephew (son of second brother) (patient 121) was diagnosed with CNS haemangioblastoma at 12Y, multiple pancreatic lesions at 18Y, pheochromocytoma at 18Y. This cohort was also published in a paper by Wang et al., 2018 (PMID: 24581539) and Wang et al., 2014 (PMID: 29330336). Wang et al., 2018 reported 8 patients from 1 family who were genetically confirmed with this germline mutation. Wang et al., 2014 reported 4 patients from 1 family who were genetically confirmed with the mutation. It is likely that all three papers published the same family.

A retrospective cohort study included all the VHL patients diagnosed at Peking University First Hospital (Beijing, China) prior to June 1 2016. A total of 291 patients from 115 different families were included for analysis. This frameshift mutation was found in the VHL gene of family #15. The proband (patient 42) has four family members confirmed with the mutation, and two family members who passed away before diagnosis/ genetic confirmation of the mutation. Family members confirmed with the mutation include: the proband's daughter (patient 43) who had retinal angioma (age 17), renal cell carcinoma (21), multiple pancreatic cysts/tumors (18), and a pheochromocytoma (19). His son (patient 44) who was asymptomatic at the time of assessment. His niece 1 (patient 47) who had a retinal angioma and renal cell carcinoma (22). And his niece 2 (patient 28) who had a CNS hemangioblastoma (19) and a retinal angioma (18). Family members who passed away before diagnosis and were not genetically confirmed include: his sister (patient 45) who had a CNS hemangioblastoma (22), and his brother (patient 46) who had a retinal angioma and renal cell carcinoma at 52. This cohort was also published in a paper by Wang et al., 2018 (PMID: 29330336). It is likely that the same family was published in both papers, with two additional members added in the 2018 paper.

A retrospective cohort study included all the VHL patients diagnosed at Peking University First Hospital (Beijing, China) prior to June 1 2016. A total of 291 patients from 115 different families were included for analysis. A female proband, patient 111 from kindred 39, was found with this germline splice site mutation (c.340+5G>C). She was diagnosed with a CNS hemangioblastoma at 35Y. This kindred was likely published in a later paper by Wang et al., 2018 (PMID: 29330336).

A retrospective cohort study included all the VHL patients diagnosed at Peking University First Hospital (Beijing, China) prior to June 1 2016. A total of 291 patients from 115 different families were included for analysis. A male patient was confirmed for nonsense mutation in the VHL gene (c.217C>T; p.Gln73*). He was diagnosed with CNS haemangioblastoma at 66Y, retinal angioma at 66Y, renal cell carcinoma at 37Y, and multiple pancreatic cysts or tumours at 54Y. This cohort was also published in a paper by Wang et al., 2018 (PMID: 24581539) and Wang et al., 2014 (PMID: 29330336). It is likely that all three papers reported on the same individual.

A retrospective cohort study included all the VHL patients diagnosed at Peking University First Hospital (Beijing, China) prior to June 1 2016. A total of 291 patients from 115 different families were included for analysis. One proband was genetically confirmed with this germline mutation: a 46 year old female from kindred #89 who presented with a renal cell carcinoma at that age. This patient was also published in a later paper by Wang et al., 2018 (PMID: 29330336).

A retrospective cohort study included all the VHL patients diagnosed at Peking University First Hospital (Beijing, China) prior to June 1 2016. A total of 291 patients from 115 different families were included for analysis. This mutation was confirmed in kindred 56. A female proband, patient 153, presented with a retinal angioma at 26 and multiple pancreatic cysts/tumors at 31. No family information was given. This cohort was also published in a paper by Wang et al., 2018 (PMID: 29330336).

150 index patients with pheochromocytoma/paraganglioma were evaluated. Phenotypic data were collected and germline mutations in five susceptibility genes (RET, VHL, SDHB, SDHD and SDHC) were tested. Out of 30 patients with familial presentation, 13 were clinically diagnosed as VHL syndrome. A 20 year old Asian Indian female presented with bilateral pheochromocytoma with pancreatic cysts. Using PCR and Sanger sequencing through peripheral blood, genetic testing confirmed a mutation at c.588_588dupA of the VHL gene. Family history is positive.

A case describes a 22Y male presenting with a believed novel sporadic mutation labeled as 'NT474INST' by the authors (c.474insT resulting in a frameshift mutation). He complained of a shadow in his right eye and was diagnosed with retinal detachment secondary to a retinal hemangioblastoma. MRI of the brain revealed a hyper vascular lesion in the medulla and a smaller lesion at the medullary-spinal cord junction. Weighted abdominal MRI revealed innumerable pancreatic cysts and multiple benign-appearing renal cysts. He also presented with an epididymal cyst. After surgical resection of the retinal hemangioblastoma, the patient remained asymptomatic. The patient has a no family history of VHL but family genotype is not described.

Of 38 PPGL (pheochromocytoma and paraganglioma) cases studied, two were associated with an individual carrying the germline mutation c.308C>A (ENSP00000256474.2 p.Pro103His) of the VHL gene. This mutation was identified using germline clinical sequencing on fresh and formalin-fixed and paraffin-embedded (FFPE) tissue and confirmed with sanger sequencing. The authors identified the reference sequence used as NM_000551.3. The first patient (ID: WM01) presented with pheochromocytoma said to be MAX-Like (7yo), and the second (ID: WM04) with a pheochromocytoma (13yo). No family history, or relation, was presented.

Screening of 92 unrelated patients with VHL disease revealed 61 DNA variants. No variants were found in 96 control cases. Direct sequencing on peripheral blood was systematically performed on exon 1 PCR products. Southern blotting was used to search for larger alterations to the gene. The V62Cfs*5 (c.180del) mutation was found in a VHL type 1 (no pheochromocytoma) patient (patient no. V9). The variant was reported as c.179del and mapped to c.180del.

Screening of 92 unrelated patients with VHL disease revealed 61 DNA variants. No variants were found in 96 control cases. Direct sequencing on peripheral blood was systematically performed on exon 1 PCR products. Southern blotting was used to search for larger alterations to the gene. The E46* (c.136G>T) missense mutation was found in a VHL type 1 (no pheochromocytoma) patient (patient no. V65).

Screening of 92 unrelated patients with VHL disease revealed 61 DNA variants. No variants were found in 96 control cases. Direct sequencing on peripheral blood was systematically performed on exon 1 PCR products. Southern blotting was used to search for larger alterations to the gene. The L188P (c.563T>C) missense mutation was found in a VHL type 1 (no pheochromocytoma) patient (patient no. V269). Five unaffected family members were negative for the variant.

Analysis of peripheral blood samples from 5 Japanese families with PCR and SSCP revealed 4 germline mutations. This splice site mutation NM_000551.3:c.464-1G>T was found in a family of 10 individuals. Please note that the paper states the mutation as c.464-1G>T because it uses a legacy reference sequence. Three VHL patients were identified with hemangioblastomas of the central nervous system and retinal angiomas (family C). The eldest patient also had renal cell carcinoma (listed as C-1 in publication).

3 unrelated patients with unexpected or non-syndromic phenotypes, accompanied by a VHL component tumor received germline gene panel testing as part of the PROACTIVE study at the Dana-Farber Cancer Institute, each detected with a low penetrant variant. At age 72, patient 2 (female) presented to genetics due to personal of cervical cancer at age 41, which was treated surgically, as well as due to family history of cancer. Her mother and maternal grandmother had colon cancer, while her uncle and cousin on the maternal side had prostate cancer. At age 79, she was diagnosed with high-grade papillary urothelial carcinoma, as well as metastasis in a right supraclavicular lymph node. Her treatment included neoadjuvant chemotherapy, radical cystectomy, and adjuvant chemotherapy. Somatic testing of the lymph node revealed several variants and and copy number alterations. As part of the PROACTIVE study, peripheral blood was collected. The germline panel of 133 genes (including MAX/NF1/RET/SDHA SHDB/SHDC/SDHD/SDHAF2/TMEM127) revealed a heterozygous germline VHL p.Tyr98His(c.292T>C) variant, along with other somatic VUS variants in other genes. Patient 2 died from cancer at age 82, a few months after receiving her genetic results. ACMG: PP4

Seven Hungarian families (35 members) with VHL and 37 unrelated patients with apparently sporadic pheochromocytoma had their VHL gene molecularly analyzed. Peripheral blood samples were isolated, amplified by PCR, and subsequently underwent direct sequencing using either SequiTherm Excel II DNA Sequencing Kit-LC (Epicentre Technologies, Madison, WI, USA) and an automated Li-COR IR2 sequencer (Li-COR Inc., Lincoln, NE, USA) or the Big Dye Terminator Cycle Sequencing kit v3.1 (Applied Biosystems, Foster City, CA, USA), and run on an automated sequencer 310 Genetic Analyzer from Applied Biosystems.The missense mutation, c.500G>A, was found in family F and was molecularly confirmed in 2 family members. The proband was a 27 year old man who presented with retinal hemangiomas, unilateral pheochromocytoma, and horseshoe kidneys. The mother of the proband was also confirmed to have the VHL mutation and was diagnosed with renal cell carcinoma at age 49.

During the period from 2009 to 2012, 39 families with VHL disease were diagnosed with hereditary VHL disease at Peking University First Hospital Department of Urology were described as part of a broader study examining telomere length and anticipation. In these families, 19 families had two or more than two patients in two or more than two generations, including one family with obscure onset age. Therefore, a total of 57 patients with VHL disease from 18 families were enrolled in this study. Using peripheral blood and southern blotting, DNA sequencing was completed on the following patients. Family 10 involved two individuals with a c.533T>G mutation leading to a p.Leu178Arg change. The first individual, proband 10, a male, presented with hemangioblastoma at 38, retinal capillary hemangioma at 57, renal cell carcinoma at 57, and pancreatic cysts at 57. The second individual, the proband’s son, presented with renal cell carcinoma at 30. ACMG Evidence Code: 'PP4' because the patients' phenotypes and family histories are highly specific for VHL disease.

During the period from 2009 to 2012, 39 families with VHL disease were diagnosed with hereditary VHL disease at Peking University First Hospital Department of Urology were described as part of a broader study examining telomere length and anticipation. In these families, 19 families had two or more than two patients in two or more than two generations, including one family with obscure onset age. Therefore, a total of 57 patients with VHL disease from 18 families were enrolled in this study. Using peripheral blood and southern blotting, DNA sequencing was completed on the following patients. Family 3 involved two members with a c.280G>T change leading to a p.Glu94Stop protein change. The first member, a male, presented with hemangioblastoma at age 54 and renal cell carcinoma and pheochromocytoma at age 53. The second member, the proband’s son, presented with papillary cystadenoma of the epididymis at 23, renal cell carcinoma at 29, and pancreatic cysts at 29. Family 12 was additionally found with this mutation. This included a male proband found with hemangioblastomas at 31Y. His mother was also found with hemangioblastoma at 30Y, however she died before diagnosis; thus no DNA sample available. ACMG Evidence Code: 'PP4' because the patients' phenotypes and family histories are highly specific for VHL disease.

During the period from 2009 to 2012, 39 families with VHL disease were diagnosed with hereditary VHL disease at Peking University First Hospital Department of Urology were described as part of a broader study examining telomere length and anticipation. In these families, 19 families had two or more than two patients in two or more than two generations, including one family with obscure onset age. Therefore, a total of 57 patients with VHL disease from 18 families were enrolled in this study. Using peripheral blood and southern blotting, DNA sequencing was completed on the following patients. Family 5 involved four members with a c.499C>T change leading to a p.Arg167Trp protein change. The first member, the father, presented with renal cell carcinoma at 63. His 3 daughters also presented. The first daughter presented with hemangioblastoma, renal cell carcinoma, ovarian cyst, and pancreatic cysts at 37, as well as pheochromocytoma at 39. The second daughter presented with pancreatic cysts at 36 and hemangioblastoma at 38. The third presented with retinal capillary hemangioma at 34. The first born son did not present with any symptoms nor the mutation. ACMG Evidence Code: 'PP4' because the patients' phenotypes and family histories are highly specific for VHL disease.

93 families examined at the Clinical Center of the National Institutes of Health (Bethesda, MD) underwent genetic testing for germline mutations of the VHL gene. Whole blood was extracted and genomic DNA was amplified by PCR and products were sequences using a cycle sequencing kit with dye-labeled terminators and automatically sequenced. This missense mutation was found in a VHL type 1 family (lineage 128). The variant was reported as c.470C>G (AF010238.1) and was mapped to P86R (c.257C>G) (NM_000551.3).

93 families examined at the Clinical Center of the National Institutes of Health (Bethesda, MD) underwent genetic testing for germline mutations of the VHL gene. Whole blood was extracted and genomic DNA was amplified by PCR and products were sequences using a cycle sequencing kit with dye-labeled terminators and automatically sequenced. This missense mutation was found in a VHL type 1 family (lineage 132). The variant was reported as c.440T>C (AF010238.1) and was mapped to F76S (c.227T>C) (NM_000551.3).

93 families examined at the Clinical Center of the National Institutes of Health (Bethesda, MD) underwent genetic testing for germline mutations of the VHL gene. Whole blood was extracted and genomic DNA was amplified by PCR and products were sequences using a cycle sequencing kit with dye-labeled terminators and automatically sequenced. This mutation was found in 2 VHL type 1 families (lineages 136, 183). The variant was reported as c.439_441delTTC (AF010238.1) and was mapped to F76del (c.226_228del) (NM_000551.3). Family 183 is stated as being previously reported in PMID 7728151 (2 families reported with this mutation) and/or 8956040 (7 families reported with this mutation).

In a study of 114 unrelated VHL families, 85 germline mutations were found. VHL mutations were detected in affected family members, but not in unaffected family members or 96 normal individuals. This nonsense mutation was found in a VHL type 1 family of 6 individuals (Family ID 3770), using PCR and SSCP sequencing. The mutation of c.556G>T was referred to in the paper by c.769G>T due to a difference in transcript sequence. All 6 individuals were identified with CNS hemangioblastomas, 4 with retinal hemangioblastomas, and 5 with renal cell carcinoma. Supporting evidence of pathogenicity because of the introduction of an early stop codon in a gene where loss of function is an established mechanism of disease (ACMG code: PVS1).

In a study of 114 unrelated VHL families, 85 germline mutations were found. VHL mutations were detected in affected family members, but not in unaffected family members or 96 normal individuals. This mutation was found in 2 family members with VHL disease (family ID 4402), using PCR and SSCP sequencing. The mutation of c.470C>T was referred to in the paper by c.683C>T due to a difference in transcript sequence. One carried pheochromocytoma and both carried retinal hemangioblastoma.

In a study of 114 unrelated VHL families, 85 germline mutations were found. VHL mutations were detected in affected family members, but not in unaffected family members or 96 normal individuals. This missense mutation was detected in a VHL type 2B patient (family ID 4422), using PCR and SSCP sequencing. The mutation of c.482G>A was referred to in the paper by c.695G>A due to a difference in transcript sequence. The patient had  retinal hemangioblastoma, renal cell carcinoma, and pheochromocytoma. Only supporting evidence for pathogenicity because patient's phenotyoe is highly specific for a disease with a single genetic etiology (ACMG code: PP4).

In a study of 114 unrelated VHL families, 85 germline mutations were found. VHL mutations were detected in affected family members, but not in unaffected family members or 96 normal individuals. This missense mutation was found in 4 VHL type 2 families of 51 individuals altogether (family IDs 2338, 3490, 4477, 4405). 36 of 51 individuals had retinal hemangioblastomas, 17 had CNS hemangioblastomas, 16 with renal cell carcinoma, and 36 with pheochromocytomas. This mutation was also detected in an additional family with 1 single member, with retinal angiomas (family ID 3699). Supporting evidence of pathogenicity because cosegregation with disease in multiple affected family members in a gene definitively known to cause the disease (ACMG code: PP1).

In a study of 114 unrelated VHL families, 85 germline mutations were found. VHL mutations were detected in affected family members, but not in unaffected family members or 96 normal individuals. This frameshift mutation was found in 4 family members with VHL type 1 phenotype (Family ID 3616), using PCR and SSCP sequencing. The mutation of c.462delA was referred to in the paper by c.675delA due to a difference in transcript sequence. Two of 4 members carried retinal hemangioblastomas and 2 with renal cell carcinoma. Supporting evidence of pathogenicity because of the introduction of an early stop codon in a gene where loss of function is an established mechanism of disease (ACMG code: PVS1).

In a study of 114 unrelated VHL affected families, germline mutations in the VHL gene were identified in 85 of the families. VHL mutations were detected in affected family members, but not in unaffected family members or 96 normal individuals, however it seems unaffected family members were not tested genetically for the mutations. Mutations in the VHL gene were detected by SSCP and DNA sequencing. Haplotypes were determined with four microsatellite polymorphic probes. The missense mutation L184P (c.551T>C) (In the paper noted as nucleotide position 764 and codon position 255 due to old reference transcript), was found in 2 individuals in family 3708. Both individuals were identified with renal cell carcinoma and one also presented with a CNS hemangioblastoma. Additional information regarding the family information, including a pedigree, were not provided. ACMG evidence codes: 'flag for PP4' since the patients phenotype and family history is consistent with VHL, 'flag for PP1' because there is co-segregation of variant with the phenotype in two members of the family.

14 Chinese patients from 10 families were diagnosed with Von Hippel-Lindau disease based on clinical criteria. Genetic testing using single-strand conformation polymorphism (SSCP) was conducted and aberrant bands were purified and sequenced. Patient 8, a single individual belonging to family IV was found to harbour a c.388G>T mutation in the VHL gene (stated as c.601G>T in the paper which used the Latif sequence as reference). They presented with hemangioblastoma a pancreatic tumor.

Seventeen patients (age range 3–73) with VHL, including 12 probands and 5 relatives, that were observed between 2005 and 2020 were recruited for a retrospective analysis in relation to pancreatic neuroendocrine tumors (PanNETs). A total of 13/17 patients had PanNETs. In 4 patients, PanNETs were found at VHL diagnosis and in 9 patients they were found during follow-up after a median time of 12 years. Six of these patients underwent surgery and seven were conservatively treated. Four of these patients had lymph node metastases and a tumordiameter significantly larger than the nonmetastatic PanNETs. Five patients who underwent surgery had stable disease after a 3 year follow up, while one patient showed liver metastases. Similarly, six non-resected PanNETs were stable after a 2 year, while one patient developed a novel small PanNET and slight growth of the pre-existing PanNET. Genomic DNA from blood was extracted and high-coverage sequencing for VHL gene was performed using an AmpliSeq custom panel that targets all exons, as well as intron–exon junctions of genes that are frequently mutated in PanNETs (VHL; RET; MEN1). A total of nine different VHL mutations were identified. Three patients were found to be de novo, but authors did not specify who. This proband (Patient 9) presenting with Renal Cell Carcinoma, pancreatic cyst, kidney cyst and no PanNET was found to have a germline nonsense variant R161*(c.481C>T) in exon 3 of the VHL gene.

The patient was a 40 year old Caucasian male with a germline mutation c.257C>T (stated as c.470C>T in the paper which used the Latif sequence as reference) in the VHL gene. The patient presented with renal cell carcinoma, papillary cystadenoma of the epididymis, renal cyst, pancreatic cysts, and hypertension for three years. He had a significant family history of cancer. His father died of renal cell carcinoma at 49, while his brother had a spinal hemangioblastoma that was removed at 19 years of age. His mother had uterine cancer and his paternal grandmother died from pancreatic cancer at age 92. His paternal grandfather died at age 52 from lung cancer and the paternal grandfather's nephew has been diagnosed with a brain tumor. His brother and brother's son were confirmed to have the same VHL gene mutation. The brother presented with spinal hemangioblastoma at age 19 and the son was asymptomatic. However, his mother and twin brothers did not carry the same mutation. The mutation was identified through polymerase chain reaction and direct sequence analysis. Genetic evaluation was performed at The Genetic Diagnostic Laboratory and University of Pennsylvania School of Medicine.

Of 65 VHL families from central Europe, 53 were identified with germline mutations using SSCP analysis of PCR products by peripheral blood. This missense mutation was found in a German, VHL type 2B family. Only one patient was identified with retinal angiomas, hemangioblastomas of the central nervous system, pheochromocytoma, renal cysts, and renal cell carcinoma. The R161G (c.481C>G) mutation was detected in the patient, referred to in the paper by c.694C>G due to a difference in transcript sequence. This patient's phenotype is highly specific for a disease with a single etiology (ACMG code: PP4).

Of 65 VHL families from central Europe, 53 were identified with germline mutations using SSCP analysis of PCR products by peripheral blood. This mutation was found in 2 German, VHL type 1, family members with hemangioblastomas of the central nervous system, renal cell carcinoma, renal cysts, and pancreatic cysts. Hepatic adenomas were also listed as an additional complication (family VHL 44). The L129fs (c.384delT) mutation was detected in the patient, referred to in the paper by c.597delT due to a difference in transcript sequence. This study contains very strong evidence of pathogenicity in the form of a frameshift variant in a gene where loss-of-function is a known mechanism of disease (ACMG code: PVS1).

Patients with ELSTs were identified and included in the International Endolymphatic Sac Tumor Registry. The registry contained 93 patients as of July 2014, and of those, 25 were sporadic, and 68 associated with VHL. Patients with ELSTs and clinically confirmed diagnoses of VHL were included in this study. Statistics of this VHL subgroup are as follows: mean age 30, 62% female, 91% symptomatic, 54% with retinal angioma, 72% with CNS hemangioblastoma, 46% with renal cell carcinoma, 19% with pheochromocytoma, and 12% with pancreatic neuroendocrine tumour. In the data from this study, only endolymphatic sac tumours can be confirmed as a phenotype for this mutation (c.500G>C), which was found in 1 individual from France. 


Patients with ELSTs were identified and included in the International Endolymphatic Sac Tumor Registry. The registry contained 93 patients as of July 2014, and of those, 25 were sporadic, and 68 associated with VHL. Patients with ELSTs and clinically confirmed diagnoses of VHL were included in this study. Statistics of this VHL subgroup are as follows: mean age 30, 62% female, 91% symptomatic, 54% with retinal angioma, 72% with CNS hemangioblastoma, 46% with renal cell carcinoma, 19% with pheochromocytoma, and 12% with pancreatic neuroendocrine tumour. In the data from this study, only endolymphatic sac tumours can be confirmed as a phenotype for this mutation (c.256C>G), which was found in 1 individual from Italy.


Patients with ELSTs were identified and included in the International Endolymphatic Sac Tumor Registry. The registry contained 93 patients as of July 2014, and of those, 25 were sporadic, and 68 associated with VHL. Patients with ELSTs and clinically confirmed diagnoses of VHL were included in this study. Statistics of this VHL subgroup are as follows: mean age 30, 62% female, 91% symptomatic, 54% with retinal angioma, 72% with CNS hemangioblastoma, 46% with renal cell carcinoma, 19% with pheochromocytoma, and 12% with pancreatic neuroendocrine tumour. In the data from this study, only endolymphatic sac tumours can be confirmed as a phenotype for this mutation (c.583C>T), which was found in 2 individuals from France.


A cohort of 36 PCCs and four functional PGL tumours were analysed by exome-seq and high-density SNP-array analysis using tumor paired blood. This specified germline VHL mutation (tumor ID: VCB-PH-05T) belonged to a female diagnosed with recurrent phaeochromocytoma at age 12Y, surgery at 27Y. She also had a personal history of angioma of the right eye, laparotomy & numerous thyrocotomies, and a history of hypertension. The p.T157I (c.470C>T) mutation was also accompanied by a Germline SDHA mutation: c.113A>T (p.Asp38Val). Family history was unknown. Additionally, This patient was previously published in PMID: 26796762.

Molecular and clinical analysis of 126 French VHL families tested for renal involvement revealed 92 different mutations. 90 of these families had renal involvement. The p.L178_D179insRVKPEL (c.531_532insCTGAGAGTAAAGCCTGAA) germline variant of the VHL gene was detected in a family (ID 31). Member 2656 had unknown renal involvement. Members 2511, 4325, 2744 and 2668 presented with mild renal disease (RCC). Members 2588, 2746 and 4630 presented with moderate renal disease (RCC). Members 2060 and 2279 presented with severe renal disease (RCC). The paper states that this family is previously reported in PMID 9829912.

Molecular and clinical analysis of 126 French VHL families tested for renal involvement revealed 92 different mutations. 90 of these families had renal involvement. The p.P86S (c.256C>T) germline variant of the VHL gene was detected in a family (ID 11). Member 319 presented with moderate renal disease (RCC). Members 600, 327, 1066 and 601 presented with severe renal disease (RCC). Members 334 and 332 had unknown renal involvement. Member 286 had no renal involvement. The paper states that this family is previously reported in PMID 12202531 and/or 9829912. Another family (ID 503) was found with this variant. Member 10602 presented with moderate renal disease (RCC) and member 10603 presented with mild renal disease. This family was not previously reported.

Molecular and clinical analysis of 126 French VHL families tested for renal involvement revealed 92 different mutations. 90 of these families had renal involvement. The p.R82P (c.245G>C) germline variant of the VHL gene was detected in a family (ID 12). Members 697, 752 and 2682 presented with moderate renal disease (RCC). Member 4321 presented with severe renal disease (RCC). Members 2062, 606, 607, 609, 444, 3974 and 2523 presented with unknown renal involvement. Member 5357 had no renal involvement. Member 5037 had mild renal disease and member 605 had moderate renal disease (RCC). The paper states that this family is previously reported in PMID 12202531 and/or 9829912.

Molecular and clinical analysis of 126 French VHL families tested for renal involvement revealed 92 different mutations. 90 of these families had renal involvement. The p.Y175* (c.525C>G) germline variant of the VHL gene was detected in a family (ID 63). Member 2656 had unknown renal involvement. Members 2511, 4325, 2744 and 2668 presented with mild renal disease (RCC). Members 2588, 2746 and 4630 presented with moderate renal disease (RCC). Members 2060 and 2279 presented with severe renal disease (RCC). The paper states that this family is previously reported in PMID 12202531 and/or 9829912.

Molecular and clinical analysis of 126 French VHL families tested for renal involvement revealed 92 different mutations. 90 of these families had renal involvement. The p.S80N (c.239G>A) germline variant of the VHL gene was detected in a family (ID 19). Members 697, 752 and 2682 presented with moderate renal disease (RCC). Members 759, 4070, 2596, 6632, 2367, 3255 and 3267 presented with unknown renal involvement. Member 4101 presented with severe renal disease (RCC). Members 2727 and 2728 had no renal involvement. The paper states that this family is previously reported in PMID 12202531 and/or 9829912.

Molecular and clinical analysis of 126 French VHL families tested for renal involvement revealed 92 different mutations. 90 of these families had renal involvement. The p.Y175* (c.525C>A) germline variant of the VHL gene was detected in a family (ID 7). Members 6548 and 6362 had unknown renal involvement. Member 6364 presented with mild renal disease. Member 6728 presented with moderate renal disease (RCC). The paper states that this family is previously reported in PMID 12202531. Another family (ID 320) was found with this variant. Member 7851 presented with severe renal disease (RCC) and member 7547 presented with moderate renal disease (RCC). This family has not been previously reported.

A case report revealed a female proband with a germline mutation on the VHL gene (p.Pro86Ser). Genotyping and sampling methods were unclear. The proband had pancreatic cysts at 34Y and 35Y; renal cysts at 35Y and cerebellar hemangioblastoma at 47Y. Her 28Y son was asymptomatic but additional screening revealed the same germline VHL mutation and a cerebellar hemangioblastoma, pancreatic endocrine tumor, renal cell carcinoma, and pheochromocytoma. The proband's father died from a myocardial infraction at 61Y and her mother was alive with no relevant disease. ACMG: PP4, PP2

Germline sequencing of the VHL gene was performed on patients who presented with VHL syndromic hemangioblastomas, and germline results were compared to sequencing results from matched tumor tissue samples. Patients were only identified by their tumor sample number. Sample 1 (patient 1) had a Gln96_Pro102 deletion (c.287_307del) in the VHL gene. Genetic analysis was conducted as follows: peripheral blood samples were collected from patients, and genomic DNA was extracted, amplified, and purified using ExoSAP IT (ExoSAP IT (Thermo Scientific Inc., MA, USA). Sanger sequencing was performed on samples (Bigdye direct cycle sequencing v3.1, Life Technologies, CS, USA) and capillary electrophoresis was performed on an ABI Genetic Analyzer (ABI 3130xl, Life technologies, CS, USA). Deletion and duplication analysis of the VHL gene spanning the chr3p.25.3 locus was tested by the Multiplex Ligation-dependant Probe Amplification (MLPA) procedure. VHL gene locus-specific probes (P016-C2 VHL) were purchased from MRC Holland (MRC Holland, Amsterdam, Netherlands). ACMG evidence codes: PP4- Patient’s phenotype or family history is highly specific for a disease with a single genetic etiology.

Germline sequencing of the VHL gene was performed on patients who presented with VHL syndromic hemangioblastomas, and germline results were compared to sequencing results from matched tumor tissue samples. Patients were only identified by their tumor sample number. Sample 3 (patient 3) had a Glu94Lys missense mutation (c.280G>A) in the VHL gene. Genetic analysis was conducted as follows: peripheral blood samples were collected from patients, and genomic DNA was extracted, amplified, and purified using ExoSAP IT (ExoSAP IT (Thermo Scientific Inc., MA, USA). Sanger sequencing was performed on samples (Bigdye direct cycle sequencing v3.1, Life Technologies, CS, USA) and capillary electrophoresis was performed on an ABI Genetic Analyzer (ABI 3130xl, Life technologies, CS, USA). Deletion and duplication analysis of the VHL gene spanning the chr3p.25.3 locus was tested by the Multiplex Ligation-dependant Probe Amplification (MLPA) procedure. VHL gene locus-specific probes (P016-C2 VHL) were purchased from MRC Holland (MRC Holland, Amsterdam, Netherlands). ACMG evidence codes: PP4- Patient’s phenotype or family history is highly specific for a disease with a single genetic etiology.

41 non-related patients with retinal capillary hemangioma presenting in the Benjamin Franklin University Eye Clinic between 1988 to 1999 Germany were evaluated for the presence of other VHL lesions. VHL disease was diagnosed in 33 patients based on the presence of other severe VHL lesions (spinal hemangioblastoma, cerebellar hemangioblastoma, renal cysts, pancreatic cyst, renal carcinoma, and pheochromocytoma), molecular genetic testing, and/or family history. VHL germline mutations were identified in 27 out of 29 VHL families screened with molecular genetics. Blood samples were genetically tested using PCR-SSCP sequencing. One patient with VHL-related retinal angiomatosis was found to have germline splicing mutation (c.464-1G>C) mutation in the VHL gene. This variant was reported as c.677-1G>C (AF010238.1) and mapped to c.464-1G>C (NM_000551.3).

41 non-related patients with retinal capillary hemangioma presenting in the Benjamin Franklin University Eye Clinic between 1988 to 1999 Germany were evaluated for the presence of other VHL lesions. VHL disease was diagnosed in 33 patients based on the presence of other severe VHL lesions (spinal hemangioblastoma, cerebellar hemangioblastoma, renal cysts, pancreatic cyst, renal carcinoma, and pheochromocytoma), molecular genetic testing, and/or family history. VHL germline mutations were identified in 27 out of 29 VHL families screened with molecular genetics. Blood samples were genetically tested using PCR-SSCP sequencing. One patient with VHL-related retinal angiomatosis was found to have germline splicing (c.463+3A>G) mutation in the VHL gene. 1 other affected relative was evaluated and found to have the variant but no phenotypes were provided. This variant was reported as c.676+3A>G (AF010238.1) and mapped to c.463+3A>G (NM_000551.3).

75 individuals pre-diagnosed with pheochromocytoma and/or paraganglioma were referred for molecular genetic testing. Genomic DNA of each participant was isolated from peripheral blood samples to look for germline mutations in pheochromocytoma and paraganglioma targeted susceptibility genes. 6 targeted genes (IF1B, RET, SDHB, SDHD, TMEM127, and VHL) were sequenced and identified variants were confirmed by sanger sequencing. Sequencing identified a germline S68P(c.202T>C) missense variant in the VHL gene of 3 patients: 4, 5 and 6. Patient 4 (29, M) was pre-diagnosed with pheochromocytoma (bilateral renal and eyes, pancreatic cysts) and had two affected family members, his mother (without any described phenotype or genotype) and his brother (patient 5). Patient 5 (32Y, M) was pre-diagnosed with pheochromocytoma (right kidney-clear cell carcinoma). Lastly, patient 6 was a 33Y male pre-diagnosed with paraganglioma and liver cysts. This patient's cousin was an affected family member described (without genotype or phenotypes details).

75 individuals pre-diagnosed with pheochromocytoma and/or paraganglioma were referred for molecular genetic testing. Genomic DNA of each participant was isolated from peripheral blood samples to look for germline mutations in pheochromocytoma and paraganglioma targeted susceptibility genes. 6 targeted genes (IF1B, RET, SDHB, SDHD, TMEM127, and VHL) were sequenced and identified variants were confirmed by sanger sequencing. Patient 2 was pre-diagnosed with bilateral pheochromocytoma and sequencing identified a germline R161*(c.481C>T) nonsense variant in the VHL gene. Patient 2 had two affected family members, his father and sister. It does not describe how they were affected nor any diagnosis or variant.

3-year-old white male child presented with hematuria of two days and a CT scan showed a right renal mass determined to be renal cell carcinoma. A nephrectomy was done and the tumour tissue was sent for karyotyping. Peripheral blood samples were also taken and sent for sequence analysis of the VHL gene. Genetic testing confirmed 2 mutations: a translocation between chromosome X and 1 t(X;1)(p11.2; p34.3) and a germline missense mutation, c.562C>G (p.Leu188Val) in the VHL gene. Family history was remarkable for a renal tumour in the paternal grandmother at age 65, treated by nephrectomy in 1998. Furthermore, a maternal uncle had a history of thyroid cancer, and a maternal cousin had a history of breast cancer. The patient’s parents and a 13-year-old half-brother were healthy.

A case report reveals a 3Y girl with right adrenal malignant pheochromocytoma, with the mother diagnosed with von Hippel-Lindau syndrome. The mother was 38Y when diagnosed with bilateral pheochromocytoma that was resected when she was 18Y. She presented several relapses, the most recent one at 32Y when she was also diagnosed with cerebral angioma. Genetic tests revealed the presence of the VHL c 244 C>G (p. Arg 82 Gly) heterozygote mutation in the mother, as well as in the child. Genetic tests for mutations in RET, SDHB, SDHC and SDHD in the mother were negative. After 6 months from the complete resection of the tumor, the daughter is without any clinical symptoms, with normal blood pressure, normal ophthalmoscopy, no tumor markers and no evidence of tumor on cerebral or abdominal MRI.

15 Dutch VHL patients from 5 families were taken to measure the expression of vascular endothelial growth factor (VEGF). Nine patients were found to harbour the VHL germline mutation V170D, referred to in the paper by V241D due to a difference in transcript sequence. Patient 1, a 57 year old female, presented with pancreatic cysts. Patient 4, a 31 year old male, presented with a kidney cyst and a brain hemangioblastoma and no signs of eye lesions. Patient 5, a 28 year old female, presented with a kidney cyst, brain hemangioblastoma, renal cell carcinoma and a left eye angioma. Patient 7, a 28 year old female, presented with a kidney cyst and a right eye angioma. Patient 8, a 28 year old female, presented with pancreatic cysts and a right eye angioma. Patient 9, a 61 year old male, presented with pancreatic and liver cysts, a brain hemangioblastoma and a left eye angioma. Patient 11, a 38 year old male, presented with pancreatic, liver and kidney cysts, brain and spinal cord hemangioblastomas and a right eye angioma. Patient 12, a 50 year old male, presented with kidney & pancreatic cysts, a pheochromocytoma and a brain hemangioblastoma as well as no sign of eye lesions. Patient 13, a 34 year old male, presented with pancreatic cysts, brain and spinal cord hemangioblastomas and right and left eye angiomas. ACMG: 'PP1' since we have cosegregation with the disease in multiple affected family members, 'PP4' since the patient presents phenotypes highly specific for a disease with a single genetic etiology.

53 patients with a molecularly confirmed VHL mutation (by PCR through EDTA-anticoagulated blood) and a pancreatic neuroendocrine tumor (ICT) were collected from the German NET-Registry and the German VHL-registry. An additional 296 patients were stated in this paper with a VHL associated germline mutation, but without pancreatic neuroendocrine tumors. 2 patients were found to have the above mutation (c.208G>T). 1 was a VHL patient with an ICT (pancreatic neuroendocrine tumor) taken from the VHL-registry. The other patient was stated to be in this study with the given mutation, however no ICT. Other clinical manifestations outside of a pancreatic neuroendocrine tumor are possible, but were not discussed. ACMG evidence codes: 'PVS1' because they observe a nonsense variant in a gene where loss of function is a known mechanism of disease.

53 patients with a molecularly confirmed VHL mutation (by PCR through EDTA-anticoagulated blood) and a pancreatic neuroendocrine tumor (ICT) were collected from the German NET-Registry and the German VHL-registry. An additional 296 patients were stated in this paper with a VHL associated germline mutation, but without pancreatic neuroendocrine tumors. 2 patients were found to have the above mutation (c.472C>G). 1 was a VHL patient with an ICT (pancreatic neuroendocrine tumor) taken from the VHL-registry. The other patient was stated to be in this study with the given mutation, however no ICT. Other clinical manifestations outside of a pancreatic neuroendocrine tumor are possible, but were not discussed. ACMG evidence codes: 'PP2' because they observe a missense variant in a gene where missense variants are a common mechanism of disease.

53 patients with a molecularly confirmed VHL mutation (by PCR through EDTA-anticoagulated blood) and a pancreatic neuroendocrine tumor (ICT) were collected from the German NET-Registry and the German VHL-registry. An additional 296 patients were stated in this paper with a VHL associated germline mutation, but without pancreatic neuroendocrine tumors. 5 patients were found to have the above mutation (c.548C>A). 1 was a VHL patient with an ICT (pancreatic neuroendocrine tumor) taken from the VHL-registry. The other 4 patients were stated to be in this study with the given mutation, however no ICT. Other clinical manifestations outside of a pancreatic neuroendocrine tumor are possible, but were not discussed. ACMG evidence codes: 'PVS1' because they observe a nonsense variant in a gene where loss of function is a known mechanism of disease .

53 patients with a molecularly confirmed VHL mutation (by PCR through EDTA-anticoagulated blood) and a pancreatic neuroendocrine tumor (ICT) were collected from the German NET-Registry and the German VHL-registry. An additional 296 patients were stated in this paper with a VHL associated germline mutation, but without pancreatic neuroendocrine tumors. 1 patient was found to have the above mutation (c.277G>C). Other clinical manifestations outside of a pancreatic neuroendocrine tumor are possible, but were not discussed. ACMG evidence codes: 'PP2' because they observe a missense variant in a gene where missense variants are a common mechanism of disease.

37 Danish patients diagnosed with VHL before 18 years from a prospective cohort study from Denmark's National vHL Research Database (Binderup et al., 2013) were assessed. One family with 1 patient (Family ID8) were found with 2 germline missense mutations in the VHL gene: (L118P; c.353T>C) and (R107G; c.319C>G). Patient 1 was diagnosed at 15Y old and had 6 CNS hemangioblastomas at 15Y, and 2 retinal hemangioblastomas at 15Y. The patient was 35Y by the end of the study.

A cohort of VHL patients from the Greater Toronto Area in Canada were retrospectively reviewed. Patients ID 3 and 25 had this germline mutation (p.D92V). Patient 3 had: CNS hemangioblastoma. Patient 25 pheochromocytoma/paraganglioma (recorded here as a neuroendocrine neoplasm). Whether patients were related, age, and sex data was not reported. ACMG PP4

A cohort of VHL patients from the Greater Toronto Area in Canada were retrospectively reviewed. Patient ID 12 had this germline mutation (c.463+2T>G) and clear cell renal cell carcinoma and pancreatic endocrine tumor. Whether patients were related, age, and sex data was not reported. ACMG PP4

A 17-year-old girl was referred to the Department of Endocrinology at La Rabta University Hospital for the investigation of a newly diagnosed diabetes mellitus and a severe arterial hypertension. The patient is the first child of consanguineous parents and has a family history of type 2 diabetes and dyslipidemia, but no conditions that may suspect VHL disease. Blood tests disclosed hypercalcemia and abdominal computed tomography showed bilateral adrenal tumors. A clinical diagnosis of multiple endocrine neoplasia type 2a (MEN2A) was originally suspected but genetic analysis (not further detailed) revealed the missense mutation c.191G > C (p.Arg64Pro) in exon 1 of the VHL gene. The authors concluded that the presence of pheochromocytomas and the missense mutation in VHL gene suggests type 2 VHL disease. She was further diagnosed with dyslipidemia and primary hyperparathyroidism. The patient underwent a bilateral adrenalectomy marked by normalization of blood pressure, blood glucose, calcium, and PTH levels.

Fourty-four of 389 patients with VHL (between 1988-1999) screened positive for pancreatic neuroendocrine tumours (PNETs) either by pathologic analysis of tissue specimens or by characteristic radiographic appearance on CT and MRI. VHL gene germline mutations were determined by quantitative Southern blotting to detect deletions of the gene and gene rearrangements as well as complete gene sequencing. Five patients presented with the c.500G>A VHL gene mutation (stated as 713 G to A in the paper which used the Latif sequence as reference) causing a change of Arg to Gln at protein position 167. From family 5960, patient 38 (41-year-old male) presented with a single radiologic PNET and patient 39 (39-year-old female) presented with a single pathologic PNET. From family 5069, patient 41 (48-year-old male) presented with five pathologic PNETs, and patient 42 (16-year-old male) presented with a three pathologic PNETs. Unrelated patient 40 (31-year-old female) presented with a single pathologic PNET. No ACMG code.

216 patients with clinically expected VHL disease due to family history or presence of VHL typical tumours were routinely examined in an eye centre in Germany between January 2019 and January 2020, majority of which were of European descent (~95%). Since data on the genotype–phenotype correlation of the onset of Retinal Hemangioblastoma (RH) is scarce in relation to VHL disease, this study wanted to inversitage the relationship between VHL gene variants (truncating vs single amino acid substitutions) and the ocular phenotype of the disease. 42 different rare VHL gene variants in 148 patients were detected - some of whom had RH and some who did not. Although date of onset of RH, as well as data on central nervous system haemangioblastomas (CNSH), pheochromocytoma/paraganglioma (PPGL), clear cell renal cell carcinoma (ccRCC) and pancreatic neuroendocrine neoplasm (PNEN) were collected, it was not reported on a per patient basis and therefore could not be discussed per variant. Out of the 3 patients with a p.L89P (c.266T>C) mutation, 67% (2/3) of individuals had RH while the other 33% (1/3) did not. No family information was provided.

216 patients with clinically expected VHL disease due to family history or presence of VHL typical tumours were routinely examined in an eye centre in Germany between January 2019 and January 2020, majority of which were of European descent (~95%). Since data on the genotype–phenotype correlation of the onset of Retinal Hemangioblastoma (RH) is scarce in relation to VHL disease, this study wanted to inversitage the relationship between VHL gene variants (truncating vs single amino acid substitutions) and the ocular phenotype of the disease. 42 different rare VHL gene variants in 148 patients were detected - some of whom had RH and some who did not. Although date of onset of RH, as well as data on central nervous system haemangioblastomas (CNSH), pheochromocytoma/paraganglioma (PPGL), clear cell renalcell carcinoma (ccRCC) and pancreatic neuroendocrine neoplasm (PNEN) were collected, it was not reported on a per patient basis and therefore could not be discussed per variant. There was 1 patient reported with c.499C>T (p.R167W). This member presented with retinal hemangiomas. No family information was provided.

301 were enrolled in a prospective clinical protocol to evaluate the natural history of vHL-associated pancreatic lesions (NCT00062166). Of these, 69 patients with pancreatic manifestations of vHL disease and missense VHL mutations were included in the current study. Individual patient phenotype was not described, but overall of the 60 patients, 57 had RCC, 41 had CNS hemangioblastomas, 28 had retinal hemangioblastoma, 23 had pheochromocytoma. 36/69 patients were female and the mean age was 50.5 years old. Four patients were listed with this missense mutation, c.256C>T. All patients underwent genetic testing using Sanger sequencing and Southern Blotting.

Of 246 patients affected with von Hippel-Lindau disease, 64 patients (26%) from 38 families had pheochromocytoma, including 33 newly diagnosed during screening studies and 31 treated before screening at NIH. Germline mutations in the VHL gene was determined from peripheral blood. Family 4339 were found with p.Arg176fs mutation (legacy notation c.739delA converted to NM_000551.3) in the VHL gene. Family 4339: 1/4 had pheochromocytoma, 4/4 had renal cell carcinoma, 1/4 had CNS hemangioblastoma, 1/4 had retinal angiomas, 1/4 had cysts. Individual patient phenotype-genotype was not described in the paper.

Of 246 patients affected with von Hippel-Lindau disease, 64 patients (26%) from 38 families had pheochromocytoma, including 33 newly diagnosed during screening studies and 31 treated before screening at NIH. Germline mutations in the VHL gene was determined from peripheral blood. Family 4783 were found with a p.Leu128Phe mutation (legacy notation c.595C>T converted to NM_000551.3) in the VHL gene. Family 4783: 3/3 had pheochromocytoma, 1/3 had retinal angiomas and pancreatic neuroendocrine tumor was also found in the family (number of affected members with PNET was not described). Individual patient phenotype-genotype was not described in the paper.

A study investigated paraganglioma/pheochromocytoma tumors in a retrospective cohort and a prospective cohort. Tumour tissue samples were sequenced by next-generation sequencing using a custom gene panel. In some cases, Sanger sequencing was used to detect germline mutations. Some of these tumors were found with germline VHL mutations. Patient ID 100, from the prospective cohort, presented with multiple paragangliomas and a germline mutation in the VHL gene c.191G>C. Other phenotypic and demographic information was not available. The paper performed carbonic anhydrase IX immunohistochemistry and immunostaining with CA9 was observed in patient 100.

A study investigated paraganglioma/pheochromocytoma tumors in a retrospective cohort and a prospective cohort. Some of these tumors were found with germline VHL mutations. Tumour ID 47 was bilateral PCC and a germline c.467A>C mutation in the VHL gene. Other phenotypic and demographic information was not available.

A study investigated paraganglioma/pheochromocytoma tumors in a retrospective cohort and a prospective cohort. Some of these tumors were found with germline VHL mutations. Tumour ID 40 and ID 41 (both from a 15 years old patient presenting with multiple benign tumors), and ID 42 (from a 40 years old patient presenting with multiple benign tumors) were PCC and found with a germline mutation in the VHL gene p.Leu178Pro. It is not clear if tumour samples 40 and 41 were taken from the same patient at 2 different time points or different patients. Other phenotypic and demographic information was not available.

The study assessed the long-term prognosis of VHL and non-VHL patients among 110 consecutive patients with haemangioblastoma (HB) of the CNS treated between 1953 and 1993 at one neurosurgical unit. 14 patients had VHL disease, but only the genotype of the patients was only described for case 8 (NM_000551.3:c.278G>C) and 11 (NM_000551.3:c.293A>G). One of these two patients had pheochromocytoma in a family member, but it is unclear which patient. Case 8 had a germline p.Gly93Ala mutation. Genotyping methods included PCR and direct sequencing. Additional phenotypes and family history were not provided on an individual level for VHL patients.

This report studied the prevalence of germline variants in Japanese PPGL patients. Of the 370 PPGL original Japanese probands, this cohort consisted of 15 patients with pheochromocytoma and/or paragangliomas with germline VHL varaints. Blood samples were used to examine 7 genes: MAX, SDHB, SDHC, SDHD, TMEM127, VHL, and RET, using Sanger sequencing. Of these 15 VHL patients: 10 had bilateral PCC, 4 had PCC with abdominal/thoracic PGL, and one had metastatic PPGL. The variant R167W (c.499C>T) was one of the 15 found in a patient with PPGL. Their family history or further manifestations were not detailed.

This paper analyzed the VHL gene of 21 patients who had clinical a clinical diagnosis of VHL, familial pheochromocytoma, or sporadic pheochromocytoma. 7 patients were from 3 families, while the remaining 14 were unrelated. Mutations were detected by realtime quantitative PCR (RQ-PCR) assays using peripheral venous blood. The mutation (c.499C>T) was found in 1 individual (33Y, M), without family history, who presented with pheochromocytoma, hypertension and diabetes mellitus. This case was also later presented in PMID: 24735130 (Pai et al, 2014). ACMG evidence codes: 'PP2' because they observe a missense variant in a gene where missense variants are a common mechanism of disease.

Peripheral blood from unrelated patients with pheochromocytoma was tested for mutations of proto-oncogene RET, tumor suppressor gene VHL, succinate dehydrogenase subunit D (SDHD) gene, and the succinate dehydrogenase subunit B (SDHB) gene. Of 271 patients with nonsydromic pheochromocytoma and no family history of the disease, 30 mutations were found in VHL. Multiple pheochromocytomas were statistically more frequent among patients with mutations than among patients without mutations (21 of 66 vs. 5 of 205, P<0.001). The mutation R161P (c.482G>C) was detected in the patient, referred to in the paper by c.695G>C due to a difference in transcript sequence. This missense mutation was found in the VHL gene and one patient with pheochromocytoma.

The case report details a 26-year-old male (case 4) presenting with cerebellar hemangioblastoma. Four years later, he was diagnosed with pancreatic endocrine tumour, pheochromocytoma, and pancreatic cysts. Mutation analysis reveals a c.500G>A change. The patient had family history of VHL. ACMG code "PS1", as this is the same mutation as a previously described pathogenic variant.

The medical records of 78 Korean patients with pheochromocytoma (PCC) and paragangliomas (PGL) were examined, 61 patients were diagnosed with PCC and the other 17 patients with PGL. 57 patients were included in the study. After excluding patients, 57 patients underwent sequencing. 14 patients received Sanger sequencing in pre-NGS period, while the other 43 patients underwent genetic testing by either direct sequencing (n=2) or targeted NGS (n=41) in the post-NGS period. 28 patients were found to have germline mutations in 7 PPGL-related genes (SDHB, RET, VHL, NF1, MAX, SDHA, and SDHD). Out of the 28 patients, 6 patients had germline mutations in VHL (5 presented with Pheochromocytoma and 1 with Paraganglioma). This patient was diagnosed with PCC due to bilateral adrenal mass and was found to have a R161Q(c.482G>A) missense variant in the VHL gene, which was detected by targeted NGS.

Tumor registries from four paediatric referral centers (Lucile Salter Packard Children’s Hospital, Childrens Hospital of Los Angeles, the Johns Hopkins Hospital, and Childrens Hospital of Buffalo) were screened for all patients < / = 21 years at diagnosis of a cerebellar hemangioblastoma. Peripheral blood leukocytes were sampled and partial or entire deletions of the VHL gene or gene rearrangements were detected by quantitative Southern blot analysis. Genomic DNA was amplified by PCR, and exons 1, 2, and 3 of the VHL gene directly sequenced using SSCP. One patient with no family history for VHL was found with this c.341-1G>A mutation. The patient was 16.1Y old at cerebellar hemangioblastoma diagnosis, and also presented with retinal angioma, cervical spinal hemangioblastoma, pancreatic cysts, and renal cysts.

43 Italian patients were molecularly analyzed for Von Hippel-Lindau disease based on clinical suspicion. Peripheral blood samples were directly sequenced on an automatic sequencer (ABI 3100, Applied Biosystems, Foster City, CA) and real-time quantitative PCR (RQ-PCR) was used to detect partial or complete VHL gene deletions. One of the 43 patients (Patient ID 1) was found to have the mutation c.337C>T leading to an early stop codon. The patient had multiple hemangioblastomas and renal cell carcinoma. This patient had family history of the disease and was classified as A1 by the authors (families meeting current VHL diagnostic criteria with involvement of at least three VHL tumors in two distinct organs).

Eighteen VHL patients who underwent pancreatectomy for one or several endocrine tumors at Beaujon hospital from 1997 to 2008 were retrieved from the file of VHL patients. All patients had VHL germline mutations. Age is at the time of surgery. Patients #15 (male) was found with this germline VHL mutation (p.R82P (c.245G>C)) and pancreatic endocrine tumours. The last known age of the patient was 48Y. Other phenotypes and demographics were unknown. ACMG PP4

Eighteen VHL patients who underwent pancreatectomy for one or several endocrine tumors at Beaujon hospital from 1997 to 2008 were retrieved from the file of VHL patients. All patients had VHL germline mutations. Age is at the time of surgery. Patients #2 (female) and #12 (male) were found with this germline VHL mutation and pancreatic endocrine tumours. The last known age of the patients was 28Y, and 43Y respectively. Other phenotypes and demographics were unknown. ACMG PP4

Study of a German family of 16 family members with cosegregation of the pheochromocytoma-only phenotype with two concurrent germline mutations in the VHL protein alpha and beta domains. These mutations were c.562 C>T (stated as 775C>G in the paper which used the Latif sequence as reference) and c.241C>T (stated as 454C>T in the paper). Patient 1 was unaffected and had the wild-type allele. His spouse, patient 2, presented with pheochromocytoma and had both mutations. They had four children; one had a pheochromocytoma and died, two (patients 5 and 7) had both mutations and presented with pheochromocytomas, and one child (patient 6) was unaffected and carried neither mutation. Patient 7 had a child (patient 9) who also carried both mutations and presented with pheochromocytoma. In the other branch of the family, patient 3 who carried both mutations and presented with pheochromocytoma had a child with patient 4 who was unaffected. Their child (patient 8) carried both mutations and presented with pheochromocytoma. Patient 8 had two children; one who did not carry the mutation and was unaffected (patient 10) and another that carried both mutations but was disease-free (patient 11). DNA was isolated with whole blood and denaturing high-performance liquid chromatography (DHPLC) and dye terminator method on an ABI Prism 310 genetic analyzer (Applied Biosystems, Weiterstadt, Germany) were used for sequencing. Cosegregation with disease in multiple affected family members in a gene definitively known to cause the disease (ACMG code: PP1).