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.F136S(c.407T>C) was identified in patient #45. This mutation was noted as c.620T>C in the paper due to a difference in reference transcript used (see Table S1). No family information is available for this patient.

A case report of a 42-year-old woman genetically confirmed with c.500G>A (p.Arg167Gln) in the VHL gene and type 2A VHL disease presented with two CNS hemangioblastomas (brain and spinal cord), left pheochromocytoma, pancreatic neuroendocrine tumors, and presented with a parotid mucoepidermoid carcinoma at age 42. The R167Q mutation was previously diagnosed using a blood test at age 34. Using the TruSeq Amplicon Cancer Panel to sequence the patient's tumor specimen, it was found that the patient had a second somatic VHL mutation at codon 105 (p.T105M). The patient had a positive family history, with her mother and children having VHL disease; no additional information on phenotype and genotype for family members was provided.

100 patients (age: 13–95 years) who had either one or two non-syndromic pheochromocytoma without a positive familial history, without any other endocrine tumours, without any clinical signs for von Hippel–Lindau disease, MEN type 2 or neurofibromatosis type 1, were analyzed. Two patients in the study (Patient number 1 and 5) were found to both have a germ line mutation in the VHL gene, p.Tyr156Cys. The specific nucleotide change was not specified. Genetic analysis was conducted in search of a germ-line mutation in VHL by PCR-sequencing on both strands of DNA. Patient 1 was a 13-year old male who had a bilateral pheochromocytoma and Patient 5 was a 43-year old female who had a unilateral pheochromocytoma.

At 46Y, the male proband underwent surgery for pheochromocytoma associated with malignant hypertension. 20Y later at 66Y old, the proband experienced gait disturbance due to cerebellar dysfunction for 2 years and was found with meningioma and cerebellar hemangioblastoma. Later he also developed RCC and died from multiple cerebral infarctions and RCC. Genetic analysis included SSCP analysis of peripheral blood and revealed a germline VHL mutation NM_000551.3:c.472C>G. Please note that the paper describes it as c.685C>G because it uses a legacy reference system. His older sister passed away at 30Y old due to a cerebellar tumour. The proband's son was also confirmed with the same VHL mutation and had cerebellar hemangioblastoma.

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 (R167G (c.499C>G)) was taken from the Cambridge, Department of Pathology database, with an unknown number of members belonging to Family ID 137CA. It was reported in the paper as c.712C>G 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.

A case report of a male VHL patient presenting with retinal hemangioblastoma at the age of 15 was described. Further clinical examination revealed cerebellar hemangioblastoma and a pituitary adenoma. Genetic testing revealed the presence of the c.340G>C (p.Gly114Arg) germline mutation of the VHL gene. Family history revealed 3 additional family members with a clinical diagnosis of VHL. Clinical manifestations present in these members included: cerebellar hemangioblastoma, retinal hemangioblastoma, renal cell carcinoma, and pancreatic cysts.

A 35 year old woman was admitted with long-standing dyspepsia and pancreatic cysts. There were no other symptoms. The past medical and family histories were not of significance. Imaging studies showed the presence of multiple cysts in the head and tail of the pancreas suggesting pancreatic serous cystadenomas. Cysts were also found within the left kidney and spleen. MRI of the brain and entire spine found no hemangioblastomas and abdominal MRI scan excluded renal carcinoma. Pheochromocytoma, angiomas of the retina and endolymphatic sac tumors were also excluded. Genetic testing revealed a germline missense mutation N90I (c.269A>T) in the VHL gene. This variant was also found in her 7 year old daughter. Phenotypic information for the daughter was not described. Authors state this variant as being likely de novo in the proband.

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. Eight patients presented with the c.499C>T VHL gene mutation (stated as 712 C to T in the paper which used the Latif sequence as reference) causing a change of Arg to Trp at protein position 167. From family 3407, patients 30 (50-year-old male) and 31 (33-year-old male) both presented with a single radiologic PNET. From family 2301, patient 32 (34-year-old female) presented with two pathologic PNETs with metastases, and patient 33 (20-year-old female) presented with a single pathologic PNET with metastases. Unrelated patients 34 (43-year-old female), patient 35 (20-year-old female), and patient 36 (27-year-old female), all presented with three pathologic PNETs. Unrelated patient 37 (46-year-old female) presented with a single pathologic PNET. ACMG evidence codes: 'PP1' since we have co-segregation with the disease in multiple affected family members (family 3407, 5069, and 2301), 'PP4' since the patients' present phenotypes highly specific for a disease with a single genetic etiology.

Fourty-four of 389 patients with VHL (between 1988-1999) screened positive for 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. Patient 18, a 19 year old female, was found to harbour the c.257C>G mutation in the VHL gene (stated as 470 C to G in the paper which used Latif sequence as reference) leading to an amino acid change of Pro to Arg at protein position 86. She presented with a single radiologic pancreatic neuroendocrine tumor. ACMG evidence codes: 'PP4' since the patient presents phenotypes highly specific for a disease with a single genetic etiology.

Fourty-four of 389 patients with VHL (between 1988-1999) screened positive for 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. Patient 29 (29-year-old male) was found to harbour the c. 486C>G mutation in the VHL gene (stated as 699 C to G in the paper which used the Latif sequence as reference) and presented with two pathologic pancreatic neuroendocrine tumors. ACMG evidence codes: 'PP4' since the patient presents phenotypes highly specific for a disease with a single genetic etiology.

Fourty-four of 389 patients with VHL (between 1988-1999) screened positive for pancreatic neuroendocrine tumors (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. Patient 43 was found to have the gemline VHL mutation c.562C>G (stated as 775 C to G in the paper which used the Latif sequence as reference) leading to a Leu to Val change at protein position 188. She presented with four pathologic pancreatic neuroendocrine tumors at age 35. She was also found to harbour the c.241C>T (stated in the paper as 454 C to T) VHL gene mutation leading to a Pro to Ser change at protein position 81. ACMG evidence codes: 'PP4' since the patient presents phenotypes highly specific for a disease with a single genetic etiology.

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 W117* (c.351G>A) was referred to in the paper by c.564G>A due to a difference in transcript sequence.  This nonsense mutation found in a VHL family without PC (family ID 97). Only moderate evidence for pathogenicity because protein length changes as a result of stop-loss variant (ACMG code: PM4).

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 c.214delT was referred to in the paper by c.427delT due to a difference in transcript sequence. This deletion mutation was found in a VHL family without PC (family ID 112). Only moderate evidence for pathogenicity because protein length changes as a result of in-frame deletion in a nonrepeat region or stop-loss variant (ACMG code: PM4).

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 R177* (c.499C>T) was referred to in the paper by c.712C>T due to a difference in transcript sequence. This missense mutation was found in one of the VHL families (family ID 153).

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 V166F (c.496G>T) was referred to in the paper by c.709G>T due to a difference in transcript sequence. This missense mutation was found in a VHL family with PC (family ID 50).

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 splice mutation c.340+5G>C was referred to in the paper by cc.553+5G>C due to a difference in transcript sequence. This mutation was found in a VHL family without PC (family ID 139). The mutation has a large allele frequency meaning this is likely non disease causing (not deleterious).

A case report of a woman with VHL presenting with a malignant pheochromocytoma at the age of 22 was described. 34 years later, she presented to the emergency department in 2008 with a hypertensive crisis characterized by 3 days of severe headache, malignant hypertension and seizures complicated by bilateral humeral fractures. Computed tomography and magnetic resonance imaging (MRI) scans showed large bilateral renal angiomyolipomas (AMLs), a right renal artery aneurysm, a bulky left adrenal gland with no discrete mass lesion, and a 15 mm calcified lesion within the left anterior mediastinum (not related to the sympathetic chain). Histological examination confirmed a pheochromocytoma metastasis within a lymph node. After the operation, blood pressure improved significantly. Further investigations of the patient have revealed mild sensorineural hearing loss. No evidence of hemangioblastomas via fundoscopy or on MRI scanning of the brain and spinal cord. Recent imaging has shown no change in the patient's presumed bilateral AMLs indicating the absence of renal cell carcinoma. The patient was found to carry the germline mutation Gly144Arg of the VHL gene. Two of her three children have inherited the mutation, they have had no disease manifestations.

Tissue analysis from 61 VHL patients, 30 sporadic renal cell carcinoma (RCC) tumor samples, and 6 sporadic RCC cell lines revealed 22 variants within VHL patients, 10 variants in tumor samples and variants in all cell lines. All mutations cluster to the 3’ end of the VHL gene open reading frame, implicating this region as important for the tumor-suppressor function of the VHL protein. Northern blot, SSCP and RT-PCR analysis from normal tissue found a germline p.L198R (c.593T>G) mutation, stated in the paper as c.806T>G due to an older transcript. It was seen in family [pedigree No] 7, with an unknown number of members described as ‘VHL-affected’ with RCC. Other manifestations are possible but not clear.

Tissue analysis from 61 VHL patients, 30 sporadic renal cell carcinoma (RCC) tumor samples, and 6 sporadic RCC cell lines revealed 22 variants within VHL patients, 10 variants in tumor samples and variants in all cell lines. All mutations cluster to the 3’ end of the VHL gene open reading frame, implicating this region as important for the tumor-suppressor function of the VHL protein. RT-PCR analysis in RCC cell lines found a p.G104Afs*14 (c.310delG) mutation, stated in the paper as c.523del1 due to an older transcript. It was seen in cell line 786-O. Any family information or other manifestations apart from RCC were unclear.

Tissue analysis from 61 VHL patients, 30 sporadic renal cell carcinoma (RCC) tumor samples, and 6 sporadic RCC cell lines revealed 22 variants within VHL patients, 10 variants in tumor samples and variants in all cell lines. All mutations cluster to the 3’ end of the VHL gene open reading frame, implicating this region as important for the tumor-suppressor function of the VHL protein. Northern blot, SSCP and RT-PCR analysis from normal tissue found a germline p.S183* (c.548C>A) mutation, stated in the paper as c.761C>A due to an older transcript. It was seen in family [pedigree No] 22, with an unknown number of members described as ‘VHL-affected’ with RCC. Other manifestations are possible but not clear.

Tissue analysis from 61 VHL patients, 30 sporadic renal cell carcinoma (RCC) tumor samples, and 6 sporadic RCC cell lines revealed 22 variants within VHL patients, 10 variants in tumor samples and variants in all cell lines. All mutations cluster to the 3’ end of the VHL gene open reading frame, implicating this region as important for the tumor-suppressor function of the VHL protein. Northern blot, SSCP and RT-PCR analysis from normal tissue found a germline c.Y98H (c.292T>C) mutation, stated in the paper as c.505T>C due to an older transcript. It was seen in 2 families: [pedigree No] 14 and 15, with an unknown number of members described as ‘VHL-affected’ with RCC. Other manifestations are possible but not clear.

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 (L129_V130insE (c.386_387insAGA)) was taken from the National Cancer Institute database, with an unknown number of members belonging to Family ID 4410NCI. It was reported in the paper as c.599insE 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 (c.340+5G>C) was taken from the Cambridge, Department of Pathology database, with an unknown number of members belonging to Family ID 139CA. It was reported in the paper as c.553+5G>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 (Y112H (c.334T>C)) was taken from the National Cancer Institute database, with an unknown number of members belonging to two family IDs: Family ID 3738NCI and 4874NCI. It was reported in the paper as c.547T>C 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 PMIDs: 7728151 and 8863170.

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 (I180V (c.538A>G)) was taken from the Cambridge, Department of Pathology database, with an unknown number of members belonging to Family ID 79CA. It was reported in the paper as c.751A>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 (Q164* (c.490C>T)) was taken from the Munich, university of Freiburg and Institute of Pathology database, with an unknown number of members belonging to Family ID 65CE. It was reported in the paper as c.703C>T 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 (Q73* (c.217C>T)) was taken from the Cambridge, Department of Pathology database as well as the National Cancer Institute database, with an unknown number of members belonging to Family ID 38CA, 74CA, and 2753NCI. It was reported in the paper as c.430C>T 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. Later publications reference this cohort as being reported in PMIDs: 7728151 and 9829911.

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 (F136S (c.407T>C)) was taken from the Neuro Olcologie database in Paris, and the Cambridge, Department of Pathology database. It had an unknown number of members belonging to Family ID 53F, reported in Table 3, as being found in VHL families without pheochromocytoma, and with renal cell carcinoma. Additionally, it was reported in table 5, specified as families without detailed clinical information, under family ID 27CA. It was reported in the paper as c.620T>C due to a difference in transcript reference.

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 (c.464-2A>G) was taken from the Bristol Myers Squibb database, and the Munich, university of Freiburg and Institute of Pathology database. It had an unknown number of members belonging to Family ID 31CE, reported in Table 3, as being found in VHL families without pheochromocytoma, and with renal cell carcinoma. Additionally, it was reported in table 5, specified as families without detailed clinical information, under family ID 28BMS. It was reported in the paper as c.677-2A>G due to a difference in transcript reference.

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 (L188V (c.562C>G)) was taken from the Munich, university of Freiburg and Institute of Pathology database, with an unknown number of members belonging to two family IDs: Family ID 37CE, and 38CE. It was reported in the paper as c.775C>G 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.

A study retrospectively reviewed medical records of 710 patients diagnosed with PC/PGL for associated renal neoplasia and primary renal PGLs, at Mayo Clinic, Rochester, MN. Three patients that met inclusion criteria were found with VHL mutations, only 2 of which had documented pathogenic mutations in the VHL gene. One patient (Patient 13) was a 33 year old male with a germline c.191G>C transversion resulting in a p.Arg64Pro protein change presenting with bilateral pheochromocytoma, and ccRCC.

Germline mutations were found in 20 Brazilian, VHL probands and their families. The E94* (c.280G>T) germline nonsense mutation was found in a VHL type 1 family of 3 affected individuals. All three family members had retinal angiomas and 2 also had hemangioblastomas of the central nervous system (VHL family 4).

Germline mutations were found in 20 Brazilian, VHL probands and their families. This missense mutation was found in 2 VHL family members (VHL family 3). One had hemangioblastomas of the central nervous system and one had retinal angiomas. This variant was not found in 120 alleles from normal subjects in the Brazilian population.

Germline mutations were found in 20 Brazilian, VHL probands and their families. The S80R (c.238A>C) germline missense mutation was found in a VHL type 1 family of 10 affected individuals (VHL family 2). Seven had hemangioblastomas of the central nervous system, 3 had retinal angiomas, and 3 had renal carcinoma. This variant was not found in 120 alleles from normal subjects in the Brazilian population.

A 68Y male patient presented with cerebellar hemangioblastoma, retinal hemangioblastoma, clear cell renal cell carcinoma, and serous cystadenoma. At 73Y he developed adrenal pheochromocytoma and at 74 he developed a clear cell renal cell carcinoma metastasis in the thyroid. Genetic testing confirmed a mutation NM_000551.3:c.245G>C (p.Arg82Pro) in the VHL gene which was confirmed in 3 other family members. Please note that no specific method of genetic testing was described in the paper. One was his 10Y grandson which was asymptomatic, his 46Y son that had renal cell carcinoma and pancreatic cysts or serous cystadenoma, a 47Y male distant relative had CNS hemangioblastoma, renal cell carcinoma, symptomatic pheochromocytoma, and pancreatic cysts/serous cystadenoma, and a deceased 37Y female distant relative had retinal hemangioblastoma. The paper also asserted that 7 other relatives were carriers of the mutation; 4 other relatives had CNS hemangioblastoma. It should be noted that genetic testing was not conducted on these seven relatives.

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 V166F (c.496G>T) was one of the 15 found in a patient with PPGL. Their family history or further manifestations were not detailed.

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.V166F (c.496G>T) germline variant of the VHL gene was detected in a family (ID 49).Member 2046 had no renal involvement. Member 1258 presented with mild renal disease. 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.V165Lfs*5 (c.492del) germline variant of the VHL gene was detected in an individual (ID 9572, Family 405) who presented with mild renal disease (simple renal cyst, multicystic kidney[s], small lesion [less than 10 mm in size] of unknown significance, or small solid tumor without indication for surgical removal [less than 30 mm in size]).

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.F76del (c.226_228del) germline variant of the VHL gene was detected in a family (ID 44). Member 4975 presented with moderate renal disease (RCC), members 4978, 4977 and 4976 presented with mild renal disease and member 5283 had unknown renal involvement.

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.C162W (c.486C>G) germline variant of the VHL gene was detected in 3 individuals (ID 1114/Family 236, ID452/Family 345 and ID 9035/Family 379) who all presented with renal cell carcinoma. The paper states that these individuals were all 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.Q73* (c.217C>T) germline variant of the VHL gene was detected in a family (ID 64). Members 2339, 2333 and 2338 presented with severe renal disease (RCC). Members 2331, 2329 and 1345 had unknown renal involvement. Member 2340 had mild renal disease. Member 2341 had no renal involvement. The paper states that this family is previously reported in PMID 12202531, where 4 patients with this mutation are reported but relation between patients is unknown. This variant was also found in another family (ID 84). Member 4605 presented with mild renal disease, member 4580 presented with moderate renal disease (RCC) and members 4581 and 4603 had unknown renal involvement. 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.R161* (c.481C>T) germline variant of the VHL gene was detected in a family (ID 41). Members 4732 and 1426 had unknown renal involvement. Member 1825 had no renal involvement. Members 1206 and 3600 presented with mild renal disease. Members 4616, 4731, 4446 and 1937 presented with moderate renal disease (RCC). Another family (ID 46) was found with this variant. Member 5024 had unknown renal involvement. Member 5025 had no renal involvement. Another member (no ID provided) presented with mild renal disease (RCC) as well as member 5080. Member 5078 presented with moderate renal disease (RCC). The paper states that these families are previously reported in PMID 12202531 and/or 9829912.

A retrospective series of case reports were analyzed to detect and compare changes in VHL disease-associated tumors before and after sunitinib treatment. Sanger sequencing of blood identified the mutations discussed. One case includes a 56 year old female with a c.232A>T mutation in the VHL gene, leading to a p.Asn78Tyr protein change. Her presentations included pancreatic endocrine tumour, renal cysts (identified at bilateral RCC), and unidentified back tumor. Her sister was identifies with the same point mutation of VHL and found with pancreatic NET. She died of the hemorrhagic stroke at age of 40 years. Other family members include a brother and nephew who each were identified with the same point mutation of VHL, but refused for further examinations for VHL-related lesions. The mother died of stroke at early of 50 years.

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. Two of the 43 patients (patients 19 and 13) were found to have the missense mutation, c.500G>A in the VHL gene. Patient 19 had multiple hemangioblastomas of the central nervous system and spinal cord as well as bilateral renal cell carcinoma. Patient 13 had hemangioblastoma (either of CNS or spinal cord-not indicated), ovarian cysts, pancreatic cysts, and renal cell carcinoma. Both patients had no family history and were classified as A2 (sporadic patients with involvement of at least two distinct organs).

27 index patients suspected of having VHL disease from unrelated Chinese families and their relatives were studied. The nucleotide were numbered according to the sequence numbering by Latif et al, 1993 and this was converted to NM_000551.3. The paper did not differentiate between a pancreatic cysts and tumor and presented these together as a 'pancreatic lesion'. The paper summarized the phenotypes of the families together. Family 27 was not recorded with a documented VHL mutation. In the paper it is recorded as “?”. The proband is a Chinese male with multiple CNS hemangioblastomas and pancreatic cysts. His mother had multiple retinal angiomas, but also no recorded VHL mutation. No further information was given.

27 index patients suspected of having VHL disease from unrelated Chinese families and their relatives were studied. The nucleotide were numbered according to the sequence numbering by Latif et al, 1993 and this was converted to NM_000551.3. The paper did not differentiate between a pancreatic cysts and tumor and presented these together as a 'pancreatic lesion'. The paper summariezed the phenotypes of the families together. Family 2 was found with p.Glu70* mutation in the VHL gene: 1/2 members had CNS hemangioblastoma. 1/2 had retinal angioma, and 1/2 members had a pancreatic lesion. ACMG PP4

27 index patients suspected of having VHL disease from unrelated Chinese families and their relatives were studied. The nucleotide were numbered according to the sequence numbering by Latif et al, 1993 and this was converted to NM_000551.3. The paper did not differentiate between a pancreatic cysts and tumor and presented these together as a 'pancreatic lesion'. The paper summarized the phenotypes of the families together. Family 9 and 10 were found with p.Pro86Leu mutation in the VHL gene. Family 9: the mutation was detected in a 53 years old woman (proband) with asynchronous bilateral renal clear cell carcinomas, multiple renal cysts and pancreatic cysts, an asympomatic member and with (initially) no positive family history. Subsequently, genetic analysis and clinical evaluations showed that the same mutation was also detected in her niece who had no detectable VHL lesions and in her young brother who had pancreatic cysts. Family 10: 1/4 members had CNS hemangioblastoma, 1/4 had retinal angioma, 2/4 had RCC, and 1/4 had a pancreatic lesion. ACMG PP4

27 index patients suspected of having VHL disease from unrelated Chinese families and their relatives were studied. The nucleotide were numbered according to the sequence numbering by Latif et al, 1993 and this was converted to NM_000551.3. The paper did not differentiate between a pancreatic cysts and tumor and presented these together as a 'pancreatic lesion'. The paper summariezed the phenotypes of the families together. Family 14 was found with p.Ser111Asn mutation in the VHL gene: 4/5 members had CNS hemangioblastoma, 1/5 had retinal angioma, 3/5 had RCC, and 1/5 had a pancreatic lesion. ACMG PP4

At age 9, a male caucasian patient was admitted with hypertension and tachicardia. His history included café-au-lait spots since birth, four months of night sweats and three weeks of fevers, bifrontal headaches and fatigue. An MRI scan showed a 3cm left adrenal mass and an MIBG showed increased activity in the left adrenal gland. One month later the patient underwent a successfull adrenalectomy. Four years post-surgery, the patient presented with bitemporal headaches, fatigue and low-grade fevers but both MRI and MIGB are inconclusive. One year later, symptoms worsened, MRI results were negative. At 18Y he later relapsed with another adrenal pheochromocytoma which was resected. DNA analysis showed a germline mutation Val84Leu in VHL. This mutation could correspond to either c.250G>C or c.250G>T. Neither of the parents carried mutations of the VHL gene. ACMG evidence codes: 'PS2' since both parents are tested and do not present with the variant (de novo), 'PP4' since the patient presents phenotypes highly specific for a disease with a single genetic etiology.

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 #86. The proband (patient 229) was a male who was diagnosed with CNS hemangioblastoma at 39Y, retinal angioma at 57Y, renal cell carcinoma at 57Y, and multiple pancreatic cysts/tumors at 57Y. His sister (patient 230) was diagnosed with CNS hemangioblastoma at 40Y. His first brother (brother 1, patient 231) was diagnosed with retinal angioma at 25Y and renal cell carcinoma at 54Y. His second brother (patient 232), who passed away before he could be genetically confirmed with the mutation, was diagnosed with CNS hemangioblastoma at 35Y. His nephew (son of second brother, patient 233) was diagnosed with a renal cell carcinoma at 30Y. This kindred 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. One female patient, patient 112, was found with this germline mutation from family #40. Shee was diagnosed with a CNS hemangioblastoma at 34Y, renal cell carcinoma at 44Y, and multiple pancreatic cysts/tumors at 44Y. This kindred was likely 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. One male patient was found with this germline mutation from family #45. He was diagnosed with CNS hemangioblastoma at 30Y. One male patient was found with this germline mutation from family #46. He was diagnosed with CNS hemangioblastoma at 30Y, RCC at 31Y, pheochromocytoma at 30Y, and multiple pancreatic lesions at 30Y. No other family information was published for these two patients. Both kindreds were also likely published in a paper by Wang et al., 2018 (PMID: 29330336). Kindred #46 was also previously published in Wang et al., 2014 (PMID: 24581539). Wang et al., 2018 reported 4 patients from 2 families (corresponding with kindred #45 and #46) who were genetically confirmed with this germline mutation. Wang et al., 2014 reported one patient who was genetically confirmed with this germline mutation likely corresponding with family #46.

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. Genotyping confirmation was performed by PCR-direct sequencing on peripheral blood samples. This mutation was confirmed in families #4, #5, and #5. Only the proband of family #4 (male, patient #7) was genetically confirmed with the mutation. He was diagnosed with renal cell carcinoma and multiple pancreatic cysts/tumors at age 33. Only the proband of family #5 was genetically confirmed with the mutation. He presented with a CNS hemangioblastoma at age 21 and multiple pancreatic cysts and tumors at age 30. His grandfather, father, and uncle presented with a CNS hemangioblastoma at ages 30, 36, and 33, respectively, and all passed away before they could be diagnosed with VHL disease. Family #6 is large: the proband (F, renal cell carcinoma at age 36 and pheochromocytoma at age 37) had 8 family members genetically confirmed with the mutation and two family members who passed away before diagnosis. The 8 genetically confirmed family members include patient 13 (asymptomatic,) 14 (CNS hemangioblastoma at age 59,) 15 (CNS hemangioblastoma age 45, renal cell carcinoma at 59, multiple pancreatic cysts/tumors at 65), 18 (CNS hemangioblastoma at age 22, renal cell carcinoma at 30, multiple pancreatic cysts/tumors at 31,) 19 (asymptomatic), 20 (asymptomatic), 21 (CNS hemangioblastoma and retinal angioma at 53,) and 22 (multiple pancreatic cysts/tumors at 29). The 2 family members who passed away before diagnosis are patients 16 (CNS hemangioblastoma at 22) and 17 (CNS hemangioblastoma, renal cell carcinoma, and multiple pancreatic cysts/tumors at age 51). 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. One proband was genetically confirmed with this germline mutation: an 18 year old male from kindred #88 who was diagnosed with CNS hemangioblastoma, retinal angioma, renal cell carcinoma, and multiple pancreatic cysts/tumors. This patient was also published in a later paper by Wang et al., 2018 (PMID: 29330336).

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 2 patients (Family ID7) was found with a germline missense mutation in the VHL gene (R167W; c.499C>T). Patient 1 was diagnosed at 4Y and had 3 retinal hemangioblastomas at 8Y and 16Y. Patient 1 was 40 by the end of the study. Patient 2 was diagnosed at age 13Y and had no manifestations by the end of the study when they were 36Y.

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. Parts of the Danish cohort have been previously assesed in the following papers PMID: 19863552, 25834951, 27966541, 27539272. Peripheral blood samples were isolated and sequenced coding exons and exon-intron boundaries using MLPA (Multiplex Ligation-dependent Probe Amplification). One family with one patient (Family ID14) was found with a germline mutation in the VHL gene (R161*; c.481C>T). The patient was diagnosed at 10Y and had 2 epididymal cystadnomas at 10Y. The patient was 46 by the end of the study.

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 ID 4) was found with a germline missense mutation in the VHL gene (F136S; c.407T>C). Patient 1 was diagnosed at 7Y and had 2 retinal hemangioblastomas at 7Y and 14Y. The patient passed at 53Y by the end of the study.

A case report revealed a 47Y female patient with two synchronous kidney tumors on the right side that were clear cell renal cell carcinoma and cystic renal cell carcinoma, and a clear cell pancreatic neuroendocrine tumor. She also presented with multiple pancreatic and renal cysts. Genetic analysis confirmed a c.257C>G (p.Pro86Arg) mutation in the VHL gene. The patient had a positive family history with their mother and brother having VHL disease, but genotypic and phenotypic information for family members was not available.

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. There was 1 patient reported with the c.262T>A (p.W88R) mutation. This member presented with retinal hemangiomas. 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 the c.386T>C (p.L129P) mutation. This member presented with retinal hemangiomas. 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 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.Q164* (c.490C>T) mutation, 67% (2/3) of individuals had RH while 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.562C>G (p.L188V). This member presented with retinal hemangiomas. 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 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. There were 2 patients reported with the p.C162W (c.486C>G) mutation. One member presented with retinal hemangiomas, while the other did not - however no other clinical manifestations were discussed. Additionally, 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 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. There were 2 patients reported with the p.F136S (c.407T>C) mutation. One member presented with retinal hemangiomas, while the other did not - however no other clinical manifestations were discussed. Additionally, 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 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. There was 1 patient reported with the c.238A>C (p.S80R) mutation. This member presented with retinal hemangiomas. 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 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. There were 3 patients reported with the p.S183* (c.548C>A) mutation. All 3 members presented with retinal hemangiomas, while no other clinical manifestations were discussed. Additionally, no family information was provided.

A case report of a 31 year old female patient diagnosed with VHL disease revealed a c.481C>T mutation in the VHL gene, leading to the subsequent protein change Arg161*. Tissue resection was analyzed through DNA amplification. There was no family history of illness. Over an 8 year follow-up period the 31Y old patient presented with bilateral RCC, bilateral renal cysts, cardiomegaly, multiple brain hemangioblastomas, pancreatic neuroendocrine carcinoma, pancreatic cysts, kidney cysts, and ovarian cysts.

Medical records of 45 Italian patients with pheochromocytoma/paraganglioma referred to the hypertension centers of the University of Brescia and the University of Turin were reviewed. Germline DNA was extracted through peripheral blood, and PCR products were purified. Only familial and sporadic cases of isolated PHEO/PGL were considered. Patient number 10 was found with a germline VHL mutation (p.P81L) and left para-adrenal paraganglioma, apparently sporadic. This patient was reported to have a negative family history. ACMG: 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 a VHL type 2B family of 7 individuals (Family ID 4414), using PCR and SSCP sequencing. The mutation of c.357C>G was referred to in the paper by c.570C>G due to a difference in transcript sequence. Two of seven individuals were identified with retinal hemangioblastomas, 3 with CNS hemangioblastomas, one with renal cell carcinoma, and one with pheochromocytomas. 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 missense mutation was found in 3 VHL type 1 family members (family ID 3738), using PCR and SSCP sequencing. The mutation of c.334T>C was referred to in the paper by c.547T>C due to a difference in transcript sequence. All had pheochromocytomas and one with retinal hemangioblastomas.

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 7 individuals (family ID 2753), using PCR and SSCP sequencing. The mutation of c.217C>T was referred to in the paper by c.430C>T due to a difference in transcript sequence. Six of 7 individuals were identified with retinal hemangioblastomas, 3 with cerebellar hemangioblastomas and one with renal cell carcinoma. This was possibly previously published in PMID: 9829911. 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).

Direct sequencing and multiplex ligation dependent probe amplification were performed on thirteen unrelated subjects with VHL mutations. The clinical manifestations and family histories of the subjects were evaluated. A 14 year old Korean male patient presented with central nervous system hemangioblastoma, retinal hemangioblastoma, multiple pancreatic lesions, and bilateral renal cysts. Genetic testing confirmed a non-familial missense mutation at c.233A > G of the VHL gene. The patient had no family history of VHL.

Direct sequencing and multiplex ligation dependent probe amplification were performed on thirteen unrelated subjects with VHL mutations. The clinical manifestations and family histories of the subjects were evaluated. A 21 year old Korean female patient presented with pheochromocytoma at age 24 and multiple pancreatic lesions at age 21. Genetic testing confirmed a familial frameshift mutation at c.592delC of the VHL gene. The patient has a mother who presented with pheochromocytoma at age 27, multiple pancreatic lesions at age 48, and developmental venous anomaly or meningioma.

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 C162R (c.484T>C) germline mutation of the VHL gene was found in a VHL type 1 family of 6 presenting with CNS hemangioblastoma (1/6), retinal angioma (2/6), renal cell carcinoma (3/6) and pancreatic tumor or cyst (unspecified) (1/6) (family ID 4). 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 V155M (c.463G>A) germline missense mutation was found in a VHL type 1 family of 9 affected individuals who presented with CNS hemangioblastoma (5/9), retinal angioma (1/9), renal cell carcinoma (9/9) and pancreatic tumor or cyst (unspecified) (2/9) (family ID 32). This family was previously reported in PMID 8634692.

Screening of 92 unrelated patients with VHL disease revealed 61 DNA variants. Blood samples were isolated and sequenced by direct sequencing using prism-ready reaction dye primer cycle sequencing kits (Applied Biosystem Roissy CDG France) and an ABI 373A automated sequencer (AppliedBiosystem Roissy CDG France). No variants were found in 96 control cases. This splice site mutation, c.340+1G>A (stated as IVS1+1G→A in the paper), was found in a VHL type 2B patient (no. V283). Type 2B VHL families are described as having pheochromocytoma and presence of renal and pancreatic manifestations. No family information was provided.

Screening of 92 unrelated patients with VHL disease revealed 61 DNA variants. Blood samples were isolated and sequenced by direct sequencing using prism-ready reaction dye primer cycle sequencing kits (Applied Biosystem Roissy CDG France) and an ABI 373A automated sequencer (AppliedBiosystem Roissy CDG France). No variants were found in 96 control cases. This stop-gain mutation, c.525C>G, was found in a VHL type 1 patient (patient no. V63). No patient information was provided and no additional family members were screened.

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 Y156C (c.467A>G) missense mutation was found in a VHL type 1 (no pheochromocytoma) patient (patient no. V265). One unaffected family member was negative for this mutation.

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 V130L (c.388G>C) missense mutation was found in a VHL type 1 (no pheochromocytoma) patient (patient no. V91). No unaffected family members were tested.

Screening of 92 unrelated patients with VHL disease revealed 61 DNA variants. Blood samples were isolated and sequenced by direct sequencing using prism-ready reaction dye primer cycle sequencing kits (Applied Biosystem Roissy CDG France) and an ABI 373A automated sequencer (AppliedBiosystem Roissy CDG France).No variants were found in 96 control cases. This missense mutation, c.486C>G, was found in a VHL type 2B patient (patient no. V345). No patient information was provided and no additional family members were screened.

Medical records of 31 Asian Indian genetically proven VHL patients with pheochromocytoma/paraganglioma were studied. A 30 year old male (Patient 23) presented with unilateral pheochromocytoma (30Y). Genetic testing (PCR and Sanger sequencing) confirmed a missense mutation at c.509T>C of the VHL gene. He has a negative family history.

A retrospective study reviewed cases of 23 patients with advanced and complicated VHL eye disease who underwent pars plana vitrectomy in the years 1999 to 2012 that were a part of 223 VHL patients consulted at the International Hereditary Cancer Center and Polish VHL Registry. Genetic molecular analysis was performed in 22/23 patients through PCR and MLPA, and 24 eyes from 23 patients were studied. A germline c.481C>T transition was would in 3 individuals, and results in a p.Arg161* protein change and premature stop codon. A 35 year old (Patient 3) had a previous history of HbCNS. Patient was examined in a study of advanced VHL eye disease and had multiple retinal capillary hemangioblastomas requiring surgery. A 35 year old patient (Patient 7) had a previous history of HbCNS and RCC. The patient was examined in a study of advanced VHL eye disease and had multiple retinal capillary hemangioblastomas requiring surgery. An 18 year old (Patient 9) had a previous history of HbCNS, ELST, and RCC. Patient was examined in a study of advanced VHL eye disease and had multiple retinal capillary hemangioblastomas requiring surgery. Since patients were listed 'by eye' it is impossible to determine how many patients listed above actually represent unique patients.

This study reports 1,548 germline and somatic mutations from 945 VHL families, including 30 previously unpublished kindreds from The Netherlands (six novel mutations). This mutation (c.245G>T) was detected by direct sequencing and MLPA in a VHL kindred with pheochromocytoma (case no. 393). ACMG evidence codes: 'PP2' because they observe a missense variant in a gene where missense variants are a common mechanism of disease. 

This study reports 1,548 germline and somatic mutations from 945 VHL families, including 30 previously unpublished kindreds from The Netherlands (six novel mutations). This exon was detected in a VHL kindred with hemangioblastomas of the central nervous system, retinal hemangioblastomas and Grawitz tumor (also known as RCC) (case no. 2034 p.F76del(c.227_229delTCT)), by direct sequencing and MLPA. This case is published in later publication: PMID: 32179488.

This study reports 1,548 germline and somatic mutations from 945 VHL families, including 30 previously unpublished kindreds from The Netherlands (six novel mutations). This mutation (c.266T>C) was detected by direct sequencing and MLPA in 2 VHL kindreds. It was found in a kindred with hemangioblastomas of the central nervous system, renal cell carcinoma, pancreatic and renal cysts (case no. 23530). ACMG evidence codes: 'PP2' because they observe a missense variant in a gene where missense variants are a common mechanism of disease, 'PP4' because the patient's phenotypes or family history are highly specific for a disease with single genetic etiology.

This study reports 1,548 germline and somatic mutations from 945 VHL families, including 30 previously unpublished kindreds from The Netherlands (six novel mutations). This mutation (c.430G>T) was detected by direct sequencing and MLPA. This nonsense mutation was found as sporadic germline in a VHL case with bilateral renal cell carcinoma and hemangioblastoma (case no. 442). This was suggested to be de novo but it could not be confirmed. There is very strong evidence of pathogenicity in the form of a nonsense variant in a gene where loss-of-function is known mechanism of disease (ACMG code: PSV1).

Of 46 cases of RCC in patients younger than 25 years, one patient (case 45) was found to harbour a germline VHL splicing mutation G>C at position 340+5, stated in the paper as c.553+5 due to a difference in transcript sequence. Genomic DNA was extracted using DNeasy Blood and Tissue Kit, from neoplastic and non-neoplastic tissue samples and was analyzed for polymerase chain reaction-based loss of heterozygosity (LOH) where results were viewed by silver nitrate staining. This patient presented with a papillary RCC (stage/diameter: pT1N0M0) on her right kidney at age 16. She presented no evidence of disease at her 36 month follow-up after undergoing a radical nephrectomy. She had a known family history for VHL disease and her mother has cerebellar hemangioblastoma. Genetic data on the mother was not described. ACMG evidence codes: 'PP4' since the patient presents phenotypes highly specific for a disease with a single genetic etiology.

188 individuals presenting with a single hemangioblastoma, no family history of VHL disease, and no evidence of retinal or abdonimal manigestations of the disease at the time of diagnosis, were were investigated for mutations in the VHL gene. Patient B5 presented with a cerebellar hemangioblastoma at age 39. Genetic testing via direct sequencing on a DNA blood sample found a c.598C>T (p.Arg200Trp) mutation in their VHL gene (noted as c.811C>T in the paper due to their use of an older VHL transcript used for mapping).Two first-degree relatives (FDRs) tested positive for the mutation: a 28 year old FDR and a 38 year old FDR, neither of whom showed any features of VHL disease. Patient B4 presented with a cerebellar hemangioblastoma at age 36. Genetic testing via direct sequencing on a DNA blood sample also found a c.598C>T (p.Arg200Trp) mutation in their VHL gene (noted as c.811C>T in the paper due to their use of an older VHL transcript used for mapping). No family germline testing was performed, and no features of VHL disease were noted in their family history.

188 individuals presenting with a single hemangioblastoma, no family history of VHL disease, and no evidence of retinal or abdonimal manigestations of the disease at the time of diagnosis, were investigated for mutations in the VHL gene. Patient B3 presented with a cerebellar hemangioblastoma at age 40, and a phaeochromocytome at age 41. Genetic testing via direct sequencing on a DNA blood sample found a c.500G>A (Arg167Gln) mutation in their VHL gene (noted as c.713G>A in the paper due to an older VHL transcript used for mapping). Two of their first degree relatives (FDRs) also had the mutation. The first presented with a hemangioblastoma at 18, and a retinal angioma at 18. The second FDR presented with a renal cyst at 17.

540 patients from 187 unrelated Chinese VHL families at the Peking University First Hospital were all screened for VHL clinical features and germline mutations. Of these, clinical characteristics were observed in 258 patients from 80 unrelated Chinese families. These were reported in this study, grouped by the 21 mutations found. PCR and MLPA on peripheral blood detected the VHL mutations. Possessing the p.R167Q (c.500G>A) mutation, 24 members total from 10 families were described, and phenotypic information was given for this group. With ages of onset ranging from 14-53Y, hemangioblastomas were observed in 15/24 members, retinal capillary hemangioma in 4, renal cell carcinoma in 10, pancreatic cysts in 5, and pheochromocytoma in 9 members. This cohort has been previously published in PMID 24581539 (as family #11); 29330336 (as 7 families of 18 people); 28388566 (family #78, 79, 80, 81, 82, 83).

540 patients from 187 unrelated Chinese VHL families at the Peking University First Hospital were all screened for VHL clinical features and germline mutations. Of these, clinical characteristics were observed in 258 patients from 80 unrelated Chinese families. These were reported in this study, grouped by the 21 mutations found. PCR and MLPA on peripheral blood detected the VHL mutations. Possessing the p.W117G (c.349T>G) mutation, 11 members total from 2 families were described, and phenotypic information was given for this group. With ages ranging from 12-61Y, hemangioblastomas were observed in 8/11 members, retinal capillary hemangioma in 5, renal cell carcinoma in 6, pancreatic cysts in 8, pheochromocytoma in 1 member and and in a papillary cystadenoma of the epididymis genital system tumor (epididymis or broad ligament) 1 member. See more information as this cohort has been previously published in 24581539 (as family 6 of 10 members); 29330336; and 28388566.

540 patients from 187 unrelated Chinese VHL families at the Peking University First Hospital were all screened for VHL clinical features and germline mutations. Of these, clinical characteristics were observed in 258 patients from 80 unrelated Chinese families. These were reported in this study, grouped by the 21 mutations found. PCR and MLPA on peripheral blood detected the VHL mutations. Of these, 21 patients from two families were found to harbour the VHL germline mutation c.262T>C (p.Trp88Arg). From family 1, a total of 17 family members including the proband were identified. Three of these did not present VHL clinical features. The 32 year old proband (patient 15) presented with pancreatic neuroendocrine tumor or pancreatic cyst and renal cell carcinoma. Her maternal grandfather presented with renal cell carcinoma at age 65. Her maternal great-uncle (patient 3), great-aunt (patient 4), mother (patient 6) and 6 first cousins once removed (patients 7, 8, 9, 10, 13, 14) each presented with cerebellar hemangioblastomas at ages 62, 37, 35, 41, 40, 35, 29 37 and 37 respectively. One of these cousins (9) also presented with renal cell carcinoma at age 42. Two other 1st cousins once removed (patients 11 and 12) presented with renal cell carcinoma at ages 35 and 40, as did her sister at the age of 27. From family 2, the proband presented with no clinical features, however, her mother (2) presented with pancreatic cancer at age 45. Both her maternal uncle (patient 3) and grandmother (patient 1) presented with cerebellar hemangioblastomas at ages 50 and 48 respectively. This same uncle also had pheochromocytoma, also identified at the age of 50. ACMG evidence codes: 'PP1' since we have cosegregation with the disease in multiple affected family members (both families 1 and 2), 'PP4' since the patient presents phenotypes highly specific for a disease with a single genetic etiology.

17 patients with genetic variants in SDHB, VHL, EPAS1, or RET with pheochromocytoma were identified retroactively for this study. 5 pathogenic germline variants were confirmed in patients with clinical criteria of von Hippel Lindau syndrome. Next generation sequencing (NGS) re-confirmed these germline mutations. One of which was a p.R107S (c.319C>A) mutation in patient 3, a 26Y male with pheochromocytoma. No further manifestation details were provided, nor family information.

A case report describes a 35 years old women presenting with osteoporosis. Her past medical history revealed retinal angioma at 16 years, cerebellar hemangioblastoma at 21 years, spinal hemangioblastoma at 28 years, pheochromocytoma at 32 years, and amenorrhea (started menstruating at age 12 and stopped at 16). She had a germline p.Arg167Gln mutation in the VHL gene. She had a negative family history for VHL disease and osteoporosis, but gentoype of family members was not described.

A retrospective analysis of pheochromocytoma tissue samples from the Department of Pathology of the Erasmus MC (Rotterdam) revealed an 18Y female patient (ID #F119) with a confirmed germline mutation in the VHL gene, p.Arg64Pro. The patient had pheochromocytoma but other phenotypes were not described. Family history was not described. Pheochromocytoma is highly specific to VHL disease (PP4) and disease-causing missense variants are common in VHL disease (PP2).

The authors presented a cased of a 28-year-old woman with neurological symptoms from two haemangioblastomas (cerebellar and cerebro-spinal). Refined clinical work-up revealed pancreatic cysts and angiomatosis retinae (retinal hemangioblastoma). Clinical work-up revealed no pheochromocytoma was found in the patient. PCR/SSCP analysis and fluorescence-labelled based excision sequence scanning (F-BESS) were used to detect mutation in the VHL gene. The L140Pfs*4 (c.418dup) mutation was detected in the patient, referred to in the paper by c.631_632insC due to a difference in transcript sequence. No signs of VHL disease was detected in the patient's family members (mother, brother, sister), and genetic analysis also did not reveal any VHL mutation in the family members. The patient’s father died from a myocardial infarction at the age of 58 years and was not tested for VHL. ACMG: 'PP4' because the patients' phenotypes are highly specific for a disease with single genetic etiology.

This paper analyzed VHL sequences in the renal cell carcinomas of patients exposed to high, cumulative doses of trichloroethylene. Whole blood and tumour tissues were isolated and mutations were identified, respectively, by direct sequencing of purified PCR products. For fast analysis of the presence of a frequent mutation at nt 454 (C>T), a restriction endonuclease-based assay (enzyme AciI based assay) was used.Four patients (1654, 1656, 1658, and 1695) were found to have the germline P81S (c. 241C>T) (denoted as nt454 C>T mutation in the paper) VHL mutation in DNA isolated from tumor-adjacent normal kidney parenchyma. All four patients presented with renal cell carcinoma and were diagnosed respectively at ages 59, 70, 66, 38 (1654, 1656, 1658, 1695). Three of the four patients (1654,1658, 1695) had multiple somatic VHL mutations and LOH. Patient 1656 only had the one germline mutation in the VHL gene.

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: V-PH-35T) belonged to a male diagnosed with right adrenal phaeochromocytoma at age 39Y, surgery the same year. He also had a personal history including hemangioblastomas, and hypertension. The VHL mutation R161* (c.481C>T) was present. This patient was said to be the singling of ID: V-PH-35T, who possessed the germline VHL mutations: p.Arg161Ter and p.R200fs (c.598del). Additionally, this patient was likely previously published in PMID: 26796762 (ID: WM02).

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. Two patients with VHL-related retinal angiomatosis were found to have germline missense (c.344A>G) mutation in the VHL gene. 1 other affected relative was also evaluated for one of the patients and was found to have the mutation but phenotypes were not described. This variant was reported as c.557A>G (AF010238.1) and mapped to p.H115R (c.344A>G) (NM_000551.3).

Of 31 Brazilian patients, one was found to harbour the c.227_229delTCT (p.Phe76del) mutation of the VHL gene. Genomic DNA was extracted and all VHL coding regions were analyzed by Sanger sequencing. MLPA was performed to investigate large deletions using SALSA® MLPA® P016 VHL probe mix. Data analysis was performed with Genescan 3.7. This patient, age 44, presented with renal cell carcinoma, CNS hemangioblastoma and retinal hemangioblastoma. It is not clear if this mutation is familial.

200 patients with intracranial aneurysms were molecularly analyzed for VHL to determine if it is a candidate gene for aneurysm formation using peripheral blood. Eight patients were analyzed in detail and determined to have the above SNP mutation (c.463+43A>G). The patients were only recorded to have an intracranial aneurysm (or dilatation of the cerebral artery). Any relation is unknown. This mutation has a large allele frequency and therefore a polymorphism that is likely non disease causing

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 splice site variant, c.464-1G>A, was found in a 15-year-old VHL patient with retinal hemangioblastomas (index case 227). The patient did not have hemangioblastoma, pancreatic cyst, pheochromocytoma, clear cell renal cell carcinoma, renal cysts. This study contains very strong evidence for pathogenicity in the form of a splice site variant in a gene where loss-of-function is a known mechanism of disease.

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 V170G (c.509T>G) germline missense mutation of the VHL gene was found in a 37 yr old VHL patient with retinal hemangioblastomas and pheochromocytoma (index case 53).

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 P59S (c.175C>T) germline variant of the VHL gene was found in a 62 year old patient (ID 364) presenting with pheochromocytoma. Transmission of the variant could not be determined (n/d) since both parents and family history were not available.

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 C162W (c.486C>G) germline missense mutation of the VHL gene was found in a 29 year old VHL patient with pancreatic cysts or tumors (index case 78).

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 C162F (c.485G>T) germline missense mutation of the VHL gene was found in a 34 yr old VHL patient with multiple pancreatic cysts or tumors, pheochromocytoma, renal cysts, and central nervous system and retinal hemangioblastomas (index case 74).

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 missense mutation, c.191G>C, was found in a 26 year old VHL patient with pheochromocytoma and no family history (index case 123). Only supporting evidence for pathogenicity because patient's phenotype is highly specific for a disease with a single genetic etiology.

A 40-year-old woman was found with clear cell renal cell carcinoma, pancreatic endocrine tumours, and pancreatic-islet cell hyperplasia. Genetic analysis using peripheral blood revealed a germline mutation c.499C>T (p.Arg167Trp) in the VHL gene. The patient had negative family history for VHL disease. 46 months later the patient had no evidence of recurrence.

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.340G>A). 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. 13 patients were found to have the above mutation (c.482G>A). 5 were VHL patients with ICTs (pancreatic neuroendocrine tumors) taken from the VHL-registry. The other 8 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: 'PP2' because they observe a missense variant in a gene where missense variants are a common mechanism of disease.

One hundred and sixteen unrelated VHL disease patients have been investigated by molecular genetic analysis. Twenty-two of these were previously shown to have germline deletions on Southern analysis, and in the present study the remaining 94 patients were screened for intragenic mutations via SSCP and heteroduplex analysis of 855 bp of the VHL coding sequence and 800 bp of the 3' UTR. An intragenic mutation was identified in 55 (59%) out of 94 patients without germline deletions on Southern analysis. In all patients with multiple affected family members the missense mutation segregated with the disease. Large deletions or intragenic mutations predicted to cause a truncated protein were found in 36 of 53 families without pheochromocytoma (VHL type 1) and 2 of 12 families with pheochromocytoma (VHL type 2). This study also investigated the frequency of different mutation classes between kindreds with and without pheochromocytoma. The p.Y185 (c.555C>G) variant (noted as p.Y256 (c.768C>G) in the paper due to a difference in reference transcript used) was found in at least one patient in kindreds 82 and 88. Pheochromocytoma was not observed in any members of either kindred (implying the affected patient(s) had VHL disease type 1). No other familial phenotypic information could be inferred from the paper.

Pathogenic variants from 10,389 individuals across 33 cancer types were examined from The Cancer Genome Atlas (TCGA) cohort. 12 of these individuals had a germline mutation in the VHL gene. The W117* (c.351G>A) nonsense VHL mutation was found in a white 23Y0M male diagnosed with pheochromocytoma and paraganglioma. No family information was provided.

Pathogenic variants from 10,389 individuals across 33 cancer types were examined from The Cancer Genome Atlas (TCGA) cohort. 12 of these individuals had a germline mutation in the VHL gene. The E134*(c.400G>T) nonsense VHL mutation was found in a white 29Y0M female diagnosed with clear cell renal cell carcinoma. No family information was provided.

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 (c.584_585delAG) was found in a VHL type 1 patient with retinal angiomas (at 45Y old), hemangioblastomas of the cerebellum (at 24Y old), and renal cell carcinoma (at 45Y old).

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). A missense mutation, S65W(c.194C>G), affecting residue within the pVHL protein core, was found in a VHL family with 8 affected individuals including 6 with retinal hemangioblastomas, 5 with cerebellar hemangioblastomas, and 1 with renal cell carcinoma. ACMG codes as follows: supporting evidence for pathogenicity because of cosegregation of disease with multiple affected family members in a gene definitively known to cause the disease (ACMG code: PP1).

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). This missense mutation (c.496G>T), affecting surface residue of pVHL protein core, was found in 2 VHL type 2A families with 5 affected individuals altogether. Two of 5 patients had retinal angioma, 2 had cerebellar hemangioblastoma, and 4 had pheochromocytoma. This mutation was reported in the paper with an incorrect cDNA written as c.498A>G, however correct translation should be deduced to c.496G>T, as the Latif et al. 1993 (AFO10238) sequence given was as c.709G>T with p.V166F.

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). This missense mutation (c.538A>G), affecting residue within the pVHL protein core, was found in a VHL patient with renal cell carcinoma diagnosed at 27Y. ACMG codes as follows: Supporting evidence of pathogenicity because Missense variant in a gene that has a low rate of benign missense variation and where missense variants are a common mechanism of disease (PP2).

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 (c.555C>G) was found in 3 VHL families with 4 patients altogether. All four had retinal hemangioblastomas and cerebellar hemangioblastomas. Two had renal cell carcinoma.

Medical records of 26 patients with germline mutations of the VHL gene who had been diagnosed with VHL disease in Severance Hospital (Seoul, Republic of Korea) and Samsung Medical Center (Seoul, Republic of Korea) between 2003 and 2012 were reviewed. Peripheral blood leukocytes were isolated and directly sequenced. 8 unrelated patients diagnosed with Type 1 VHL disease shared the same VHL germline missense mutation (c.208G>A; p.Glu70Lys). The patients' phenotypes were as follows: 55 year old male with multiple CNS hemangioblastomas (Patient 3), 55Y male with multiple CNS hemangioblastomas (Patient 4), 43Y male with multiple CNS hemangioblastomas and renal cysts (other phenotypic data not available), (Patient 5), 11Y male with bilateral retinal hemangioblastoma and renal cysts (Patient 6), 48Y female with CNS hemangioblastoma (Patient 7) , 64Y female with CNS hemangioblastoma (Patient 8), 16Y F with retinal hemangioblastoma (Patient 9), 41Y female with CNS hemangioblastoma (Patient 10). All patients were negative for VHL family history. Patient 3 was previously reported in PMID 19270817. Patient 5 was previously reported in either PMID 19270817 and/or 19949673.

Medical records of 26 patients with germline mutations of the VHL gene who had been diagnosed with VHL disease in Severance Hospital (Seoul, Republic of Korea) and Samsung Medical Center (Seoul, Republic of Korea) between 2003 and 2012 were respectively reviewed. Peripheral blood leukocytes were isolated and directly sequenced using the BigDye Terminator Cycle Sequencing Ready Reaction kit on an ABI Prism 3130 genetic analyzer (Applied Biosystems). One 34Y male patient (Patient 15) presented with multiple and/or bilateral RCC and a CNS hemangioblastoma. He had a germline missense mutation in the VHL gene (c.233A>G p.Asn 78 Ser). The patient had a negative family history of VHL disease. The phenotypes of this patient were revised from a previous report in PMID 19270817 and/or 19949673.

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.221T>G) was recorded in the paper as c.434T>G due to a difference in transcript sequence. It was found in 3 patients with hemangioblastomas of the central nervous system, one of which had primary asymptomatic tumors. 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 mutation (p.W117Lfs*15) was recorded in the paper as c.559insT 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 missense mutation (c.292T>C) was recorded in the paper as c.505T>C due to a difference in transcript sequence. It was found in 21 patients with hemangioblastomas of the central nervous system, 13 of which were stated to be primary asymptomatic. Any family information or relations 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 splice site mutation (c.340+1G>A) was recorded in the paper as c.553+1G>A 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.

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.395A>C) was recorded in the paper as c.608A>C due to a difference in transcript sequence. It was found in 2 patients with hemangioblastomas of the central nervous system, one of which had primary asymptomatic tumors. Any family information or relations is unknown. No other phenotypes described.

Screening of 68 patients, who had been operated on for apparently sporadic pheochromocytomas, for germline mutations in the VHL gene. This missense mutation was found in a 72 year-old female with unilateral, benign, adrenal pheochromocytoma (patient no. 19). The tumor was operated on at age 58. The variant reported as I147T (c.653T>C), legacy nomenclature (AF010238.1), and was mapped to p.Ile147Thr (c.440T>C) in accordance with reference sequence NM_000551.3.

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 13 had genetic testing and was found to have a germline mutation c.486C>G (Cys162Trp). This missense mutation is noted as c.699C>G in the paper due to a difference in the VHL reference sequence notation used. Patient 13 was a 48-year-old male with a medical history of synchronous bilateral solid renal tumors, multiple renal cysts, CNS hemangioblastoma. AMCG evidence codes: ’flag for PP4 review’ because the patients phenotype is highly specific for a mutation in the VHL gene.

Two VHL syndrome pedigrees were investigated in this study; family 1 was a three-generation pedigree including 6 individuals. In family 1, the proband (F1-II-1), a 31-year-old male patient, presented with headache, paroxysmal hypertension and dizzy spells in 2009 with a 4-year history. Ultrasound and MRI scans confirmed single masses, multiple tumors and multiple cysts in the right adrenal glands, bilateral kidney and pancreas, respectively. Right pheochromocytoma and multiple right renal cell carcinoma were confirmed by histopathological examination. In 2012, the patient was diagnosed with bilateral epididymal head multiple nodules. Bilateral epididymal tubercle resection was performed and epididymal papillary adenoma was confirmed by histopathological examination. Additionally, the patient's younger brother (F1-II-3, 30 years old) presented with multiple right renal masses, epididymal nodules and multiple pancreatic cysts. Radical right nephrectomy was performed and histopathological analysis revealed right RCC with hemorrhagic cyst. In family 1 they identified a heterozygous variant, c.233A>G (p.N78S), within exon 1 of the VHL gene in two affected cases and one clinically normal family member (F1-III-1, the proband's 5-year-old son). Peripheral blood samples were collected for mutation analysis of the RET proto-oncogene and VHL tumor suppressor gene. PCR amplification of all exons and the flanking splice junctions of RET and VHL was performed, followed by direct bidirectional DNA sequencing. One hundred unrelated controls with a similar ethnic background were also included in this study. ACMG evidence codes: ‘PP4’ because the family history is highly specific for a disease with a single genetic etiology, ‘PP1’ because there is co-segregation of the variant with VHL disease in the family.

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.R161Q (c.482G>A) was found in 3 VHL patients. All 3 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 frameshift mutation p.H191Tfs*11 (c.571delC) was found in a VHL patient with retinal hemangioblastomas. Other manifestations are not clearly stated. No family information 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 was found in a 1 patient, without retinal hemangioblastomas, but other manifestations are not clearly stated. No family information or relation was provided. Recorded in paper as c.386insCT, this mutation automaps to NM_000551.3:c.385_386dupCT (p.V130Wfs*30). 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.R167W (c.499C>T) was found in 15 VHL patients. 5 had retinal hemangioblastomas, and the other 10 were said to not have. Other manifestations are not clearly stated. No family information or relation was provided. Some may be 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, the missense mutation (c.481C>T (p.Arg161Ter)) was found in 8 patients total - 3 of which are stated to have retinal HBs and 5 are said to not. Other manifestations are possible but not clear. This is possibly previously published in PMID: 15300849

A 19-year-old female patient was referred to a psychiatric inpatient service due to a suspicion of anorexia nervosa (AN) following severe weight loss, low food intake, and a lack of a menstrual cycle for at least 1 year. The patient is the second child of a non-consanguineous relationship and there were no abnormalities in developmental milestones, but her weight never exceeded 35 kg. Apart from weight loss and weakness, physical and neurological examinations showed no alterations. The patient frequently complained of epigastric burning sensation, nocturnal headaches, postural hypotension and difficulty swallowing. The patient did not meet the necessary criteria for a diagnosis of AN or another psychiatric disorder. The patient experienced episodes of nausea during fasting after waking up, followed by unprovoked vomiting, which is highly uncommon in AN. Blood tests, urinalysis and chest X-ray were all normal. Abdominal CT scan revealed cystic lesions throughout the parenchymatous portion of the pancreas, suggestive of VHL. After 3 weeks of hospitalization, the patient progressively developed gait abnormality and bilateral dysmetria, which was worse on the left side. A brain MRI examination revealed a massive lesion suggestive of a hemangioblastoma (HB) in the fourth ventricle causing hypertensive hydrocephalus. HB and pancreatic cysts support the diagnosis for VHL. Whole blood was obtained for sanger sequencing of all 3 exons of the VHL gene, revealing a nonsense mutation in the VHL gene (c.481C>T; p.Arg161*) that results in a truncated protein. In silico analyses by mutation analyzer 2.0.28 suggests a pathogenic variant. Surgery inserting a ventriculoperitoneal shunt to relieve the hypertensive hydrocephalus only temporarily relieved her appetite; yet she no longer had fasting nausea and vomiting, or nocturnal headaches. Fractionated radiotherapy did not improve appetite or nausea but was successful in controlling tumor growth as an MRI revealed no significant alterations in the size of the lesion six months post-radiotherapy.

Genomic DNA was extracted from whole blood samples for genetic testing of SDHB, SDHC, SDHD, and VHL. 27 patients with urinary bladder paragangliomas, revealed 17 germline mutations, three of which were in the VHL gene. PCR–based bidirectional sequencing was performed along with MLPA on large deletions. This missense mutation (p.T124I) was found in a 12-year-old male (patient no. 22), with his urinary bladder paraganglioma diagnosis at 6Y of age. Family history of pheo or paragangliomas was said to be positive.

A case report of a 30 year old young Indian female who presented with hypertension and was subsequently diagnosed to have VHL disease with a c.293A>C mutation in the VHL gene, causing the substitution of Tyr with Ser at amino acid 98. Patient presented with adrenal pheochromocytoma, pancreatic mass, and cerebellar hemangioblastoma. Family history is unavailable other than the delivery of a healthy child. ACMG: PP2, PP4

Of 38 PPGL (pheochromocytoma and paraganglioma) cases studied, three were associated with an individual carrying the germline mutation c.481C>T (ENSP00000256474.2 p.Arg161*) of the VHL gene. This mutation was identified using germline clinical sequencing on 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: RNSH06) presented with bilateral adrenal phechromocytoma (34yo), ID: RNSH09 with right adrenal phechromocytoma (39yo), and ID: RNSH10 with left adrenal phechromocytoma (40yo). No family history, or relation, was presented. RNSH09 and RNSH10 were likely previously published in PMID: 25545346.

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.277G>A germline mutation of the VHL gene. They presented with a retinal hemangioblastoma, spinal hemangioblastoma, renal cyst, pheochromocytoma and pancreatic cysts. Their age at the last follow-up was 60. 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.241C>T germline mutation of the VHL gene. They presented with a retinal hemangioblastoma, cerebellar hemangioblastoma, spinal hemangioblastoma, renal cell carcinoma, pancreatic neuroendocrine tumor, renal cyst and pheochromocytoma. Their age at the last follow-up was 59. Previously published in: PMID 24132471.

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 4 involved three affected members, however only two of which were known to have a c.269A>T change leading to a p.Asn90Ile protein change. The male proband presented with hemangioblastoma at 22 and retinal capillary hemangioma at 32. His mother presented with renal cell carcinoma at 63. The proband’s brother died before diagnosis; thus no DNA sample available, however he was affected with hemangioblastoma at age 19Y. Of family 9, the male proband presented with hemangioblastoma at 28, renal cell carcinoma at 38, and pancreatic cysts at 38. His mother, aunt and cousin all died before diagnosis; thus no DNA sample available. The mother was last observed at 33Y with CNS hemangioblastoma; the aunt at 32Y with hemangioblastoma, and the cousin at 30Y with hemangioblastoma. ACMG Evidence Code: 'PP4' because the patients' phenotypes and family histories are highly specific for VHL disease.

A 49-year-old woman had episodes of vertigo 12 years ago and abdominal pain 10 years ago. Ultrasonography and abdominal MRI revealed the presence of pancreatic cysts. Seven years ago, an abdominal computed tomography scan showed a solid mass in the right kidney. A right partial nephrectomy was performed and pathological examination revealed renal cell carcinoma of the right kidney. That same year, the patient complained of altered visual acuity and a CT scan of the skull and orbit revealed thickening of the left optic nerve and the right optic nerve had a conserved aspect. These results were suggestive of a hemangioblastoma (Hb) in the optic nerve of the left eye. Peripheral blood was sampled to extract genomic DNA for VHL gene sequencing and the detection of VHL copy number variants using multiplex ligation-dependent probe amplification (MPLA). A heterozygous W88* (c.263G>A) nonsense variant was identified in the VHL gene. The patient's family history revealed siblings with VHL manifestations. The patient's mother was the index case and died at age 63 of clear cell renal carcinoma. The patient has one child, who is 27-years-old and complains of recurrent headaches, but refuses to undergo VHL screening. The patient's 5-year-old grandson does not have symptoms. The patient has five brothers and two sisters. One sister underwent partial nephrectomy due to carcinoma in the right kidney, another sister was operated on due to CNS Hb and had pancreatic cysts, and one brother had two Hbs removed that were detected in the brain and spine. One of the patient's nephews was diagnosed with a CNS Hb at age 12 and died 4 years later due to complications with the brain tumor that was not removed. Another niece was diagnosed at age 21 of cerebral Hb, and had the tumor resected. Upon follow-up, the patient still possesses pancreatic and renal cysts, but without renal tumor recurrence or metastases. The right optic nerve is intact, and there are no CNS lesions. She has slight decrease in visual acuity in the left eye due to the optic nerve Hb. Genetic testing of any of the patient's family members was not reported.

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 1 family of 2. Both patients had hemangioblastomas of the central nervous system, renal cysts, pancreatic cysts, and cystadenomas of the epididymis (family VHL 60). Also, 1/2 patients had retinal angioma, renal cell carcinoma and liver angiomas. The H115R (c.344A>G) mutation was detected in the patient, referred to in the paper by c.557A>G due to a difference in transcript sequence. This study contains supporting evidence of pathogenicity due to cosegregation with disease in multiple affected family members in a gene definitively known to cause the disease (ACMG code: PP1).

Of 65 VHL families from central Europe, 53 were identified with germline mutations using SSCP analysis of PCR products by peripheral blood. This splice mutation was found in a Italian, VHL type 1 family. Only one affected patient was identified with retinal angiomas, hemangioblastomas of the central nervous system, renal cell carcinoma, pancreatic cysts and cystadenomas of the epididymis. (family VHL 31). The c.464-2A>G mutation was detected in the patient, referred to in the paper by c.677-2A>G due to a difference in transcript sequence.

Of 65 VHL families from central Europe, 53 were identified with germline mutations, using SSCP analysis of PCR products by peripheral blood. The mutation of c.464-1G>A was referred to in the paper by c.677-1G>A due to a difference in transcript sequence. This splice mutation was found in a German, VHL type 1 patient with hemangioblastomas of the central nervous system (family VHL 48). This study contains very strong evidence of pathogenicity in the form of a canonical -1 splice site variant in a gene where loss-of-function is a known mechanism of disease (ACMG code: PVS1).

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. 683C>T) which coincides with legacy variant nomenclature (AF010238.1) and was mapped to c.470C>T (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, renal cell carcinoma (RCC), and pheochromocytoma was available. Case 45 had a p.Thr157Ile (c.470C>T) mutation. 2 members were affected: one had RCC and the other had RCC and pheochromocytoma. Other demographics of the members were not described.

In the analysis of 63 VHL patients, 55 patients were found with pancreatic involvement. Genetic tests on 35 of 55 VHL patients revealed VHL mutations in 28 patients. Hemangioblastomas of the central nervous system were found in 47 patients, renal cell carcinoma in 25, and pheochromocytoma in 11 patients. Peripheral blood samples were extracted and PCR sequencing was used to identify point mutations and multiplex ligation-dependent probe amplification (MLPA, P016-C2, MRC-Holland, Amsterdam, Netherlands) analysis was used to detect large rearrangements of the VHL gene. This missense mutation, c.256C>T, was found in a 26-year-old female with VHL type 2B phenotype (case no. 6). This patient presented with serous cystadenoma of the pancreas and pancreatic endocrine tumor. The patient was followed up after 35.8 months. The progression of VHL disease continued even after surgical resection.

In the analysis of 63 clinically diagnosed VHL patients, 55 patients were found with pancreatic involvement. Genetic tests on 35 of 55 VHL patients revealed VHL variants in 28 patients. Of the 55 enrolled patients, hemangioblastomas of the central nervous system were previously documented in 47 patients, renal cell carcinoma in 25, and pheochromocytoma in 11 patients. Peripheral blood samples were extracted and PCR sequencing was used to identify point mutations and multiplex ligation-dependent probe amplification (MLPA, P016-C2, MRC-Holland, Amsterdam, Netherlands) analysis was used to detect large rearrangements of the VHL gene. This missense mutation, c.208G>A, was found in 37-year-old female (case no. 1) who presented with pancreatic endocrine tumour and underwent a pylorus-preserving pancreaticoduodenectomy. The patient was followed up 27.7 months later.

A case report of a 16-year-old male with VHL is described. This patient presented at the age of 10 with complaints of headache, diplopia and was found to have intractable hypertension and bilateral pheochromocytoma as well as developing bilateral retinal hemangioblastoma at age 16. Family history revealed the proband’s mother had pheochromocytoma at age 23. These findings taken together prompted genetic screening for VHL. The above mutation was found in both the proband and mother. The variant was reported as “c.695G>A” which was mapped to p.Arg161Gln (c.482G>A) in accordance with reference sequence NM_000551.3.

Two patients and their families were presented in this paper with PKD1 and PKD2 mutations. Case 2, with 3 familial generations ADPKD, was diagnosed soon after birth with multiple small cortical cysts in both kidneys. The 4 year old male inherited a PKD1 and PKD2 mutation from each parent: c.7288C>T p.(Arg2430), and c.124_125ins52bp p.(Ala42fs66). A heterozygous variant in the VHL gene (NM_000551.3 (c.556G>A p.[Glu186Lys]) was additionally identified in this individual through a gene panel based Next Generation Sequencing approach. Patient is an offspring of a family with 3 generations of autosomal dominant polycystic kidney disease (ADPKD). Father had mild liver cysts and found to be a carrier of the truncating mutation in PKD1 at 18Y0M. Mother had mildly enlarged kidneys with small cysts bilaterally, as well as liver cysts and found to be a carrier of a truncating mutation in PKD2 at 19Y0M. No information provided on whether the parents possess the VHL mutation.

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.All patients underwent genetic testing using Sanger sequencing and Southern Blotting. Four patients were listed with this missense mutation, c.445G>T.

A cohort of VHL patients from the Greater Toronto Area in Canada were retrospectively reviewed. 7 patients were found with this germline mutation (c.496_506del), belonging to two pedigrees (but no data stated to determine which individuals belong together). Patient ID 44 had CNS hemangioblastoma and clear cell renal cell carcinoma; Patient ID 65 was found with the same mutation and only CNS hemangioblastoma; Patient ID 82 had retinal capillary hemangioma and hemangioblastoma; Patient ID 85 had hemangioblastoma, clear cell renal cell carcinoma and retinal capillary hemangioma; And patient ID 69, 83, 84 were all asymptomatic. Patients' age, and sex data was not reported.

17 patients from 10 families affected with VHL syndrome, particularly with retinal capillary hemangioblastomas (RCHs) were examined. Details about this patient are not specified in the study, but patient 15 from family 9 was found to have several tumours, including CNS hemangioblastoma. Sanger sequencing of a whole blood sample revealed a germline S65W(c.194C>G) missense variant in exon 1 of the VHL gene. Patient 15 has a family member (patient 16) who also has several tumors and the same germline S65W(c.194C>G) mutation, but the familial relationship to patient 15 is not noted. Neither patient had RCH.

Tissue specimens of apparent sporadic pheochromocytoma were retrieved from the archives of the Department of Pathology of the Erasmus MC (Rotterdam), the Radboud University Nijmegen Medical Center, and the University Hospital Zurich (Switzerland). Mutation analysis was performed using SSCP analysis and BigDye® Terminator V3.1 cycle sequencing kit (Applied Biosystems, Foster City, CA). 136 VHL cases and six germline VHL mutations were found. A c.440T>C, p.Ile147Thr mutation (stated as c.653T>C in the paper which used the Latif sequence as reference) was found in one 58Y female patient with benign adrenal pheochromocytoma. No other phenotypes, patient demographics, or family history was described.

Tissue specimens of apparent sporadic pheochromocytoma were retrieved from the archives of the Department of Pathology of the Erasmus MC (Rotterdam), the Radboud University Nijmegen Medical Center, and the University Hospital Zurich (Switzerland). Mutation analysis was performed using SSCP analysis and BigDye® Terminator V3.1 cycle sequencing kit (Applied Biosystems, Foster City, CA). 136 VHL cases and six germline VHL mutations were found. A c.188T>C, p.Leu63Pro mutation (stated as 401T>C in the paper which used the Latif sequence as reference) was found in one 26Y male patient (patient ID 37) with benign adrenal pheochromocytoma. No other phenotypes, patient demographics, or family history was described.

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.V130F(c.388G>T) was identified in two patients: patient #7 and patient #10. This mutation was noted as c.601G>T in the paper due to a difference in reference transcript used (see Table S1). No family information is available for either patient, and there is no indication these patients are related.