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

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

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

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

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

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

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

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

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

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 andd SSCP analysis. This missense mutation (c.264G>C) was recorded in the paper as c.477G>C due to a difference in transcript sequence. It was found in one patient with hemangioblastomas of the central nervous system. No other phenotype described.

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

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

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

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

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

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 (c.211insT, p.P71Lfs*61) was found in a 1 patient, without retinal hemangioblastomas, but 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, the missense mutation (E52K (c.154G>A)) was found in a single patient with no VHL retinal manifestation.

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

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

41 non-related patients with retinal capillary hemangioma presenting in the Benjamin Franklin University Eye Clinic between 1988 to 1999 Germany were evaluated for the presence of other VHL lesions. VHL disease was diagnosed in 33 patients based on the presence of other severe VHL lesions (spinal hemangioblastoma, cerebellar hemangioblastoma, renal cysts, pancreatic cyst, renal carcinoma, and pheochromocytoma), molecular genetic testing, and/or family history. VHL germline mutations were identified in 27 out of 29 VHL families screened with molecular genetics. Blood samples were genetically tested using PCR-SSCP sequencing. One patient with VHL-related retinal angiomatosis was found to have germline missense (c.452T>C) mutation in the VHL gene. 3 other affected relatives were also evaluated and found to have the variant but no phenotypes were provided. This variant was reported as c.665T>C (AF010238.1) and mapped to p.I151T (c.452T>C) (NM_000551.3).

41 non-related patients with retinal capillary hemangioma presenting in the Benjamin Franklin University Eye Clinic between 1988 to 1999 Germany were evaluated for the presence of other VHL lesions. VHL disease was diagnosed in 33 patients based on the presence of other severe VHL lesions (spinal hemangioblastoma, cerebellar hemangioblastoma, renal cysts, pancreatic cyst, renal carcinoma, and pheochromocytoma), molecular genetic testing, and/or family history. Blood samples were taken and analyzed by PCR and single-strand conformational polymorphism (SSCP). VHL germline mutations were identified in 27 out of 29 VHL families screened with molecular genetics. Group 20, consisted of a proband with retinal capillary hemangioma and a relative who also harboured the germline VHL mutation, R161 (c.481C>T). Group 21, consisted of one proband with retinal capillary hemangioma who also had the R161 mutation. It should be noted that this VHL mutation was stated as c.694C>T in the paper which used the Latif sequence as reference. No other affected family members were described for group 21. Other VHL lesions harbored by the patients were not specified.

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

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

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

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

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

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

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

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

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

53 patients with a molecularly confirmed VHL mutation (by PCR through EDTA-anticoagulated blood) and a pancreatic neuroendocrine tumor (ICT) were collected from the German NET-Registry and the German VHL-registry. An additional 296 patients were stated in this paper with a VHL associated germline mutation, but without pancreatic neuroendocrine tumors. 1 patient was found to have the above mutation (c.393C>A). This was a VHL patient with an ICT (pancreatic neuroendocrine tumor) taken from the VHL-registry. 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.

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

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

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

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

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

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

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. Three individuals (one proband and two family members) were genetically confirmed to have the p.N78S(c.233A>G) familial mutation. The proband (patient #36) was a male from kindred (family) #12 who was diagnosed with a CNS hemangioblastoma, renal cell carcinoma, and multiple pancreatic cysts/tumors at the age of 34. His sister (patient #37) also possessed this germline mutation and presented with multiple pancreatic cysts/tumors at age 27. The proband's father (patient #38) also had this mutation and presented with renal cell carcinoma at age 55. A fourth individual, the proband's uncle, passed away before diagnosis. He presented with a CNS hemangioblastoma at age 19, but was not genetically confirmed to have the mutation. 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 female patient, patient 172, from kindred #68, was found with this germline mutation p.C162Y (c.485G>A). She was diagnosed with a CNS hemangioblastoma, renal cell carcinoma, and multiple pancreatic cysts/tumors at 43Y. This individual was also likely published in a later paper by Wang et al., 2018 (PMID: 29330336). No family information was described.

A retrospective cohort study included all the VHL patients diagnosed at Peking University First Hospital (Beijing, China) prior to June 1 2016. A total of 291 patients from 115 different families were included for analysis. A 33Y male patient (patient #40, family #13) was confirmed for a missense mutation in the VHL gene (c.239G>T; p.Ser80Ile). He was diagnosed with renal cell carcinoma at 31Y, multiple pancreatic cysts or tumours at 31Y, pheochromocytoma at 31Y, and retinal angioma at 33Y. No family history is reported. This patient was also published in a paper by Wang et al., 2018 (PMID: 29330336).

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

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

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.V155M(c.463G>A) was identified in patient #4. This mutation was noted as c.676G>A in the paper due to a difference in reference transcript used (see Table S1). No family information is available for this patient.

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

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

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

93 families examined at the Clinical Center of the National Institutes of Health (Bethesda, MD) underwent genetic testing for germline mutations of the VHL gene. Whole blood was extracted and genomic DNA was amplified by PCR and products were sequences using a cycle sequencing kit with dye-labeled terminators and automatically sequenced. This nonsense mutation was found in a VHL type 1 family (lineage 20). The variant was reported as c.430C>T (AF010238.1) and was mapped to Q73* (c.217C>T) (NM_000551.3). This family is stated as being previously reported in PMID 7728151 and/or 8956040. Both these reports include a family presenting with this variant and it is unclear if one or both coincide with the current family.

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

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

PatientID: 27

KindredID: 11

Case: Sex unknown, Age unknown, French

DiseaseAssertion: VHL

FamilyInfo: None

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

CaseHPOFreeText: N/A

CaseNotHPOs: N/A

CaseNotHPOFreeText: N/A

CasePreviousTesting: N/A

PreviouslyPublished: N/A

SupplementalData: N/A

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

LegacyVariant: N/A

CaseProblemVariantFreeText: N/A

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

CAID: N/A

gnomAD: N/A

VariantEvidence:N/A

MutationType: missense_variant;transition

CivicName: P86S (c.256C>T)

MultipleGeneVariants: N/A

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

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

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

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

GroupID/ KindredID: 102CA

DiseaseAssertion: VHL

GroupSize: N/A

GroupNumberMales: N/A

GroupNumberFemales: N/A

GroupEthnicity: N/A

GroupAgeRange: N/A

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

GroupPresentingHPOs: HP:0005548 (renal cell carcinoma)

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

GroupNotHPOs: HP:0002666 (pheochromocytoma)

GroupNotHPOFreeText: N/A

GroupPreviousTesting: N/A

PreviouslyPublished: N/A

SupplementalData: N/A

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

LegacyVariant: FS (c.588insC)

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

ClinVarID: N/A

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

gnomAD: N/A

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

MutationType: duplication; plus_1_frameshift_variant

CivicName: D126fs (c.375dupC)

MultipleGeneVariants: N/A

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

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

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

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

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 (C162W (c.486C>G)) was taken from two databases: the Munich, university of Freiburg and Institute of Pathology database and the National Cancer Institute database. There was an unknown number of members belonging to Family IDs: 42CE and 3618NCI. It was reported in the paper as c.699C>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. Later publications reference this cohort as being reported in PMIDs: 10458336; and 7728151.

Germline mutational data on VHL was gathered from various countries to assess variation in germline mutations that produce disease within different populations. Families were grouped according to three possible outcomes of disease: renal cell carcinoma without pheochromocytoma, renal cell carcinoma with pheochromocytoma, or pheochromocytoma alone. SSCP and denaturing gradient gel electrophoresis identified mutations in 300/469 (63%) of families affected with VHL. This germline familial VHL mutation (V74G (c.221T>G)) was taken from the Munich, university of Freiburg and Institude of Pathology database (reported in Table 3, found in VHL family without pheochromocytoma, and with renal cell carcinoma) with an unknown number of members belonging to Family ID 53CE, as well as Family ID 9CE (reported in Table 4, found in VHL families with pheochromocytoma). It was reported in the paper as c.434T>G due to a difference in transcript reference. Other phenotypes are likely, however not detailed here.

Germline mutational data on VHL was gathered from various countries to assess variation in germline mutations that produce disease within different populations. Families were grouped according to three possible outcomes of disease: renal cell carcinoma without pheochromocytoma, renal cell carcinoma with pheochromocytoma, or pheochromocytoma alone. SSCP and denaturing gradient gel electrophoresis identified mutations in 300/469 (63%) of families affected with VHL. This germline familial VHL mutation (L184P (c.551T>C)) was taken from the National Cancer Institute database, with an unknown number of members belonging to Family ID 3708NCI. It was reported in the paper as c.764T>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. Later publications reference this cohort as being reported in PMID: 7728151.

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

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

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

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

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. The E70* (c.208G>T) mutation was detected in the patient, referred to in the paper by c.421G>T due to a difference in transcript sequence. This nonsense variant was found in a VHL type 1 patient with CNS hemangioblastomas (family ID 4479).

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 insertion mutation was detected in a VHL type 1 patient (family ID 4410), using PCR and SSCP sequencing. The mutation of c.386_387insAGA was referred to in the paper by c.599insAGA due to a difference in transcript sequence. The patient had retinal hemangioblastomas, CNS hemangioblastomas, and renal cell carcinoma. Only supporting evidence for pathogenicity because patient's phenotype is highly specific for a disease with a single genetic etiology (ACMG code: PP4).

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

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

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

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

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

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

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.R167Q (c.500G>A) germline variant of the VHL gene was detected in multiple families. Family 36: Member 3359 presented with mild renal disease. Member 3354 presented with severe renal disease (RCC). Family 262: Member 5426 presented with severe renal disease (RCC). Member 5510 had no renal involvement. Family 333: Member 9428 presented with mild renal disease. Member 13001 presented with moderate renal disease (RCC). The paper states that these families are previously reported in PMID 12202531 and/or 9829912. Family 238: Member 712 presented with mild renal disease (RCC) and member 6802 had unknown renal involvement. Family 411: Member 9710 presented with mild renal disease (RCC) and members 13888 and 12271 had unknown renal involvement. Family 509: Member 11209 presented with mild renal disease (RCC), member 10939 had unknown renal involvement and members 10901 and 10902 had no renal involvement. These families were not previously reported.

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

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

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

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

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 4 patients with a p.P86H (c.257C>A) mutation, 50% (2/4) of individuals had RH while the other 50% (2/4) 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 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 p.Q132P (c.395A>C) mutation. This member did not present with retinal hemangiomas, but no other clinical manifestations were discussed. Additionally, no family information was provided.

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

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. Two patients were listed with this missense mutation, c.500G>T.

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

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

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

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

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

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

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

This 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 T124P (c.370A>C) was one of the 15 found in a patient with PPGL. Their family history or further manifestations were not detailed.

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

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

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

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

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

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

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

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

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

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

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

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

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


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


Patients with ELSTs were identified and included in the International Endolymphatic Sac Tumor Registry. The registry contained 93 patients as of July 2014, and of those, 25 were sporadic, and 68 associated with VHL. Patients with ELSTs and clinically confirmed diagnoses of VHL were included in this study. Statistics of this VHL subgroup are as follows: mean age 30, 62% female, 91% symptomatic, 54% with retinal angioma, 72% with CNS hemangioblastoma, 46% with renal cell carcinoma, 19% with pheochromocytoma, and 12% with pancreatic neuroendocrine tumour. In the data from this study, only endolymphatic sac tumours can be confirmed as a phenotype for this mutation (c.485G>A), which was found in 1 individual from France.

74 patients (38 males, 36 females; 24 with prior sequencing data) with pheochromoctyoma were first analyzed by denaturing high performance liquid chromatography (dHPLC). PCR of all exons of VHL was performed on leukocyte DNA extracted from stored blood samples, and those showing variance were sequenced. In addition, 35 positive controls were used to confirm this method. This patient was a positive control who has been previously sequenced (not referenced). One patient was found with the R167Q (c.500G>A) VHL mutation. No family history was mentioned or clearly stated of their phenotypes.

74 patients (38 males, 36 females; 24 with prior sequencing data) with pheochromoctyoma were first analyzed by denaturing high performance liquid chromatography (dHPLC). PCR of all exons of VHL was performed on leukocyte DNA extracted from stored blood samples, and those showing variance were sequenced. These patients had presented with clinical features and/or family history suggestive of a germline mutation, leading to their referral for genetic testing. 2 patients found with the V84L (c.250G>T) VHL germline mutation were listed.
 No family history was mentioned.

Mutational analysis for 16 patients with clinically diagnosed VHL disease, revealed 12 germline mutations. This nonsense mutation was found in 2 unrelated families. One family of 3 had a asymptomatic 27-year-old female, a 27-year-old male with epididymal cystadenoma, and a 55-year old with hemangioblastomas of the central nervous system, renal cell carcinoma, and pancreatic cysts (proband no. 10). The other VHL family had 2 individuals with hemangioblastomas of the central nervous system (proband no. 11).

Mutational analysis for 16 patients with clinically diagnosed VHL disease, revealed 12 germline mutations. This missense mutation was found in a 45-year-old VHL type 1 patient with hemangioblastomas of the central nervous system, renal cell carcinoma, and pancreatic cysts (proband no. 4) and no family history. HPO terms: hemangioblastoma, renal cell carcinoma, multiple pancreatic cysts.

Data was collected from 82 VHL mutation carriers in the Dutch VHL surveillance program. One patient was found with this splice varaint in the VHL gene. The patient was 32Y at the last follow-up and presented with: retinal hemangioblastoma, cerebellar hemangioblastoma, spinal cord hemangioblastoma, renal cell carcinoma, renal cysts and pancreatic cysts. Family history was not described. This cohort was published later in PMID 31087189.

This paper presents the first case of VHL associated with acute myeloid leukemia. The patient is reported to have the 407 C/T (F136S) mutation of the VHL gene. He was admitted at the time of the study (38Y) with significant anemia, increased number of leukocytes and pneumonia. He had a history of renal cell carcinoma and a hemangioblastoma in his brainstem (medulla oblongata). Three years later he had recurrent renal cell carcinoma followed by a massive hematuria, dilated cardiomyopathy, complex valves defect, severe central sleep apnea, anemia, leukocytosis, moderate thrombocytopenia and erosive gastritis. Physical examination revealed a heart murmur and splenomegaly (confirmed by ultrasound). Prolonged bone marrow aplasia resulted in severe interstitial pneumonia and septicemia. The patient died from respiratory failure. The autopsy revealed a pulmonary edema, massive hyperemia of the left lower lobe with pneumonia confluens.

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

2 affected; 2 rHAB, 1 cHAB, 1 pheo between 1994 and 2001, the Dutch VHL working group conducted DNA analysis on 146 probands with VHL-associated manifestations. DNA of probands was extracted from peripheral blood samples. Exons 1, 2 and 3 of the VHL gene and their flanking untranslated and intronic coding sequences were amplified by PCR and subjected to semi-automated sequence analysis. Screening for structural rearrangements, including deletions, involving the VHL gene was performed by Southern blot analysis and the multiplex ligation-dependent probe amplification (MLPA) assay. Deletions encompassing the entire VHL gene were confirmed by FISH analysis on lymphocytes from affected individuals. VHL mutations were identified in 43 families. Family 24 has 2 affected members. Genetic analysis reveals mutation Q195*(c.583C>T) in the VHL gene. The patients have a family history of VHL disease. Of the 2 affected members, 2 members had retinal capillary hemangioma, 1 member had CNS hemangioblastoma, 1 member had pheochromocytoma. ACMG: 'PP4' because the patient's phenotype and family history are highly specific for a disease with single genetic etiology.

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

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

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 c.464-1G>T germline mutation of the VHL gene was found in a VHL type 1 family of 7. All seven had hemangioblastomas of the central nervous system, 3 had retinal angiomas, one had pancreatic cysts or tumor (unspecified), and 3 had renal cell carcinoma (family ID 3). This family was previously reported in PMID 8634692. ACMG codes as follows: A canonical -1 splice-site variant in a gene where loss-of-function is a known mechanism of disease (PVS1). Cosegregation with disease in multiple affected family members in a gene definitively known to cause the disease (PP1).

Screening of 3 patients affected with familial PC for mutations in the RET proto-oncogene (mutations related to MEN 2 syndrome) and the VHL tumor suppressor gene (mutations related to VHL disease) with the use of PCR, SSCP, heteroduplex analysis, and direct sequencing, revealed missense VHL gene mutations. Please note that there was no clinical diagnosis of VHL disease. This missense mutation c.499C>T was found in a Polish family with 3 siblings (two male siblings and their mother) with no evidence of RET; the older brother presented with unilateral pheochromocytoma at 15Y, the younger brother presented with bilateral pheochromocytoma at 11Y, and the mother presented with bilateral pheochromocytoma at 41Y. Additional phenotypic information was not provided. Please note that the paper states the mutation as c.712C>T because it uses a legacy reference sequence.

20 unrelated VHL families were retrospectively identified in SickKids Genetics and Cancer Genetics Program SHIRE genetics database, with data entered between 30 August 1984 and 8 August 2016. Family 9 had one patient with the missense mutation c.257C>T in the VHL gene (legacy notation c.470C>T). The proband (male) was diagnosed with CNS hemangioblastoma at 16Y and was 23Y at the end of the study. This patient was later published in 31368132- updated phenotypes available.

20 vHL families were retrospectively identified in SickKids Genetics and Cancer Genetics Program SHIRE genetics database, with data entered between 30 August 1984 and 8 August 2016. 6 members of family 17 tested positive for the c.496_506del germline variant of the VHL gene. The proband (ID 1), cousin (ID 8), and second cousin (ID 25) were asymptomatic at 3Y, 4Y and 9Y respectively. The mother (ID 4) presented with CNS hemangioblastoma (23 and 25Y), retinal angioma (22Y) and renal and pancreatic cyst or tumor (unspecified) (25Y). An aunt (ID 5) presented with retinal angioma (16Y). A first cousin once removed (ID 22) presented with CNS hemangioblastoma (36Y). 2 members were assumed to have the variant, due to personal and family history, but were not genetically confirmed (ID 20 and ID 11). A grandmother (ID 11) and great aunt (ID 20) both presented with CNS hemangioblastoma at 43Y and 40Y respectively. ID 11 also presented with pancreatic and renal cysts or tumors (unspecified) but the age of presentation was unknown.

Of 14 caucasian patients with retinal vascular proliferation and known family history of VHL disease, 12 patients were enrolled in an approved study protocol at the National Cancer Institute. 11 of these patients underwent genetic testing and were found to have mutations in the VHL gene. Patient 11, age 19, was diagnosed only with retinal vascular proliferation. She was found to carry the R167Q (c.500G>A) mutation. Presurgical and postsurgical optical coherence tomography in patient 11 demonstrated the fibrovascular complex as a hyperreflective layer that was limited to the vitreoretinal surface. There was traction on the underlying retinal tissue, resulting in thickening and obliteration of the foveal contour. Postsurgically, excision of the complex restored the overall shape of the foveal contour, leaving behind only a small disruption in the outer retinal lamination and a small residual macular epiretinal membrane. ACMG evidence codes: 'PP4' since the patient presents phenotypes highly specific for a disease with a single genetic etiology.

Of 14 patients with retinal vascular proliferation and known family history of VHL disease, 12 patients were enrolled in an approved study protocol at the National Cancer Institute. 11 of these patients underwent genetic testing and were found to have mutations in the VHL gene. Patient 6, age 34, had retinal vascular proliferation, CNS hemangioblastoma, pancreatic and renal abnormalities, and an epididymal cyst. She was found to carry the N78D (c.232A>G) mutation. ACMG evidence codes: 'PP4' since the patient presents phenotypes highly specific for a disease with a single genetic etiology.

Using polymerase chain reaction (PCR)-direct sequencing and universal primer quantitative fluorescent multiplex–PCR, mutations in VHL gene in 19 VHL disease families recruited from the Department of Urology, Peking University First Hospital in the period from 2009 to 2012 were examined. One proband (family 8) was found to carry the mutation c.350G>C in exon 2 of the VHL gene. The proband presented with hemangioblastoma at age 18 and renal cell carcinoma and abnormality of the pancreas at age 24. The patient had negative VHL family history. This patient was previously published in 29330336.

Using polymerase chain reaction (PCR)-direct sequencing and universal primer quantitative fluorescent multiplex–PCR, mutations in VHL gene in 19 VHL disease families recruited from the Department of Urology, Peking University First Hospital in the period from 2009 to 2012 were examined. One proband (family 5) was found to carry the mutation c.217C>T in exon 1 of the VHL gene. The proband presented with renal cell carcinoma at age 37 and pancreatic cysts or tumours at age 54. The patient had positive VHL family history but this was not confirmed by genetic testing. The proband's nephew had renal cell carcinoma at age 34 and cysts or tumours of the pancreas at age 37. The proband's niece had renal cell carcinoma and cysts or tumours of the pancreas at age 35.This family was previously published in 29330336.

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

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

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.340+1G>A germline mutation of the VHL gene. They presented with a retinal hemangioblastoma, cerebellar hemangioblastoma, spinal hemangioblastoma, renal cell carcinoma, renal cyst and pancreatic cysts. Their age at the last follow-up was 32. Previously published in: PMID 24132471.

The 32-year-old male patient was admitted to Xiangya Hospital in 2006 and was found with multiple CNS hemangioblastoma of the pons, medulla oblongata, cerebellum and spinal cord. Clinical workup also identified a solid renal tumor. He was followed-up for 5 years with no other complications. The patient and his 7 family members (his parents, sister, son, sister’s son, brother, and brother’s son) were assessed by RQ-PCR assay, and no large deletions were found. Direct sequencing identified the p.Phe76del (c.227_229delTCT) mutation in the proband and this mutation (or any other abnormality) was not found in any of the family members. All members were asymptomatic. This is supportive of a de novo mutation ACMG PM6.

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

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

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

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

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. Mutation G93C (c.277G>T) was identified in proband 43. Proband 43 presented with retinal capillary hemangioma and pheochromocytoma. Proband and mother were found to carry the V84L (c.250G>C) germline mutation of the VHL gene. No phenotype data was provided for the mother.

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. Two patients had a germline missense mutation in the VHL gene (c.383T>C; Leu128Pro). Patient 30, 27Y female, and Patient 31, 42Y female, both presented with CNS hemangioblastoma, renal cell carcinoma and a negative family history of VHL disease. The phenotypes of patient 30 were revised from a previous report (PMID 19270817).

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. 1 patient presented with the c.250G>T/p.Val84Leu germline variant of the VHL gene. Patient 16, 60Y female previously published in PMID 16142346, presented with multiple pheochromocytomas and a positive family history for VHL. She was diagnosed with VHL Type 2C.

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. Three unrelated probands were genetically confirmed with a germline missense mutation at c.499C>T of the VHL gene. Patients 42 and 44, 10Y and 30Y males respectively, both presented with multiple and/or bilateral pheochromocytoma and CNS hemangioblastomas. Patient 43, 30Y male, presented with multiple and/or bilateral pheochromocytoma and retinal hemangioblastoma. Patients 42 and 43 had a family history negative for VHL disease while patient 44 had a positive family history of VHL disease. Patient 43s phenotypes were revised from a previous report (PMID 19949673). Patient 44 was previously reported in PMID 16142346 as well as Kim et al., 2005 (reference 15 in the current paper). All of these patients were diagnosed with VHL Type 2A.

42 CNS hemangioblastoma samples were obtained from 31 patients, who were consecutively treated between 1993 and 1997 at the Department of Neurosurgery of the Freiburg University Medical Centre were analyzed for VHL mutations. Two patients of 17 presenting with VHL germline mutations presented with the L89P (c.266T>C) mutation (stated as c.479T>C in the paper which used the Latif sequence as reference). These patients were related but their relation is not made clear. Single strand conformation polymorphism and southern blotting for variant identification in the DNA found in blood samples. The patients were established as having VHL disease by standard of Neumann at al., 1987. Both patients presented with a single cerebellum tumors.

42 CNS hemangioblastoma samples were obtained from 31 patients, who were consecutively treated between 1993 and 1997 at the Department of Neurosurgery of the Freiburg University Medical Centre were analyzed for VHL mutations. Three patients of 17 presenting with VHL germline mutations presented with the c.292T>C mutation (stated as c.505C>T in the paper which used the Latif sequence as reference). These patients were related but their relation is not made clear. Single strand conformation polymorphism and southern blotting for variant identification in the DNA found in blood samples. The patients were established as having VHL disease by standard of Neumann at al., 1987. The first two of these patients (patients 11 and 13) each presented with a single cerebellum tumor. The third of these patients presented with a medulla oblongata tumor and a spinal tumor at C8.

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

Germline mutations were found in 20 Brazilian, VHL probands and their families. The P102Rfs*57 (c.305del) germline mutation was found in a VHL type 1 patient with hemangioblastomas of the central nervous system, pancreatic cystadenoma, and renal carcinoma (VHL family 6). The patient had no family history. 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. Peripheral blood samples were taken and genomic DNA was sequenced using PCR and Southern Blotting. The three exons of the VHL gene were amplified by PCR, sequenced in both strands with Big Dye terminators (Applied Biosystems), and collected and analysed on an ABI Prism 377 (Applied Biosystems). Germline rearrangements involving part or the entire VHL gene were detected by quantitative Southern blotting using g7 cDNA11 and a β globin probe. This splice site mutation, c.463+1G>T (stated as IVS2+1G>A in the paper), was found in a VHL type 1 South American family of five affected individuals (family 14). Three family members had hemangioblastoma of central nervous system, one had a retinal capillary hemangioma and another had renal cell carcinoma. One or more members of the family had pancreatic cyst-adenoma. No ACMG code available.

Two groups were analyzed for germline VHL mutation where Group 1 (84 individuals) presented with only one hemangioblastoma and no other clinical VHL features and those in Group 2 (4 individuals) presented with multiple hemangioblastomas and no other VHL clinical features. Peripheral blood analysis was conducted of all exons and flanking regions (90 nucleotides upstream and 45 downstream) of the VHL gene (numbered using Genbank accession number L15409). Southern blot, PCR and SSCP were performed. Patient B1 of Group 1 presented with a singular cerebellar hemangioblastoma at age 29 and was found to be harbouring a 7bp frameshift T124del (c.369_371del), referred to in the paper as del 582 GACACAC due to a difference in transcript sequence. She developed pancreatic cysts at age 34.

Through Next Generation Sequencing (NGS), PCR and automated Sanger sequencing, genetic analysis of 21 patients with known germline mutations in an inherited pheochromocytoma/paraganglioma gene was done. Ten of which were in the VHL gene. This missense mutation (c.292C>T) was found in 8 unrelated patients with pheochromocytoma (tumor IDs P5, P6, P7, P8, P9, P10, P11, P12). Family history is not stated.

33 patients diagnosed with VHL disease, according to (Maher et al. 1990), were tested for germline VHL mutations. Genetic testing was performed on high-molecular-weight DNA from peripheral blood using Southern analysis and PCR. One patient was found to harbour the 217C>T (Gln73Ter) VHL gene mutation, recorded in the paper as c.430C>T due to a difference in transcript sequence. This patient presented with renal cell carcinoma. ACMG evidence codes: 'PP4' since the patient presents phenotypes highly specific for a disease with a single genetic etiology.

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

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

This paper analyzes a large family with Von-Hippel Lindau disease. Clinical manifestations consistent with VHL were found in 13 members of this family and all 5 affected family members who were genetically tested were positive for the Tyr112Asn (T547A; assumed to be reported according to AF010238.1 and mapped to c.334T>A [NM_000551.3]) germline mutation of the VHL gene. The other affected individuals were unavailable for testing or died before testing could be completed. Interestingly, 3 family members tested positive without clinical manifestations, but may be too young to present with features. Clinical manifestations in this family included: CNS hemangioblastoma, renal cell carcinoma (7/13 members), retinal hemangioblastoma, renal cysts, and pheochromocytoma. ACMG evidence codes: 'PP1' based on segregation results, 'PP2' because they observe a missense variant in genes where missense variants are common mechanisms of disease, 'PP4' because the patient's phenotypes and family history are highly specific for a disease with single genetic etiology.

The proband (II/3), of italian descent, was found to have the c.499C>T change (referred to in the paper by 8702C->T due to a difference in transcript sequence) causing R167W of the VHL gene. This variant was detected using direct sequencing of PCR products obtained from genomic DNA extracted from the patients peripheral blood leukocytes using the DNA extractor BIO ROBOT EZ 1 (Qiagen). The c.499C>T change was detected in the exon 3 of VHL gene. He had retina angioma with renal carcinoma. His daughter (III/3) also had the mutation and was symptomatic but specific phenotypes were not specified. His son (III/4) was asymptomatic, but it is unclear if he was tested or not for the genetic variant. His father (I/1), had the mutation and presented with a haemangioblastoma. His brother (II/2), also had the mutation and presented with renal carcinoma, central nervous system haemangioblastoma, pheochromacytoma and pancreatic cysts. ACMG evidence codes: 'PP1' since we have cosegregation with the disease in multiple affected family members, 'PP4' since the patient presents phenotypes highly specific for a disease with a single genetic etiology.

This abstract reported the case of unilateral single familial pheochromocytoma diagosed in a father and his son. They both carried a single mutation in the VHL gene, c.235C>G (p.R79G). The son was diagnosed and treated at age 9, while the father was intervened at age 40. ACMG: 'PP4' because the patients' phenotypes and family history are highly specidic for a disease with single genetic etiology.

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). The p.V66* (c.196del) nonsense mutation (noted as c.409delG (p.137TER) in the paper due to a difference in reference transcript used) was found in at least one member of this family (kindred no. 43) affected with VHL disease type 1 (no phenochromocytoma). Pheochromocytoma was not observed in any members of this patient's family. ACMG evidence codes: none.

A case report of a woman presenting with bilateral adrenal pheochromocytoma and  multiple pancreatic massess (paraganglioma) at 18 years of age is presented. Molecular analysis on peripheral leukocytes confirmed the presence of the above mutation (c.499C>T). Additional clinical manifestations in this patient include retinal and cerebellar hemangioblastomas (at 19Y and 22Y), and a neuroendocrine tumour (at 24Y). Family history shows 3 additional family members who likely have VHL, but have not been genetically tested. Clinical manifestations in these individuals included: pheochromocytoma, pancreatic cysts, and paraganglioma. 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 patients' phenotype or family history are highly specific for a disease with single genetic etiology.

The National Cancer Institute has a frozen tumour bank of ccRCC tumours that have been removed from VHL patients. Illumina's Human Whole-Genome Sequencing service was used to sequence blood, tumour tissue, and normal kidney tissue. One 28Y male patient with ccRCC was confirmed with a germline missense mutation (p.Asn78Ser) in the VHL gene. No family history or other phenotype was provided.

Through PCR purification and Sanger sequencing of peripheral blood (using an alcohol precipitation method), 121 consecutive, unrelated, index PCC/PGL patients underwent genetic testing for five PCC/PGL susceptibility genes (RET, VHL, SDHB, SDHD and SDHC) and were evaluated for clinical diagnosis of neurofibromatosis type1. Three unrelated probands were genetically confirmed with a familial mutation at c.499C>T of the VHL gene. The first was an 11 year old male who presented with bilateral pheochromocytoma and retinal capillary hemangioma. The second was an 11 year old male who presented with bilateral pheochromocytoma and PNETs. The third was an 18 year old female who presented with bilateral pheochromocytoma and sympathetic paraganglioma, CNS hemangioblastoma and PNETs. Family histories are unknown but positive.

This paper reports a VHL family harboring a novel missense mutation (N78D). Genetic testing was performed using peripheral whole blood, on 9 members of this kindred, which revealed 3 patients carried the above mutation. The 6 members who did not carry the mutation were asymptomatic, while the 3 carriers had manifestations. In addition to the 3 carriers, there was 3 additional kin who had clinical features of VHL, but died before testing could be performed. The clinical manifestations in this kindred included: spinal hemangioblastoma, cerebellar hemangioblastoma, endolymphatic sac tumor, pancreatic cysts, and renal cysts. ACMG evidence codes: 'PP1' because of segregation results, '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.

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

A 2.75 year old patient (32 month-old girl) presented with a pheochromocytoma. Genetic sequencing (not specified) revealed a c.491A>G (p.Gln164Arg) heterozygous germline mutation in exon 3 of the VHL gene. Sequencing of her mother, father, a sibling, and her paternal grandparents revealed that the mutation was present only in her father, and was de-novo in her father. Upon screening, her father (a 31 year old male) presented with a retinal angioma, an adenoma of right adrenal gland, a right-sided renal cyst, an epidydymal cyst, a contrast-enhancing lesion in the pancreas, and bilateral pheochromocytomas.

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 21 and 25 were each one member (proband) each and were found with complete VHL deletions. Proband 21 had only hemangioblastoma and genetic testing of her parents confirmed a de novo mutation. Proband 25 was found with hemangioblastoma, retinal angioma and pancreatic lesion. Proband 25 had no family history of VHL but parents were not available for genetic testing so de novo mutation cannot be assessed. ACMG PVS1

Two Swedish families with von Hippel-Lindau disease (VHL) were analyzed. Family B consists of 4 members across 2 generations. Patient I:1 (proband) is a female with renal cell carcinoma and cerebellar hemangioblastomas. Genetic analysis of blood samples revealed a frameshift mutation N174fs (c.520insAA, originally written in paper was legacy notation c.732insAA) with a downstream premature stop codon at position 201. Genetic analysis of her two children also confirmed the mutation: her son, patient II:1, had cerebellar hemangioblastoma and her daughter, patient II:2, was asymptomatic. The proband's partner (I:2) had no mutation and no VHL phenotype. ACMG: PP4