Case3: Male, 35 years report, 5 years onset, Algeria, African

CasePresentingHPOs: HP:0003323, HP:0003749, HP:0002355, HP:0007340, HP:0008956, HP:0003484, HP:0003236, HP:0003198 (Progressive muscle weakness, Pelvic girdle muscle weakness, Difficulty walking, Lower limb muscle weakness, Proximal lower limb amyotrophy, Upper limb muscle weakness, Increased serum CK, Myopathy)

HPOsFreeText: At grade 3 of Vignos and Walton scales, CK rate was 5 times the normal value and AST and ALT were normal, EMG was compatible with a myopathic disorder, GAA activity measured in leukocytes was significantly decreased

CaseNotHPOs: HP:0003115, HP:0003116, HP:0001270, HP:0001290, HP:0002240, HP:0001640, HP:0005952, HP:0002540 (Abnormal ECG, Abnormal echocardiography, Motor delay, Generalized hypotonia, Hepatomegaly, Cardiomegaly, Decreased pulmonary function, Non-ambulatory)

Case12: Male, 35 years onset, 50 years diagnosis, 61 years report, France

CasePresentingHPOs: HP:0002540, HP:0002878, HP:0004887 (Inability to walk, Respiratory failure, Respiratory failure requiring assisted ventilation)

HPOsFreeText: Lost ambulation at 48 years, Started invasive ventilation at 35 years, Started ERT given every other week at a dose of 20 mg/kg at 56 years for 49 months, Peak Anti-rhGAA antibodies at 1:1600, FVC at 0 (L / % expected), MIP at 22 (cm H2O), MEP at 28 (cm H2O), Time off ventilation stayed constant at 0 min, 6MWT stayed constant at 0 meters

CaseNotHPOs: HP:0030148 (Heart murmur)

NotHPOsFreeText: No adverse reaction to ERT was recorded

CaseAddInfo: Late Onset Pompe Disease

Case3: Male, 28 years onset, 30 years diagnosis, 69 years report, France

CasePresentingHPOs: HP:0002540, HP:0002878, HP:0004887, HP:0002093 (Inability to walk, Respiratory failure, Respiratory failure requiring assisted ventilation, Respiratory insufficency)

HPOsFreeText: Lost ambulation at 58 years, Started invasive ventilation at 28 years, Started ERT given every other week at a dose of 20 mg/kg at 68 years for 11 months, Peak Anti-rhGAA antibodies at 1:1600, FVC at 0.45/13 (L / % expected), MIP at 17 (cm H2O), MEP at 29 (cm H2O), Time off ventilation stayed constant at 0, Total Motor Function Measure from 31 to 27

CaseNotHPOs: HP:0030148 (Heart murmur)

NotHPOsFreeText: No adverse reaction to ERT was recorded

CaseAddInfo: Late Onset Pompe Disease, Patient died before publication

Case25: Male, 2.7 yr onset, 3.5 yr diagnosis, 5.9 yr report

HPOsFreeText: FVC z-score sitting 0.34 (104% pred), FVC z-score supine 0.04 (100% pred), 87% MRC Scale, 908 CK U/l, 2.8 GAA activity (nmol/h/mg)^h from cultured fibroblasts

NotHPOsFreeText: No wheelchair used, No respiratory support

Case21: Male, 16 yr diagnosis, 17.1 yr report

HPOsFreeText: FVC z-score sitting 1.29 (115% pred), FVC z-score supine -0.25 (97% pred), 1506 CK U/l, Deficent GAA activity (nmol/h/mg)^h from cultured fibroblasts

NotHPOsFreeText: No wheelchair used, No respiratory support

Case1: Male, 0.5 yr onset, 2.5 yr diagnosis, 10.04 yr report

CasePresentingHPOs: HP:0001371 (Contractures)

HPOsFreeText: Wheelchair used, FVC z-score sitting -0.25 (97% pred), FVC z-score supine -1.12 (87% pred), 65% MRC Scale, 530 CK U/l, 9.1 GAA activity (nmol/h/mg)^h from cultured fibroblasts

NotHPOsFreeText: No respiratory support

Case9: Male, 2 yr onset, 1 yr diagnosis, 6.6 yr report

HPOsFreeText: FVC z-score sitting -0.97 (89% pred), FVC z-score supine -1.4 (84% pred), 94% MRC Scale, 763 CK U/l, 11 GAA activity (nmol/h/mg)^h from cultured fibroblasts

NotHPOsFreeText: No wheelchair used, No respiratory support

Case9: Male, 5 yr onset, 10.8 yr diagnosis, 10.8 yr report

HPOsFreeText: FVC z-score sitting 0.2 (102% pred), FVC z-score supine -0.29 (97% pred), 97% MRC Scale, 588 CK U/l, 11.9 GAA activity (nmol/h/mg)^h from cultured fibroblasts

NotHPOsFreeText: No wheelchair used, No respiratory support

Case19: Male, 13.1 yr diagnosis, 13.1 yr report

HPOsFreeText: FVC z-score sitting -0.69 (92% pred), FVC z-score supine -1.77 (80% pred), 100% MRC Scale, 614 CK U/l, Deficent GAA activity (nmol/h/mg)^h from cultured fibroblasts

NotHPOsFreeText: No wheelchair used, No respiratory support

Case7: Male, 1 yr onset, 2 yr diagnosis, 13.7 yr report

CasePresentingHPOs: HP:0002650 (Scoliosis)

HPOsFreeText: Respiratory support at night, FVC z-score sitting -4.08 (54% pred), FVC z-score supine -5.4 (39% pred), 87% MRC Scale, 548 CK U/l, 17.9 GAA activity (nmol/h/mg)^h from cultured fibroblasts

NotHPOsFreeText: No wheelchair used

CaseI: Early adult onset, 49 years diagnosis

CasePresentingHPOs: HP:0003701, HP:0003388, HP:0003236, HP:0003198 (Proximal muscle weakness, Easy fatigability, Increased serum CK, Myopathy)

HPOsFreeText: Plasma CK 5–10× Normal, GAA activity from lymphocytes: 14.2%

CaseNotHPOs: HP:0002795 (Abnormal respiration)

CaseAddInfo: Late-Onset Pompe Disease

CaseIV,3: Early adult onset, 50 years diagnosis

CasePresentingHPOs: HP:0003236, HP:0003546 (Increased serum CK, Exercise intolerance)

HPOsFreeText: Plasma CK 2.5–5× Normal

CaseNotHPOs: HP:0002795 (Abnormal respiration)

CaseAddInfo: Late-Onset Pompe Disease

CaseV,2: Adult onset, 33 years diagnosis

CasePresentingHPOs: HP:0003701, HP:0003236, HP:0003388 (Proximal muscle weakness, Increased serum CK, Easy fatigability)

HPOsFreeText: Plasma CK 5-10× Normal, 6-Minute Walking Test: 566 min, Forced Vital Capacity Seated: 3000 (90%), GAA activity from dried blood spot: 0.20%

CaseNotHPOs: HP:0002795 (Abnormal respiration)

CaseAddInfo: Late-Onset Pompe Disease

CaseIV,2: Early adult onset, 47 years diagnosis

CasePresentingHPOs: HP:0003701, HP:0003236, HP:0002795, HP:0003546 (Proximal muscle weakness, Increased serum CK, Abnormal respiration, Exercise intolerance)

HPOsFreeText: Plasma CK 2.5–5× Normal

CaseAddInfo: Late-Onset Pompe Disease

CaseA: Male, 10 months onset, 18 months diagnosis

CaseAPresentingHPOs: HP:0001270, HP:0003324 (Motor delay, Generalized weakness)

HPOsFreeText: CRIM positive

CaseNotHPOs: HP:0100543 (Cognitive impairment)

CaseAddInfo: Infantile-Onset Pompe Disease

Case4: Female, 25 years onset, 43 years diagnosis, 58 years report

CasePresentingHPOs: HP:0003325, HP:0003327, HP:0003236, HP:0002747 (Limb-girdle muscle weakness, Axial muscle weakness, Increased serum CK, Respiratory muscle weakness)

CaseAddInfo: Late-Onset Pompe Disease, Sibling of Patient 4a

Case8: Female, 18 years onset, 18.5 years diagnosis, 29 years report

CasePresentingHPOs: HP:0003325, HP:0003236 (Limb-girdle muscle weakness, Increased serum CK)

CaseAddInfo: Late-Onset Pompe Disease

Case14: Male, 28 years onset, 36 years diagnosis, 37 years report

CasePresentingHPOs: HP:0003325, HP:0003327, HP:0003236, HP:0003691 (Limb-girdle muscle weakness, Axial muscle weakness, Increased serum CK, Scapula alata)

CaseAddInfo: Late-Onset Pompe Disease

Case10: Female, 47 years onset, 50 years diagnosis, 64 years report

CasePresentingHPOs: HP:0003325, HP:0003236 (Limb-girdle muscle weakness, Increased serum CK)

CaseAddInfo: Late-Onset Pompe Disease

Patient15: Male, 21 years report, 2 years onset

CasePresentingHPOs: N/A

HPOsFreeText: GAA activity 4.7% from a dried blood spot

CaseNotHPOs: HP:0002098, HP:0001324, HP:0001640, HP:0002240 (Respiratory distress, Muscle weakness, Cardiomegaly, Hepatomegaly)

NotHPOsFreeText: No wheelchair needed, Respiratory Assistance not needed

CaseAddInfo: LOPD

Case1: Male, Caucasian, United States, Birth onset

CasePresentingHPOs: HP:0008872, HP:0002100, HP:0001508, HP:0008947, HP:0001655, HP:0003236, HP:0200136, HP:0001290, HP:0002835, HP:0002194 (Feeding difficulties in infancy, Recurrent aspiration pneumonia, Poor weight gain, Infantile muscular hypotonia, Patent foramen ovale, Increased serum CK, Oropharyngeal dysphagia, Low muscle tone, Aspiration, Delayed gross motor development)

HPOsFreeText: HyperCKemia was present since the early neonatal period (378 IU/L, day 8 of life; N: 60–305 IU/L), ERT at a dose of 20 mg/kg every 2 weeks was initiated at age 1 month, CK levels were normalized on ERT by age 4 months (80 IU/L), Flexion/abduction/ external rotation positioning of lower extremities, iliotibial (IT) band tightness, lack of independent sitting, AIMS score between 10th and 25th percentile, GMFM at 23.16%, At age 23 months, he had achieved significant motor milestones; started sitting with support at 10 months and without support at 12 months, pulled to stand at 12 months, walked at 15 months and could feed himself with an immature pincer grasp. Swallow study was normal, ALT 26 U/L (Normal 5–40 U/L), AST 45 U/L (Normal 15–41 U/L), ALP 211 U/L (Normal 24–170 U/L), Urinary Glc4 3.7 mmol/mol creatinine (Normal ≤8.3 mmol/mol creatinine)

CaseNotHPOs: HP:0011675, HP:0002910 (ECG abnormality, Elevated hepatic transaminases)

NotHPOsFreetext:

CaseAddInfo: Older brother of Patient 2, Late-Onset Pompe Disease

Case2: Male, Caucasian, United States, Birth onset

CasePresentingHPOs: HP:0001290, HP:0200136, HP:0002033, HP:0003236, HP:0001655, HP:0002058, HP:0002194, HP:0002421, HP:0000467, HP:0002015 (Generalized hypotonia, Oral-pharyngeal dysphagia, Poor suck, Increased serum CK, Patent foramen ovale, Myopathic face, Delayed gross motor development, Poor head control, Neck muscle weakness, Dysphagia)

HPOsFreeText: hyperCKemia at 435 IU/L on day of life 4, Patent foramen ovale detected at birth by ECHO but resolved spontaneously, PT evaluation at age 2 months revealed flexion/abduction/external rotation positioning of lower extremities with tightness bilaterally in hip flexors, hamstrings, and IT bands, Had a tendency to remain on toes when held in supported standing with additional support required for weight bearing through lower extremities, Had difficulty lifting his head in prone, with asymmetric head lift to 45 degrees, and an inability to maintain head in midline when supine, AIMS score at age 2 months was between 10–25th percentile, ALT 41 U/L (Normal 5–40 U/L), AST 33 U/L (Normal 15–41 U/L), ALP 239 U/L (Normal 24–170 U/L), Urinary Glc4 4.5 mmol/mol creatinine (Normal ≤8.3 mmol/mol creatinine)

CaseNotHPOs: HP:0011016 (Abnormality of urine glucose concentration)

NotHPOsFreeText:

CaseAddInfo: Younger brother of Patient 1, Late-Onset Pompe Disease

Case7: Female, Caucasian, United States, 6 months diagnosis, 8 months report

CasePresentingHPOs: HP:0001643, HP:0006699, HP:0006682, HP:0003722, HP:0008969 (Patent ductus arteriosus, Premature atrial contractions, Premature ventricular contractions, Neck flexor weakness, Leg muscle stiffness)

HPOsFreeText: At 6 months head lag during pull to sit was noted, Had predominant overall flexion position of lower extremities with a tendency towards hip flexion/abduction/external rotation, with lateral thigh grooves suggestive of bilateral IT band tightness, AIMS score was between 10th and 25th percentile for chronological age and 50th percentile for her corrected age adjusted for prematurity while GMFM score was 16.55%, ALT 37 U/L (Normal 5–40 U/L), AST 53 U/L (Normal 15–41 U/L), ALP 174 U/L (Normal 24–170 U/L)

CaseNotHPOs: HP:0003236, HP:0001712, HP:0001290 (Increased serum CK, Left ventricular hypertrophy, Decreased muscle tone)

NotHPOsFreeText: Other evaluations including pulmonary, speech, and swallow were normal

Case6: Male, Caucasian, United States, 1 month onset, 6 months diagnosis, 8 months report

CasePresentingHPOs: HP:0003236, HP:0001655, HP:0003701, HP:0006589, HP:0003093, (Increased serum CK, Patent foramen ovale, Proximal muscle weakness, Flaring of lower rib cage, Limited hip extension,

HPOsFreeText: HyperCKemia was noticed in the first month (438 U/L; Normal 60–305) but normalized spontaneously by age 6 months, flexion/abduction/external rotation positioning of lower extremities and with bilateral IT band tightness was revealed at the same time, also had decreased active use of abdominal oblique muscles with a lack of active trunk rotation, Decreased active use of lower abdominal muscles with inability to achieve a posterior pelvic tilt in supine, AIMS score was at 1st percentile for age and GMFM was at 12.3%, ALT 30 U/L (Normal 5–40 U/L), AST 90 U/L (Normal 15–41 U/L), ALP 249 U/L (Normal 24–170 U/L), Urinary Glc4 2.6 mmol/mol creatinine (Normal ≤8.3 mmol/mol creatinine)

CaseNotHPOs: HP:0001712, HP:0001290 (Left ventricular hypertrophy, Decreased muscle tone)

NotHPOsFreeText: Other evaluations including cardiac, pulmonary, speech, and swallow were normal

Case15: Female, 13 years onset, 43 years diagnosis

CaseAddInfo: Late-Onset Pompe Disease

Case20: Female, 3 years onset, 4 years diagnosis

CaseAddInfo: Late-Onset Pompe Disease

Case6: Male, 21 years report, Italy

HPOsFreeText: Height is 1.74 m, Weight is 71 kg, BMI is 23.5 kg/cm2, 7 years on ERT, Forced Vital Capacity at 99%

CaseNotHPOs: HP:0002098 (Respiratory distress)

NotHPOsFreeText: Nocturnal non-invasive ventilation

CaseAddInfo: Late Onset Pompe Disease

Case1: Female, 46 years report, Italy

CasePresentingHPOs: HP:0003551 (Difficulty climbing stairs)

HPOsFreeText: Height is 1.64 m, Weight is 50 kg, BMI is 18.6 kg/cm2, 2 years on ERT, Forced Vital Capacity at 108%

CaseNotHPOs: HP:0002098 (Respiratory distress)

NotHPOsFreeText: Nocturnal non-invasive ventilation

CaseAddInfo: Late Onset Pompe Disease

Case1: Female, 12 years report, 3 years onset, 12 years diagnosis

CasePresentingHPOs: HP:0003325, HP:0002093, HP:0002650 (Limb-girdle muscle weakness, Respiratory insufficiency, Scoliosis)

HPOsFreeText: 730 CK (U/L), Normal Heart, 0 GAA activity (µmol/L/h), Showed respiratory involvement and required nocturnal non-invasive ventilation, An ambulant patient with 284 m six minute walk distance at the time of diagnosis

CaseNotHPOs: HP:0003457, HP:0001627 (EMG abnormality, Abnormality of the heart)

NotHPOsFreeText: Muscle weakness confined to limb-girdle muscles

Case1: Male, French-Canadian, 18 years onset, 54 years diagnosis and report

CasePresentingHPOs: HP:0008994, HP:0000822, HP:0003119, HP:0004398, HP:0007185, HP:0009109, HP:0002094, HP:0012764, HP:0002098 (Proximal muscle weakness in lower limbs, High blood pressure, dyslipidemia, Peptic ulcer, Loss of consciousness, Denervation of the diaphragm, Dyspnea, Orthopnea, Shortness of breath)

HPOsFreeText: Dependent on continuous positive airway pressure or bilevel positive airway pressure when he is lying flat or sleeping because of previous intubation and tracheostomy, GAA activity measured from dried blood spot was 0.87 pmol/h/punch (Normal 4.96 - 27.3)

CaseAddInfo: Late Onset Pompe Disease, The patient’s mother had six miscarriages and died at age 75 of lung cancer and his father died of cardiac problems in his early sixties, It appears that no other family members, including his parents and two siblings, were affected, patient had a previous appendectomy, He has no allergies, stopped smoking in 2002, and denied any alcohol or drug use

RescuePatientCell: Primary fibroblasts from a SLC6A8-deficient patient.

GeneAlteration: c.1540C>T (p.Arg514X) (previously described by Salomons et al, 2001, Am J Hum Genet, 68: 1497-1500)

RescuePhenotype: Ability to transport creatine into the cell.

RescueMethod: The wild type SLC6A8 open reading frame was cloned into an expression vector with EGFP. This construct was stably transfected into primary fibroblasts from patient with creatine transporter deficiency due to a nonsense variant, c.1540C>T (p.Arg514X) in SLC6A8 (previously described in PMD 11326334). Expression of the SLC6A8 fusion protein or EGFP protein (empty vector) was confirmed by the presence of green fluorescence. Expression was also confirmed by RT-PCR. Transfected cells were incubated for 24 hours with 25 umol/L creatine, 500 umol/L creatine, or 500 umol/L creatine plus 500 umol/L guanidinoproprionate (creatine transporter inhibitor). Intracellular creatine level was measured by GC-MS.

RescueEvidence: When the wild type SLC6A8 open reading frame was expressed in primary fibroblasts from patient with creatine tarnsporter deficiency, due to a nonsense variant, c.1540C>T (p.Arg514X) in SLC6A8 (previously described in PMID 11326334), creatine uptake was restored to a level comparable with controls.

RescueVariants: ClinVarID: 11696

Case2: Male, 59 years report, 45 years diagnosis, Germany

CasePresentingHPOs: HP:0002878 (Respiratory failure)

HPOsFreeText: 6-min walk test at 204 (m)

CaseNotHPOs:

NotHPOsFreeText: Not wheelchair bound

CaseAddInfo: 6.5 years on ERT

Case1: Male, 51 years report, 50 years diagnosis, Germany

CasePresentingHPOs: HP:0002878 (Respiratory failure)

HPOsFreeText: 6-min walk test at 484 (m)

CaseNotHPOs: register NotHPOsFreeText: Not wheelchair bound

CaseAddInfo: 1.5 years on ERT

CaseExome-2:I-1: Male, Japan, Asian

CaseExome-2:I-1PresentingHPOs: HP:0001873, HP:0040185 (Thrombocytopenia, Macrothrombocytopenia)

HPOsFreeText: Platelet Count at 113 (x (10^9)/l), Platelet Size at 2.7 ± 0.9 (μm), No bleeding tendency

CaseNotHPOs:

NotHPOsFreeText:

CaseAddInfo: Father of proband, II-1

CaseSanger-3:I-2: Female, Japan, Asian

CasePresentingHPOs: HP:0001873, HP:0040185 (Thrombocytopenia, Macrothrombocytopenia)

HPOsFreeText: Platelet Count at 132 (x (10^9)/l), Platelet Size at 2.8 ± 0.8 (μm), No bleeding tendency

CaseNotHPOs:

NotHPOsFreeText:

CaseAddInfo: Mother of Proband, II-2

CaseExome-2:II-2: Male, Japan, Asian

CaseExome-2:II-2PresentingHPOs: HP:0001873, HP:0040185, HP:0000421 (Thrombocytopenia, Macrothrombocytopenia, Epistaxis)

HPOsFreeText: Platelet Count at 124 (x (10^9)/l), Platelet Size at 2.9 ± 0.9 (μm), No bleeding tendency

CaseNotHPOs:

NotHPOsFreeText:

CaseAddInfo: Brother of proband, II-1

CaseSanger-3:II-2: Female, Japan, Asian

CasePresentingHPOs: HP:0001873, HP:0040185, HP:0000421 (Thrombocytopenia, Macrothrombocytopenia, Epistaxis)

HPOsFreeText: Platelet Count at 80 (x (10^9)/l), Platelet Size at 3.0 ± 0.9 (μm), 59% (2.5 μm) aggregation ADP

CaseNotHPOs:

NotHPOsFreeText:

CaseAddInfo: Proband of Family Sanger-3

CaseExome-1:II-1: Female, Japan, Asian

CaseExome-1:II-1PresentingHPOs: HP:0001873, HP:0040185 (Thrombocytopenia, Macrothrombocytopenia)

HPOsFreeText: Platelet Count at 91 (x (10^9)/l), Platelet Size at 3.8 ± 1.4 (μm), No bleeding tendency

CaseNotHPOs:

NotHPOsFreeText:

CaseAddInfo:

CaseExome-2:II-1: Male, Japan, Asian

CaseExome-2:II-1PresentingHPOs: HP:0001873, HP:0040185 (Thrombocytopenia, Macrothrombocytopenia)

HPOsFreeText: Platelet Count at 54 (x (10^9)/l), Platelet Size at 3.1 ± 1.1 (μm), No bleeding tendency, 4.5 min bleeding time, 34% (2 μm) aggregation ADP

CaseNotHPOs:

NotHPOsFreeText:

CaseAddInfo: Proband for Family Exome-2

CaseExome-1:I-2: Female, Japan, Asian

CaseExome-1:I-2PresentingHPOs: HP:0001873, HP:0040185 (Thrombocytopenia, Macrothrombocytopenia)

HPOsFreeText: Platelet Count at 80 (x (10^9)/l), Platelet Size at 3.7 ± 1.1 (μm), No bleeding tendency

CaseNotHPOs:

NotHPOsFreeText:

CaseAddInfo: Mother of proband, II-1

CaseSanger-1:I-1: Male, Japan, Asian

CasePresentingHPOs: HP:0001873, HP:0040185 (Thrombocytopenia, Macrothrombocytopenia)

HPOsFreeText: Platelet Count at 131 (x (10^9)/l), Platelet Size at 4.2 ± 1.3 (μm), No bleeding tendency, 1 min bleeding time, 49% (2 μm) aggregation ADP

CaseNotHPOs:

NotHPOsFreeText:

CaseAddInfo: Father of proband, II-1

CaseSanger-1:II-2: Female, Japan, Asian

CasePresentingHPOs: HP:0001873, HP:0040185 (Thrombocytopenia, Macrothrombocytopenia)

HPOsFreeText: Platelet Count at 97 (x (10^9)/l), Platelet Size at 3.6 ± 1.4 (μm), No bleeding tendency, 74% (2 μm) aggregation ADP

CaseNotHPOs:

NotHPOsFreeText:

CaseAddInfo: Sister of Proband, II-1

CaseExome-3:I-2: Female, Japan, Asian

CasePresentingHPOs: HP:0001873, HP:0040185 (Thrombocytopenia, Macrothrombocytopenia)

HPOsFreeText: Platelet Count at 100 (x (10^9)/l), Platelet Size not done, No bleeding tendency

CaseNotHPOs:

NotHPOsFreeText:

CaseAddInfo: Mother of proband, II-2

CaseSanger-1:II-1: Female, Japan, Asian

CasePresentingHPOs: HP:0001873, HP:0040185 (Thrombocytopenia, Macrothrombocytopenia)

HPOsFreeText: Platelet Count at 120 (x (10^9)/l), Platelet Size at 3.9 ± 1.6 (μm), No bleeding tendency, 66% (2 μm) aggregation ADP

CaseNotHPOs:

NotHPOsFreeText:

CaseAddInfo: Proband for Family Sanger-1

CaseExome-3:II-2: Male, Japan, Asian

CasePresentingHPOs: HP:0001873, HP:0040185, (Thrombocytopenia, Macrothrombocytopenia)

HPOsFreeText: Platelet Count at 77 (x (10^9)/l), Platelet Size at 2.9 ± 0.6 (μm), No bleeding tendency, 49% (2 μm) aggregation ADP

CaseNotHPOs:

NotHPOsFreeText:

CaseAddInfo: Proband for Family Exome-3

CaseSanger-2:III-2: Female, Japan, Asian

CasePresentingHPOs: HP:0001873, HP:0040185 (Thrombocytopenia, Macrothrombocytopenia)

HPOsFreeText: Platelet Count at 77 (x (10^9)/l), Platelet Size at 3.7 ± 1.5 (μm), No bleeding tendency, 2 min bleeding time

CaseNotHPOs:

NotHPOsFreeText:

CaseAddInfo: Proband of Family Sanger-2, Had a sister, mother, and grandfather who had the disorder but were not genetically tested to confirm the presence or absence of her variant

CaseF7: Male, Italy

CasePresentingHPOs: HP:0001873 (Thrombocytopenia)

HPOsFreeText: World Health Organization bleeding score is 0 (1 member); Mean platelet count is 110 ×10^9/L; Mean platelet volume is 12.1 fL, Mean platelet diameter is 2.5 μm

CaseAddInfo: N/A

CaseF8P: Italy

CasePresentingHPOs: N/A

CaseAddInfo: Proband for F8

Case: Haagerup II:5

CasePresentingHPOs:

CasePresentingHPOsFreeText: not specified

CaseNOTHPOs:

CaseNOTHPOSFreeText: not specified

CasePreviousTesting: same as group

CaseGenotypingMethod: same as group

Variants: c926T>C and c236G>A

ClinVarID: or CAID: ClinVarID: 8426; ClinVarID: 8424

gnomAD: 2/129174 European (non-Finnish) alleles at a frequency of 0.00001548 with no homozygotes; not present in gnomAD

AddCaseInfo:

PreviouslyPublished: no

VariantEvidence: predicted but variant was proven to segregate with disease; no detectable protease activity when analyzed by western blotting (PMID: 10074491)

c.684+1G>T variant falls within this range

This variant is mentioned in reference to a previous publication that is not associated with disease.

GroupID/ KindredID: VHL 48

Case: Sex unknown, Age unknown, German

DiseaseAssertion: assumed VHL

FamilyInfo: One patient with the given mutation is stated to belong to VHL family 48

CasePresentingHPOs: HP:0010797 (hemangioblastoma)

CaseHPOFreeText: N/A

CaseNotHPOs: N/A

CaseNotHPOFreeText: N/A

CasePreviousTesting: N/A

PreviouslyPublished: N/A

SupplementalData: N/A

Variant: c.464-1G>A

CaseProblemVariantFreeText: N/A

LegacyVariant: c.677-1G>A

ClinVarID: 43603 https://www.ncbi.nlm.nih.gov/clinvar/variation/43603/

CAID: N/A

gnomAD: N/A

VariantEvidence: N/A

MutationType: splice_site_variant

CivicName: Splice Site (c.464-1G>A)

MultipleGeneVariants: N/A

Of 65 VHL families from central Europe, 53 were identified with germline mutations using SSCP analysis of PCR products by peripheral blood. The Q164 (c.490C>T) mutation was detected in the patient, referred to in the paper by Q164 (c.703C>T) due to a difference in transcript sequence. This nonsense mutation was found in an Iranian, VHL type 1 patient with retinal angiomas, hemangioblastomas of the central nervous system, renal cell carcinoma, pancreatic cysts, and pancreatic islet cell tumors (family VHL 65). This study contains very strong evidence of pathogenicity in the form of a nonsense variant in a gene where loss-of-function is a known mechanism of disease (ACMG code: PVS1).

150 index patients with pheochromocytoma/paraganglioma were evaluated. Phenotypic data were collected and germline mutations in five susceptibility genes (RET, VHL, SDHB, SDHD and SDHC) were tested. Out of 30 patients with familial presentation, 13 were clinically diagnosed as VHL syndrome. A 32 year old Asian Indian female presented with unilateral pheochromocytoma. Using PCR and Sanger sequencing through peripheral blood, genetic testing confirmed a non-familial splice mutation at c.341-2 A>G of the VHL gene. Family history is unknown.

KindredID: 15

DiseaseAssertion: VHL

GroupSize: Unknown

GroupNumberMales: N/A

GroupNumberFemales: N/A

GroupEthnicity: N/A

GroupAgeRange: N/A

GroupInfo: This is a family belonging to prefigure No. 15. They are described as ‘VHL-affected’.

GroupPresentingHPOs: HP:0005584 (renal cell carcinoma)

GroupHPOFreeText: RCC-affected; other manifestations are possible but not clear.

GroupNotHPOs: N/A

GroupNotHPOFreeText: N/A

GroupPreviousTesting: N/A

PreviouslyPublished: N/A

SupplementalData: N/A

Variant: p.Y98H (c.292T>C)

LegacyVariant: p.Y169H (c.505T>C)

CaseProblemVariantFreeText: N/A

ClinVarID: 2223 https://www.ncbi.nlm.nih.gov/clinvar/variation/2223/

CAID: N/A

gnomAD: 0.00001343, https://gnomad.broadinstitute.org/variant/3-10183823-T-C?dataset=gnomad_r2_1

VariantEvidence: N/A

MutationType: missense_variant; transition

CivicName: Y98H (c.292T>C)

MultipleGeneVariants: N/A

KindredID: 14

DiseaseAssertion: VHL

GroupSize: Unknown

GroupNumberMales: N/A

GroupNumberFemales: N/A

GroupEthnicity: N/A

GroupAgeRange: N/A

GroupInfo: This is a family belonging to prefigure No. 14. They are described as ‘VHL-affected’.

GroupPresentingHPOs: HP:0005584 (renal cell carcinoma)

GroupHPOFreeText: RCC-affected; other manifestations are possible but not clear.

GroupNotHPOs: N/A

GroupNotHPOFreeText: N/A

GroupPreviousTesting: N/A

PreviouslyPublished: N/A

SupplementalData: N/A

Variant: p.Y98H (c.292T>C)

LegacyVariant: p.Y169H (c.505T>C)

CaseProblemVariantFreeText: N/A

ClinVarID: 2223 https://www.ncbi.nlm.nih.gov/clinvar/variation/2223/

CAID: N/A

gnomAD: 0.00001343, https://gnomad.broadinstitute.org/variant/3-10183823-T-C?dataset=gnomad_r2_1

VariantEvidence: N/A

MutationType: missense_variant; transition

CivicName: Y98H (c.292T>C)

MultipleGeneVariants: N/A

Tissue analysis from 61 VHL patients, 30 sporadic renal cell carcinoma (RCC) tumor samples, and 6 sporadic RCC cell lines revealed 22 variants within VHL patients, 10 variants in tumor samples and variants in all cell lines. All mutations cluster to the 3’ end of the VHL gene open reading frame, implicating this region as important for the tumor-suppressor function of the VHL protein. Northern blot, SSCP and RT-PCR analysis from normal tissue found a germline p.Q145Hfs*28 (c.435_436delGC) mutation, stated in the paper as c.648_649delGC due to an older transcript. It was seen in family [pedigree No] 9, with an unknown number of members described as ‘VHL-affected’ with RCC. Other manifestations are possible but not clear.

PatientID: (proband)

KindredID: 4

Case: M, Age unknown, Chinese

DiseaseAssertion: pheochromocytoma 
FamilyInfo: There is a history of non-syndromic familial pheochromocytoma in this patient’s family history as well as a confirmed presence of this same germline mutation in their family. It is not perfectly clear which pedigree belongs to this family.

CasePresentingHPOs: HP:0002666 (pheochromocytoma)

CaseHPOFreeText: non-syndromic pheochromocytoma

CaseNotHPOs: N/A

CaseNotHPOFreeText: No clinical manifestations of hereditary syndromes, including MEN2A, MEN2B, VHL, and NF1 were found

CasePreviousTesting: N/A

PreviouslyPublished: N/A

SupplementalData: N/A

Variant: p.R167W (c.499C>T)

LegacyVariant: p.R167W (c.712C>T)

CaseProblemVariantFreeText: N/A

ClinVarID: 2218, https://www.ncbi.nlm.nih.gov/clinvar/variation/2218/

CAID: N/A

gnomAD: 0.000007954, https://gnomad.broadinstitute.org/variant/3-10191506-C-T?dataset=gnomad_r2_1

VariantEvidence: N/A

MutationType: missense_variant;transition

CivicName: R167W (c.499C>T)

MultipleGeneVariants: N/A

Genotype-phenotype correlations of 573 VHL patients were analyzed and confirmed that higher risk of pheochromocytoma is associated with missense mutations that result in substitution of a surface amino acid or that disrupt the structural integrity of the VHL gene product (pVHL). This missense mutation (c.496G>T), affecting surface residue of pVHL protein core, was found in 2 VHL type 2A families with 5 affected individuals altogether. Two of 5 patients had retinal angioma, 2 had cerebellar hemangioblastoma, and 4 had pheochromocytoma. This mutation was reported in the paper with an incorrect cDNA written as c.498A>G, however correct translation should be deduced to c.496G>T, as the Latif et al. 1993 (AFO10238) sequence given was as c.709G>T with p.V166F.

Genotype-phenotype correlations of 573 VHL patients were analyzed and confirmed that higher risk of pheochromocytoma is associated with missense mutations that result in substitution of a surface amino acid or that disrupt the structural integrity of the VHL gene product (pVHL). This missense mutation (c.538A>G), affecting residue within the pVHL protein core, was found in a VHL patient with renal cell carcinoma diagnosed at 27Y. ACMG codes as follows: Supporting evidence of pathogenicity because Missense variant in a gene that has a low rate of benign missense variation and where missense variants are a common mechanism of disease (PP2).

Genotype-phenotype correlations of 573 VHL patients from 200 kindreds were analyzed and confirmed that higher risk of pheochromocytoma is associated with missense mutations that result in substitution of a surface amino acid or that disrupt the structural integrity of the VHL gene product (pVHL). Age-related risks of retinal hemangoblastomas and renal cell carcinoma were higher in patients with nonsense or frameshift mutations. Southern blot and/or MLPA analysis by whole blood detected this mutation (c.555C>G) was found in 3 VHL families with 4 patients altogether. All four had retinal hemangioblastomas and cerebellar hemangioblastomas. Two had renal cell carcinoma.

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

Germline mutational data on VHL was gathered from various countries to assess variation in germline mutations that produce disease within different populations. Families were grouped according to three possible outcomes of disease: renal cell carcinoma without pheochromocytoma, renal cell carcinoma with pheochromocytoma, or pheochromocytoma alone. SSCP and denaturing gradient gel electrophoresis identified mutations in 300/469 (63%) of families affected with VHL. This germline familial VHL mutation (F136S (c.407T>C)) was taken from the Neuro Olcologie database in Paris, and the Cambridge, Department of Pathology database. It had an unknown number of members belonging to Family ID 53F, reported in Table 3, as being found in VHL families without pheochromocytoma, and with renal cell carcinoma. Additionally, it was reported in table 5, specified as families without detailed clinical information, under family ID 27CA. It was reported in the paper as c.620T>C due to a difference in transcript reference.

Germline mutational data on VHL was gathered from various countries to assess variation in germline mutations that produce disease within different populations. Families were grouped according to three possible outcomes of disease: renal cell carcinoma without pheochromocytoma, renal cell carcinoma with pheochromocytoma, or pheochromocytoma alone. SSCP and denaturing gradient gel electrophoresis identified mutations in 300/469 (63%) of families affected with VHL. This germline familial VHL mutation (c.464-2A>G) was taken from the Bristol Myers Squibb database, and the Munich, university of Freiburg and Institute of Pathology database. It had an unknown number of members belonging to Family ID 31CE, reported in Table 3, as being found in VHL families without pheochromocytoma, and with renal cell carcinoma. Additionally, it was reported in table 5, specified as families without detailed clinical information, under family ID 28BMS. It was reported in the paper as c.677-2A>G due to a difference in transcript reference.

Germline mutational data on VHL was gathered from various countries to assess variation in germline mutations that produce disease within different populations. Families were grouped according to three possible outcomes of disease: renal cell carcinoma without pheochromocytoma, renal cell carcinoma with pheochromocytoma, or pheochromocytoma alone. SSCP and denaturing gradient gel electrophoresis identified mutations in 300/469 (63%) of families affected with VHL. This germline familial VHL mutation (L188V (c.562C>G)) was taken from the Munich, university of Freiburg and Institute of Pathology database, with an unknown number of members belonging to two family IDs: Family ID 37CE, and 38CE. It was reported in the paper as c.775C>G due to a difference in transcript reference. As reported in Table 4, this mutation is found in VHL families with pheochromocytoma. Other phenotypes are likely, including renal cell carcinoma, however not detailed here.

GroupID/ KindredID: 4325NCI, 25CA

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 National Cancer Institute databas, and the Cambridge, Department of Pathology database, with an unknown number of members belonging to Family ID 4325NCI and 25CA. Table 3 lists all ID and mutations associated with individuals possessing renal carcinoma, without phechromocytoma.

GroupPresentingHPOs: HP:0005548 (renal cell carcinoma)

GroupHPOFreeText: Described as a VHL families 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.S65W (c.194C>G)

LegacyVariant: p.S65W (c.407C>G)

CaseProblemVariantFreeText: N/A

ClinVarID: 43597 https://www.ncbi.nlm.nih.gov/clinvar/variation/43597

CAID: N/A

gnomAD: N/A

VariantEvidence: N/A

MutationType: missense_variant; transversion

CivicName: S65W (c.194C>G)

MultipleGeneVariants: N/A

GroupID/ KindredID: 74CA, 2753NCI, 38CA

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 as well as the National Cancer Institute database with an unknown number of members belonging to Family ID 38CA, 74CA, 2753NCI. Table 3 lists all ID and mutations associated with individuals possessing renal carcinoma, without phechromocytoma.

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: 7728151; 9829911

SupplementalData: N/A

Variant: p.Q73* (c.217C>T)

LegacyVariant: p.Q73* (c.430C>T)

CaseProblemVariantFreeText: N/A

ClinVarID: 223164 https://www.ncbi.nlm.nih.gov/clinvar/variation/223164/

CAID: N/A

gnomAD: N/A

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

MutationType: stop_gained

CivicName: Q73* (c.217C>T)

MultipleGeneVariants: N/A

GroupID/ KindredID: 37CE, 38CE

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 Munich, university of Freiburg and Institute of Pathology database, with an unknown number of members belonging to two family IDs: Family 37CE and 38CE. Table 4 lists all ID and mutations associated with individuals with pheochromocytoma.

GroupPresentingHPOs: HP:0002666 (pheochromocytoma)

GroupHPOFreeText: Described as multiple VHL families with pheochromocytoma. Other phenotypes are likely, including renal cell carcinoma, however not detailed here.

GroupNotHPOs: N/A

GroupNotHPOFreeText: N/A

GroupPreviousTesting: N/A

PreviouslyPublished: N/A

SupplementalData: N/A

Variant: p.L188V (c.562C>G)

LegacyVariant: p.L188V (c.775C>G)

CaseProblemVariantFreeText: N/A

ClinVarID: 2225 https://www.ncbi.nlm.nih.gov/clinvar/variation/2225/

CAID: N/A

gnomAD: 0.00002121 https://gnomad.broadinstitute.org/variant/3-10191569-C-G

VariantEvidence: This paper declares this variant is predicted to compromise the Elongin-binding site by location between codons 157 and 189.

MutationType: missense_variant;transversion

CivicName: L188V (c.562C>G)

MultipleGeneVariants: N/A

GroupID/ KindredID: 88CA, 82CA

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 the Cambridge, Department of Pathology database. There was an unknown number of members belonging to Family IDs: 88CA and 82CA. 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.Y185* (c.555C>G)

LegacyVariant: p.Y185* (c.768C>G)

CaseProblemVariantFreeText: N/A

ClinVarID: 223233 https://www.ncbi.nlm.nih.gov/clinvar/variation/223233/

CAID: N/A

gnomAD: N/A

VariantEvidence: This paper declares this variant is predicted to compromise the Elongin-binding site either by truncating the protein or by location between codons 157 and 189.

MutationType: stop_gained

CivicName: Y185* (c.555C>G)

MultipleGeneVariants: N/A

Fourty-four of 389 patients with VHL (between 1988-1999) screened positive for PNETs either by pathologic analysis of tissue specimens or by characteristic radiographic appearance on CT and MRI. VHL gene germline mutations were determined by quantitative Southern blotting to detect deletions of the gene and gene rearrangements as well as complete gene sequencing. Patient 29 (29-year-old male) was found to harbour the c. 486C>G mutation in the VHL gene (stated as 699 C to G in the paper which used the Latif sequence as reference) and presented with two pathologic pancreatic neuroendocrine tumors. ACMG evidence codes: 'PP4' since the patient presents phenotypes highly specific for a disease with a single genetic etiology.

Fourty-four of 389 patients with VHL (between 1988-1999) screened positive for pancreatic neuroendocrine tumors (PNETs) either by pathologic analysis of tissue specimens or by characteristic radiographic appearance on CT and MRI. VHL gene germline mutations were determined by quantitative Southern blotting to detect deletions of the gene and gene rearrangements as well as complete gene sequencing. Patient 43 was found to have the gemline VHL mutation c.562C>G (stated as 775 C to G in the paper which used the Latif sequence as reference) leading to a Leu to Val change at protein position 188. She presented with four pathologic pancreatic neuroendocrine tumors at age 35. She was also found to harbour the c.241C>T (stated in the paper as 454 C to T) VHL gene mutation leading to a Pro to Ser change at protein position 81. ACMG evidence codes: 'PP4' since the patient presents phenotypes highly specific for a disease with a single genetic etiology.

Fourty-four of 389 patients with VHL (between 1988-1999) screened positive for PNETs either by pathologic analysis of tissue specimens or by characteristic radiographic appearance on CT and MRI. VHL gene germline mutations were determined by quantitative Southern blotting to detect deletions of the gene and gene rearrangements as well as complete gene sequencing. Patient 14, a 42 year old male, was found to harbour the c.490 C>T mutation in the VHL gene (stated as 703 C to T in the paper which used the Latif sequence for reference) leading to a premature stop codon at protein position 164 in exon 3. He presented with a single pathologic pancreatic neuroendocrine tumor. ACMG evidence codes: 'PP4' since the patient presents phenotypes highly specific for a disease with a single genetic etiology.

Fourty-four of 389 patients with VHL (between 1988-1999) screened positive for PNETs either by pathologic analysis of tissue specimens or by characteristic radiographic appearance on CT and MRI. VHL gene germline mutations were determined by quantitative Southern blotting to detect deletions of the gene and gene rearrangements as well as complete gene sequencing. One patient (15), a 26 year old female, was found to harbour the p.Val181Glyfs*20 (c.540_543delCGTC) VHL gene mutation, stated in the paper as c.753_756delCGTC according to an older transcript reference. She presented at 21Y with a single radiologic pancreatic neuroendocrine tumor. No family information was present.

GroupID: 3127

DiseaseAssertion: assumed VHL

GroupSize: 47

GroupNumberMales: N/A

GroupNumberFemales: N/A

GroupEthnicity: German

GroupAgeRange: N/A

GroupInfo: Familial VHL stated; 47 family members recorded with this mutation.

GroupPresentingHPOs: HP:0010797; HP:0009711; HP:0002666 (hemangioblastoma, retinal capillary hemangioma, pheochromocytoma)

GroupHPOFreeText:  22 members had an angioma (hemangioma), 27 had pheochromocytomas, and only 4 member had CNS hemangioblastoma

GroupNotHPOs: HP:0005584 (renal cell carcinoma)

GroupNotHPOFreeText: N/A

GroupPreviousTesting: N/A

PreviouslyPublished: 8956040 (family confirmed in Zbar 1996 as Type 2 3127NCI); 10458336

SupplementalData: N/A

Variant: p.Y98H (c.292T>C)

CaseProblemVariantFreeText: N/A

LegacyVariant: p.Y98H (c.505T>C)

ClinVarID: 2223 https://www.ncbi.nlm.nih.gov/clinvar/variation/2223/

CAID: N/A

gnomAD: 0.00001343 https://gnomad.broadinstitute.org/variant/3-10183823-T-C

VariantEvidence: N/A

MutationType: missense_variant; transition

CivicName: Y98H (c.292T>C)

MultipleGeneVariants: N/A

In a study of 114 unrelated VHL families, 85 germline mutations were found. VHL mutations were detected in affected family members, but not in unaffected family members or 96 normal individuals. This nonsense mutation was found in a VHL type 1 family of 7 individuals (family ID 2753), using PCR and SSCP sequencing. The mutation of c.217C>T was referred to in the paper by c.430C>T due to a difference in transcript sequence. Six of 7 individuals were identified with retinal hemangioblastomas, 3 with cerebellar hemangioblastomas and one with renal cell carcinoma. This was possibly previously published in PMID: 9829911. Supporting evidence of pathogenicity because cosegregation with disease in multiple affected family members in a gene definitively known to cause the disease (ACMG code: PP1).

In a study of 114 unrelated VHL families, 85 germline mutations were found. VHL mutations were detected in affected family members, but not in unaffected family members or 96 normal individuals.  This mutation was found in a VHL family of 4 individuals (family ID 4409), using PCR and SSCP sequencing. The mutation of c.340G>C was referred to in the paper by c.553G>C due to a difference in transcript sequence. 4/4 retinal angioma; 2/4 CNS hemangioblastoma; 1/4 RCC; 1/4 Pheo. Walther et al., 1999 also reports a a partial update of this family, describing that Family 4409 also had pancreatic neuroendocrine tumour but not how many members were affected by PNET. (ACMG code: PP4).

GroupID: 2753

DiseaseAssertion: assumed VHL

GroupSize: 7

GroupNumberMales: N/A

GroupNumberFemales: N/A

GroupEthnicity: N/A

GroupAgeRange: N/A

GroupInfo: Familial VHL stated; 7 family members recorded with this mutation.

GroupPresentingHPOs: HP:0010797; HP:0009711; HP:0005584 (hemangioblastoma, retinal capillary hemangioma, renal cell carcinoma)

GroupHPOFreeText: 6/7 members had a retinal angioma (hemangioma), 3 had RCC, and 1 member had CNS hemangioblastoma

GroupNotHPOs: HP:0002666 (pheochromocytoma)

GroupNotHPOFreeText: N/A

GroupPreviousTesting: N/A

PreviouslyPublished: Possibly previously published in PMID: 9829911 [Q73* (c.217C>T) family (ID 20)]

SupplementalData: N/A

Variant: p.Q73* (c.217C>T)

CaseProblemVariantFreeText: N/A

LegacyVariant: p.Q73* (c.430C>T)

ClinVarID: 223164 https://www.ncbi.nlm.nih.gov/clinvar/variation/223164

CAID: N/A

gnomAD: N/A

VariantEvidence: N/A

MutationType: stop_gained

CivicName: Q73* (c.217C>T)

MultipleGeneVariants: N/A

GroupID: 4409

DiseaseAssertion: assumed VHL

GroupSize: 4

GroupNumberMales: N/A

GroupNumberFemales: N/A

GroupEthnicity: N/A

GroupAgeRange: N/A

GroupInfo: Familial VHL stated; 4 family members recorded with this mutation.

GroupPresentingHPOs: HP:0010797; HP:0009711; HP:0005584; HP:0002666 (hemangioblastoma, retinal capillary hemangioma, renal cell carcinoma, pheochromocytoma)

GroupHPOFreeText: all 4 members had retinal angiomas (hemangiomas), 2 had CNS hemangioblastomas, 1 had RCC and 1 had pheochromocytoma

GroupNotHPOs: N/A

GroupNotHPOFreeText: N/A

GroupPreviousTesting: N/A

PreviouslyPublished: 10458336

SupplementalData: N/A

Variant: p.G114R (c.340G>C)

CaseProblemVariantFreeText: N/A

LegacyVariant: p.G185R (c.553G>C)

ClinVarID: 223193 https://www.ncbi.nlm.nih.gov/clinvar/variation/223193

CAID: N/A

gnomAD: N/A

VariantEvidence: N/A

MutationType: missense_variant; transversion

CivicName: G114R (c.340G>C)

MultipleGeneVariants: N/A

A retrospective study of metastatic (n = 10) or unresectable (n=5) PPGL patients who received Lu-DOTATATE therapy (registered at KEM Hospital, Mumbai, India) between January 2010 and December 2019 were examined. Two VHL mutations were found through genotyping based on the protocol from PMID: 27539324. One male aged 27Y when therapy started, had pheochromocytoma, pancreatic neuroendocrine tumor and sympathetic paraganglioma, which were considered progressive, along with liver metastasis. He had a Y98S (c.293A>C) VHL mutation. No family information was given.

A retrospective study of metastatic (n = 10) or unresectable (n=5) PPGL patients who received Lu-DOTATATE therapy (registered at KEM Hospital, Mumbai, India) between January 2010 and December 2019 were examined. Two VHL mutations were found through genotyping based on the protocol from PMID: 27539324. One male aged 14Y when therapy started, had pheochromocytoma and pancreatic neuroendocrine tumor which were considered progressive and inoperable. He had a R167W (c.499C>T) VHL mutation. No family information was given.

A 20Y male patient presented with pheochromocytoma and genetic analysis confirmed a germline mutation c.311G>T (p.Gly104Val) in the VHL gene. He also presented with nausea, vomiting, progressively rising blood pressure, an epididymal cyst, scrotal hydrocele, portal/gonadal vein thrombosis and a nonfunctional adrenal mass at 20Y and a history of anxiety and episodic diaphoresis. This mutation was confirmed in the patient's 48Y father who presented with a T1 vertebral body hemangioma, multiple bilateral renal cysts, left adrenal nodularity, hearing loss and an indeterminate right kidney midpole lesion. The paternal first cousin once removed died due to complications from clear cell renal cell carcinoma at 50Y, although genotype for the cousin is not provided. Mother and 2 siblings of the proband tested negative for the VHL mutation.

PatientID: 1

KindredID: 5

Case: F, 13Y0M, Ethnicity Unknown

DiseaseAssertion: VHL

FamilyInfo: Positive. The patient's mother (4) also carried this mutation and had a hemangioblastoma. The proband's aunt (5) was confirmed to have the mutation and had VHL presentations. The cousin (8) had the mutation but was asymptomatic. The grandfather (13) was inferred to have the mutation and had a hemangioblastoma. See Figure 2a for family pedigree.

CohortInfo: 20 unrelated VHL families from an ethnically diverse cohort containing both affected and unaffected individuals.

CasePresentingHPOs: HP:0010797, HP:0001732 (hemangioblastoma, abnormality of the pancreas)

CaseHPOFreeText: The patient presented with a hemangioblastoma and abnormality of the pancreas at age 10.

CaseNotHPOs: HP:0009711, HP:0005584, HP:0002666, HP:0000077 (retinal capillary hemangioma, renal cell carcinoma, pheochromocytoma, abnormality of the kidney)

CaseNotHPOFreeText: N/A

CasePreviousTesting: N/A

PreviouslyPublished: 31368132

SupplementalData: Supplementary file 1.

Variant: p.F136Lfs*23 (c.408delT)

CaseProblemVariantFreeText: N/A

LegacyVariant: N/A

ClinVarID: 43601 https://www.ncbi.nlm.nih.gov/clinvar/variation/43601/

CAID: N/A

gnomAD: N/A

VariantEvidence: N/A

MutationType: minus_1_frameshift_variant

CivicName: F136Lfs*23 (c.408delT)

MultipleGeneVariants: N/A

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. Family 5 was found with this frameshift mutation, c.408delT in the VHL gene . The female proband (ID 1) was found with hemangioblastoma, abnormality of the pancreas and had a VHL age of onset of 10Y and her last known age was 13Y. Her mother (ID 4) was found with hemangioblastoma at 37Y and was 39Y at the time of the study. Her aunt (ID 5) was found with hemangioblastoma, retinal capillary hemangioma, abnormality of the kidney, abnormality of the pancreas, had a VHL age of onset of 29Y and her last known age was 37Y. Her male cousin (ID 8) was 5Y and had no manifestations. Her grandfather (ID 13) was not genetically confirmed with the mutation, but was found with hemangioblastoma at 57Y and was 63Y at the time of the study. This family was later published in 31368132.

In a study of 426 unrelated VHL patients, 111 were discovered to have alterations in the VHL gene. 18 novel variants were identified in VHL patients, but none were present in 200 unaffected control individuals. DNA samples from peripheral blood leukocytes underwent polymerase chain reaction (PCR) amplification, denaturing high-performance liquid chromatography (DHPLC), and direct sequence analysis of the entire coding sequence and intron-exon boundaries. The C162F (c.485G>T) germline missense mutation of the VHL gene was found in a 34 yr old VHL patient with multiple pancreatic cysts or tumors, pheochromocytoma, renal cysts, and central nervous system and retinal hemangioblastomas (index case 74).

In a study of 426 unrelated VHL patients, 111 were discovered to have alterations in the VHL gene. 18 novel variants were identified in VHL patients, but none were present in 200 unaffected control individuals. DNA samples from peripheral blood leukocytes underwent polymerase chain reaction (PCR) amplification, denaturing high-performance liquid chromatography (DHPLC), and direct sequence analysis of the entire coding sequence and intron-exon boundaries. This missense mutation, c.191G>C, was found in a 26 year old VHL patient with pheochromocytoma and no family history (index case 123). Only supporting evidence for pathogenicity because patient's phenotype is highly specific for a disease with a single genetic etiology.

In a study of 426 unrelated VHL patients, 111 were discovered to have alterations in the VHL gene. 18 novel variants were identified in VHL patients, but none were present in 200 unaffected control individuals. DNA samples from peripheral blood leukocytes underwent polymerase chain reaction (PCR) amplification, denaturing high-performance liquid chromatography (DHPLC), and direct sequence analysis of the entire coding sequence and intron-exon boundaries. This splice site variant, c.463+1G>T, was found in an asymptomatic 8 year-old with family history of VHL disease (index case 242).

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 T157I (c.470C>T) germline missense mutation of the VHL gene was found in two VHL patients (23Y and 26Y). Both had retinal hemangioblastomas and pheochromocytoma (index cases 8 and 19). The 23 yr old patient also had hemangioblastomas of the central nervous system.

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 nonsense mutation, c.481C>T, was found in 3 VHL patients. The 17-year-old VHL patient only had hemangioblastomas of the central nervous system (CNS) (index case 218). The 24-year-old VHL patient had CNS hemangioblastomas and clear cell renal cell carcinoma (index case 46). The 29-year-old VHL patient had CNS and retinal capillary hemangioma and pheochromocytoma (index case 79). ACMG code as follows: There is very strong evidence for pathogenicity in the form of a nonsense variant in a gene where loss-of-function is a known mechanism of disease.

Molecular and clinical analysis of 126 French VHL families tested for renal involvement revealed 92 different mutations. 90 of these families had renal involvement. The p.R161* (c.481C>T) germline variant of the VHL gene was detected in a family (ID 41). Members 4732 and 1426 had unknown renal involvement. Member 1825 had no renal involvement. Members 1206 and 3600 presented with mild renal disease. Members 4616, 4731, 4446 and 1937 presented with moderate renal disease (RCC). Another family (ID 46) was found with this variant. Member 5024 had unknown renal involvement. Member 5025 had no renal involvement. Another member (no ID provided) presented with mild renal disease (RCC) as well as member 5080. Member 5078 presented with moderate renal disease (RCC). The paper states that these families are previously reported in PMID 12202531 and/or 9829912.

14 Chinese patients from 10 families were diagnosed with Von Hippel-Lindau disease based on clinical criteria. Genetic testing using single-strand conformation polymorphism (SSCP) was conducted and aberrant bands were purified and sequenced. Family III had 4 people belonging to it Patinet 4, 5, 6, and 7. They were found to harbour the c.340+5G>C mutation in the VHL gene (stated as c.553+5G>C in the paper which used the Latif sequence as reference). They all presented with hemangioblastomas; patient 4 presented with pancreatic tumors. This mutation c.340+5G>C has a large allele frequency and therefore a polymorphism that is likely non disease causing

A study of 103 patients with VHL retinal manifestations and 108 patients without VHL retinal manifestations extracted from the French VHL database revealed that the number of hemangioblastomas appeared higher in patients who had a substitution when compared with patients with a truncated protein. Detected by PCR and SSCP, the missense mutation (c.481C>T (p.Arg161Ter)) was found in 8 patients total - 3 of which are stated to have retinal HBs and 5 are said to not. Other manifestations are possible but not clear. This is possibly previously published in PMID: 15300849

A 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 missense mutation (p.Q132P (c.395A>C)) was found in a VHL patient without retinal hemangioblastomas. Other manifestations are not clearly stated. No family information was provided. This is possibly previously published in PMID: 15300849

A study of 103 patients with VHL retinal manifestations and 108 patients without VHL retinal manifestations extracted from the French VHL database revealed that the number of hemangioblastomas appeared higher in patients who had a substitution when compared with patients with a truncated protein. Detected by PCR and SSCP, this mutation p.V130L (c.388G>C) was found in 3 VHL patients. 2 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

PatientID: IV-12

KindredID: 1

Case: M, Age Unknown, Ethnicity Unknown

DiseaseAssertion: Assumed VHL

FamilyInfo: 38 family members had been affected at the date of 1986, and 10 had passed away. Pheochromocytoma, cerebellar, spinal and retinal hemangioblastomas, and various cysts were clinical manifestations of VHL seen throughout the family. This male (IV-12) appears to have inherited this disease from his mother however her children were unaffected at the time. See Fig. 1 for family pedigree.

CasePresentingHPOs: HP:0009711 (retinal capillary hemangioma)

CaseHPOFreeText: This member was examined in St. Johns at the Memorial University of Newfoundland. He had retinal angiomas which were not further described.

CaseNotHPOs: N/A

CaseNotHPOFreeText: N/A

CasePreviousTesting: N/A

PreviouslyPublished: N/A

SupplementalData: N/A

Variant: p.R167W (c.499C>T)

LegacyVariant: N/A

CaseProblemVariantFreeText: Although never given at the time of this publication, the germline mutation is confirmed in this family in later publications (PMID 7987306; 29437867) for this specific family. It is only here assumed this member had the VHL mutation.

ClinVarID: 2218, https://www.ncbi.nlm.nih.gov/clinvar/variation/2218/

CAID: N/A

gnomAD: 0.000007954, https://gnomad.broadinstitute.org/variant/3-10191506-C-T?dataset=gnomad_r2_1

VariantEvidence: N/A

MutationType: missense_variant;transition

CivicName: R167W (c.499C>T)

MultipleGeneVariants: N/A

PatientID: IV-21

KindredID: 1

Case: F, Age Unknown, Ethnicity Unknown

DiseaseAssertion: Assumed VHL

FamilyInfo: 38 family members had been affected at the date of 1986, and 10 had passed away. Pheochromocytoma, cerebellar, spinal and retinal hemangioblastomas, and various cysts were clinical manifestations of VHL seen throughout the family. This patient (IV-21) appears to have inherited this disease from her mother. See Fig. 1 for family pedigree.

CasePresentingHPOs: HP:0009711 (retinal capillary hemangioma)

CaseHPOFreeText: This member was examined in St. Johns at the Memorial University of Newfoundland. She had retinal angiomas which were not further described.

CaseNotHPOs: N/A

CaseNotHPOFreeText: N/A

CasePreviousTesting: N/A

PreviouslyPublished: N/A

SupplementalData: N/A

Variant: p.R167W (c.499C>T)

LegacyVariant: N/A

CaseProblemVariantFreeText: Although never given at the time of this publication, the germline mutation is confirmed in this family in later publications (PMID 7987306; 29437867) for this specific family. It is only here assumed this member had the VHL mutation.

ClinVarID: 2218, https://www.ncbi.nlm.nih.gov/clinvar/variation/2218/

CAID: N/A

gnomAD: 0.000007954, https://gnomad.broadinstitute.org/variant/3-10191506-C-T?dataset=gnomad_r2_1

VariantEvidence: N/A

MutationType: missense_variant;transition

CivicName: R167W (c.499C>T)

MultipleGeneVariants: N/A

PatientID: III-7

KindredID: 1

Case: M, Age Unknown (deceased), Ethnicity Unknown

DiseaseAssertion: Assumed VHL

FamilyInfo: 38 family members had been affected at the date of 1986, and 10 had passed away. Pheochromocytoma, cerebellar, spinal and retinal hemangioblastomas, and various cysts were clinical manifestations of VHL seen throughout the family. This male (III-7) appears to have inherited this disease from his father and has passed it onto to his kin thereafter. See Fig. 1 for family pedigree.

CasePresentingHPOs: HP:0002666 (pheochromocytoma)

CaseHPOFreeText: This patient had pheochromocytoma as well as Vitamin-D-Resistant rickets, likely inherited from his mother.

CaseNotHPOs: N/A

CaseNotHPOFreeText: N/A

CasePreviousTesting: N/A

PreviouslyPublished: N/A

SupplementalData: N/A

Variant: p.R167W (c.499C>T)

LegacyVariant: N/A

CaseProblemVariantFreeText: Although never given at the time of this publication, the germline mutation is confirmed in this family in later publications (PMID 7987306; 29437867) for this specific family. It is only here assumed this member had the VHL mutation.

ClinVarID: 2218, https://www.ncbi.nlm.nih.gov/clinvar/variation/2218/

CAID: N/A

gnomAD: 0.000007954, https://gnomad.broadinstitute.org/variant/3-10191506-C-T?dataset=gnomad_r2_1

VariantEvidence: N/A

MutationType: missense_variant;transition

CivicName: R167W (c.499C>T)

MultipleGeneVariants: N/A

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 mutation (c.548C>A) was detected in 1 member of family 22. The patient presented with CNS hemangioblastoma and retinal capillary hemangioma. The patient does not have a family history of VHL disease. The genetic information of the patient's parents are not available.

Data was collected from 82 VHL mutation carriers in the Dutch VHL surveillance program. One patient was found with this missense varaint in the VHL gene. The patient was 43Y at the last follow-up and presented with: retinal hemangioblastoma, spinal cord hemangioblastoma, renal cell carcinoma, pheochromocytoma, and renal cysts. Family history was not described. This cohort was published later in PMID 31087189.

Data was collected from 82 VHL mutation carriers in the Dutch VHL surveillance program. One patient age 20Y at the last follow-up had retinal hemangioblastoma and a germline c.208G>A (p.Glu70Lys) mutation in the VHL gene. Family history was not described. This cohort was published later in PMID 31087189.

Data was collected from 82 VHL mutation carriers in the Dutch VHL surveillance program. One patient was found with this missense varaint in the VHL gene. The patient was 27Y at the last follow-up and presented with: pancreatic neuroendocrine tumour and pheochromocytoma. Family history was not described. This cohort was published later in PMID 31087189.

Data was collected from 82 VHL mutation carriers in the Dutch VHL surveillance program. Seven patients were found with this missense variant in the VHL gene. The ages of the patients was represented as a range 7-88 (mean 45). Phenotypes were grouped together: pheochromocytoma and renal cysts, although it is unclear if only one of the seven patients had renal cysts. Family history was not described. This cohort was published later in PMID 31087189.

PatientID: 5

KindredID: N/A

Case: Sex Unknown, Age Unknown, Ethnicity Unknown

DiseaseAssertion: VHL

FamilyInfo: N/A

CohortInfo: The authors evaluated 64 VHL patients (32 women and 32 men) with renal involvement of whom 54/64 had undergone renal surgery or a radiofrequency thermal ablation procedure for RCC. In 10/64 VHL patients, at least one or more RCC were 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.

CasePresentingHPOs: HP:0005584 (Renal cell carcinoma)

CaseHPOFreeText: This is a tumor resolution study (VHL patients were investigated based on the presence of renal tumors.) No other phenotypic information is available.

CaseNotHPOs: N/A

CaseNotHPOFreeText: N/A

CasePreviousTesting: N/A

PreviouslyPublished: N/A

SupplementalData: Supplementary Table S2 describes 9 patients with nonsense mutations, including this patient.

Variant: p.S65* (c.194C>A)

LegacyVariant: c.407C>A

CaseProblemVariantFreeText: N/A

ClinVarID: 223161, https://www.ncbi.nlm.nih.gov/clinvar/variation/223161/

CAID: N/A

gnomAD: N/A

VariantEvidence: N/A

MutationType: stop_gained

CivicName: S65* (c.194C>A)

MultipleGeneVariants: N/A

The authors described the growth kinetics of 96 renal tumours in 64 VHL patients (32 males, 32 females). All patients had VHL disease with renal involvement. 54/64 had undergone renal surgery or a radiofrequency thermal ablation procedure for RCC. In 10/64 patients, at least one or more RCC was diagnosed. However, at the time of diagnosis, an indication for treatment was not yet present. For all patients, several CT/MRI scans existed for retrospective description of tumour growth kinetics. 10-ml EDTA anticoagulated blood samples were available from each patient for mutation screening of all exons of the VHL gene. Genomic DNA from blood samples was extracted using standard methods. Small deletions/insertions or point mutations were identified by DNA sequencing followed by complete mutation screening using PCR/single-strand conformation polymorphism and denaturing high-performance liquid chromatography. A germline nonsense variant p.S183*(c.548C>A) was identified in patient #57. This mutation was noted as c.761C>A in the paper due to a difference in reference transcript used (see Table S2).

PatientID: 4

KindredID: N/A

Case: Sex Unknown, Age Unknown, Ethnicity Unknown

DiseaseAssertion: VHL

FamilyInfo: N/A

CohortInfo: The authors evaluated 64 VHL patients (32 women and 32 men) with renal involvement of whom 54/64 had undergone renal surgery or a radiofrequency thermal ablation procedure for RCC. In 10/64 VHL patients, at least one or more RCC were 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.

CasePresentingHPOs: HP:0005584 (Renal cell carcinoma)

CaseHPOFreeText: This is a tumor resolution study (VHL patients were investigated based on the presence of renal tumors.) No other phenotypic information is available.

CaseNotHPOs: N/A

CaseNotHPOFreeText: N/A

CasePreviousTesting: N/A

PreviouslyPublished: N/A

SupplementalData: Supplementary Table S1 describes 19 patients with missense mutations, including this patient.

Variant: p.V155M(c.463G>A)

LegacyVariant: c.676G>A

CaseProblemVariantFreeText: N/A

ClinVarID: 223219, https://www.ncbi.nlm.nih.gov/clinvar/variation/223219/

CAID: N/A

gnomAD: N/A

VariantEvidence: N/A

MutationType: missense_variant;transition

CivicName: V155M (c.463G>A)

MultipleGeneVariants: N/A

KindredID: 33

GroupID: 43

GroupSize: Unknown

GroupNumberMales: Unknown

GroupNumberFemales: Unknown

GroupEthnicity: Unknown

GroupAgeRange: Unknown

DiseaseAssertion: VHL

GroupInfo: Kindred no.33 describes a family where at least one member (unspecified how many) had this mutation in the VHL gene. Family history indicates that pheochromocytoma is present in this kindred.

GroupPresentingHPOs: HP:0002666 (Pheochromocytoma)

GroupHPOFreeText: This study investigated the frequency of different mutation classes between kindreds with and without pheochromocytoma. No other information about disease presentation within members of a kindred could be inferred.

GroupNotHPOs: N/A

GroupNotHPOFreeText: N/A

GroupPreviousTesting: N/A

PreviouslyPublished: N/A

SupplementalData: N/A

Variant: p.R167Q (c.500G>A)

LegacyVariant: c.713G>A (p.R238Q)

CaseProblemVariantFreeText: N/A

ClinVarID: 2216, https://www.ncbi.nlm.nih.gov/clinvar/variation/2216/

CAID: N/A

gnomAD: 0.000003977, https://gnomad.broadinstitute.org/variant/3-10191507-G-A?dataset=gnomad_r2_1

VariantEvidence:

MutationType: missense_variant;transition

CivicName: R167Q (c.500G>A)

MultipleGeneVariants: N/A

PatientID: 5

KindredID: 1

Case: Sex Unknown, 35Y0M, Ethnicity Unknown

DiseaseAssertion: VHL

FamilyInfo: Significant family history. Pedigree follows 8 generations (see Figure 3). Family immigrated to Iowa, US, from Baden-Baden, Germany, in 1832; Likely sharing a common ancestor to the traced lineage of the Black Forest region of Germany.

CasePresentingHPOs: HP:0002666; HP:0009711 (pheochromocytoma; retinal capillary hemangioma)

CaseHPOFreeText: 20Y at first pheochromocytoma and ocular angioma (retinal capillary hemangioma); both bilateral. Total of 2 ocular angiomas and 2 pheochromocytomas.

CaseNotHPOs: HP:0010797; HP:0005584 (hemangioblastoma, renal cell carcinoma)

CaseNotHPOFreeText: N/A

CasePreviousTesting: N/A

PreviouslyPublished: N/A

SupplementalData: N/A

Variant: p.Y98H (c.292T>C)

LegacyVariant: p.Y98H (c.505T>C)

CaseProblemVariantFreeText: N/A

ClinVar: 2223 https://www.ncbi.nlm.nih.gov/clinvar/variation/2223/

CAID: N/A

gnomAD: 0.00001343 https://gnomad.broadinstitute.org/variant/3-10183823-T-C

VariantEvidence: N/A

MutationType: missense_variant;transition

CivicName: Y98H (c.292T>C)

MultipleGeneVariants: N/A

216 patients with clinically expected VHL disease due to family history or presence of VHL typical tumours were routinely examined in an eye centre in Germany between January 2019 and January 2020, majority of which were of European descent (~95%). Since data on the genotype–phenotype correlation of the onset of Retinal Hemangioblastoma (RH) is scarce in relation to VHL disease, this study wanted to inversitage the relationship between VHL gene variants (truncating vs single amino acid substitutions) and the ocular phenotype of the disease. 42 different rare VHL gene variants in 148 patients were detected - some of whom had RH and some who did not. Although date of onset of RH, as well as data on central nervous system haemangioblastomas (CNSH), pheochromocytoma/paraganglioma (PPGL), clear cell renal cell carcinoma (ccRCC) and pancreatic neuroendocrine neoplasm (PNEN) were collected, it was not reported on a per patient basis and therefore could not be discussed per variant. There was 1 patient reported with the c.238A>C (p.S80R) mutation. This member presented with retinal hemangiomas. No family information was provided.

216 patients with clinically expected VHL disease due to family history or presence of VHL typical tumours were routinely examined in an eye centre in Germany between January 2019 and January 2020, majority of which were of European descent (~95%). Since data on the genotype–phenotype correlation of the onset of Retinal Hemangioblastoma (RH) is scarce in relation to VHL disease, this study wanted to inversitage the relationship between VHL gene variants (truncating vs single amino acid substitutions) and the ocular phenotype of the disease. 42 different rare VHL gene variants in 148 patients were detected - some of whom had RH and some who did not. Although date of onset of RH, as well as data on central nervous system haemangioblastomas (CNSH), pheochromocytoma/paraganglioma (PPGL), clear cell renal cell carcinoma (ccRCC) and pancreatic neuroendocrine neoplasm (PNEN) were collected, it was not reported on a per patient basis and therefore could not be discussed per variant. There were 3 patients reported with the p.S183* (c.548C>A) mutation. All 3 members presented with retinal hemangiomas, while no other clinical manifestations were discussed. Additionally, no family information was provided.

PatientID: 18

GroupID/ KindredID: 18

Case: Sex Unknown, Age Unknown, Ethnicity Unknown

DiseaseAssertion: hemangioblastoma

FamilyInfo: N/A

CasePresentingHPOs: HP:0010797 (hemangioblastoma)

CaseHPOFreeText: other phenotypes unknown.

CaseNotHPOs: N/A

CaseNotHPOFreeText: N/A

CasePreviousTesting: N/A

PreviouslyPublished: N/A

SupplementalData: N/A

Variant: c.340+1G>A

LegacyVariant: c.553+1G>A

CaseProblemVariantFreeText: N/A

ClinVarID: 223190 http://www.ncbi.nlm.nih.gov/clinvar/variation/223190

CAID: N/A

gnomAD: N/A

VariantEvidence: N/A

MutationType: splice_site_variant

CivicName: Splice Site (c.340+1G>A)

MultipleGeneVariants: N/A

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.500G>A) was recorded in the paper as c.713G>A due to a difference in transcript sequence. It was found in one patient with a hemangioblastoma of the central nervous system. Any family information is unknown. No other phenotypes described.

A cohort of 141 patients with hemangioblastomas of the central nervous system was analyzed. 81 patients had germline mutations in the VHL gene, analyzed through peripheral blood by Southern blotting and SSCP analysis. This nonsense mutation (c.548C>A) was recorded in the paper as c.761C>A due to a difference in transcript sequence. It was found in 4 patients with hemangioblastomas of the central nervous system. Any family information or relations is unknown. No other phenotypes described.

A cohort of 141 patients with hemangioblastomas of the central nervous system was analyzed. 81 patients had germline mutations in the VHL gene, analyzed through peripheral blood by Southern blotting and SSCP analysis. This missense mutation (c.241C>T) was recorded in the paper as c.454C>T 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.

GroupID: 15

DiseaseAssertion: hemangioblastoma

GroupSize: 21

GroupNumberMales: N/A

GroupNumberFemales: N/A

GroupEthnicity: N/A

GroupAgeRange: N/A

GroupInfo: 21 out of the 81 patients listed in this hemangioblastoma cohort were found with this c.292T>C mutation. Any relation is unknown.

GroupPresentingHPOs: HP:0010797 (hemangioblastoma)

GroupHPOFreeText: all patients had hemangioblastoma of the CNS; other phenotypes are unknown. 13 patents were said to be found with primary asymptomatic tumors.

GroupNotHPOs: N/A

GroupNotHPOFreeText: N/A

GroupPreviousTesting: N/A

PreviouslyPublished: N/A

SupplementalData: N/A

Variant: p.Y98H (c.292T>C)

LegacyVariant: p.Y98H (c.505T>C)

CaseProblemVariantFreeText: N/A

ClinVarID: 2223 https://www.ncbi.nlm.nih.gov/clinvar/variation/2223/

CAID: N/A

gnomAD: 0.00001343 https://gnomad.broadinstitute.org/variant/3-10183823-T-C

VariantEvidence: N/A

MutationType: missense_variant; transition

CivicName: Y98H (c.292T>C)

MultipleGeneVariants: N/A

GroupID: None

DiseaseAssertion: VHL

GroupSize: 82 GroupNumberMales: N/A GroupNumberFemales: N/A GroupEthnicity: N/A GroupAgeRange: N/A

GroupInfo: Pennsylvania kindred with VHL Type 2A.

GroupPresentingHPOs: HP:0002666; HP:0009711 (Pheochromocytoma, Retinal capillary hemangioma)

GroupHPOFreeText: 18 living individuals with only pheochromocytoma. 7 living individuals with only retinal angioma. 6 living individuals with both. 51 other first-degree relatives. 5 additional relatives with pheochromocytomas, including a 69Y woman and her 42Y son described previously as non-penetrant individuals (PMID 8239848). Family classified as a VHL Type 2A (pheochromocytoma, retinal angioma, CNS hemangioblastoma, and little to no RCC).

GroupNotHPOs: HP:0005584 (Renal cell carcinoma)

GroupNotHPOFreeText: N/A

GroupPreviousTesting: N/A

PreviouslyPublished: 8239848 (no genetic information; only phenotypes); 21713522

SupplementalData: N/A

Variant: p.Y112H (c.334T>C)

LegacyVariant: c.547T>C

CaseProblemVariantFreeText: N/A

ClinVarID: 2222, https://www.ncbi.nlm.nih.gov/clinvar/variation/2222/

CAID: N/A

gnomAD: N/A

VariantEvidence: All 31 individuals (including 6 archival specimens) with a diagnosis of pheochromocytoma, retinal angioma or both had the identical mutation.

MutationType: missense_variant;transition

CivicName: Y112H (c.334T>C)

MultipleGeneVariants: N/A

A study describing phenotypic and genotypic characteristics of VHL in an Indian population of 31 subjects, from 15 families diagnosed with VHL. Multi-cystic pancreas and CNS hemangioblastomas were the most common manifestations in this population. This missense mutation was found in a male VHL patient with retinal angiomas and pheochromocytoma (family ID D).

GroupID/ KindredID: VHL 34

DiseaseAssertion: assumed VHL

GroupSize: 2

GroupNumberMales: N/A

GroupNumberFemales: N/A

GroupEthnicity: German

GroupAgeRange: N/A

FamilyInfo: 2 affected members belong to this VHL family. Diagnosis of VHL was clinically confirmed in the index person and VHL type could not be determined due to the small size of the pedigree. One representative affected member was analyzed with this mutation, and VHL was diagnosed clinically in this index person. It is unclear as to whether the additional patients had this precise mutation, although assumed.

GroupPresentingHPOs: HP:0010797; HP:0005584 (hemangioblastoma, renal cell carcinoma)

GroupHPOFreeText: both (2) individuals had RCC, where as only one had a CNS hemangioblastoma.

GroupNotHPOs: HP:0009711; HP:0002666; HP:0001737; HP:0009715 (retinal capillary hemangioma, pheochromocytoma, pancreatic cysts, papillary cystadenoma of the epididymis)

GroupNotHPOFreeText: No patients had the above tumors

GroupPreviousTesting: N/A

PreviouslyPublished: 11134110, 9681856, 11850829

SupplementalData: N/A

Variant: p.Y98H (c.292T>C)

LegacyVariant: p.Y98H (c.505T>C)

CaseProblemVariantFreeText: N/A

ClinVar: 2223 https://www.ncbi.nlm.nih.gov/clinvar/variation/2223/

CAID: N/A

gnomAD: 0.00001343 https://gnomad.broadinstitute.org/variant/3-10183823-T-C

VariantEvidence: N/A

MutationType: missense_variant;transition

CivicName: Y98H (c.292T>C)

MultipleGeneVariants: N/A

All patients that were operated on between 1995 and 2010 in the University Medical Center Groningen from who frozen hemangioblastoma tissue was available in the tissue bank of the Department of Pathology were eligible. Through PCR (MSP) and MLPA, one patient sample was confirmed with a germline mutation (c.490C>T) in the VHL gene. The patients' manifestations was thus only stated as hemangioblastoma; No other phenotypic or demographic information was available.

5 cases treated for thoracic paraganglioma in Freiburg, Germany aged 6–70 at the time of diagnosis were identified and had germline VHL mutations identified using constitutional genomic DNA from blood samples. The mutation p.Tyr98His (legacy notation c.505T>C converted to c.292T>C) was found in case 1 and case 2. Case 1 was a 30Y female patient with thoracic paraganglioma, data on family members was not available. Case 2 was a 35Y female who also developed bilateral adrenal pheochromocytomas, pancreatic islet cell tumor, and basophilic pituitary adenoma and died. Case 2 (index) did not have blood available for testing, but the mutation was confirmed in 2 of her living sisters and her daughter. Her daughter developed retinal angioma, and the status of the two sisters is not known. ACMG: PP4

5 cases treated for thoracic paraganglioma in Freiburg, Germany aged 6–70 at the time of diagnosis were identified and had germline VHL mutations identified using constitutional genomic DNA from blood samples. The mutation p.Gly93Ser (legacy notation c.490G>A converted to c.277G>A) was found in case 3, a 6Y boy with thoracic paraganglioma. His parents were negative for the mutation, supporting de novo germline mutatation. ACMG: PM6

In a study of 138 unrelated VHL families, 109 germline mutations were found. Missense mutations were more frequently found in families with pheochromocytoma (PC). Deletions, nonsense, and frameshift mutations were more frequently found in families without PC. Large deletions and mutations predicted to cause truncated protein were associated with lower risk of PC. The splice mutation c.340+5G>C was referred to in the paper by cc.553+5G>C due to a difference in transcript sequence. This mutation was found in a VHL family without PC (family ID 139). The mutation has a large allele frequency meaning this is likely non disease causing (not deleterious).

15 Dutch VHL patients from 5 families were taken to measure the expression of vascular endothelial growth factor (VEGF). Two patients were found to harbour the VHL germline mutation R167Q, referred to in the paper by R238Q due to a difference in transcript sequence. Patient 10, a 40 year old male, presented with pancreatic and kidney cysts, renal cell carcinoma and brain and spinal cord hemangioblastomas as well as no signs of eye lesions. Patient 15, a 29 year old female, presented with a brain cyst. ACMG evidence codes: 'PP4' since the patient presents phenotypes highly specific for a disease with a single genetic etiology.

37 Danish patients diagnosed with VHL before 18 years from a prospective cohort study from Denmark's National vHL Research Database (Binderup et al., 2013) were assessed. One family with 1 patient (Family ID 4) was found with a germline missense mutation in the VHL gene (F136S; c.407T>C). Patient 1 was diagnosed at 7Y and had 2 retinal hemangioblastomas at 7Y and 14Y. The patient passed at 53Y by the end of the study.

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 4 patients (Family ID 3) were found with a germline missense mutation in the VHL gene (G93D; c.278G>A). Patient 1 was diagnosed at 7Y and had no manifestations by the end of the study at 18Y old. Patient 2 and 3 were both diagnosed at 9Y and and both developed pheochromocytoma at age 15Y. They were both 20Y at the end of the study. Patient 4 was diagnosed at 10Y and had 2 pheochromocytomas at age 12Y, and 1 retinal hemangioblastoma at age 10Y. They were 21Y at the end of the study.

A case series covered 2 cases of pheochromocytoma and VHL in pregnancy. The proband (case 2) was a hispanic female with pheochromocytoma at 25Y and retinal angioma at 11Y. Genetic analysis revealed a germline mutation on the VHL gene (c.499C>T; p.Arg167Trp). Unclear on the methods behind genetic testing. The proband’s father, brother, and 8-year-old son have been diagnosed with VHL, although phenotypes were not specified. Her other infant son also tested positive for the same mutation. Her 5-year-old daughter is unaffected. ACMG: PP4

All patients included in the study were VHL mutation carriers of 18 years or older examined between 1972 and 2012 in the VHL expertise centers University Medical Center Groningen (UMCG) and University Medical Center Utrecht (UMCU). Clinical data of 75 VHL mutation carriers were retrospectively retrieved from electronic and written patient record. Of these patients, one was found to harbour the c.241C>T germline mutation of the VHL gene. They presented with a retinal hemangioblastoma, cerebellar hemangioblastoma, spinal hemangioblastoma, renal cell carcinoma, pancreatic neuroendocrine tumor, renal cyst and pheochromocytoma. Their age at the last follow-up was 59. Previously published in: PMID 24132471.

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.208G>A germline mutation of the VHL gene. They presented with a retinal hemangioblastoma. Their age at the last follow-up was 20. Previously published in: PMID 24132471.

26 VHL patients from 18 families, who had undergone a partial adrenalectomy for pheochromocytoma were retrospectively analyzed. 1 patient from a single family was confirmed to have the mutation (p.L128F (c.381_382delinsTT)). Additional manifestations aside from pheochromocytoma are possible, but were not discussed.

79 patients with head and neck paragangliomas were analyzed for SDHA, SDHB, SDHC, SDHD, SDHAF2, VHL, MAX, and TMEM127 genes by PCR/MLPA sequencing. Via peripheral blood leukocytes, 1 patient was found to have the above mutation (p.P81L (c.242C>T)), and presented with a right (non-secreting) carotid body paraganglioma. No family information was presented.