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

Case#: 23015, Age 74, Male, Blood Type A+ (Table S3)

Disease Assertion: VWD1

FamilyInfo: The patient is the only member of Family# 2310.

Parental Genotype: Not listed

CasePresentingHPOs: N/A

CaseHPOFreeText:VWF:Ag: 18, VWF:RCo:17.3, FVIII:C:19, VWF:CB:13

CaseNotHPOs: N/A

CaseNotHPOFreeText: Bleeding Score 3

Protein Expression: N/A

CasePreviousTesting: 48,48 Access-Array amplification. NGS Molecular analysis was performed. Sanger sequencing was used to validate selected mutations

Gene: VWF

Variant: NM_000552.4:c.2546+132G>A(p.=) and NM_000552.4:c.3614G>A (p.Arg1205His)

ClinVar: 308 ClinVar

CAID: CA232299449 and CA114160 CAID1 CAID2

gnomAD: MAF of 0.001179 ( 1/848 alleles) in the Latino population (CA232299449). gnomAD

Case#: 01010, Age 55, Male, Blood Type O+ (Table S3)

Disease Assertion:VWD1

FamilyInfo: Not listed

Parental Genotype:Not listed

CasePresentingHPOs:

CaseHPOFreeText: Bleeding score 5, VWF:Ag 27, VWF:RCo: 30, FVIII:C 51, VWF:CB: 32

CaseNotHPOs:

CaseNotHPOFreeText:

Protein Expression:

CasePreviousTesting: 48,48 Access-Array amplification. NGS Molecular analysis was performed. Sanger sequencing was used to validate selected mutations.

Gene: VWF

Variant:NM_000552.4(VWF):c.4751A>G (p.Tyr1584Cys)

ClinVar: 310

ClinVarID

CAID: CA114162

CAID

gnomAD: MAF of 0.004024 (519/128972 alleles) in the European (non-Finnish) population.

gnomAD

Case#: 32005, Age 28, Female, Blood Type O- (Table S3)

Disease Assertion: VWD 1

FamilyInfo: The patient is the only member of Family# 3205.

Parental Genotype: Not listed

CasePresentingHPOs: N/A

CaseHPOFreeText: VWF:Ag: 30, VWF:RCo: 25, FVIII:C: 25, VWF:CB: 32

CaseNotHPOs: N/A

CaseNotHPOFreeText: Bleeding Score 4

Protein Expression: N/A

CasePreviousTesting: 48,48 Access-Array amplification. NGS Molecular analysis was performed. Sanger sequencing was used to validate selected mutations.

Gene: VWF

Variant: NM_000552.4:c.2546+132G>A (p.=) and NM_000552.4:c.4751A>G (p.Tyr1584Cys)

ClinVar: 310

CAID: CA232299449 and CA114162 CAID1 CAID2

gnomAD: MAF of 0.001179 (1/848 alleles) in the Latino population (CA232299449) gnomAD

MAF of 0.004024 (519/ 128972 alleles) in the European (non-Finnish) population. gnomAD

Case#: 02023, Age 28, Female, Blood type O+ (Table S3)

Disease Assertion: VWD 1

FamilyInfo: Family# 0206. 3 family members were included in the study. Everyone in the family have the same variant and all have type 1 VWD.

Parental Genotype: Not listed

CasePresentingHPOs:

CaseHPOFreeText: Bleeding score 18, VWF:Ag: 5.5, VWF:RCo: 8, FVIII:C: 11, VWF:CB: 4.2

CaseNotHPOs:

CaseNotHPOFreeText:

Protein Expression:

CasePreviousTesting: 48,48 Access-Array amplification. NGS Molecular analysis was performed. Sanger sequencing was used to validate selected mutations.

Gene: VWF

Variant: NM_000552.4:c.3614G>A (p.Arg1205His)

ClinVar: 308 ClinVar

CAID: CA114160 CAID

gnomAD: N/A

Case#: 29002, Age 38, Female, Blood Type O+ (Table S3)

Disease Assertion: VWD 1

FamilyInfo: The patient is the only member of Family#2902.

Parental Genotype: Not listed

CasePresentingHPOs: N/A

CaseHPOFreeText: Bleeding score 7, VWF:Ag: 22, VWF:RCo: 17, FVIII:C: 54, VWF:CB:17

CaseNotHPOs: N/A

CaseNotHPOFreeText: N/A

Protein Expression: N/A

CasePreviousTesting: 48,48 Access-Array amplification. NGS Molecular analysis was performed. Sanger sequencing was used to validate selected mutations.

Gene: VWF

Variant: NM_000552.4:c.3223-7_3236dup (p.Pro1079_Tyr1080insLeuGlnValAspProGluPro) and NM_000552.4(VWF):c.5277C>T (p.Asp1759=)

ClinVar: 310061 ClinVar

CAID: CA1139532066 and CA6402333 CAID1 CAID2

gnomAD: MAF of 0.01423 (1837/129090 alleles) in the European (non-Finnish) population (CA6402333). gnomAD

Case#: 30008, Age 24, Female, Blood Type A+ (Table S3)

Disease Assertion: VWD 1

FamilyInfo: The patient is one of two members of Family #3006. Both patients have the same variant.

Parental Genotype: Not listed

CasePresentingHPOs: N/A

CaseHPOFreeText: Bleeding Score 12, VWF:Ag:18, VWF:RCo:19.3, FVIII:C: 35, VWF:CB: 21

CaseNotHPOs: N/A

CaseNotHPOFreeText: N/A

Protein Expression: N/A

CasePreviousTesting: 48,48 Access-Array amplification. NGS Molecular analysis was performed. Sanger sequencing was used to validate selected mutations.

Gene: VWF

Variant: NM_000552.4:c.6536C>T (p.Ser2179Phe)

ClinVar: 100442 ClinVar

CAID: CA228746 CAID

gnomAD: N/A

Case#: 35008, Age 64, Male, Blood Type A+ (Table S3)

Disease Assertion: VWD 1

FamilyInfo: The patient is the only member of Family# 3503.

Parental Genotype: Not listed

CasePresentingHPOs: N/A

CaseHPOFreeText: VWF:Ag: 5.3, VWF:RCo:5, FVIII:C: 10, VWF:CB: 4.9

CaseNotHPOs: N/A

CaseNotHPOFreeText: Bleeding Score 2

Protein Expression: N/A

CasePreviousTesting: 48,48 Access-Array amplification. NGS Molecular analysis was performed. Sanger sequencing was used to validate selected mutations.

Gene: VWF

Variant: NM_000552.4:c.1625C>G (p.Ala542Gly), NM_000552.4:c.5198T>C (p.Leu1733Pro), and NM_000552.4:c.6187C>T (p.Pro2063Ser)

ClinVar: 381621 and 100436 ClinVar1 ClinVar2

CAID: CA6403319, CA383496267, and CA202650 CAID1 CAID2 CAID3

gnomAD: MAF of 0.001355 (48/ 35418 alleles) in the Latino population (CA6403319). gnomAD

MAF of 0.04854 (1486/30616 alleles) in the South Asian population (CA202650). gnomAD

Case#: 07003, Age 29, Female, Blood Type O+ (Table S3)

Disease Assertion: VWD1

FamilyInfo: The patient is the only member of Family#0703.

Parental Genotype: Not listed

CasePresentingHPOs: N/A

CaseHPOFreeText: VWF:Ag: 25, VWF:RCo:24.9, FVIII:C: 50, VWF:CB:28

CaseNotHPOs: N/A

CaseNotHPOFreeText: Bleeding Score 5

Protein Expression: N/A

CasePreviousTesting: 48,48 Access-Array amplification. NGS Molecular analysis was performed. Sanger sequencing was used to validate selected mutations

Gene: VWF

Variant:NM_000552.4:c.5170+10C>T and NM_000552.4:c.8254-5T>G

ClinVar: 100418 ClinVar1

CAID: CA228702 and CA923726359 CAID1 CAID2

gnomAD: MAF of 0.01298 (326/ 25116 alleles) in the European (Finnish) population (CA228702).

gnomAD1

Case#: 35011, Age 46, Male, Blood Type O+ (Table S3)

Disease Assertion: VWD 1

FamilyInfo: The patient is one of 3 members of Family#3505. All have VWD type 1 and patient# 35013 has an additional intronic variant that the other two family members don't have.

Parental Genotype: Not listed

CasePresentingHPOs: N/A

CaseHPOFreeText: VWF:Ag: 18, VWF:RCo: 14, FVIII:C: 42, VWF:CB: 12

CaseNotHPOs: N/A

CaseNotHPOFreeText: Bleeding Score 2

Protein Expression: N/A

CasePreviousTesting: 48,48 Access-Array amplification. NGS Molecular analysis was performed. Sanger sequencing was used to validate selected mutations.

Gene: VWF

Variant: NM_000552.4:c.5198T>C (p.Leu1733Pro) and NM_000552.4(VWF):c.6187C>T (p.Pro2063Ser)

ClinVar: 100436 ClinVar

CAID: CA383496267 and CA202650 CAID1 CAID2

gnomAD: MAF of 0.04854 (1486/30616 alleles) in the South Asian population (CA202650). gnomAD

Case#: 05008, Age 30, Male, Blood Type A+ (Table S3)

Disease Assertion: VWD1

FamilyInfo: Patient is the only member of Family #0507

Parental Genotype: Not listed

CasePresentingHPOs: N/A

CaseHPOFreeText: Bleeding Score 2, VWF:Ag: 5.8, VWF:RCo:5, FVIII:C: 7.4, VWF:CB:6.1

CaseNotHPOs: N/A

CaseNotHPOFreeText: N/A

Protein Expression: N/A

CasePreviousTesting: 48,48 Access-Array amplification. NGS Molecular analysis was performed. Sanger sequencing was used to validate selected mutations

Gene: VWF

Variant: NM_000552.4:c.3614G>A (p.Arg1205His)

ClinVar: 308 ClinVar

CAID: CA114160 CAID

gnomAD: N/A

Case#: 22001, Age 41, Female, Blood Type O+ (Table S3)

Disease Assertion: VWD 1

FamilyInfo: The patient is the only member of Family# 2201.

Parental Genotype: Not listed

CasePresentingHPOs: N/A

CaseHPOFreeText: Bleeding Score 14, VWF:Ag:18, VWF:RCo:17.5, FVIII:C: 35, VWF:CB: 17

CaseNotHPOs: N/A

CaseNotHPOFreeText: N/A

Protein Expression: N/A

CasePreviousTesting: 48,48 Access-Array amplification. NGS Molecular analysis was performed. Sanger sequencing was used to validate selected mutations

Gene: VWF

Variant: NM_000552.4:c.5170+10C>T (p.=) and NM_000552.4(VWF):c.7730-1G>C (p.=)

ClinVar: 100418 and 100485 ClinVar1 ClinVar2

CAID: CA228702 and CA228816 CAID1 CAID2

gnomAD: 0.01298 (326/25116 alleles) in the European (Finnish) population. (CA228702) gnomAD

Case#: 13018, Age 5, Male, Blood type not determined (TableS3)

Disease Assertion: VWD 1

FamilyInfo: The patient is the only member of Family#1314.

Parental Genotype: Not listed

CasePresentingHPOs: N/A

CaseHPOFreeText: Bleeding Score 0, VWF:Ag:18, VWF:RCo:21, FVIII:C:39, VWF:CB: 15

CaseNotHPOs: N/A

CaseNotHPOFreeText: N/A

Protein Expression:N/A

CasePreviousTesting: 48,48 Access-Array amplification. NGS Molecular analysis was performed. Sanger sequencing was used to validate selected mutations

Gene: VWF

Variant: NM_000552.4:c.4238C>T (p.Pro1413Leu)

ClinVar: 100342 ClinVar

CAID: CA228565 CAID

gnomAD: MAF of 0.00003266 ( 1/30616 alleles) in the South Asian population. gnomAD

Case#: 02059, Age 38, Male, Blood type A+ (Table S3)

Disease Assertion: VWD 1

FamilyInfo: Patient is a member of family # 02059. There is one other member of the family (patient 02060). 02060 has the same exonic variant (c.3614G>A), but has a different intronic variant ( c.7438-169G>A).

Parental Genotype: Not listed

CasePresentingHPOs: N/A

CaseHPOFreeText: Bleeding score 12, VWF:Ag: 5.4, VWF:RCo: 6.1, FVIII:C:11, VWF:CB: 4.5

CaseNotHPOs: N/A

CaseNotHPOFreeText: N/A

Protein Expression: N/A

CasePreviousTesting: 48,48 Access-Array amplification. NGS Molecular analysis was performed. Sanger sequencing was used to validate selected mutations

Gene: VWF

Variant: NM_000552.4:c.3614G>A (p.Arg1205His) and

ClinVar: 308 ClinVar

CAID: CA114160 CAID1

gnomAD:

Case#: 02001, Age 63, Female, Blood Type A+ (Table S3)

Disease Assertion: VWD 1

FamilyInfo:( Family 0201) 8 family members were included in the study. All of them had type VWD type 1. All had the same variant besides patient 02019.

Parental Genotype: Not listed

CasePresentingHPOs: N/A

CaseHPOFreeText: Bleeding Score 14, VWF:Ag: 9.7, VWF:RCo: 2.6, FVIII:C: 19, VWF:CB: 6

CaseNotHPOs: N/A

CaseNotHPOFreeText: N/A

Protein Expression: N/A

CasePreviousTesting: 48,48 Access-Array amplification. NGS Molecular analysis was performed. Sanger sequencing was used to validate selected mutations.

Gene: VWF

Variant: NM_000552.4:c.3614G>A (p.Arg1205His)

ClinVar: 308 ClinVar

CAID: CA114160

CAID

gnomAD: N/A

Case#: 02069, Age 45, Female, Blood Type O+ (Table S3)

Disease Assertion: VWD 1

FamilyInfo: The patients was 1 of 3 members of family #0226.

Parental Genotype: Not listed

CasePresentingHPOs: N/A

CaseHPOFreeText: Bleeding Score 5, VWF:Ag: 30, VWF:RCo:27, FVIII:C: 81, VWF:CB: 40

CaseNotHPOs: N/A

CaseNotHPOFreeText: N/A

Protein Expression: N/A

CasePreviousTesting: 48,48 Access-Array amplification. NGS Molecular analysis was performed. Sanger sequencing was used to validate selected mutations

Gene: VWF

Variant: NM_000552.4:c.3467C>T (p.Thr1156Met) and NM_000552.3:c.-3151T>G

ClinVar: 100264 ClinVar

CAID: CA228414 CAID

gnomAD: N/A

Case#: 27030, Age 23, Male, Blood Type O+ (Table S3)

Disease Assertion: VWD 1

FamilyInfo: The patient is one of 6 members in Family #2703. All members have the same variant.

Parental Genotype: Not listed

CasePresentingHPOs: N/A

CaseHPOFreeText: VWF:Ag:26, VWF:RCo:28.6, FVIII:C:81, VWF:CB: 30

CaseNotHPOs: N/A

CaseNotHPOFreeText: Bleeding Score 2

Protein Expression: N/A

CasePreviousTesting: 48,48 Access-Array amplification. NGS Molecular analysis was performed. Sanger sequencing was used to validate selected mutations.

Gene: VWF

Variant: NM_000552.4:c.1533+15G>A (p.=)

ClinVar: 100182 ClinVar

CAID: CA228275 CAID

gnomAD: MAF of 0.002649 ( 93/ 35104 alleles) in the Latino Population. gnomAD

Case#: 01037, Age 16, Male, Blood Type O- (Table S3)

Disease Assertion: VWD 1

FamilyInfo: Not listed

Parental Genotype: Not listed

CasePresentingHPOs:

CaseHPOFreeText: Bleeding type 14, VWF:Ag: 22, VWF:RCO: 32, FVIII:C: 28, VWF:CB: 22

CaseNotHPOs: N/A

CaseNotHPOFreeText: N/A

Protein Expression: N/A

CasePreviousTesting: 48,48 Access-Array amplification. NGS Molecular analysis was performed. Sanger sequencing was used to validate selected mutations.

Gene: VWF

Variant: NM_000552.4:c.3426T>C (p.Cys1142=) and NM_000552.4:c.3485_3486inv (p.Pro1162Leu)

ClinVar: 256668 and 619934

ClinVar1 ClinVar2

CAID: CA6402757 and CA658683073

CAID1 CAID2

gnomAD: MAF of 0.2228 (2240/10052 alleles) in the African population (CA6402757).

gnomAD1

Case#: 02047, Age 71, Male, Blood Type: O+ (Table S3)

Disease Assertion:VWD 1

FamilyInfo: Family# 0218; 3 Family members were included in the study. All 3 members had VWD type 1. All 3 have the same variant as patient 02047.

Parental Genotype: Not listed

CasePresentingHPOs: N/A

CaseHPOFreeText: Bleeding Score 9, VWF:Ag: 9.3, VWF:RCo:6.4, FVIII:C: 30, VWF:CB: 6.1

CaseNotHPOs:N/A

CaseNotHPOFreeText:N/A

Protein Expression: N/A

CasePreviousTesting: 48,48 Access-Array amplification. NGS Molecular analysis was performed. Sanger sequencing was used to validate selected mutations.

Gene: VWF

Variant: NM_000552.4:c.5471C>A (p.Pro1824His)

ClinVar: 100430 ClinVar

CAID:CA228725

CAID

gnomAD: N/A

Case#: 32010, Age 21, Male, Blood Type not determined (Table S3)

Disease Assertion: VWD 1

FamilyInfo: The patient is one of two members of Family#3210. The second member is a carrier of the variant c.3931C>T.

Parental Genotype: Not listed

CasePresentingHPOs: N/A

CaseHPOFreeText: Bleeding Score 12, VWF:Ag:7.2, VWF:RCo: 5.7, FVIII:C: 21, VWF:CB: 6.8

CaseNotHPOs: N/A

CaseNotHPOFreeText: N/A

Protein Expression: N/A

CasePreviousTesting: 48,48 Access-Array amplification. NGS Molecular analysis was performed. Sanger sequencing was used to validate selected mutations.

Gene: VWF

Variant: NM_000552.4:c.3931C>T (p.Gln1311Ter) and NM_000552.4:c.55G>A (p.Gly19Arg)

ClinVar: 100307 and 100432 ClinVar1 ClinVar

CAID: CA228496 and CA228729 CAID1 CAID2

gnomAD: MAF of 0.0003921 (12/ 30604 alleles) in the South Asian population (CA228496). gnomAD

MAF of 0.0008819 (27/ 30616 alleles) in the South Asian population.

gnomAD

Case#: 06002, Age 39, Female,Blood Type O+ (Table S3)

Disease Assertion: VWD1

FamilyInfo: Two total members in Family #0602. Patient 06003 has the c.2685G>C variant and a seperate intronic variant.

Parental Genotype: Not listed

CasePresentingHPOs: N/A

CaseHPOFreeText: Bleeding score 8, VWF:Ag: 25, VWF:RCo: 19.1, FVIII:C:88, VWF:CB:24

CaseNotHPOs: N/A

CaseNotHPOFreeText: N/A

Protein Expression: N/A

CasePreviousTesting: 48,48 Access-Array amplification. NGS Molecular analysis was performed. Sanger sequencing was used to validate selected mutations

Gene: VWF

Variant: NM_000552.4:c.2685G>C (p.Gln895His) and NM_000552.4:c.4751A>G (p.Tyr1584Cys)

ClinVar: 310

ClinVar

CAID: CA383519910 and CA114162

CAID1 CAID2

gnomAD: MAF of 0.004024 (519/128972 alleles) in the European (non-Finnish) population.

gnomAD

Case#: 06009, Age 60, Female, Blood Type O+ (Table S3)

Disease Assertion: VWD1

FamilyInfo: The patient is the only member of family#0607.

Parental Genotype: Not listed

CasePresentingHPOs: N/A

CaseHPOFreeText: Bleeding Score 2, VWF:Ag:23, VWF:RCo: 25, FVIII:C 83, VWF:CB: 23

CaseNotHPOs: N/A

CaseNotHPOFreeText: N/A

Protein Expression: N/A

CasePreviousTesting: 48,48 Access-Array amplification. NGS Molecular analysis was performed. Sanger sequencing was used to validate selected mutations

Gene: VWF

Variant: NM_000552.4:c.2435del (p.Pro812fs)

ClinVar: 303 ClinVar

CAID: CA114153 CAID

gnomAD: MAF of 0.0003601(9/ 24994 alleles) in the European (Finnish) population.

gnomAD

Case#: 13006, Age 66, Male, Blood Type A-

Disease Assertion: VWD 1

FamilyInfo: The patient is the only member of Family# 1304.

Parental Genotype: Not listed

CasePresentingHPOs: N/A

CaseHPOFreeText: Bleeding Score 8, VWF:Ag:10, VWF:RCo: 6, FVIII:C:16, VWF:CB:6.1

CaseNotHPOs: N/A

CaseNotHPOFreeText: N/A

Protein Expression: N/A

CasePreviousTesting: 48,48 Access-Array amplification. NGS Molecular analysis was performed. Sanger sequencing was used to validate selected mutations

Gene: VWF

Variant: NM_000552.4:c.3614G>A (p.Arg1205His)

ClinVar: 308 ClinVar

CAID: CA114160 CAID

gnomAD: N/A

Case#: 13014, Age20, Male, Blood type O+ (TableS3)

Disease Assertion: VWD 1

FamilyInfo: The patient is the only member of Family#1311.

Parental Genotype: Not listed

CasePresentingHPOs: N/A

CaseHPOFreeText: Bleeding Score 6, VWF:Ag:24, VWF:RCo: 22.5, FVIII:C:40, VWF:CB: 20

CaseNotHPOs: N/A

CaseNotHPOFreeText: N/A

Protein Expression: N/A

CasePreviousTesting: 48,48 Access-Array amplification. NGS Molecular analysis was performed. Sanger sequencing was used to validate selected mutations

Gene: VWF

Variant: NM_000552.4:c.4751A>G (p.Tyr1584Cys)

ClinVar: 310 ClinVar

CAID: CA114162 CAID

gnomAD: MAF of 0.004024 (519/ 128972 alleles) in the European (non-Finnish) population

Case#: 27012, Age 24, Female, Blood Type O+ (Table S3)

Disease Assertion: VWD 1

FamilyInfo: The patient is the only member of Family #2707.

Parental Genotype: Not listed

CasePresentingHPOs: N/A

CaseHPOFreeText: VWF:Ag: 33, VWF:RCo: 29, FVIII:C: 91, VWF:CB: 32

CaseNotHPOs: N/A

CaseNotHPOFreeText: Bleeding score 0

Protein Expression: N/A

CasePreviousTesting: 48,48 Access-Array amplification. NGS Molecular analysis was performed. Sanger sequencing was used to validate selected mutations.

Gene: VWF

Variant: NM_000552.4:c.3467C>T (p.Thr1156Met) and c.-3151T>G

ClinVar: 100264 ClinVar

CAID: CA228414 CAID

gnomAD:

Case#: 22011, Age 67, Male, Blood Type A- (Table S3)

Disease Assertion: VWD 1

FamilyInfo: The patient is the only member of Family#2207.

Parental Genotype: N/A

CasePresentingHPOs: N/A

CaseHPOFreeText: Bleeding score 7, VWF:Ag: 18, VWF:RCo:16, FVIII:C:46, VWF:CBNM_0:14

CaseNotHPOs: N/A

CaseNotHPOFreeText: N/A

Protein Expression: N/A

CasePreviousTesting: 48,48 Access-Array amplification. NGS Molecular analysis was performed. Sanger sequencing was used to validate selected mutations

Gene: VWF

Variant: NM_000552.4:c.6799-47G>A (p.=), NM_000552.4:c.7464C>T (p.Gly2488=), NM_000552.4:c.998-46C>T (p.=)

ClinVar: 619755 ClinVar1

CAID: CA6401901, CA232284628, CA6403602 CAID1 CAID2 CAID3

gnomAD: MAF of 0.0005876 (17/ 28930 alleles) in the South Asian population (CA6401901). gnomAD1

MAF of 0.000008799 (1/ 113644 alleles) in the European (non-Finnish) population (CA232284628). gnomAD2

MAF of 0.00007581 (8/105528 alleles) in the European (non-Finnish) population (CA6403602) gnomAD3

Case#: 37006, Age 45, Female, Blood Type B+ (Table S3)

Disease Assertion: VWD 1

FamilyInfo: The patient is the only member of Family#3706.

Parental Genotype: Not listed

CasePresentingHPOs: N/A

CaseHPOFreeText: VWF:Ag:7.6, VWF:RCo:5, FVIII:C: 8.9, VWF:CB: 6.9

CaseNotHPOs: N/A

CaseNotHPOFreeText: Bleeding Score 5

Protein Expression: N/A

CasePreviousTesting: 48,48 Access-Array amplification. NGS Molecular analysis was performed. Sanger sequencing was used to validate selected mutations.

Gene: VWF

Variant: NM_000552.4:c.3426T>C (p.Cys1142=), NM_000552.4:c.3614G>A (p.Arg1205His), NM_000552.4:c.7771-49G>A , NM_000552.4:c.3485_3486delinsTG (pPro1162Leu)

ClinVar: 256668 , 308, 619934 ClinVar1 ClinVar2 ClinVar3

CAID: CA6402757, CA114160, CA1139532197, and CA658683073 CAID1 CAID2 CAID3 CAID4

gnomAD: MAF of 0.2228 (2240/ 10052 alleles) in the African population (CA6402757). gnomAD

Case#: 35004, Age 16, Male, Blood Type A+ (Table S3)

Disease Assertion: VWD 1

FamilyInfo: The patient is one of 4 members of Family# 3501. All members have the same variants, and patient 35001 has an additional variant.

Parental Genotype: Not listed

CasePresentingHPOs: N/A

CaseHPOFreeText: Bleeding Score 6, VWF:Ag: 5.6, VWF:RCo: 7.9, FVIII:C: 19, VWF:CB: 7.8

CaseNotHPOs: N/A

CaseNotHPOFreeText: N/A

Protein Expression: N/A

CasePreviousTesting: 48,48 Access-Array amplification. NGS Molecular analysis was performed. Sanger sequencing was used to validate selected mutations.

Gene: VWF

Variant: NM_000552.3:c.5198T>C (p.Leu1733Pro) and NM_000552.4:c.6187C>T (p.Pro2063Ser)

ClinVar: 100436 ClinVar

CAID: CA383496267 and CA202650 CAID1 CAID2

gnomAD: MAF of 0.04854 (1486/ 30616 alleles) in the South Asian population (CA202650). gnomAD

Case#: 02068, Age 58, Female, Blood Type O+ (Table S3)

Disease Assertion: VWD 1

FamilyInfo: Patient is the only member of family #0225

Parental Genotype: Not listed

CasePresentingHPOs: N/A

CaseHPOFreeText: Bleeding Score 13,VWF:Ag: 11, VWF:RCo: 9.2, FVIII:C: 46, VWF:CB:14

CaseNotHPOs: N/A

CaseNotHPOFreeText: N/A

Protein Expression: N/A

CasePreviousTesting: 48,48 Access-Array amplification. NGS Molecular analysis was performed. Sanger sequencing was used to validate selected mutations

Gene: VWF

Variant:NM_000552.4:c.6352C>T (p.Arg2118Trp) and NM_000552.4:c.8155+3G>C

ClinVar: 100490

ClinVar

CAID: CA6402006 and CA228825

CAID1

CAID2

gnomAD: MAF of 0.0001504 (3/19950 alleles) in the East Asian population (CA6402006).

gnomAD

Case#: 23013, Age 46, Female, Blood Type O+ (Table S3)

Disease Assertion: VWD 1

FamilyInfo: The patient is the member of Family #2309.

Parental Genotype: Not listed

CasePresentingHPOs: N/A

CaseHPOFreeText: Bleeding Score 0, VWF:Ag: 33, VWF:RCo: 27.5, FVIII:C:56, VWF:CB:26

CaseNotHPOs: N/A

CaseNotHPOFreeText: N/A

Protein Expression: N/A

CasePreviousTesting: 48,48 Access-Array amplification. NGS Molecular analysis was performed. Sanger sequencing was used to validate selected mutations

Gene: VWF

Variant: NM_000552.4:c.6911G>A (p.Cys2304Tyr)

ClinVar: 100451 ClinVar

CAID: CA228762 CAID

gnomAD: N/A

Case#: 10003, Age 33, Male, Blood Type A+ (Table S3)

Disease Assertion: VWD1

FamilyInfo: The patient and one other patient are members of Family#1002. The second member has the same exonic variant and also has an intronic variant.

Parental Genotype: Not listed

CasePresentingHPOs: N/A

CaseHPOFreeText: Bleeding Score 1, VWF:Ag:7.4, VWF:RCo: 7.2, FVIII:C:11, VWF:CB: 6.3

CaseNotHPOs: N/A

CaseNotHPOFreeText: N/A

Protein Expression: N/A

CasePreviousTesting: 48,48 Access-Array amplification. NGS Molecular analysis was performed. Sanger sequencing was used to validate selected mutations

Gene: VWF

Variant: NM_000552.4:c.3614G>A (p.Arg1205His)

ClinVar: 308 ClinVar

CAID: CA114160 CAID

gnomAD: N/A

Case#: 36001, Age 5, Male, Blood Type A+ (Table S3)

Disease Assertion: VWD 1

FamilyInfo: The patient is the only member of Family#3601.

Parental Genotype: Not listed

CasePresentingHPOs: N/A

CaseHPOFreeText: VWF:Ag: 33, VWF:RCo: 26.4, FVIII:C: 88, VWF:CB: 58

CaseNotHPOs: N/A

CaseNotHPOFreeText: Bleeding Score 2

Protein Expression: N/A

CasePreviousTesting: 48,48 Access-Array amplification. NGS Molecular analysis was performed. Sanger sequencing was used to validate selected mutations.

Gene: VWF

Variant: NM_000552.4(VWF):c.5170+10C>T (p.=) and NM_000552.4(VWF):c.7730-1G>C (p.=)

ClinVar: 100418 and 100485 ClinVar1 ClinVar2

CAID: CA228702 and CA228816 CAID1 CAID2

gnomAD: MAF of 0.01298 (326/ 25116 alleles) in the European (Finnish) population (CA228702). gnomAD

Case#: 22006, Age 58, Male, Blood Type O+ (Table S3)

Disease Assertion: VWD 1

FamilyInfo: The patient is one of 2 patients in Family# 2206. Patient 22007's variant information was not described.

Parental Genotype: Not listed

CasePresentingHPOs: N/A

CaseHPOFreeText: Bleeding Score 7, VWF:Ag: 27, VWF:RCo: 20, FVIII:C: 12, VWF:CB:22

CaseNotHPOs: N/A

CaseNotHPOFreeText: N/A

Protein Expression: N/A

CasePreviousTesting: 48,48 Access-Array amplification. NGS Molecular analysis was performed. Sanger sequencing was used to validate selected mutations

Gene: VWF

Variant:NM_000552.4:c.1001G>A (p.Gly334Glu) and NM_000552.4(VWF):c.2561G>A (p.Arg854Gln)

ClinVar: 626947 and 296 ClinVar1 ClinVar2

CAID: CA232322015 and CA114139 CAID1 CAID2

gnomAD: MAF of 0.000008824 (1/11326 alleles) in the European (non-Finnish) population (CA232322015). gnomAD1

MAF of 0.005692 (143/25122 alleles) in the European (Finnish) population (CA114139). gnomAD2

Case#: 13010, Age 37, Female, Blood Type O+ (Table S3)

Disease Assertion: VWD1

FamilyInfo: The patient is one of two members of Family#1308. Patient 13011 has one of the same exonic variant but has one different exonic variant (c.546G>A).

Parental Genotype: Not listed

CasePresentingHPOs: N/A

CaseHPOFreeText: Bleeding Score 18, VWF:Ag:16, VWF:RCo:15.1, FVIII:C:45, VWF:CB14

CaseNotHPOs: N/A

CaseNotHPOFreeText:N/A

Protein Expression: N/A

CasePreviousTesting: 48,48 Access-Array amplification. NGS Molecular analysis was performed. Sanger sequencing was used to validate selected mutations

Gene: VWF

Variant: NM_000552.4:c.1472G>C(p.Arg491Pro) and NM_000552.4:c.546G>A (p.=)

ClinVar: 256684 ClinVar

CAID: CA383500718 and CA6403759 [CAID1] (http://reg.clinicalgenome.org/redmine/projects/registry/genboree_registry/by_caid?caid=CA383500718)

CAID2

gnomAD: MAF of 0.002398 (85/35440 alleles) in the Latino population (CA6403759).

gnomAD

Case#: 13012, Age 42, Male, Blood Type A+ (Table S3)

Disease Assertion: VWD 1

FamilyInfo: The patient is the only member of Family#1309.

Parental Genotype: Not listed

CasePresentingHPOs: N/A

CaseHPOFreeText: Bleeding score 7, VWF:Ag:14, VWF:RCo:9, FVIII:C: 20, VWF:CB:11

CaseNotHPOs: N/A

CaseNotHPOFreeText: N/A

Protein Expression: N/A

CasePreviousTesting: 48,48 Access-Array amplification. NGS Molecular analysis was performed. Sanger sequencing was used to validate selected mutations

Gene: VWF

Variant: NM_000552.4:c.3614G>A (p.Arg1205His) and

ClinVar: 308 ClinVar

CAID: CA114160 and CA6403769 CAID1 CAID2

gnomAD: MAF of 0.0005784 (20/34578 alleles) in the Latino population (CA6403769). gnomAD

Case#: 21023, Age 43, Female, Blood Type O+ (Table S3)

Disease Assertion: VWD 1

FamilyInfo: The patient is the only member of Family #2120.

Parental Genotype: Not listed

CasePresentingHPOs: N/A

CaseHPOFreeText: VWF:Ag:7.6, VWF:RCo:6, FVIII:C: 11, VWF:CB:3.9

CaseNotHPOs: N/A

CaseNotHPOFreeText: Bleeding Score 2

Protein Expression: N/A

CasePreviousTesting: 48,48 Access-Array amplification. NGS Molecular analysis was performed. Sanger sequencing was used to validate selected mutations

Gene: VWF

Variant: NM_000552.4:c.1110-26T>A (p.=) , NM_000552.4:c.2103C>T (p.Cys701=), and NM_000552.4:c.6248G>A (p.Gly2083Asp)

ClinVar: 310076 ClinVar1

CAID: CA6403542 , CA6403175, and CA383492072 CAID1 CAID2 CAID3

gnomAD: MAF of 0.008911 (222/ 24914 alleles) in the African population (CA6403542). MAF of 0.005928 (148/ 24968 alleles) in the African population (CA6403175). gnomAD1

Case#: 35003, Age 75, Female, Blood Type A- (Table S3)

Disease Assertion: VWD 1

FamilyInfo: The patient is one of two members in Family# 3502.

Parental Genotype: Not listed

CasePresentingHPOs: N/A

CaseHPOFreeText: VWF:Ag: 6.6, VWF:RCo: 8.5, FVIII:C:13, VWF:CB: 11

CaseNotHPOs: N/A

CaseNotHPOFreeText: Bleeding Score 2

Protein Expression: N/A

CasePreviousTesting: 48,48 Access-Array amplification. NGS Molecular analysis was performed. Sanger sequencing was used to validate selected mutations.

Gene: VWF

Variant: NM_000552.4:c.3614G>A (p.Arg1205His)

ClinVar: 308 ClinVar

CAID: CA114160 CAID

gnomAD: N/A

Case#: 02074, Age 37, Female, Blood Type A+ (Table S3)

Disease Assertion: VWD 1

FamilyInfo: Patient was the only member of family #0228.

Parental Genotype: Not listed

CasePresentingHPOs: N/A

CaseHPOFreeText: Bleeding Score 3, VWF:Ag: 24, VWF:RCo: 21, FVIII:C: 111, VWF:CB: 29

CaseNotHPOs: N/A

CaseNotHPOFreeText: N/A

Protein Expression: N/A

CasePreviousTesting: 48,48 Access-Array amplification. NGS Molecular analysis was performed. Sanger sequencing was used to validate selected mutations

Gene: VWF

Variant: NM_000552.4(VWF):c.3426T>C (p.Cys1142=), NM_000552.4:c.6254G>T (p.Cys2085Phe), and NM_000552.4:c.3485_3486inv (p.Pro1162Leu)

ClinVar: 256668 and 619934 ClinVar ClinVar2

CAID: CA6402757, CA383492060, and CA658683073

CAID1

CAID2

gnomAD: MAF of 0.2228 (2240/10052 alleles) in the African population (CA6402757).

Case#: 02064, Age 32, Male, Blood type O- (Table S3)

Disease Assertion: VWD 1

FamilyInfo: Patient is a part of family # 0224. There are 3 other members of the family (02065, 02066, 02067). All members of the family have the same variant.

Parental Genotype: Not listed

CasePresentingHPOs: N/A

CaseHPOFreeText: Bleeding score 13, VWF:Ag: 4.7, VWF:RCo: 5, FVIII:C: 16, VWF:CB:6

CaseNotHPOs: N/A

CaseNotHPOFreeText: N/A

Protein Expression: N/A

CasePreviousTesting: 48,48 Access-Array amplification. NGS Molecular analysis was performed. Sanger sequencing was used to validate selected mutations

Gene:VWF

Variant: NM_000552.4:c.3614G>A (p.Arg1205His)

ClinVar: 308

ClinVar

CAID: CA114160

CAID1

gnomAD: N/A

Case#: 27024, Age 56, Female, Blood Type A+ (Table S3)

Disease Assertion: VWD 1

FamilyInfo: The patient is the only member of Family#2713.

Parental Genotype: Not listed

CasePresentingHPOs: N/A

CaseHPOFreeText: VWF:Ag:5.3, VWF:RCo:6, FVIII:C:38, VWF:CB:4.2

CaseNotHPOs: N/A

CaseNotHPOFreeText: Bleeding Score 3

Protein Expression: N/A

CasePreviousTesting: 48,48 Access-Array amplification. NGS Molecular analysis was performed. Sanger sequencing was used to validate selected mutations.

Gene: VWF

Variant: NM_000552.4:c.3467C>T (p.Thr1156Met) and c.-3151T>G

ClinVar: 100264 ClinVar

CAID: CA228414 CAID

gnomAD: N/A

Case#: 06024, Age 36, Female, Blood Type O+ (Table S3)

Disease Assertion: VWD1

FamilyInfo: The patient is the only member of Family#0617.

Parental Genotype: Not listed

CasePresentingHPOs: N/A

CaseHPOFreeText: Bleeding Score 1, VWF:Ag:35, VWF:RCo:25, FVIII:C:69, VWF:CB: 27

CaseNotHPOs: N/A

CaseNotHPOFreeText: N/A

Protein Expression:N/A

CasePreviousTesting: 48,48 Access-Array amplification. NGS Molecular analysis was performed. Sanger sequencing was used to validate selected mutations

Gene: VWF

Variant: NM_000552.4:c.7300C>T (p.Arg2434Ter)

ClinVar: 100464 ClinVar

CAID: CA228781 CAID

gnomAD: MAF of 0.0001298 (2/15414 alleles) in the European (non-Finnish) population. gnomAD

Case#: 02058, Age 64, Female, Blood type A+ (Table S3)

Disease Assertion: VWD1

FamilyInfo: Patient is the only member of family #0221.

Parental Genotype: Not listed

CasePresentingHPOs: N/A

CaseHPOFreeText: Bleeding score 19, VWF:Ag: 7.3, VWF:RCo: 5.9, FVIII:C: 19, VWF:CB: 6.3

CaseNotHPOs: N/A

CaseNotHPOFreeText: N/A

Protein Expression: N/A

CasePreviousTesting: 48,48 Access-Array amplification. NGS Molecular analysis was performed. Sanger sequencing was used to validate selected mutations.

Gene: VWF

Variant: NM_000552.4:c.3614G>A (p.Arg1205His)

ClinVar: 308 ClinVar

CAID: CA114160 CAID

gnomAD: N/A

Case#: 03035, Age 51, Male (Table S3)

Disease Assertion: VWD1

FamilyInfo: A part of family #03150 along with two other patients.

Parental Genotype: Not listed

CasePresentingHPOs: Not listed

CaseHPOFreeText: Bleeding Score 4, VWF:Ag: 34, VWF:RCo: 26, FVIII:C: 43, VWF:CB 33

CaseNotHPOs: N/A

CaseNotHPOFreeText: N/A

Protein Expression:N/A

CasePreviousTesting: 48,48 Access-Array amplification. NGS Molecular analysis was performed. Sanger sequencing was used to validate selected mutations

Gene: VWF

Variant: NM_000552.4:c.3467C>T (p.Thr1156Met) and NM_000552.3:c.-3151T>G

ClinVar: 100264 ClinVar

CAID: CA228414 CAID

gnomAD: N/A

Case#: 32002, Age 49, Female, Blood Type +

Disease Assertion: VWD 1

FamilyInfo: The patient is the only member of Family# 3202.

Parental Genotype: Not listed

CasePresentingHPOs: N/A

CaseHPOFreeText: VWF:Ag: 34, VWF:RCo: 30, FVIII:C: 53, VWF:CB: 29

CaseNotHPOs: N/A

CaseNotHPOFreeText: Bleeding Score 3

Protein Expression: N/A

CasePreviousTesting: 48,48 Access-Array amplification. NGS Molecular analysis was performed. Sanger sequencing was used to validate selected mutations.

Gene: VWF

Variant: NM_000552.4:c.7408C>T (p.Gln2470Ter)

ClinVar: 100469 ClinVar

CAID: CA228790 CAID

gnomAD: MAF of 0.00002891 (1/34590 alleles) in the Latino population.

Case#: 18001, Age 50, Male, Blood Type B+ (Table S3)

Disease Assertion: VWD 1

FamilyInfo: The patient one of 3 members of Family#1801. All of the patients have the same variant.

Parental Genotype: N/A

CasePresentingHPOs: N/A

CaseHPOFreeText: VWF:Ag:10, VWF:RCo:6.1, FVIII:C:17, VWF:CB:8.2

CaseNotHPOs: N/A

CaseNotHPOFreeText: N/A

Protein Expression: N/A

CasePreviousTesting: 48,48 Access-Array amplification. NGS Molecular analysis was performed. Sanger sequencing was used to validate selected mutations

Gene: VWF

Variant: NM_000552.4:c.3614G>A (p.Arg1205His) and NM_000552.4:c.6989T>G (p.Val2330Gly)

ClinVar: 308 ClinVar1

CAID: CA114160 and CA6401808 CAID1 CAID2

gnomAD: MAF of 0.000008823 (1/113342 alleles) in the European (non-Finnish population) (CA6401808) gnomAD

Case#: 36008, Age 34, Female, Blood Type O+ (Table S3)

Disease Assertion: VWD 1

FamilyInfo: The patient is the only member of Family#3606.

Parental Genotype: Not listed

CasePresentingHPOs: N/A

CaseHPOFreeText: Bleeding Score 8, VWF:Ag: 34, VWF:RCo:14, FVIII:C: 37, VWF:CB: 25

CaseNotHPOs: N/A

CaseNotHPOFreeText: N/A

Protein Expression: N/A

CasePreviousTesting: 48,48 Access-Array amplification. NGS Molecular analysis was performed. Sanger sequencing was used to validate selected mutations.

Gene: VWF

Variant: NM_000552.4:c.2771G>A(p.Arg924Gln) and NM_000552.4:c.3390C>T (p.Cys1130=)

ClinVar: 627186 ClinVar

CAID: CA228368 and CA478101454 CAID1 CAID2

gnomAD: MAF of 0.01866 (2411/ 129182 alleles) in the European (non-Finnish) population (CA228368). gnomAD

Case#: 02052, Age 25, Male, Blood type A+ (Table S3)

Disease Assertion: VWD 1

FamilyInfo: Patient is the only family member of family # 0220

Parental Genotype: Not listed

CasePresentingHPOs:N/A

CaseHPOFreeText: Bleeding Score: 11, VWF:Ag: 15, VWF:RCO:12, FVIII:C:20, VWF:CB: 16

CaseNotHPOs: N/A

CaseNotHPOFreeText: N/A

Protein Expression: N/A

CasePreviousTesting: 48,48 Access-Array amplification. NGS Molecular analysis was performed. Sanger sequencing was used to validate selected mutations.

Gene: VWF

Variant: NM_000552.4:c.1625C>G (p.Ala542Gly) and NM_000552.4:c.6187C>T (p.Pro2063Ser)

ClinVar: 381621 and 100436 ClinVar1

ClinVar2

CAID: CA6403319 and CA202650 CAID1

CAID2

gnomAD: MAF of 0.001355 (48/ 35418 alleles) in the Latino population (CA6403319). MAF of 0.04854 (1486/ 30616 alleles) in the South Asian Population (CA202650).

gnomAD1

gnomAD2

CaseIII-3: female

DiseaseAssertion: LVNC, Ebstin anomaly, congenital heart disease

FamilyInfo: all mutation carriers have LVNC, with some having Ebstein anomaly and/or other CHD

CasePresentingHPOs:

CaseHPOFreeText:

CaseNotHPOs:

CaseNotHPOFreeText:

CasePreviousTesting:

GenotypingMethod:

Variant: NM_000257.4:c.847T>G

ClinVar: 66081

CAID: CA016893

gnomAD:

Case#: Subject number 1, male, 22 years (report), 9 years (onset), 11 years (diagnosis)

DiseaseAssertion: Pompe disease

FamilyInfo: N/A

CasePresentingHPOs: HP:0002650, HP:0001638 (Scoliosis, cardiomyopathy)

CaseHPOFreeText: on BiPap, ambulatory

CaseNotHPOs: HP:0002617, HP:0000365, HP:0000360, HP:0000939, HP:0009830, HP:0100647, HP:0010788, HP:0000821 (aneurysm, hearing loss, tinnitus, osteoporosis, peripheral neuropathy, Graves disease, Testicular cancer, hypothyroidism)

CaseNotHPOFreeText: No infusion reaction, no tracheostomy

CasePreviousTesting: “documented deficiency of acid alpha-glucosidase enzyme activity”; on enzyme replacement therapy

Variant 1: NM_000152.5(GAA):c.1655T>C (p.Leu552Pro)

Variant 1 ClinVarID:279811

Variant 1 CAid:CA8815429

Variant 1 gnomAD MAF: 0.00007053; 8/113426 (European non-Finnish)[https://gnomad.broadinstitute.org/variant/17-78085800-T-C?dataset=gnomad_r2_1]

Variant 2: NM_000152.5(GAA):c.2238G>C (p.Trp746Cys)

Variant 2 ClinVarID:265160

Variant 2 CAid:CA9301331

Variant 2 gnomAD MAF: 0.0005673; 73/128684 (European non-Finnish)[https://gnomad.broadinstitute.org/variant/17-78090815-G-C?dataset=gnomad_r2_1]

Zygosity: Compound heterozygous

ParentalGenotype: N/A

PreviouslyPublished: N/A

Case: P7, M/41

DiseaseAssertion: VWD2A

FamilyInfo: Positive family history, no details provided.

ParentalGenotype: NA

CaseHPOFreeText: The bleeding score was 7, aPTT was 36 seconds, FVIII:C was 36%, VWF:RCo was not determined, and VWF:Ag was 25 IU/dL. The VWF:RCo/VWF:Ag ratio was not applicable. The VWF multimer pattern showed a mild loss of high molecular weight multimers, RIPA was 4 ohms, and the platelet count was 301 × 10⁹/L.

CasePreviousTesting: NA

GenotypingMethod: Direct sequencing of all exons, flanking intronic sequences, and the promoter of VWF was performed.

Gene: VWF

Variant: c.4789C>T, p.R1597W — heterozygous

HGVS: NM_000552.5(VWF):c.4789C>T (p.Arg1597Trp)

ClinVar: 285

CAID: CA114117

gnomAD: https://gnomad.broadinstitute.org/variant/chr12-6018629-G-A?dataset=gnomad_r4

Case: P22, F/5

DiseaseAssertion: VWD2A

FamilyInfo: Positive family history, no details provided.

ParentalGenotype: NA

CaseHPOFreeText: The bleeding score was 3, aPTT was 43 seconds, FVIII:C was 47%, VWF:RCo was 122 IU/dL, and VWF:Ag was 20 IU/dL, with a VWF:RCo/VWF:Ag ratio of 0.1. RIPA was normal, VWF multimer analysis was 2, and platelet count was 270 × 10⁹/L.

CasePreviousTesting: NA

GenotypingMethod: Direct sequencing of all exons, flanking intronic sequences, and the promoter of VWF was performed.

Gene: VWF

Variant: c.4825G>A, p.G1609R — heterozygous

HGVS: NM_000552.5(VWF):c.4825G>A (p.Gly1609Arg)

ClinVar: 100395

CAID: CA228665

gnomAD: https://gnomad.broadinstitute.org/variant/chr12-6018593-C-T?dataset=gnomad_r4

Case: P21, F/5

DiseaseAssertion: VWD2M

FamilyInfo: Positive family history, no details provided.

ParentalGenotype: NA

CaseHPOFreeText: The bleeding score was 2, aPTT was 53 seconds, FVIII:C was 30%, and VWF:RCo was 25 IU/dL. The VWF:RCo/VWF:Ag ratio indicated a mild loss of high molecular weight multimers. RIPA was 4, VWF multimer analysis was 301, and platelet count was not specified.

CasePreviousTesting: NA

GenotypingMethod: Direct sequencing of all exons, flanking intronic sequences, and the promoter of VWF was performed.

Gene: VWF

Variant: c.4121G>T, p.R1374L — heterozygous

HGVS: NM_000552.5(VWF):c.4121G>T (p.Arg1374Leu)

ClinVar: 100331

CAID: CA228543

gnomAD: https://gnomad.broadinstitute.org/variant/chr12-6019297-C-A?dataset=gnomad_r4

Case: P20, F/33

DiseaseAssertion: VWD1

FamilyInfo: No family history

ParentalGenotype: NA

CaseHPOFreeText: The bleeding score was 4, aPTT was 25 seconds, FVIII:C was 6%, VWF:RCo was 19 IU/dL, and VWF:Ag was 19 IU/dL, with a VWF:RCo/VWF:Ag ratio of 1.0. RIPA was normal, VWF multimer analysis was not tested, and platelet count was 475 × 10⁹/L.

CasePreviousTesting: NA

GenotypingMethod: Direct sequencing of all exons, flanking intronic sequences, and the promoter of VWF was performed.

Gene: VWF

Variant: c.1135T>G, p.C379G, c.7390C>T, p.R2464C. Compound heterozygous

HGVS: NM_000552.5(VWF):c.1135T>G (p.Cys379Gly), NM_000552.5(VWF):c.7390C>T (p.Arg2464Cys)

ClinVar: 100467

CAID: CA6403534 , CA228786

gnomAD: https://gnomad.broadinstitute.org/variant/chr12-6071318-A-C?dataset=gnomad_r4, https://gnomad.broadinstitute.org/variant/chr12-5976158-G-A?dataset=gnomad_r4

Case: P18, F/39

DiseaseAssertion: VWD1

FamilyInfo: No family history

ParentalGenotype: NA

CaseHPOFreeText: The bleeding score was 9, aPTT was 40 seconds, FVIII:C was 15%, VWF:RCo was 6 IU/dL, and VWF:Ag was 8 IU/dL, with a VWF:RCo/VWF:Ag ratio of 0.8. The VWF multimer pattern was absent, RIPA was 2 ohms, and the platelet count was 273 × 10⁹/L.

CasePreviousTesting: NA

GenotypingMethod: Direct sequencing of all exons, flanking intronic sequences, and the promoter of VWF was performed.

Gene: VWF

Variant: c.3943C>T, p.R1315C — heterozygous

HGVS: NM_000552.5(VWF):c.3943C>T (p.Arg1315Cys)

ClinVar: 100310

CAID: CA228502

gnomAD: https://gnomad.broadinstitute.org/variant/chr12-6019475-G-A?dataset=gnomad_r4

Case: P17, F/30

DiseaseAssertion: VWD2A

FamilyInfo: No family history

ParentalGenotype: NA

CaseHPOFreeText: The bleeding score was 8, aPTT was 42 seconds, FVIII:C was 23%, VWF:RCo was 20 IU/dL, and VWF:Ag was 11 IU/dL, with a VWF:RCo/VWF:Ag ratio of 1.8. The VWF multimer pattern showed a loss of intermediate and high molecular weight multimers, RIPA was not tested, and the platelet count was 276 × 10⁹/L.

CasePreviousTesting: NA

GenotypingMethod: Direct sequencing of all exons, flanking intronic sequences, and the promoter of VWF was performed.

Gene: VWF

Variant: c.3814T>C, p.C1272R — heterozygous

HGVS: NM_000552.5(VWF):c.3814T>C (p.Cys1272Arg)

ClinVar: 301

CAID: CA114149

gnomAD: absent from gnomad v4.1.0

Case: P9, M/26

DiseaseAssertion: VWD1

FamilyInfo: No family history

ParentalGenotype: NA

CaseHPOFreeText: The bleeding score was 1, aPTT was 44 seconds, FVIII:C was 9%, VWF:RCo was 13 IU/dL, and VWF:Ag was 6 IU/dL, with a VWF:RCo/VWF:Ag ratio of 2.0. The VWF multimer pattern was normal, RIPA was 3 ohms, and the platelet count was 217 × 10⁹/L.

CasePreviousTesting: NA

GenotypingMethod: Direct sequencing of all exons, flanking intronic sequences, and the promoter of VWF was performed.

Gene: VWF

Variant: c.3614G>A, p.R1205H — heterozygous

HGVS: NM_000552.5(VWF):c.3614G>A (p.Arg1205His)

ClinVar: 308

CAID: CA114160

gnomAD: https://gnomad.broadinstitute.org/variant/chr12-6021960-C-T?dataset=gnomad_r4

Case: P10, M/2

DiseaseAssertion: VWD1

FamilyInfo: No family history

ParentalGenotype: NA

CaseHPOFreeText: The bleeding score was 1, aPTT was 33 seconds, FVIII:C was 96%, VWF:RCo was 30 IU/dL, and VWF:Ag was 36 IU/dL, resulting in a VWF:RCo/VWF:Ag ratio of 0.8. The VWF multimer pattern was normal, RIPA was not tested, and the platelet count was 222 × 10⁹/L.

CasePreviousTesting: NA

GenotypingMethod: Direct sequencing of all exons, flanking intronic sequences, and the promoter of VWF was performed.

Gene: VWF

Variant: c.3835G>A, p.V1279I — heterozygous

HGVS: NM_000552.5(VWF):c.3835G>A (p.Val1279Ile)

ClinVar: 100288

CAID: CA228459

gnomAD: https://gnomad.broadinstitute.org/variant/chr12-6019583-C-T?dataset=gnomad_r4

Case: p.R1374C, Male, Age 39, Caucasian

Disease Assertion: VWD2M

FamilyInfo: 25 family members tested, 1 with high bleeding score, some affected by VWD (number not defined)

Parental Genotype: Not listed

CasePresentingHPOs:

CaseHPOFreeText: VWF:RCo 12 IU/dL, VWF:GPIbM 18 U/dL, VWF:CB3 19 IU/dL, VWF:CB4 14 U/dL, VWF:RCo/VWF:Ag ratio 0.55

CaseNotHPOs: HP:0040240

(Increased Ratio of VWF Propeptide to VWF antigen)

CaseNotHPOFreeText:bleeding score 2, normal multimer distribution,VWF:Ag 22 IU/dL

Protein Expression:

CasePreviousTesting: VWF sequencing

Gene: VWF

Variant: NM_000552.4(VWF):c.4120C>T (p.Arg1374Cys)

ClinVar: 100329 ClinVarID

CAID: CA228539

CAID

gnomAD: MAF of 0.000008824 (1/113322 alleles) in the European (non-Finnish) population.

gnomAD

Patient28: Male

CasePresentingHPOs: HP:0004887, HP:0011471, HP:0001270, HP:0002546, HP:0000365, HP:0001371

HPOsFreeText: CRIM positive, Anti-rh GGA antibodies maximum titer = 1:3200, infusion associated reaction, tracheostomy, heart normalization

CaseNotHPOs:

NotHPOsFreeText:

CaseAddInfo: variants in trans, symptom onset at 2yo, diagnosis at 4yo, ERT at 9yo

CasePreviousTesting: same as group

CaseGenotypingMethod: same as group

Variant1: NM_000152.4(GAA):c.2237G>A (p.Trp746Ter)

ClinVar: 280063, https://www.ncbi.nlm.nih.gov/clinvar/variation/280063/

gnomAD: 0.000003988, http://gnomad.broadinstitute.org/variant/17-78090814-G-A

Variant2: NM_000152.4(GAA):c.1655T>C (p.Leu552Pro)

ClinVar: 279811, https://www.ncbi.nlm.nih.gov/clinvar/variation/279811/

gnomAD: 0.00003188, http://gnomad.broadinstitute.org/variant/17-78085800-T-C

Patient24: Female

CasePresentingHPOs: HP:0000365

HPOsFreeText: CRIM positive, heart normalization

CaseNotHPOs: HP:0004887, HP:0025156, HP:0002355, HP:0002546, HP:0001371

NotHPOsFreeText: infusion associated reaction

CaseAddInfo: variants in trans, symptom onset at 2yo, diagnosis at 3yo, ERT at 4yo

CasePreviousTesting: same as group

CaseGenotypingMethod: same as group

Variant1: NM_000152.4(GAA):c.1933G>A (p.Asp645Asn)

ClinVar: 188728, https://www.ncbi.nlm.nih.gov/clinvar/variation/188728/

gnomAD: 0.000008250, http://gnomad.broadinstitute.org/variant/17-78086719-G-A

Variant2: NM_000152.3:c.1822C>T

CAID: CA8815488, http://reg.clinicalgenome.org/redmine/projects/registry/genboree_registry/allele?hgvs=NM_000152.3%3Ac.1822C%3ET

gnomAD: 0.00001806, http://gnomad.broadinstitute.org/variant/17-78086444-C-T

Case#: Patient 4, Female, 10 days old at diagnosis, African American

DiseaseAssertion: Pompe disease

FamilyInfo:N/A

CasePresentingHPOs: HP:0009062, HP:0001638

(Infantile axial hypotonia, Cardiomyopathy)

CaseHPOFreeText: Before ERT patient withdraws extremities to stimulation, contractures of large joint of upper and lower extremities, weak grasp. Also patient was invasively ventilated. Baseline – G-tube feeds and continues

Currently patient is off ventilator 10-12 hours a day. Sits with support; and has minimal capacity for weight bearing on lower extremities. G tube feed.

CaseNotHPOs: N/A

CaseNotHPOFreeText: N/A

CasePreviousTesting: No GAA measurement

Variant 1: M_000152.5(GAA):c.525del (p.Glu176fs)

ClinVar: 4033

CAID: CA220406

gnomAD: N/A

Variant 2: NM_000152.5(GAA):c.2560C>T (p.Arg854Ter)

ClinVar: 4034

CAID: CA340130

gnomAD: Highest population MAF= 0.001891 (African)

ParentalGenotype:N/A

PreviouslyPublished:N/A

Case#: P 10, Male, 7.0 months old when started ERT (Died at 21.1 months old)

DiseaseAssertion: Pompe disease

FamilyInfo: N/A

CasePresentingHPOs: HP:0001639 , HP:0001252 , HP:0003701, HP:0006532

(Hypertrophic cardiomyopathy, Muscular hypotonia, Proximal muscle weakness, Recurrent pneumonia )

CaseHPOFreeText: Patient required tube feeding

Recurrent pneumonia was a complication during ERT

Patient did not achieved motor function

CaseNotHPOs: N/A

CaseNotHPOFreeText: Patient was not ventilated

CasePreviousTesting: CRIM status was unknown

Antibody titer was not available

Variant: NM_000152.5(GAA):c.2560C>T (p.Arg854Ter)

ClinVar: 4034

CAID: CA340130

gnomAD: Highest population MAF = 0.001891 (African)

ParentalGenotype: Both the paternal and maternal allele carried the c.2560C>T (p.Arg854Ter) Variant

PreviouslyPublished: N/A

Patient12: Male, 19 years report, 13 years onset

CasePresentingHPOs: HP:0001324 (Muscle weakness)

HPOsFreeText: GAA activity 9.9% from leukocyte

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

NotHPOsFreeText: No wheelchair needed, Respiratory Assistance not needed

CaseAddInfo: LOPD

ExperimentName: Factor VIII binding assays for variants exogenously expressed in COS-7 cells

NonPatientCell: COS-7 cell line (CLO:0002597, EFO:0002009)

GeneFunction: GO:0005515 (protein binding, specifically to Factor VIII)

GeneAlteration: Site-directed mutagenesis was used to introduce a mutation encoding "His54Gln" (corresponding to the p.His817Gln substitution from the variant) into the mature (cleaved) VWF protein.

FunctionalEvidence: The variant did not disrupt multimer formation (Figure 5) but reduced Factor VIII binding to 75% of wild-type levels (Figure 6). In combination with another missense mutation in cis that reduced Factor VIII binding by an additional 21-23%, and a frameshift mutation in trans predicted to cause the absence of a gene product, the variant appears likely to be contributing to pathogenicity.

FAVariants: A construct equivalent to the c.2451T>A (p.His817Gln) variant was used (ClinVar:100224), although it expressed the proteolytically processed version of the protein product and referred to the variant as "His54Gln".

Case#: Case III-2, female, 64 yr, early childhood onset

DiseaseAssertion: None

FamilyInfo: Family C, her mother (II-1) and maternal grandmother (I-1) had late-onset leg weakness (Fig. 1C), her brother (III-4) had severe generalized weakness and was diagnosed with an unspecified muscular dystrophy

CasePresentingHPOs: Gait disturbance, lower limb muscle weakness, hypertension, osteoarthritis, hyperlipidemia, glaucoma, neck flexor weakness, weakness of facial musculature, weakness of long finger extensor muscles, foot dorsiflexor weakness (HP:0001288, HP:0007340, HP:0000822, HP:0002758, HP:0003077, HP:0000501, HP:0003722, HP:0030319, HP:0009077, HP:0009027)

CaseHPOFreeText: mild heel cord contractures, mild eye closure weakness, unable to stand on heels, limb weakness

CaseNotHPOs: Scoliosis, scapular winging, generalized muscle hypertrophy (HP:0002650, HP:0003691, HP:0003720)

CaseNotHPOFreeText: hand or foot deformities, normal knee bend and stand on tiptoes, normal reflexes and sensory

CasePreviousTesting: N/A

CaseGenotypingMethod: neurological examination, serum creatine kinase, nerve conduction studies, concentric needle electromyography, electrocardiogram, echocardiogram, forced vital capacity, muscle biopsy, immunohistochemical analyses, CK 156 unit/L (n < 143), FVC 2.13 L (68%), ECG and echo normal, left ventricular ejection fraction 65%, normal NCS except low-amplitude peroneal and tibial compound muscle action potential, abnormal increase of small polyphasic MUAP in tibialis anterior, no fibrillation potentials, no positive sharp waves, no myotonic discharges

Variant: NM_182476.3(COQ6):c.115G>A (p.Val39Met) & NM_000257.4(MYH7):c.4844_4846AGA[2] (p.Lys1617del)

ClinVar: 214236 https://www.ncbi.nlm.nih.gov/clinvar/variation/214236/ & 190401 https://www.ncbi.nlm.nih.gov/clinvar/variation/190401/

CAID: N/A

gnomAD: 0.00006720 African https://gnomad.broadinstitute.org/variant/14-74417150-G-A & absent

CaseB: Female, 2 months onset, 6 months diagnosis

CasePresentingHPOs: HP:0001270, HP:0003324, HP:0002540, HP:0006532, HP:0003202, HP:0004887, HP:0001638 (Motor delay, Generalized weakness, Inability to walk, Pulmonary infection, Muscle atrophy, Respiratory failure requiring assisted ventilation, Cardiomyopathy)

HPOsFreeText: CRIM positive, Has a tracheostoma, Needs ventilatory support for almost 24 h/day

CaseNotHPOs: HP:0100543 (Cognitive impairment)

CaseAddInfo: Infantile-Onset Pompe Disease

Case#: Patient 4, Gender unknown, age unknown, ethnicity unknown

DiseaseAssertion: Pompe disease

FamilyInfo: N/A

CasePresentingHPOs: HP:0001638, HP:0003756

(Cardiomyopathy, Skeletal myopathy)

CaseHPOFreeText: N/A

CaseNotHPOs: N/A

CaseNotHPOFreeText: N/A

CasePreviousTesting: Sequence analysis of the GAA gene was performed in a CLIA and CAP certified laboratory. Cross Reactive Immune reaction was indeterminate in peripheral blood mononuclear cells (PBMCs)

Cross Reactive Immune reaction was not available in Dried blood spots.

Variant: NM_000152.5(GAA):c.2560C>T (p.Arg854Ter)

ClinVar: 4034

CAID:CA340130

gnomAD: Highest population MAF= 0.001891 (African)

ParentalGenotype: N/A

Case#: Basani_1996_LeM, female, Caucasian, >1 y.o.

DiseaseAssertion: GT

FamilyInfo: nonconsanguineous parents

ParentalGenotype: one obligatory carrier

CasePresentingHPOs: HP:0000967 Petechiae, HP:0000979 Purpura,

PlateletAggregation: absence of platelet aggregation in response to adenosine diphosphate (ADP), epinephrine, and collagen, but a normal response to ristocetin.

CaseHPOFreeText: Platelet Count (1 day old): 181,000/mm^3

CaseNotHPOs:

CaseNotHPOFreeText:

ProteinExpression: Flow cytometry: platelets expressed <5% of the normal amount of αIIbβ3 when stained with the β3-specific MoAb SSA6,24 the heterodimer-dependent MoAb A2A923,24 or the activation-dependent MoAb PAC1.25 On the other hand, LeM and control platelets bound comparable amounts of the anti-GPIb MoAb AP1, confirming that the defect in the patient's platelets was confined to αIIbβ3

CasePreviousTesting:

Method: PCR using VENT polymerase confirmed with SSCPA

PreviouslyPublished:

Gene: ITGA2B

Variant: exon 13 sequence containing the 6 bp deletion 5’-GTGACA-3’ (deletion of Val425-Asp426) (heterozygous)

HGVS: NM_000419.5:c.1366_1371del (p.Val456_Asp457del)

ClinVar: 2898

CAID: CA8603160

gnomAD: 0.000008791 European (non-Finnish)

III-1, Female 10 years old

disease assertion: VWD

**positive family history **

Parental Genotype: each has heterozygous, 3251G>A

CasePresentingHPOs: CaseHPOFreeText: BS 4, VWF Ag:0.38, *VWF:GPIbM:0.15,VWF:CB:0.13, VWFpp :0.76, FVIII:C:0.10

CaseNOTHPOs:* N.a.

CaseNOTHPOFreeText: n.a.

CasePreviousTesting: *GenotypingMethod: Sanger seqencing;

*PreviouslyPublished:n.a.

Gene:VWF Variant:c.3251G>A*

*HGVS:NM_000552.5:c.3251G>A

• *ClinVarID:Variation ID: 626952

*CAID:n.a.

*gnomAD:n.a.

Case#: 3G

DiseaseAssertion:

FamilyInfo: Her father (3F), who reported mild bruising symptoms, and both asymptomatic younger male siblings (3H and 3J) were also investigated. All family members were found to be heterozygous for a previously described mutation in exon 30:c.5191 T → A, resulting in Ser1731Thr.

CaseHPOFreeText4: In family 3, the female proposita (3G) presented in childhood with epistaxis and easy bruising. VWF:Ag and VWF:RCo were borderline low in 3F and 3G but normal in the asymptomatic brothers, 3H and 3J. Multimer analysis was normal in all family members tested (Figure 1). VWF:CB binding to type I collagen was reduced in all family members (CB/Ag ratio 0.4-0.7), whereas only one (3G) showed a reduction in CB:Ag ratio with type III collagen.

CasePreviousTesting: Fifty-one primer sets were used to amplify the essential regions of VWF (52 exons, flanking intronic regions, 5′ promoter region, and the 3′ untranslated region).

Gene: VWF

Variant: heterozygous for a previously described mutation in exon 30:c.5191 T → A, resulting in Ser1731Thr

HGVS: NM_000552.5(VWF):c.5191T>A (p.Ser1731Thr)

ClinVar: 100420

CAID: CA228704

gnomAD: 0.004289 in middle eastern pop

Case#: 2D

DiseaseAssertion:

FamilyInfo: Her son 2E had mild bruising and epistaxis.

CaseHPOFreeText: The proposita (2D) in family 2 was investigated after a postpartum hemorrhage requiring blood transfusion. She gave a longstanding history of menorrhagia, and her calculated bleeding score was 10, the highest of all the persons in this study. VWF:Ag, VWF:RCo, and multimer analysis were all normal. VWF:CB was significantly reduced with both types I and III collagen, resulting in CB:Ag ratios of 0.2 to 0.4. Closure times in the PFA-100 were normal in 2D with both cassettes (Coll/Epi, 111 seconds; Coll/ADP, 98 seconds). The discrepant reduction in VWF:CB persisted after DDAVP administration to 2D despite normalization of both VWF:Ag and VWF:CB (samples taken 90 minutes after DDAVP administration showed VWF:Ag 191 IU/dL, VWF:CB 67 IU/dL, CB:Ag ratio = 0.35).

CasePreviousTesting: Fifty-one primer sets were used to amplify the essential regions of VWF (52 exons, flanking intronic regions, 5′ promoter region, and the 3′ untranslated region).

Gene: VWF

Variant: Her and son were shown to be heterozygous for a novel mutation in exon 31:c.5347 T → G resulting in Ser1783Ala

HGVS: NM_000552.5(VWF):c.5347T>G (p.Ser1783Ala)

ClinVar: 31012

CAID: CA228715

gnomAD: 0.0006918 in south asian pop

Case#: 1A

DiseaseAssertion:

FamilyInfo: Family 1. Her 2 offspring, 1B (son) and 1C (daughter), reported minor bleeding symptoms that had not required treatment. 1B was found to be heterozygous for Trp1745Cys only and 1C was heterozygous for Arg760His only, confirming that these mutations are in trans in 1A.

CaseHPOFreeText4: was an elderly woman with lifelong bleeding episodes, including epistaxis, ecchymosis, menorrhagia, and prolonged bleeding after dental extractions, requiring multiple medical interventions. The coexistence of both mutations appears to produce a more significant bleeding disorder as the bleeding score in 1A (8) was higher than that in either of her children (4 and 2). The position of Trp1745 in the A3 domain is shown in Figure 2. Both persons with this mutation (1A and 1B) had markedly reduced ratios of VWF collagen-binding activity to VWF antigen (CB:Ag) with type III collagen (0.1 and 0.3, respectively) and type I collagen (0.2 and 0.3, respectively). The ratio of VWF:RCo to VWF:Ag (RCo:Ag) was near unity in both persons, and VWF multimer analysis (Figure 1B) and RIPA were normal. Closure times in the PFA-100 were mildly prolonged with the ADP cartridge (1A = 173 seconds, 1B = 189 seconds, NR < 112 seconds). With the epinephrine cartridge, the closure time was prolonged in 1A only (1A = 271 seconds, 1B = 90 seconds, NR < 179 seconds).

CasePreviousTesting: Fifty-one primer sets were used to amplify the essential regions of VWF (52 exons, flanking intronic regions, 5′ promoter region, and the 3′ untranslated region).

Gene: VWF

Variant: 1A demonstrated compound heterozygosity for 2 mutations in the VWF gene. The first is a novel mutation in exon 30:c.5235 G → T resulting in Trp1745Cys and the second is c.2279 G → A resulting in Arg760His.

HGVS: NM_000552.5(VWF):c.5235G>T (p.Trp1745Cys), NM_000552.5(VWF):c.2279G>A (p.Arg760His)

ClinVar: 100421, 31011

CAID: CA228706, CA228316

gnomAD: 8.474e-7 in the european non finnish pop, 0.00002669 in the African American population

Case#: MK0251

DiseaseAssertion: Type 2 VWD

CaseHPOFreeText: 51 year old, female, VWF:AG LEVELS = 60

Gene: VWF

Variant: compound heterozygous for 4751A>G (p.Y1584C) and c.7483del (p.Leu2495CysfsTer8) variants

ClinVar: 310 (p.Y1584C) and 631687 (p.Leu2495CysfsTer8)

gnomAD: absent grpmax (p.Leu2495CysfsTer8), grpmax of 0.004019 based on european non finnish pop (p.Y1584C)

CaseII.1: 67 year old female, proband

DiseaseAssertion: progressive asymmetric distal limb weakness - Welander distal myopathy

FamilyInfo: 66-year-old sister has a mild distal myopathy, supraventricular tachycardia and hypertrophic cardiomyopathy. Both sisters carry the only known pathogenic TIA1 mutation and a heterozygous MYH7 variant (c.5459G > A; p.Arg1820Gln). Another sibling with isolated distal myopathy carries only the TIA1 mutation.

CasePresentingHPOs: HP:0002460, HP:0003805

"distal limb weakness, rimmed vacuoles"

CaseHPOFreeText: congophilic inclusions

CaseNotHPOFreeText: cardiac involvement

CasePreviousTesting: Electromyography - showed myopathic changes in distal upper and lower limb muscles with fibrillation potentials and rare myotonic discharges. Neurological examination - asymmetric weakness limited to distal muscles.

ANO5, BAG3, CAV3, CRYAB, DES, DNAJB6, DYSF, FHL1, FLNC, GNE, LDB3, MATR3, MYH7, MYOT, TCAP, TIA1, TTN and VCP were tested, all coding regions plus 20 bp flanking non-coding regions were analyzed through a combination of targeted next generation sequencing and Sanger sequencing

MultipleGeneVariants:

(1) MYH7Variant: NM_000257.4:c.5459G>A

ClinVar: 419278

CAID: CA046848

gnomAD:https://gnomad.broadinstitute.org/variant/14-23884304-C-T

Allele Frequency: East Asian 0.00005437

(2) TIA1Variant: NM_022173.4:c.1150G>A

ClinVar: 41480

CAID: CA1697383

gnomAD: https://gnomad.broadinstitute.org/variant/2-70439862-C-T?dataset=gnomad_r2_1

Total Allele Frequency: Other 0.0001629

GenotypingMethod: Targeted next generation sequencing and Sanger sequencing

Case#: HCM562, female, 36 yrs, smoker

DiseaseAssertion: HCM

FamilyInfo: None

CasePresentingHPOs: 36mm LAD, 0mm LVST, 36mm LVEDD, 66% LVEF, 21mm Maxi LVWT, 70mmHg Maxi LVOTG, 119176332 raw RNA-seq reads, 113518840 HQ reads (95.25%), 96.63% mapping rate, 782848 total variant, 34382 exonic variant

CaseHPOFreeText: N/A

CaseNotHPOs: N/A

CaseNotHPOFreeText: N/A

CasePreviousTesting: N/A

CaseGenotypingMethod: RNA-seq, read mapping, variant calling, identify functional variants, differential expression analysis, miRWalk miRNA target prediction, LncRNA annotation, Co-expression network analysis, PPI network analysis, gene over-representation analysis,

Variant: NM_000257.4(MYH7):c.427C>T (p.Arg143Trp)

ClinVar: 164401 https://www.ncbi.nlm.nih.gov/clinvar/variation/164401/

CAID: N/A

gnomAD: 0.0001087 East Asian https://gnomad.broadinstitute.org/variant/14-23901923-G-A

Case1: 7-year-old girl

DiseaseAssertion: HCM

FamilyInfo: clinical history was negative. Unable to obtain clinical information on first-degree relatives. Both parents carried MYH7 mutation.

CasePresentingHPOs: HP:0025490, HP:0100598

"Myocardial bridging, Pulmonary edema"

CaseHPOFreeText: In the last weeks, patient had frequently been visited by a pediatrician to treat seasonal allergies. Full-body CT-scan was performed to exclude abuse or trauma and did not show any anomaly. The heart (weight: 240 g, longitudinal diameter: 11 cm, transverse diameter: 10 cm, thickness of the anterior wall of the right ventricle: 0·3 cm, thickness of the left ventricle: 1·0 cm) did not reveal morphologic or structural anomalies of ventricular walls and valves. The coronary circulation was right-dominant and presented myocardial bridging (length: 1·1 cm, thickness: 0·5 cm) involving the middle segment of the left anterior descending coronary artery. No other significant elements were found upon macroscopic observation, with the exception of pulmonary edema. Histopathological examination showed patchy interstitial fibrosis, myocyte disarray and waving of the myofibers in the tissue surrounding the tunneled artery. Several areas of interstitial and subendocardic fibrosis in the other samples of the left ventricular myocardium were also observed.

CaseNotHPOs:

CaseNotHPOFreeText:

CasePreviousTesting: The low value of serum tryptase (6·12 µg/L) excluded a fatal anaphylaxis. Toxicology tests were negative. NGS analysis used a custom resequencing panel which included 147 genes associated with cardiomyopathies and channelopathies. Authors identified five rare genetic variants in heterozygous state (Table 4).

GenotypingMethod: Genomic DNA was extracted from postmortem whole blood of the proband. Whole blood samples of the proband mother, father and sister were obtained to extract genomic DNA and thus to perform Sanger sequencing to analyze the segregation of each genetic variant identified by NGS. Authors performed Sanger sequencing on the DNA extracted from the mother, the father and the sister of the proband. The father carried both variants identified in the DPP6 gene, whereas the mother carried the rare variants identified in MYH7, SCN2B and NOTCH1. The sister carried all the genetic variants.

Variant: NM_000257.4:c.2585C>T

ClinVar: 36636

CAID: CA012680

gnomAD: https://gnomad.broadinstitute.org/variant/14-23894072-G-A

0.00008014 African/African-American

Case10: Male, 50 years onset, 52 years diagnosis, Italy

HPOsFreeText: See Group Phenotypes, ERT started at 53, GAA Activity at ~13 pmol/min/mg before ERT and ~15 after

GroupGenotypingMethod: PCR Amplification and GAA Gene Sequencing

Variant1: NM_000152.4:c.-32-13T>G

ClinVar: 4027 https://www.ncbi.nlm.nih.gov/clinvar/variation/4027/

gnomAD: Ashkenazi Jewish 5.541e-3, Overall 3.401e-3 http://gnomad.broadinstitute.org/variant/17-78078341-T-G

Variant2: NM_000152.4:c.784G>A (p.Glu262Lys)

ClinVar: 188806 https://www.ncbi.nlm.nih.gov/clinvar/variation/188806/

gnomAD: East Asian 5.016e-5, Overall 1.417e-5 http://gnomad.broadinstitute.org/variant/17-78081447-G-A

CaseAddInfo: Late Onset Pompe Disease

Case: 19

DiseaseAssertion: hypertrophic cardiomyopathy, Left ventricular hypertrophy

FamilyInfo: Finnish, no family info

CasePresentingHPOs: HP:0001639, HP:0001712

"hypertrophic cardiomyopathy, Left ventricular hypertrophy"

CaseHPOFreeRext:

CaseNOTHPOs:

CaseNOTHPOFreeText:

CasePreviousTesting: 24 h Holter registration and bicycle ergometer test performed in majority of patients. Cardiac magnetic resonance imaging was performed to verify the diagnosis of HCM

GenotypingMethod: Targeted next-generation sequencing of exons, UTRs and 10 bp flanking regions of 59 cardiomyopathy associated genes, filtered variants confirmed by Sanger sequencing

Variant:

MYH7 Variant: NM_000257.4:c.3158G>A

ClinVarID: 155814

gnomAD: http://gnomad.broadinstitute.org/variant/14-23891476-C-T

Total Allele Frequency: 0.00006010 European Finnish: 0.0006368

PreviouslyPublished:

SupplementalData: Table S3: list of all 382 patients with HCM included in the FinHCM Genetic Study, and the rare variants (GMAF <1%) detected in each patient.

Case: 8

DiseaseAssertion: hypertrophic cardiomyopathy, Left ventricular hypertrophy

FamilyInfo: Finnish, no family info

CasePresentingHPOs: HP:0001639, HP:0001712

"hypertrophic cardiomyopathy, Left ventricular hypertrophy"

CaseHPOFreeRext:

CaseNOTHPOs:

CaseNOTHPOFreeText:

CasePreviousTesting: 24 h Holter registration and bicycle ergometer test performed in majority of patients. Cardiac magnetic resonance imaging was performed to verify the diagnosis of HCM

GenotypingMethod: Targeted next-generation sequencing of exons, UTRs and 10 bp flanking regions of 59 cardiomyopathy associated genes, filtered variants confirmed by Sanger sequencing

MultipleGeneVariants:

(1) LAMA4 Variant: c.2593C>G (unknown HGVS descriptor)

ClinVarID: unknown

gnomAD: unknown

(2) MYH7 Variant: NM_000257.4:c.3158G>A

ClinVarID: 155814

gnomAD: http://gnomad.broadinstitute.org/variant/14-23891476-C-T

Total Allele Frequency: 0.0000601 European (Finnish): 0.0006368

PreviouslyPublished:

SupplementalData: Table S3: list of all 382 patients with HCM included in the FinHCM Genetic Study, and the rare variants (GMAF <1%) detected in each patient.

Case: 2

DiseaseAssertion: hypertrophic cardiomyopathy, Left ventricular hypertrophy

FamilyInfo: Finnish, no family info

CasePresentingHPOs: HP:0001639, HP:0001712

"hypertrophic cardiomyopathy, Left ventricular hypertrophy"

CaseHPOFreeRext:

CaseNOTHPOs:

CaseNOTHPOFreeText:

CasePreviousTesting: 24 h Holter registration and bicycle ergometer test performed in majority of patients. Cardiac magnetic resonance imaging was performed to verify the diagnosis of HCM

GenotypingMethod: Targeted next-generation sequencing of exons, UTRs and 10 bp flanking regions of 59 cardiomyopathy associated genes, filtered variants confirmed by Sanger sequencing

MultipleGeneVariants:

(1) MYH7 Variant: NM_000257.4:c.3158G>A

ClinVarID: 155814

gnomAD: http://gnomad.broadinstitute.org/variant/14-23891476-C-T

Total Allele Frequency: 0.00006010 European Finnish: 0.0006368

(2) MYBPC3 Variant: NM_000256.3:c.440C>T

ClinVarID: 181035

gnomAD: http://gnomad.broadinstitute.org/variant/11-47371630-G-A

Total Allele Frequency: 0.0005896 European (Finnish): 0.005563

(3) MYOZ2 Variant: NM_016599.4:c.738A>G

ClinVarID: 30509

gnomAD: http://gnomad.broadinstitute.org/variant/4-120107298-A-G

Total Allele Frequency: 0.0023 European (Finnish): 0.02188

(4) PSEN2 Variant: NM_012486.2:c.211C>T

ClinVarID: 192130

gnomAD: http://gnomad.broadinstitute.org/variant/1-227071475-C-T

Total Allele Frequency: 0.003711 European (Finnish): 0.01791

(5) SCN5A Variant: NM_000335.4:c.1715C>A

ClinVarID: 67683

gnomAD: http://gnomad.broadinstitute.org/variant/3-38645378-G-T

Total Allele Frequency: 0.005175 European (Finnish): 0.03855

PreviouslyPublished:

SupplementalData: Table S3: list of all 382 patients with HCM included in the FinHCM Genetic Study, and the rare variants (GMAF <1%) detected in each patient.

Case: 8

DiseaseAssertion: hypertrophic cardiomyopathy, Left ventricular hypertrophy

FamilyInfo: Finnish, no family info

CasePresentingHPOs: HP:0001639, HP:0001712

"hypertrophic cardiomyopathy, Left ventricular hypertrophy"

CaseHPOFreeRext:

CaseNOTHPOs:

CaseNOTHPOFreeText:

CasePreviousTesting: 24 h Holter registration and bicycle ergometer test performed in majority of patients. Cardiac magnetic resonance imaging was performed to verify the diagnosis of HCM

GenotypingMethod: Targeted next-generation sequencing of exons, UTRs and 10 bp flanking regions of 59 cardiomyopathy associated genes, filtered variants confirmed by Sanger sequencing

MultipleGeneVariants:

(1) LAMA4 Variant: c.2593C>G (unknown HGVS descriptor)

ClinVarID: unknown

gnomAD: unknown

(2) MYH7 Variant: NM_000257.4:c.3158G>A

ClinVarID: 155814

gnomAD: http://gnomad.broadinstitute.org/variant/14-23891476-C-T

Total Allele Frequency: 0.0000601 European (Finnish): 0.0006368

PreviouslyPublished:

SupplementalData: Table S3: list of all 382 patients with HCM included in the FinHCM Genetic Study, and the rare variants (GMAF <1%) detected in each patient.

Case: 2

DiseaseAssertion: hypertrophic cardiomyopathy, Left ventricular hypertrophy

FamilyInfo: Finnish, no family info

CasePresentingHPOs: HP:0001639, HP:0001712

"hypertrophic cardiomyopathy, Left ventricular hypertrophy"

CaseHPOFreeRext:

CaseNOTHPOs:

CaseNOTHPOFreeText:

CasePreviousTesting: 24 h Holter registration and bicycle ergometer test performed in majority of patients. Cardiac magnetic resonance imaging was performed to verify the diagnosis of HCM

GenotypingMethod: Targeted next-generation sequencing of exons, UTRs and 10 bp flanking regions of 59 cardiomyopathy associated genes, filtered variants confirmed by Sanger sequencing

MultipleGeneVariants:

(1) MYH7 Variant: NM_000257.4:c.3158G>A

ClinVarID: 155814

gnomAD: http://gnomad.broadinstitute.org/variant/14-23891476-C-T

Total Allele Frequency: 0.00006010 European Finnish: 0.0006368

(2) MYBPC3 Variant: NM_000256.3:c.440C>T

ClinVarID: 181035

gnomAD: http://gnomad.broadinstitute.org/variant/11-47371630-G-A

Total Allele Frequency: 0.0005896 European (Finnish): 0.005563

(3) MYOZ2 Variant: NM_016599.4:c.738A>G

ClinVarID: 30509

gnomAD: http://gnomad.broadinstitute.org/variant/4-120107298-A-G

Total Allele Frequency: 0.0023 European (Finnish): 0.02188

(4) PSEN2 Variant: NM_012486.2:c.211C>T

ClinVarID: 192130

gnomAD: http://gnomad.broadinstitute.org/variant/1-227071475-C-T

Total Allele Frequency: 0.003711 European (Finnish): 0.01791

(5) SCN5A Variant: NM_000335.4:c.1715C>A

ClinVarID: 67683

gnomAD: http://gnomad.broadinstitute.org/variant/3-38645378-G-T

Total Allele Frequency: 0.005175 European (Finnish): 0.03855

PreviouslyPublished:

SupplementalData: Table S3: list of all 382 patients with HCM included in the FinHCM Genetic Study, and the rare variants (GMAF <1%) detected in each patient.

PatientID: 16

KindredID: 6006

Case: F, 24Y0M, Ethnicity Unknown

DiseaseAssertion: VHL

FamilyInfo: N/A

CasePresentingHPOs: HP:0030405 (pancreatic endocrine tumour)

CaseHPOFreeText: She presented with one pathologic pancreatic endocrine tumour.

CaseNotHPOs: N/A

CaseNotHPOFreeText: N/A

CasePreviousTesting: N/A

PreviouslyPublished: N/A

SupplementalData: N/A

Variant: p.Y185* (c.555C>G)

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

CaseProblemVariantFreeText: N/A

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

CAID: CA357075 http://reg.clinicalgenome.org/redmine/projects/registry/genboree_registry/allele?hgvsOrDescriptor=NM_000551.3%3Ac.555C%3EG

gnomAD: N/A

VariantEvidence: N/A

MutationType: stop_gained

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

MultipleGeneVariants: N/A

Case#: 32 year old female

DiseaseAssertion: VWD Type 2M

CaseHPOFreeText4: Testing was repeated multiple times, with initial results being untrustworthy. Follow up testing is as follows: VWF:RCo/VWF:Ag ratio = 0.61, VWF:CB = 10. Normal multimers

CasePreviousTesting: Molecular analysis by next generation gene sequencing analysis of VWF, F8, GPIBA and GPIBB.

Gene: VWF

Variant: variant in VWF at exon 31 (c.5347 T > G) that leads to a VWF protein change of p.Ser1783Ala

HGVS: NM_000552.5(VWF):c.5347T>G (p.Ser1783Ala)

ClinVar: 31012

CAID: CA228715

gnomAD: 0.0006918 in South Asian pop

PMID 16322474

Notes Recombinant full length p.Ile1628Thr expressed in HEK293 and purified from growth media was NOT digested by recombinant ADAMTS13 in non-denaturing conditions (Table 1, Fig 5a). However, the the A1-A3 p.Ile1628Thr truncations were digested by ADAMTS13 in non-denaturing conditions while WT recombinant VWF truncations were not (Fig 6).

Case28: Female, 6 yr onset, 7 yr diagnosis, 9.9 yr report

HPOsFreeText: Wheelchair partially used beginning at 9 yr, Respiratory support used at night beginning at 8 yr, FVC z-score sitting -6.36 (30% pred), FVC z-score supine -6.88 (25% pred), 82% MRC Scale, 979 CK U/l, 0.3 GAA activity (nmol/h/mg)^h from cultured fibroblasts

Patient1: Male, Birth onset

CasePresentingHPOs: HP:0001644, HP:0012420, HP:0002098, HP:0001640, HP:0002375, HP:0002789, HP:0001649, HP:0030148, HP:0001252, HP:0001712, HP:0025168, HP:0009128, HP:0002878 (Dilated cardiomyopathy, Meconium stained amniotic fluid, Respiratory difficulty, Cardiomegaly, Hypokinesia, Tachypnea, Tachycardia, Heart murmur, Muscle hypotonia, Left ventricular hypertrophy, Left ventricular diastolic dysfunction, Absent/underdeveloped muscles of extremities, Respiratory failure)

HPOsFreeText: Apgar scores were 2155, birth weight was 3,764 g, length was 50.5 cm, Mild diffuse pulmonary markings, Had a grade II ejection systolic murmur at the left upper sternal border with intermittent gallop, A cardiac catheterization with myocardial biopsy was performed on day of life 30 which revealed marked vacuolization of cardiac myocytes, Analysis of plasma GAA activity revealed a significant deficiency (4 pmoles/hr/spot, reference range 24–94), Moderately elevated urine glucose tetrasaccharide (Hex4) of 29.6 mmol/mol creatinine (reference range: <19 for age <6 months)

NotHPOsFreeText: The ECG showed sinus rhythm with a ventricular right axis deviation without pathognomonic changes of Pompe disease, Extremities were freely moveable, Newborn screening was normal

CaseAddInfo: Non‐consanguineous product of a term pregnancy born to a G3P2 mother, Pregnancy was complicated by limited prenatal care, cannabinoid, and amphetamine use, Family history was negative for neuromuscular diseases but significant for multiple sclerosis and seizure disorders

PMID: 26527198

Gene: ABCA4

HGNC ID: 34

Case#: 12, Somali, onset 5 years old

DiseaseAssertion: STGD

FamilyInfo: Parents were heterozygous for Arg212Cys, no family history of IRD

CasePresentingHPOs: HP:0011504, ORPHA:827, HP:0000608

CaseHPOFreeText: Bull's eye maculopathy, macular degredation, red-green color defecit, reduced cone function

CaseNotHPOs: n/a

CaseNotHPOFreeText: n/a

Genotyping Method: Whole exome sequencing

PreviouslyPublished: N/a

Variant:

ClinVar: NM_000350.3:c.5882G>A p.(Gly1961Glu) ; NM_000350.3:c.634C>T p.(Arg212Cys)

ClinVar: 7888; 7898

CAID: CA119132, CA203216

SupplementalData: N/a

Case#: Somali origin, 11 years of age, onset age 8

DiseaseAssertion: STGD

FamilyInfo: Parents were heterozygous for Arg212Cys, asymptoamtic 32 year old father was found to be homozygous for Gly1961Glu

CasePresentingHPOs: HP:0011504, ORPHA:827, HP:0000608

CaseHPOFreeText: BCVA 20/70 and 20/80, atrophic zone of outer retinal and RPE atrophy, Bull's Eye Maculopathy, Macular atrophy

CaseNotHPOs: N/a

CaseNotHPOFreeText: N/a

Genotyping Method: Whole genome sequencing

PreviouslyPublished: N/a

Variant: NM_000350.3:c.5882G>A p.(Gly1961Glu) ; NM_000350.3:c.634C>T p.(Arg212Cys)

ClinVar: 7888; 7898

CAID: CA119132, CA203216

SupplementalData: N/a

Case#: 55 year old female

DiseaseAssertion: Stargardt

FamilyInfo: no significant ocular disease shown.

CasePresentingHPOs: difficulty reading materials 6 inches from her eyes, decreased visual acuity from age seven. BCVA was 20/150 OU. Posterior segment exam and autofluorescence was significant for bilateral central atrophy and pisciform fleck atrophy involving the peripapillary, macular, and peripheral regions.

CaseHPOFreeText: NR

CaseNotHPOs: NR

CaseNotHPOFreeText: NR

Genotyping Method: ABCR400 microarray

PreviouslyPublished: NR

Variant: NM_000350.3:c.4139C>T, NM_000350.3(ABCA4):c.6089G>A

ClinVar: 7904, 99428

CAID: CA129033

SupplementalData: NR

Case#: a 46-year-old male

DiseaseAssertion: Stargardt Disease

FamilyInfo:visual acuity loss by his brother and father

CasePresentingHPOs: HP:0007663

CaseHPOFreeText: visual acuity measured 20/400 bilaterally

CaseNotHPOs: NR

CaseNotHPOFreeText:NR

Genotyping Method: ABCA4 microarray (ABCR5000 chip)

PreviouslyPublished: NR

Variant: c.5714+5G>A

ClinVar: 99403

CAID: CA227338

SupplementalData:NR

Case#: Family MD-0084 Proband, 7yo at onset

DiseaseAssertion: AR Stargardt

FamilyInfo:

CasePresentingHPOs:

CaseHPOFreeText: STGD diagnosed based on "bilateral central vision loss; fundus presenting with a beaten-bronze appearance and/or the presence of orange-yellow flecks in the retina from the posterior pole to the mid-periphery; fluorescein angiography showing typical dark choroid; and normal to subnormal electroretinogram (ERGs)." VA loss, VF loss, BCVA=<0.05/<0.05

CaseNotHPOs:

CaseNotHPOFreeText:

PreviouslyPublished: PMID: 16917483 (variant not found in this paper); PMID: 19959634

Variant: c.6410G>A (p.Cys2137Tyr) homozygous, found by ABCR400 + dHPLC + HRM + MLPA

ClinVar: 2202779

CAID: CA341277358

SupplementalData:

PMID: 25375877

Gene: ABCA4, RPGR

HGNC ID: 34, 10295

PMID: 22060670

Gene: ABCA4

HGNC ID: 34

Case#: two affected sisters

DiseaseAssertion: Stargardt Disease

FamilyInfo: compound heterozygotes for the mutations. Unaffected family members did not carry either or had one of the two mutations.

CasePresentingHPOs: NR

CaseHPOFreeText: NR

CaseNotHPOs: NR

CaseNotHPOFreeText: NR

Genotyping Method: ABCA4 408 microsatellite

PreviouslyPublished: NR

Variant: NM_000350.3(ABCA4):c.5018+2T>C , NM_000350.3(ABCA4):c.655A>T

ClinVar: 265008, 632118

CAID: CA10588304, CA645372240

SupplementalData: NR

This variant is found in table S2A to have an OR of infinity with a CI (25.35-infinity), so PS4 is applicable