Case#: 22-year-old man

DiseaseAssertion: NCCM and mildly reduced LV function

FamilyInfo: Mother and brothers have also been diagnosed with NCCM

ParentalTesting: Mother had cardio magnetic resonance imaging done

CasePresentingHPOs: not reported

CaseHPOFreeText:

CaseNotHPOs: HP:0001681, HP:0001279 Angina pectoris andSyncope

CaseNotHPOFreeText: no arrhythmic events

CasePreviousTesting: electrocardiogram, transthoracic echocardiography, cardiac-magnetic resonance imaging

Genotyping Method: next generation sequencing

FunctionalAnalysis: not reported

Variant: NM_001018005.1:c.20_22del; p.Lys7del

ClinVar:181682

CAID:CA018708

gnomAD:15-63042841-CAAG-C

AdditionalInfo:

Case#: male; age 44

DiseaseAssertion: Hypertrophic, restrictive, and dilated cardiomyopathy

FamilyInfo: The patient’s unaffected 6-year-old daughter was tested for the variant. His brother and parents were healthy

ParentalTesting: Parents refused genetic testing

CasePresentingHPOs: HP:0001639 HP:0001723 HP:0001644

CaseHPOFreeText: Hypertrophic cardiomyopathy; restrictive left-ventricular filling pattern; diastolic dysfunction

CaseNotHPOs: NR

CaseNotHPOFreeText: NR

CasePreviousTesting: NR

Genotyping Method: 25-gene Ion AmpliSeq panel with Ion PGM semiconductor sequencing; Sanger sequencing

FunctionalAnalysis: NR

Variant: TPM1 c.292G>A (p.Glu98Lys; p.E98K; rs730881131); missense variant; heterozygous

ClinVar: 1472862

CAID: CA392720843

gnomAD: NR

Case#: 29 year old female; diagnosed with DCM 8 years before death; died suddenly at home on 22 February 2017; forensic autopsy case

DiseaseAssertion: Dilated cardiomyopathy (DCM) with sudden cardiac death; MYBPC3-related

FamilyInfo: Family from Central-South China with a history of SCD. Father (I:1) reportedly died of a heart attack at 37, not genetically tested. Mother (I:2) and sister (II:2) are unaffected and do not carry the variant. No other affected relatives reported.

ParentalTesting: Not performed, both parents were not genotyped for the variant; the father was deceased before the study. Maternal non-carriage confirmed by Sanger, so the variant is either paternal or de novo; phase not established.

CasePresentingHPOs: HP:0001644; HP:0001645

CaseHPOFreeText: Diagnosed with DCM 8 years before death, with no other abnormalities. Found unresponsive at home at noon and expired; no prodromal symptoms documented. Autopsy: enlarged heart with valve thickening, myocardial fibrosis, and multi-organ congestion; no other fatal injury or disease. HE staining of left ventricle showed no hypertrophy or other anomalies. Toxicological screening for ethanol, benzodiazepines, barbiturates, amphetamine, methamphetamine, ketamine, morphine, MDMA, and THC-acid was negative; serum IgE normal (27 IU/ml); eosinophils 4.7% with no mast-cell degranulation.

CaseNotHPOs: HP:0001639; HP:0001634; HP:0003124

CaseNotHPOFreeText: No cardiomyocyte hypertrophy on histology, distinguishing this from HCM. No coronary, valvular, or congenital structural disease identified. No toxicological cause of death.

CasePreviousTesting: Whole-exome sequencing on postmortem blood, retaining synonymous variants; filtering against gnomAD and CMDB; in silico prediction with MutationTaster, PolyPhen-2, and SIFT; NetGene2-2.42 used to predict splice effects. Five variants in cardiomyopathy-related genes were found; only the MYBPC3 variant was classified likely pathogenic.

Genotyping Method: WES with Sanger confirmation on ABI 3100; family members genotyped by Sanger.

FunctionalAnalysis: Minigene assay pcDNA3.1-MYBPC3 construct spanning exon 1-intron 1-exon 2, transfected into HEK293 cells: wild-type and the c.10G>A negative control both produced a ~1,500 bp full-length band and a ~300 bp spliceosome band; the c.24A>C construct produced no spliceosome band, indicating abnormal splicing with intron 1 retention and frameshift. Immunohistochemistry showed decreased MYBPC3 protein in the proband's left ventricle versus a control heart.

Variant: MYBPC3 NM_000256.3:c.24A>C (p.Pro8Pro); heterozygous synonymous variant at the penultimate base of exon 1; disrupts the splice donor site; novel

ClinVar: 164098

CAID: CA11138

gnomAD: 0.00001541

AdditionalInfo: Classified likely pathogenic with PS3, PM2, PP1, PP3; note the paper's Discussion relabels the prediction code as PP4, an internal inconsistency. The authors describe it as the first reported deleterious synonymous MYBPC3 variant, acting by intron retention and reduced protein. The same splice site is affected by the known HCM variants c.24+1G>A and c.24+3A>C. Four other variants were found and classified likely benign or VUS: LDB3 c.2131T>C (VUS), BBS2 c.422A>G, EVC2 c.2643G>C, and BBS4 c.1548_1549del.

Case#: female; age 65

DiseaseAssertion: chemotherapy-induced cardiomyopathy with severe left ventricular systolic dysfunction

FamilyInfo: NR

ParentalTesting: NR

CasePresentingHPOs: HP:0001644 HP:0001635 HP:0001640

CaseHPOFreeText: Dilated left ventricle; left ventricular dysfunction; cardiomegaly; bilateral pleural effusion; dyspnea

CaseNotHPOs: NR

CaseNotHPOFreeText: NR

CasePreviousTesting: Previous anthracycline-associated cardiomyopathy 7 years earlier

Genotyping Method: TruSight One Expanded 6700-gene sequencing panel on an Illumina NextSeq 550

FunctionalAnalysis: NR

Variant: c.452G>A (p.Arg151Gln; rs730881101); missense variant; heterozygous

ClinVar: 181617

CAID: NR

gnomAD: NR

Case1: Caucasian. Sex and exact age not specified for the proband

DiseaseAssertion: Dilated cardiomyopathy (DCM)

FamilyInfo: Family A was notable for severe HF and sudden death in young children. Individuals I-2, III-10 and IV-7 died suddenly at ages 3 years, 13 months, and 5 months respectively, without prior evidence of heart disease. Two siblings, IV-5 and IV-8, presented with DCM and advanced HF at 5 months of age (LVEF 27% and 29%). Individuals III-3 and IV-1 developed end-stage HF refractory to medical therapy as teenagers. III-3 presented at age 13 with refractory HF, underwent mitral and tricuspid valve replacement for functional regurgitation and died weeks later of multisystem organ failure. IV-1 was asymptomatic when diagnosed with DCM at 7 years during family screening, but developed severe HF at age 17 and underwent transplant at 18 years of age. Individuals III-4, III-7 and III-12 developed mild exercise intolerance in their 5th decade.

ParentalTesting: NR

CasePresentingHPOs: HP:0001635, HP:0001644

CaseHPOFreeText: No family members with TPM1 D230N had evidence of cardiac conduction disease, and with one exception, none had evidence of skeletal myopathy

CaseNotHPOs: NR

CaseNotHPOFreeText: No family members with TPM1 D230N had evidence of cardiac conduction disease, and with one exception, none had evidence of skeletal myopathy.

CasePreviousTesting: No prior testing.

Genotyping Method: Genomic DNA was isolated from blood using standard methods in probands with DCM. Direct DNA sequence analysis of all coding regions and intron/exon boundaries was performed in 6 sarcomere genes.

FunctionalAnalysis: Functional analysis performed on recombinant proteins but not patient tissue. In vitro studies demonstrated major inhibitory effects on sarcomere function with reduced Ca2+-sensitivity, maximum activation, and Ca2+ affinity compared to wildtype TPM1.

Variant: NM_000366.5:c.688G>A (p.Asp230Asn)

ClinVar: NR

CAID: UnregisteredVariant

gnomAD: NR

AdditionalInfo: The paper does not explicitly identify which pedigree member is the Family A proband

Case2: Caucasian. Sex and exact age not specified for the proband

DiseaseAssertion: Dilated cardiomyopathy (DCM)

FamilyInfo: Young members of this family also presented with severe HF. Individual III-1 developed failure to thrive at 10 weeks of age and echocardiography revealed severe LV dilation (LVIDD Z-score 15.6) and systolic dysfunction (LVEF 15%). Transplantation was considered, but she had substantial recovery with basic medical therapy. By age 8 years, she was asymptomatic with only mild LV systolic dysfunction (LVIDD Z-score 1.3, LVEF 46%). At age 20, II-6 underwent cardiac transplantation shortly after presenting with refractory HF symptoms and severe LV systolic dysfunction (LVEF < 20%). Individual II-7 died of HF at 12 years, prior to genetic testing. In addition to DCM, he had congenital cataracts and skeletal muscle weakness. I-1 had unexplained syncope at age 56 years. Cardiac studies revealed ventricular tachycardia and DCM (LVEF 25%) which improved with medical management. Three other adults (II-2, II-3, II-4, ages 38, 36 and 33 years, respectively) had asymptomatic LV systolic dysfunction.

ParentalTesting: NR

CasePresentingHPOs: HP:0001635, HP:0001644

CaseHPOFreeText: NR

CaseNotHPOs: NR

CaseNotHPOFreeText: NR

CasePreviousTesting: No prior testing.

Genotyping Method: Genomic DNA was isolated from blood using standard methods in probands with DCM. Direct DNA sequence analysis of all coding regions and intron/exon boundaries was performed in 6 sarcomere genes.

FunctionalAnalysis: Functional analysis performed on recombinant proteins but not patient tissue. In vitro studies demonstrated major inhibitory effects on sarcomere function with reduced Ca2+-sensitivity, maximum activation, and Ca2+ affinity compared to wildtype TPM1.

Variant: NM_000366.5:c.688G>A (p.Asp230Asn)

ClinVar: NR

CAID: UnregisteredVariant

gnomAD: NR

AdditionalInfo: The paper does not explicitly identify which pedigree member is the Family B proband

Case1: 63-year-old male

DiseaseAssertion: Atrial flutter(54), VT & ICD(57); CVA(58) & DCM(58); HF(59)

FamilyInfo: There are 14 affected family members (Fig 1): six living (three sisters III-1, III-4, III-6 and three nieces IV-5, IV-15, IV-16) and eight deceased individuals (I-3, II-8, II-13, III-8, III-10, IV-1, IV-4, IV-6).

ParentalTesting:

CasePresentingHPOs:

CaseHPOFreeText: atrial flutter during evaluation for non-cardiac surgery. Subsequent evaluations led to the diagnosis of SSS. At 54 years old, echocardiogram showed mild systolic dysfunction and left ventricle dilation (EF 45%, LVEDD 58 mm). Myocardial perfusion study showed exercise intolerance but no signs of ischemia. At presentation, he felt well noting mild exercise intolerance for the past year with no palpitations or chest pain; he took no medications. At age 49, he had normal systolic function and LV dilation by echocardiogram (EF 53%, LVEDD 61 mm) and a normal myocardial perfusion study.

CaseNotHPOs: NR

CaseNotHPOFreeText: NR

CasePreviousTesting: NR

Genotyping Method: Sanger sequencing

FunctionalAnalysis: NR

Variant: NM_170707.4(LMNA):c.357-2A>G

ClinVar: 652066

CAID: CA342814936

gnomAD: NR

AdditionalInfo:

Case#: 43-year-old man family b

DiseaseAssertion: Familial dilated cardiomyopathy (FDCM)

FamilyInfo: Sister II:1 had previously been diagnosed with DCM and carried both reported variants. Brother II:2 died of sudden cardiac death.

ParentalTesting: father was negative for either varient

CasePresentingHPOs: HP:0031352, HP:0002094, HP:0002321 HP:0001644 “Dyspnea; chest tightness; dizziness; dilated cardiomyopathy

CaseHPOFreeText: Severe DCM symptoms

CaseNotHPOs: not reported

CaseNotHPOFreeText: Father I:1 and daughter III:1 were reported to have no clinical symptoms

CasePreviousTesting: Previously diagnosed with FDCM

Genotyping Method: NGS. PCR, Sanger

FunctionalAnalysis: Putative pathogenic mutations were analysed computationally using the PolyPhen-2, SIFT, and MutationTaster

Variant: MYH7 NM_000257:c.3134G>A; p.Arg1045His; heterozygous missense

ClinVar: 651054

CAID: N/A

gnomAD: 14-23422291-C-T

AdditionalInfo: two variants may have a “dose effect” and associate their combination with the proband’s severe phenotype

Case#:44-year-old woman , family 1 proband

DiseaseAssertion: Familial hypertrophic cardiomyopathy

FamilyInfo: Father, sister, and brother, died of sudden cardiac death around age 40

ParentalTesting: not reported

CasePresentingHPOs: HP:0031352, HP:0002094, HP:0005144 “Dyspnea; chest tightness, increased interventricular septum thickness”

CaseHPOFreeText: Symptoms after exercise, interventricular septum thickness 14.8 mm; left atrium 32 mm.

CaseNotHPOs: not reported

CaseNotHPOFreeText:not reported

CasePreviousTesting: no previous testing was reported

Genotyping Method: NGS. PCR, Sanger

FunctionalAnalysis: Putative pathogenic mutations were analysed computationally using the PolyPhen-2, SIFT, and MutationTaster

Variant: TNNI3 NM_000363.4:c.433C>G; p.Arg145Gly heterozygous exon 7 missense

ClinVar:12419

CAID: N/A

gnomAD: 19-55154146-G-C

AdditionalInfo: reported variant and a potential HCM hotspot

Case 1: One proband; (53-year-old woman)

DiseaseAssertion: Hypertrophic cardiomyopathy (HCM) with fasciculoventricular bypass tract (FVBT)

FamilyInfo: Autosomal dominant family with HCM and pre-excitation. Mother has an ICD for nonischemic cardiomyopathy and progressed from HCM to DCM; sister (49) carries the variant with HCM and NSVT; younger daughter (25) carries the variant with pre-excitation but no HCM. Maternal aunt died suddenly at 42; maternal uncle had a sudden cardiac arrest at 40 with subsequent ICD.

CasePresentingHPOs: HP:0001962; HP:0001639; HP:0004756; HP:0001677; HP:0001671; HP:0004308

CaseHPOFreeText: Palpitations prompted workup. ECG showed pre-excitation; Holter documented NSVT; echo showed HCM without significant outflow obstruction; EP study demonstrated an FVBT with inducible AF; CMR showed midventricular septal thickness of 22 mm with patchy enhancement involving 25% of myocardial mass.

CaseNotHPOs: HP:0001635; HP:0001279; HP:0001645

CaseNotHPOFreeText: No heart failure reported; no syncope; alive at publication.

CasePreviousTesting: ECG (pre-excitation), Holter (NSVT), echo (HCM without significant outflow obstruction), EP study (FVBT with inducible AF), CMR (midventricular septum 22 mm, patchy enhancement 25% of mass).

GenotypingMethod: HCMNext panel (Ambry Genetics); heterozygous c.655G>C (p.Val219Leu) in MYBPC3, reported pathogenic.

FunctionalAnalysis: None performed.

Variant: MYBPC3 NM_000256.3:c.655G>C (p.Val219Leu); heterozygous missense

ClinVar: 42784

CAID: CA015670

gnomAD: 0.00002

AdditionalInfo:N/A

Case1: Pedigree mother; affected carrier; HCM progressing to DCM; clinical case

DiseaseAssertion: Hypertrophic cardiomyopathy progressing to dilated cardiomyopathy, with pre-excitation; MYBPC3-related

FamilyInfo: Carries the familial MYBPC3 variant. ICD in place for nonischemic cardiomyopathy despite optimal medical therapy; never received a shock. Mother of the proband and the sister.

ParentalTesting: Not applicable, this is the transmitting generation.

CasePresentingHPOs: HP:0001639; HP:0001644; HP:0004308

CaseHPOFreeText: Serial echocardiography over 11 years documented progression from hypertrophic to dilated phenotype. ECG showed pre-excitation. ICD implanted; no appropriate or inappropriate therapy to date.

CaseNotHPOs: HP:0001279 ; HP:0001645

CaseNotHPOFreeText: No syncope reported; alive at publication.

CasePreviousTesting: Serial echocardiography; ECG; ICD interrogation.

Genotyping Method: Family screening; carrier status reported.

FunctionalAnalysis: None performed.

Variant: MYBPC3 NM_000256.3:c.655G>C (p.Val219Leu); heterozygous missense

ClinVar: 42784

CAID: CA015670

gnomAD: 0.00002

AdditionalInfo:N/A

Case#: Proband and family; distal arthrogryposis phenotype; clinical case

DiseaseAssertion: Distal arthrogryposis (DA); MYBPC1-related

FamilyInfo: Two MYBPC1 variants segregate in the family. The paper provides the pedigree and segregation data.

ParentalTesting: Not stated

CasePresentingHPOs: HP:0002804

CaseHPOFreeText: Distal arthrogryposis phenotype in affected family members, per the paper's clinical description.

CaseNotHPOs: Not stated

CasePreviousTesting: Clinical evaluation, genetic testing for MYBPC1.

Genotyping Method: Genetic testing confirmed MYBPC1 variants; specific method in the paper.

FunctionalAnalysis: Confirm whether a functional assay was performed.

Variant: MYBPC1, two variants, specific nucleotide changes need confirmation from the paper

ClinVar:N/A

CAID:N/A

gnomAD:N/A

AdditionalInfo: N/A

Case2: Sex not stated, age 8 months at diagnosis, ethnicity not stated

DiseaseAssertion: Restrictive cardiomyopathy (RCM)

FamilyInfo: Patients 2 and 3, monozygotic twins, were diagnosed with RCM at the age of 8 and 13 months, respectively. No familial history of cardiac disorders was reported in the R170W case, parental testing confirmed that the R170W variant occurred de novo (however, no parental proof was performed)

ParentalTesting: Parental testing confirmed that the R170W variant occurred de novo (however, no parental proof was performed). Parents do not carry the variant

CasePresentingHPOs: HP:0001723, HP:0002875, HP:0005110

CaseHPOFreeText: Echocardiographic examination revealed a restrictive filling pattern and enlarged atria. Both children died at the age of 12 and 18 months, respectively, due to congestive heart failure and atrial fibrillation

CaseNotHPOs: NR

CaseNotHPOFreeText: NR

CasePreviousTesting: NR

Genotyping Method: Total genomic DNA was extracted from peripheral blood using FlexiGene DNA Kit (Qiagen) according to the manufacturer’s protocol. The TNNI3 gene was screened for mutations using dye terminator sequencing. In both cases additional target sequencing including a panel of 108 cardiomyopathy- and channelopathy-associated genes was performed using Illumina MiSeq as previously described

FunctionalAnalysis: Functional analysis performed on recombinant proteins. Both cTnI-R170G/W strongly enhanced the Ca2+-sensitivity of skinned fibres, as is typical for RCM-mutations. Both mutants strongly enhanced the affinity of troponin (cTn) to tropomyosin compared to wildtype cTn. The stability of reconstituted thin filaments was reduced as revealed by electron microscopy.

Variant: NM_000363.5:c.508C>T (p.Arg170Trp)

ClinVar: 179285

CAID: CA021778

gnomAD: chr19-55154071-G-A

AdditionalInfo: Monozygotic twin

Case1: Male, age 3 at diagnosis, ethnicity not stated

DiseaseAssertion: Restrictive cardiomyopathy (RCM)

FamilyInfo: Familial screening in the R170G proband documented a hypertrophic cardiomyopathy with restrictive filling pattern in patient’s father, but the family refused additional genetic testing. No other family history of cardiac disorders stated.

ParentalTesting: The family refused additional genetic testing

CasePresentingHPOs: HP:0001723, HP:0001650, HP:0001654

CaseHPOFreeText: Patient 1 showed an impaired left ventricular function and pulmonary hypertension at three years of age. Heart biopsy showed mild cardiomyocyte hypertrophy and interstitial fibrosis. He was diagnosed with RCM and died one year later of ventricular fibrillation.

CaseNotHPOs: NR

CaseNotHPOFreeText: Target sequencing including a panel of 108 cardiomyopathy- and channelopathy-associated genes revealed no additional pathogenic or likely pathogenic variants

CasePreviousTesting: Additional target sequencing including a panel of 108 cardiomyopathy- and channelopathy-associated genes was performed using Illumina MiSeq as previously described.

Genotyping Method: Total genomic DNA was extracted from peripheral blood using FlexiGene DNA Kit (Qiagen) according to the manufacturer’s protocol. The TNNI3 gene was screened for mutations using dye terminator sequencing. In both cases additional target sequencing including a panel of 108 cardiomyopathy- and channelopathy-associated genes was performed using Illumina MiSeq as previously described

FunctionalAnalysis: Functional analysis performed on recombinant proteins. Both cTnI-R170G/W strongly enhanced the Ca2+-sensitivity of skinned fibres, as is typical for RCM-mutations. Both mutants strongly enhanced the affinity of troponin (cTn) to tropomyosin compared to wildtype cTn. The stability of reconstituted thin filaments was reduced as revealed by electron microscopy.

Variant: NM_000363.5:c.508C>G (p.Arg170Gly)

ClinVar: 165517

CAID: CA021769

gnomAD: NR

AdditionalInfo: NR

Case3: 7 week old female

DiseaseAssertion:

FamilyInfo: A cousin of the father presented with cardiac arrest due to HCM. He had a heterozygous pathogenic c.2373dup p.(Trp792fs) mutation in MYBPC3. We subsequently confirmed homozygosity for the c.2373dup variant in DNA isolated from stored fibroblasts from the deceased proband. Both parents of the proband were heterozygous for this truncating variant (Figure 2b). Cardiac re-evaluation showed only slightly reduced ECG voltages in the father (age 38) and borderline hypertrophy (septal thickness 11 mm) in the mother (age 41).

ParentalTesting:

CasePresentingHPOs: HP:0001640, HP:0002119

CaseHPOFreeText: Cardiomegaly was seen on chest X-ray, and echocardiography showed HCM with dilated ventricles. M-Mode measurements were unavailable. Noncompaction was not a clear feature, but could not be excluded. There was a small foramen ovale and mitral valve insufficiency

CaseNotHPOs: NR

CaseNotHPOFreeText: NR

CasePreviousTesting: NR

Genotyping Method: NR

FunctionalAnalysis: DNA isolated from stored fibroblasts from the deceased proband.

Variant: NC_000011.10:g.47337730dup

ClinVar: 42619

CAID: CA012139

gnomAD: 11-47359280-A-AC

AdditionalInfo: NR

Case#: II.3, 30-year-old man from an Indian family.

DiseaseAssertion: Familial hypertrophic cardiomyopathy.

FamilyInfo: Figure 1A. His father (I.1), brother (II.1), and sister (II.2) had HCM and carried TPM1 p.Gln68Arg. His mother (I.2) had a normal echocardiogram and tested negative for the variant. The authors report autosomal dominant segregation.

ParentalTesting: Father positive for TPM1 p.Gln68Arg. Mother negative.

CasePresentingHPOs: HP:0001639, HP:0001712, HP:0005144, HP:0031656, HP:0001653, HP:0010872, HP:4000004

CaseHPOFreeText: Echo showed asymmetric septal hypertrophy and trivial mitral regurgitation. The interventricular septum measured 2.1 cm and the left ventricular posterior wall measured 1.2 cm. ECG showed a left ventricular hypertrophy strain pattern with T-wave inversions in leads II, III, aVF, and V2–V6. Cardiac MRI showed patchy STIR hyperintensity and late gadolinium enhancement in the thickened septum.

CaseNotHPOs: HP:0000822 (no history of hypertension), HP:0032092 (no left ventricular outflow tract obstruction at rest)

CaseNotHPOFreeText: Asymptomatic at presentation. Both atria and the right ventricle were normal in size and function. Left ventricular ejection fraction was 63%.

CasePreviousTesting: NR (prior genetic testing details not available in the article).

Genotyping Method: Custom targeted amplicon sequencing panel of nine sarcomeric genes on the Ion Torrent PGM. Sanger sequencing confirmed the TPM1 variant.

FunctionalAnalysis: NR (functional assay results not available in the article).

Variant: NM_001018005.2(TPM1).203A>G (p.Gln68Arg), heterozygous.

ClinVar: 1490086

CAID: NR (CAID not listed in the article).

gnomAD: No variant record found in gnomAD v4.1 for 15-63044115-A-G.

AdditionalInfo: PMID 38223010. Figure 1A identifies the proband and shows the pedigree. Figures 1B–D show cardiac imaging. Figure 1E shows Sanger sequencing. Figure 1F shows conservation of the affected residue. The authors also report computational predictions of a damaging effect.

Case#: Single case; 37-year-old male construction worker; sudden unexplained death (SUD); found dead in bed ~1 hr after lunch, August 2005; molecular autopsy via WES in 2019 from autopsy tissue

DiseaseAssertion: Sudden unexplained death (SUD); no structural cardiac cause at autopsy; MYBPC3/TTN/ACADVL/RYR2-related per WES reevaluation

FamilyInfo: No family history or segregation data reported. The paper is a single molecular autopsy; no relatives were genotyped or phenotyped. No pedigree assessment performed.

ParentalTesting: Not performed, no family members tested.

CasePresentingHPOs: HP:0001698 (Sudden cardiac death); HP:0004756 (Ventricular tachycardia), the latter only as a speculative arrhythmic mechanism, not a documented clinical event

CaseHPOFreeText: Found dead in bed ~1 hr after lunch in August 2005. External examination showed no traumatic injury; toxicological blood screen negative. Clinical history unremarkable. Heart weighed 350 g (reference 331±57 g in men) with regular morphology and volume. Microscopy showed small foci of interstitial fibrosis, deemed not significant. No other organ showed significant abnormality. Case dismissed as SUD, reevaluated in 2019.

CaseNotHPOs: HP:0001639; HP:0001644; HP:0004308; HP:0001279

CaseNotHPOFreeText: No structural cardiac disease at autopsy, no hypertrophy, no dilation, normal heart morphology and volume. No evidence of any cardiomyopathy phenotype. No documented arrhythmia or syncope in life.

CasePreviousTesting: Autopsy (2005) with toxicology, histology. WES (2019) on DNA from FFPE left ventricular tissue; 562-gene cardiac panel (234 SCD/heart-disease genes + 328 heart-expressed genes); filtering against gnomAD (rare = MAF <0.1%); in silico prediction with DANN, MutationTaster, FATHMM, MutationAssessor, PolyPhen-2, SIFT, PROVEAN; ACMG 2015 classification; WebGestalt enrichment analysis.

Genotyping Method: WES on Illumina 500 (TruSeq library prep, xGen exome capture), SeqMule pipeline, ENLIS Genome Research annotation; variants confirmed by Sanger sequencing. Aligned to GRCh37.

FunctionalAnalysis: None performed by the authors, no functional assays in this paper. (ACADVL p.V383A residual activity ~20% is cited from a 2007 external study, not performed here.)

Variant: Four heterozygous missense variants, one per gene

ClinVar:926037

CAID: CA221696968

gnomAD: N/A

AdditionalInfo:N/A

Case1: Male, 24 years old at diagnosis, ethnicity not stated

DiseaseAssertion: Dilated cardiomyopathy (DCM)

FamilyInfo: Three affected family members carry the D94A mutation, mother (I.2, diagnosed age 21 with peripartum cardiomyopathy), sister (II.1, diagnosed age 29), and proband (II.2, diagnosed age 24). Sanger sequencing confirmed the MYL2 variant was present in all three affected family members (I.2, II.1, and II.2).

ParentalTesting: Mother confirmed as heterozygous carrier of D94A mutation with DCM phenotype. Father not a carrier, unaffected

CasePresentingHPOs: HP:0001644, HP:0006699

CaseHPOFreeText: The proband (II.2) and his sister (II.1) were diagnosed at ages 24 and 29 after preventive cardiovascular screening revealed mild global left ventricular systolic dysfunction. Medical therapy was initiated upon evidence of decreased ejection fraction. Patients II.1 and II.2 (proband) displayed abnormal ECG findings and systolic dysfunction as manifested by reduced EF. However, the LV and septal wall thickness were within the normal range.

CaseNotHPOs: NR

CaseNotHPOFreeText: Septal wall thickness measurements were within normal range in all affected family members. LV wall thickness within normal range.

CasePreviousTesting: No prior testing.

Genotyping Method: Twenty two variants, meeting our established criteria for exome sequencing analysis, were identified in the three affected family members of the pedigree. Exome sequencing performed; variant confirmed by Sanger sequencing. Sanger sequencing confirmed the MYL2 variant was present in all three affected family members (I.2, II.1, and II.2)

FunctionalAnalysis: Functional analysis performed on recombinant proteins, not directly on patient samples. Far-UV CD spectroscopy was used to analyze the effect of the D94A mutation on the secondary structure of the RLC. The mutation was observed to induce a reduction in the α-helical content of the RLC and imposed intra-molecular rearrangements. The mutation was seen to impair the binding of RLC to the MHC (myosin heavy chain), and its incorporation into the RLC-depleted porcine myosin. The actin-activated ATPase activity of mutant-reconstituted porcine cardiac myosin was significantly higher compared to ATPase of WT

Variant: NP_000423.2:p.Asp94Ala

ClinVar: 4710041

CAID: CA386698152

gnomAD: NR

AdditionalInfo: Three affected family members carry the variant (mother I.2, sister II.1, proband II.2)

Case#: E04

DiseaseAssertion: Early onset hypertrophic cardiomyopathy

FamilyInfo: Familiarity case reported as "yes"

ParentalTesting: Not reported

CasePresentingHPOs: HP:0001639

CaseHPOFreeText: Not reported

CaseNotHPOs: Not reported

CaseNotHPOFreeText: Not reported

CasePreviousTesting: Not reported

Genotyping Method: Next Gen Sequencing

FunctionalAnalysis: Not reported

Variant: c.2309-2A>G

ClinVar: Not found

CAID: Not found

gnomAD: 11-47337796-T-C

AdditionalInfo: known pathogenic. allele coverage: 186.

Case#: L08

DiseaseAssertion: Late onset hypertrophic cardiomyopathy

FamilyInfo: Familiarity case reported as "yes"

ParentalTesting: Not reported

CasePresentingHPOs: HP:0001639

CaseHPOFreeText: Not reported

CaseNotHPOs: Not reported

CaseNotHPOFreeText: Not reported

CasePreviousTesting: Not reported

Genotyping Method: Next Generation Sequencing

FunctionalAnalysis: Not reported

Variant: c.1624G>C (Glu542Gln), heterozygous, pathogenic. c.385C>G (Thr128Ser) heterozygous, likely benign.

ClinVar: Not found

CAID: CA407440771 (MYBPC3 CAID not found)

gnomAD: Not found

AdditionalInfo: L08 has two variants. MYBPC3 variant missense/splicing, known pathogenic; Allele coverage 188. TNNI3 variant Missense, new/likely benign; Allele coverage 400.

Case#: 31 year old man of European ancestry. Patient ID NR.

DiseaseAssertion: Nonischemic dilated cardiomyopathy.

FamilyInfo: No reported family history of dilated cardiomyopathy. Both parents had normal heart exams and echocardiograms.

ParentalTesting: Both biological parents tested negative for the TPM1 variant. Sequencing coverage was 500 to 600×. The authors thought the variant likely arose de novo. They could not rule out parental germline mosaicism.

CasePresentingHPOs: HP:0001644, HP:0025169, HP:0001635, HP:0001640, HP:0002202, HP:0002875, HP:0002094, HP:0012764, HP:0034807

CaseHPOFreeText: He first felt short of breath during activity. Later he felt short of breath at rest. He also had a persistent cough and abdominal swelling. Chest CT showed an enlarged heart and fluid around both lungs. The echo showed that both ventricles were enlarged and pumped poorly. His left ventricular ejection fraction was 12%. His heart failure worsened and he needed a heart transplant.

CaseNotHPOs: HP:4000004 (no myocardial late gadolinium enhancement on cardiac MRI).

CaseNotHPOFreeText: The biopsy and exam of the removed heart did not show evidence of active myocarditis.

CasePreviousTesting: NR

Genotyping Method: The Invitae Dilated Cardiomyopathy Panel used next generation sequencing to test 41 genes.

FunctionalAnalysis: Thin filaments with T237S needed more calcium to reach the same level of activation. At low calcium the mutant filaments were less likely to be open and myosin ATPase activity was lower. Omecamtiv mecarbil restored calcium sensitivity in the lab assay. See Figures 3 through 5 and Tables 1 and 2.

Variant: NM_001018005.2(TPM1):c.710C>G (p.Thr237Ser). Heterozygous.

ClinVar: 1067773

CAID: CA392724554

gnomAD: No variant record found in gnomAD v4.1.

AdditionalInfo: PMID 36739943. Figure 1 shows the echo and biopsy results. The clinical report called the variant a VUS. The study authors considered it likely pathogenic after lab testing.

Case1: Female, four years five months

DiseaseAssertion: RCM

FamilyInfo: “ No family history of RCM or other cardiovascular diseases was reported.”

ParentalTesting: NA

CasePresentingHPOs:HP:0002094, HP:0002875,

CaseHPOFreeText: “congestive hepatomegaly” “lower extremity edema” “increased heart shadow without calcification of the pericardium” “bi-atrial enlargement without calcification of the pericardium, thickened pericardium, or endocardium” “‘eft and right atria were enlarged, the right ventricle was slightly enlarged”

CaseNotHPOs:

CaseNotHPOFreeText: “the left ventricle was normal in size, without calcification of the pericardium, thickened pericardium, or endocardium”

CasePreviousTesting: Chest X-ray, Abdominal ultrasonography, MRI, Two-dimensional echocardiography

Genotyping Method: Sanger Sequencing

FunctionalAnalysis: NA

Variant:c.508C>T p.Arg170Trp

ClinVar: 4188245

CAID:

gnomAD:

AdditionalInfo:

Case3: Four month, 21 day old male

DiseaseAssertion: RCM

FamilyInfo: “No family history of RCM or other cardiovascular diseases was reported.”

ParentalTesting:

CasePresentingHPOs: HP:0000961

CaseHPOFreeText: “late hoarseness” “ dilation of the left and right atria”

CaseNotHPOs:

CaseNotHPOFreeText: “normal ventricular sizes, without calcification of the pericardium, thickened pericardium, or endocardium”

CasePreviousTesting: echocardiography

Genotyping Method: Sanger Sequencing

FunctionalAnalysis:

Variant: c.517C>T p.Leu173Phe

ClinVar: Not found

CAID: CA407440287

gnomAD:

AdditionalInfo: “Unfortunately, tissue velocity images were not obtained and the TTE images of this case were lost.”

Case2: Five year old female

DiseaseAssertion: RCM

FamilyInfo: “No family history of RCM or other cardiovascular diseases was reported.”

ParentalTesting:

CasePresentingHPOs:HP:0002875, HP:0002094

CaseHPOFreeText: “lower extremity edema” “congestive hepatomegaly” “ increased heart shadow without calcification of the pericardium” “bi-atrial enlargement without calcification of the pericardium, thickened pericardium, or endocardium”

CaseNotHPOs:

CaseNotHPOFreeText:

CasePreviousTesting: Abdominal ultrasonography, ECG, Chest X-ray, MRI, Two-dimensional echocardiography

Genotyping Method: Sanger Sequencing

FunctionalAnalysis:

Variant: NM_006757.4(TNNT3):c.575G>A (p.Gly192Asp)

ClinVar: 4633351

CAID:

gnomAD:

AdditionalInfo:

Case1: Female, 85 at diagnosis, ethnicity not stated

DiseaseAssertion: Hypertrophic cardiomyopathy (HCM)

FamilyInfo: An index patient’s brother (II.6) was reported with moderate HCM and another brother (II.7) with sudden cardiac death at the age of 40 years. The clinical screening led to the diagnosis of HCM in two of the index patient’s descendants, a daughter and a son. Daughter (III.4, age 59) with septal hypertrophy of 16 mm confirmed by cardiac MRI, son (III.8, age 47) with septal hypertrophy of 15 mm. Daughter (III.6, age 50) is an unaffected carrier. Five other relatives (III.1, III.5, III.6, III.7, IV.1) were healthy with normal cardiac evaluation. Pedigree shown in Figure 1.

ParentalTesting: Not preformed.

CasePresentingHPOs: HP:0001639, HP:0001279

CaseHPOFreeText: Admitted to the emergency unit with chest pain after a pre-syncopal episode at the age of 85. Electrocardiogram showed sinus rhythm, negative T waves in V1-V3 and signs of ventricular hypertrophy. Transthoracic echocardiography detected asymmetric septal hypertrophy of 25 mm with preserved ejection fraction, moderate diastolic dysfunction and mild left atrial enlargement. Twenty-four-hour Holter monitoring documented episodes of non-sustained ventricular tachycardia defined as ≥ 3 beats at a rate of ≥ 120 bpm

CaseNotHPOs: NR

CaseNotHPOFreeText: Coronary artery disease was ruled out

CasePreviousTesting: NR

Genotyping Method: The index patient’s genomic DNA was subjected to massively parallel sequencing to identify the genetic origin of the disease using a 251-gene cardiomyopathy diagnostic panel. Targeted massively parallel sequencing of the index patient’s genomic DNA was performed with an Agilent SureSelect custom library covering 251 genes. For the MYBPC3 gene: all exons including the complete sequence of introns. Sequencing performed on NovaSeq 6000 (Illumina), bioinformatics pipeline using BWA alignment and GATK variant calling, reference genome GRCh37/hg19. Sanger sequencing confirmed both variants

FunctionalAnalysis: Minigene and patient blood RNA splicing assays performed. Both the minigene and patients’ RNA splicing assays revealed that the MYBPC3 c.3331−26T>G variant promoted complete intron 30 retention in the mRNA, and also partial retention of the second half of intron 30. Normal MYBPC3 mRNA was not detected in the MUT lanes. The corresponding normal MYBPC3 fusion protein was not detected with either of the two antibodies in cells transfected with the MUT plasmid.

Variant: NM_000256.3:c.3331−26T>G

ClinVar: 1320039

CAID: CA2573146361

gnomAD: chr11-47332999-A-C

AdditionalInfo: This paper also identified two additional unrelated HCM probands carrying c.3331−26T>G in a large cohort of 9,611 HCM probands

Case1: 50‐year‐old woman

DiseaseAssertion: Ankles swelling and shortness of breath.

FamilyInfo: NR

ParentalTesting: NR

CasePresentingHPOs:

CaseHPOFreeText: dilated cardiomyopathy with end‐diastolic diameter of the left ventricle (LV) of 62 mm, severely depressed left ventricle ejection fraction (LVEF), 20%, mild hypertrophic septum (13.5 mm), trabecular aspect of lateral and inferior LV wall, thrombus in the apex, and severe tricuspid regurgitation with inferior vena cava reflow

CaseNotHPOs: NR

CaseNotHPOFreeText: NR

CasePreviousTesting: NR

Genotyping Method: next‐generation sequencing

FunctionalAnalysis: NR

Variant: NM_000256.3(MYBPC3):c.2459G>A (p.Arg820Gln), NM_032578.4(MYPN):c.3335C>T (p.Pro1112Leu)

ClinVar: 8617, 31791

CAID: CA012326, CA143763

gnomAD: 11-47359085-C-T, 10-69959174-C-T

AdditionalInfo: NR

Case#: a male in the age range of 36-40

DiseaseAssertion: Dilated cardiomyopathy and generated hiPSC-CMS

FamilyInfo: not reported

ParentalTesting: not reported

CasePresentingHPOs: HP:0002875, HP:0031352 Exertional dyspnea and chest tightness

CaseHPOFreeText: nocturnal dyspnea and reduced exercise tolerance

CaseNotHPOs:

CaseNotHPOFreeText:

CasePreviousTesting: echocardiogram and electrocardiograms

Genotyping Method: peripheral blood was extracted and use to sequence the entire genome

FunctionalAnalysis: hiPSCs were looked at in this case

Variant: NM_000256.3: c.194 C > T (p.Thr65Met)

ClinVar:407324

CAID: CA047700

gnomAD: 11-47372888-G-A

AdditionalInfo:F

Case#: Female with DCM symptoms beginning at age 30

DiseaseAssertion: Dilated cardiomyopathy (DCM) associated with a heterozygous missense variant in MYBPC3, c.3373G>A (p.Val1125Met). The authors propose autosomal dominant inheritance in this DCM case.

FamilyInfo: NR – Detailed family relationships, segregation testing, and individual relatives' phenotypes are not described for the current DCM proband.

ParentalTesting: NR

CasePresentingHPOs: HP:0001644 (Dilated cardiomyopathy); HP:0002094 (Dyspnea); HP:0001962 (Palpitations); HP:0004756 (Ventricular tachycardia)

CaseHPOFreeText: The patient experienced symptoms beginning at age 30, including frequent palpitations and dyspnea. ECG monitoring revealed multiple premature ventricular contractions and nonsustained ventricular tachycardia. Echocardiography demonstrated severe spherical left ventricular enlargement, severe systolic and diastolic dysfunction, LVEF of 10%, and mild pulmonary hypertension. She was classified as NYHA functional class II–III and received medical therapy and an implantable cardioverter-defibrillator.

CaseNotHPOs: NR

CaseNotHPOFreeText: NR

CasePreviousTesting: NR – Article describes patient's clinical evaluation, including ECG monitoring and echocardiography, before genetic analysis.

Genotyping Method: Genomic DNA was extracted from peripheral blood. Targeted next-generation sequencing was performed using a custom NimbleGen panel covering the coding regions and exon–intron boundaries of 23 genes associated with DCM. Sequencing was conducted on an Illumina HiSeq 2000 platform. Reads were aligned to the hg19 reference genome using BWA, and SAMtools was used for variant analysis. MYBPC3 variant confirmed using Sanger sequencing.

FunctionalAnalysis: In silico analyses were performed to investigate the potential functional consequences of MYBPC3 p.Val1125Met. SIFT, PolyPhen-2, and MutationTaster predicted potentially damaging effects, while CADD produced a score of 33. Multiple-sequence alignment demonstrated that Val1125 is highly conserved across species. Protein modeling using I-TASSER and Phyre2 predicted structural differences involving the region surrounding residue 1125, located within the C9 fibronectin type III domain of cardiac myosin-binding protein C (cMyBP-C). The authors proposed that the substitution could affect protein structure and function. No direct experimental functional validation of this variant was reported.

Variant: MYBPC3 NM_000256.3:c. 3373G>A (p.Val1125Met); rsID: rs121909378; missense; heterozygous

ClinVar: 8604

CAID: CA014038

gnomAD: MYBPC3 c.3373G>A (p.Val1125Met), GRCh38: 11-47332931-C-T. In gnomAD v4, the reported allele frequencies are 0.00001920 (exomes), 0.00003943 (genomes), and 0.00002111 (total). The highest ancestry-specific allele frequency identified was in the Ashkenazi Jewish population (AC = 5; AF = 0.0001695).

AdditionalInfo: Authors propose variant dosage, interactions with other variants, and genetic modifiers may contribute to differences in disease phenotype. However, the proposed autosomal dominant association with DCM was not supported by documented segregation testing in the current proband's family, and the functional analyses were computational rather than experimental.

Case#: 80-year-old male (I-1) diagnosed with HCM at approximately 50–55 years of age; familial case with his 3 adult offspring (II-2, II-3, and II-4)

DiseaseAssertion: Hypertrophic cardiomyopathy (HCM) associated with a heterozygous frameshift variant in MYBPC3.

FamilyInfo: The proband (I-1) is an elderly male with familial hypertrophic cardiomyopathy. Three adult offspring (II-2, II-3, and II-4) were evaluated and diagnosed with HCM based on interventricular septal hypertrophy. All four individuals carried the heterozygous MYBPC3 c.1042_1043insCGGCA variant. Individual II-2 experienced chest discomfort and palpitations, whereas II-3 and II-4 were asymptomatic at evaluation. These findings demonstrate segregation of the variant with HCM in the examined family members and are consistent with autosomal dominant inheritance.

ParentalTesting: NR – Genetic testing of the proband's parents was not reported but the variant was identified in his three adult offspring, supporting familial segregation.

CasePresentingHPOs: HP:0001639 (Hypertrophic cardiomyopathy); HP:0002094 (Dyspnea); HP:0001635 (Congestive heart failure)

CaseHPOFreeText: The proband experienced chest tightness, dyspnea, and recurrent heart failure. Echocardiography demonstrated marked interventricular septal hypertrophy (39 mm during diastole and 41 mm during systole), a left atrial diameter of 44 mm, and a left ventricular ejection fraction of 60%. Additional findings included premature atrial beats, intraventricular conduction block, and NYHA functional class II. A history of hypertension was reported.

CaseNotHPOs: HP:0001279 (Syncope); HP:0001681 (Angina pectoris); HP:0001695 (Cardiac arrest)

CaseNotHPOFreeText: The proband had no reported history of syncope, angina pectoris, or cardiac arrest. No surgical myectomy was reported.

CasePreviousTesting: NR – Previous genetic testing was not reported but the proband underwent clinical cardiovascular evaluation, including electrocardiography, echocardiography, and BNP measurement.

Genotyping Method: Whole-genome sequencing (WGS) was performed using peripheral blood DNA on an Illumina NovaSeq 6000 platform with 150-bp paired-end reads and approximately 30× sequencing depth. Reads were aligned to the hs37d5 reference genome using BWA, and variants were annotated using ANNOVAR.

FunctionalAnalysis: Cardiac tissue from the proband was analyzed using RT-qPCR and Western blotting. MYBPC3 mRNA expression and cardiac myosin-binding protein C (cMyBP-C) abundance were significantly reduced (P < 0.05), supporting a haploinsufficiency mechanism. These findings do not independently establish the functional effect of the specific variant, and the Western blot analysis lacked biological replicates.

Variant: MYBPC3 NM_000256.3.1038_1042dup (p.Met348ThrfsTer4). Five-base-pair duplication resulting in a frameshift and premature termination codon. Reported in Zhang et al. (2025) as c.1042_1043insCGGCA.

ClinVar: No exact match identified for MYBPC3 c.1042_1043insCGGCA using the reported HGVS notation. ClinVar Variation ID not established.

CAID: CA273429

gnomAD: No exact matching entry identified in gnomAD v4 for MYBPC3 NM_000256.3:c.1038_1042dup (CA273429). Population allele frequency could not be established.

AdditionalInfo: The variant was identified in the proband and three adult offspring with interventricular septal thickening. The study proposed MYBPC3 haploinsufficiency as a disease mechanism based on reduced transcript and protein abundance. The available familial and functional evidence supports an association with HCM, although the specific variant's pathogenicity requires interpretation alongside the limitations of the study.

Case#: AD-FDC27, II:11, male, age 35

DiseaseAssertion: Familial dilated cardiomyopathy

FamilyInfo: second multigenerational DCM family

CasePresentingHPOs: HP:0001644 HP:4000141

CaseHPOFreeText: Dyspnea on exertion; left ventricular dilation; severe systolic dysfunction

Genotyping Method: Whole-exome sequencing using Agilent SureSelect 50 Mb exome; HiSeq 2000 sequencing followed by Sanger sequencing confirmation

Variant: TNNT2 c.517C>T (p.Arg173Trp); missense single-nucleotide variant; heterozygous

ClinVar: 228409; Pathogenic

CAID: CA10576372

Case#: AD-FDC1, IV:14, male, age 45

DiseaseAssertion: Familial dilated cardiomyopathy

FamilyInfo: multigenerational DCM family

CasePresentingHPOs: HP:0001644 HP:4000141 HP:0012664

Genotyping Method: Whole-exome sequencing using Agilent SureSelect 50 Mb exome; HiSeq 2000 sequencing followed by Sanger sequencing confirmation

Variant: TNNT2 c.517C>T (p.Arg173Trp); missense single-nucleotide variant; heterozygous

ClinVar: 228409

CAID: CA10576372

Case#: Patient 3; male; 60 years old at first observation; 14 mm LV wall thickness; family history of HCM

DiseaseAssertion: Hypertrophic cardiomyopathy (HCM) associated with the heterozygous MYBPC3 intronic splice-altering variant c.1224-80G>A, classified as likely pathogenic by the study authors.

FamilyInfo: Patient was reported to have positive family history of HCM. Echocardiography demonstrated a maximum left ventricular wall thickness of 14 mm without left ventricular outflow tract obstruction. The study did not identify specific affected relatives or provide a family pedigree.

ParentalTesting: NR – Patient had a positive family history of HCM but parental genotypes and variant segregation were not documented.

CasePresentingHPOs: HP:0001639 (Hypertrophic cardiomyopathy); HP:0001712 (Left ventricular hypertrophy)

CaseHPOFreeText: Echocardiography demonstrated a maximum left ventricular wall thickness of 14 mm without left ventricular outflow tract (LVOT) obstruction. NYHA functional classification was not reported.

CaseNotHPOs: HP:0032092 (Left ventricular outflow tract obstruction)

CaseNotHPOFreeText: Echocardiographic evaluation showed no left ventricular outflow tract (LVOT) obstruction.

CasePreviousTesting: Patient 3 previously underwent targeted next-generation sequencing using the Illumina TruSight Cardio panel, which included 174 genes associated with inherited cardiac conditions. The initial genetic analysis was negative. Subsequent reanalysis of existing sequencing data identified the MYBPC3 intronic variant c.1224-80G>A.

Genotyping Method: Genomic DNA obtained from peripheral blood was analyzed using the Illumina TruSight Cardio targeted next-generation sequencing panel on an Illumina MiniSeq platform. Previously generated sequencing data were reexamined using an intronic hotspot analysis approach, with variant visualization in Integrative Genomics Viewer (IGV). The MYBPC3 c.1224-80G>A variant was subsequently confirmed by Sanger sequencing.

FunctionalAnalysis: The MYBPC3 intronic variant c.1224-80G>A creates a cryptic splice acceptor site within intron 13, resulting in retention of 78 intronic nucleotides upstream of exon 14. This produces an in-frame insertion of 26 codons. Previous functional studies demonstrated altered RNA splicing and an approximately 50% reduction in MYBPC3 transcript abundance. SpliceAI predicted a strong splice-altering effect (Δscore = 0.94). The current study did not perform patient-specific RNA functional experiments. Instead, researchers identified the variant through reanalysis of existing next-generation sequencing data and confirmed its presence using Sanger sequencing.

Variant: MYBPC3 NM_000256.3.1224-80G>A.

ClinVar: 693982

CAID: CA221697326

gnomAD: The MYBPC3 variant c.1224-80G>A (GRCh38: 11-47343342-C-T) is present at a low frequency in gnomAD v4; Exome allele frequency: 0.00001637; Genome allele frequency: 0.00005261; Total allele frequency: 0.00002005; The highest ancestry-specific allele frequency reported was in the African/African American population, with an allele count (AC) of 8 and allele frequency (AF) of 0.0001113.

AdditionalInfo: Although Patient 3 carried the same variant as Patient 2, their reported cardiac phenotypes differed. Patient 2 exhibited a maximum left ventricular wall thickness of 27 mm and LVOT obstruction with a gradient of 50 mmHg. These differences are consistent with variable clinical expression of MYBPC3-associated HCM, although the study did not determine their underlying causes.

Case#: Patient 1; female; 56 years old at first observation with diagnosis of HCM; 13 mm LV wall thickness

DiseaseAssertion: Hypertrophic cardiomyopathy (HCM) associated with the heterozygous MYBPC3 intronic splice-altering variant c.1224-80G>A, classified as likely pathogenic by the study authors.

FamilyInfo: The study reports a negative family history of HCM. No family pedigree or detailed clinical information about relatives was provided.

ParentalTesting: NR

CasePresentingHPOs: HP:0001639 (Hypertrophic cardiomyopathy); HP:0001712 (Left ventricular hypertrophy); HP:0031656 (Systolic anterior motion of the mitral valve)

CaseHPOFreeText: Hypertrophic cardiomyopathy and a maximum left ventricular wall thickness of 13 mm. Echocardiography demonstrated systolic anterior motion (SAM) of the mitral valve without left ventricular outflow tract (LVOT) obstruction. The patient had no reported family history of hypertrophic cardiomyopathy. NYHA functional classification was NR.

CaseNotHPOs: HP:0032092 (Left ventricular outflow tract obstruction)

CaseNotHPOFreeText: No left ventricular outflow tract (LVOT) obstruction was reported despite the presence of systolic anterior motion of the mitral valve.

CasePreviousTesting: Patient 1 previously underwent targeted next-generation sequencing using the Illumina TruSight Cardio panel covering 174 genes associated with inherited cardiac conditions. Initial genetic analysis was negative. Subsequent reanalysis of existing sequencing data identified the MYBPC3 intronic variant c.1224-80G>A.

Genotyping Method: Genomic DNA from peripheral blood was analyzed using the Illumina TruSight Cardio targeted next-generation sequencing panel on an Illumina MiniSeq platform. Existing sequencing data were reexamined using an intronic hotspot analysis approach and Integrative Genomics Viewer (IGV). The MYBPC3 c.1224-80G>A variant was subsequently confirmed by Sanger sequencing.

FunctionalAnalysis: The MYBPC3 intronic variant c.1224-80G>A creates a cryptic splice acceptor site within intron 13, resulting in the retention of 78 intronic nucleotides upstream of exon 14. This produces an in-frame insertion of 26 codons in the resulting transcript. Previous functional studies demonstrated abnormal RNA splicing and approximately 50% reduction in MYBPC3 transcript abundance. SpliceAI predicted a strong splice-altering effect (Δscore = 0.94). The current study did not perform patient-specific RNA functional experiments. Instead, the researchers identified the variant through reanalysis of existing next-generation sequencing data and confirmed its presence using Sanger sequencing.

Variant: MYBPC3 NM_000256.3.1224-80G>A.

ClinVar: 693982

CAID: CA221697326

gnomAD: The MYBPC3 variant c.1224-80G>A (GRCh38: 11-47343342-C-T) is present at a low frequency in gnomAD v4; Exome allele frequency: 0.00001637; Genome allele frequency: 0.00005261; Total allele frequency: 0.00002005; The highest ancestry-specific allele frequency reported was in the African/African American population, with an allele count (AC) of 8 and allele frequency (AF) of 0.0001113.

AdditionalInfo: Unlike Patients 2 and 3, Patient 1 had no reported family history of HCM. Echocardiography demonstrated systolic anterior motion of the mitral valve without left ventricular outflow tract obstruction, and maximum left ventricular wall thickness was 13 mm. Although all three patients carried the same splice-altering variant, their reported cardiac phenotypes differed. This observation is consistent with variable clinical expression of MYBPC3-associated HCM, although the study did not establish the underlying causes of these differences.

Case#: Patient 2; male; 74 years old at first observation; 27 mm LV wall thickness; family history of HCM

DiseaseAssertion: Hypertrophic cardiomyopathy (HCM) associated with the MYBPC3 intronic splice-altering variant c.1224-80G>A (authors classify as likely pathogenic).

FamilyInfo: Reported family history of hypertrophic cardiomyopathy. Echocardiographic evaluation demonstrated a maximum left ventricular wall thickness of 27 mm and left ventricular outflow tract (LVOT) obstruction, with a gradient of 50 mmHg. The patient was classified as NYHA functional class II.

ParentalTesting: NR – Patient had a positive family history of HCM but parental genotypes and variant segregation were not documented.

CasePresentingHPOs: HP:0001639 (Hypertrophic cardiomyopathy); HP:0032092 (Left ventricular outflow tract obstruction); HP:0001712 (Left ventricular hypertrophy)

CaseHPOFreeText: Echocardiography demonstrated a maximum left ventricular wall thickness of 27 mm and left ventricular outflow tract (LVOT) obstruction with a measured gradient of 50 mmHg. The patient was classified as New York Heart Association (NYHA) functional class II, indicating mild limitation of physical activity.

CaseNotHPOs: NR

CaseNotHPOFreeText: NR

CasePreviousTesting: The patient previously underwent targeted next-generation sequencing using the Illumina TruSight Cardio panel covering 174 genes associated with hereditary cardiac diseases. The initial analysis produced a negative molecular result. Subsequent reanalysis of the existing sequencing data identified the MYBPC3 intronic variant c.1224-80G>A.

Genotyping Method: Genomic DNA was extracted from peripheral blood and sequenced using the Illumina TruSight Cardio panel on a MiniSeq platform. Initial bioinformatics analysis was performed using Illumina BaseSpace software. Raw sequencing data were subsequently reanalyzed using an intronic-variant hotspot approach, with variant visualization in Integrative Genomics Viewer (IGV). The identified MYBPC3 variant was validated by Sanger sequencing.

FunctionalAnalysis: The MYBPC3 intronic variant c.1224-80G>A creates a novel splice acceptor site within intron 13, resulting in the retention of 78 intronic nucleotides upstream of exon 14. This produces an in-frame insertion of 26 codons and is associated with approximately 50% reduction in MYBPC3 transcript abundance. Previous studies demonstrated the splice-altering effect through functional analyses, while SpliceAI predicted a strong effect on splicing (Δscore = 0.94). The current study identified the variant through reanalysis of existing NGS data and confirmed its presence using Sanger sequencing; it did not perform new patient-specific RNA functional experiments.

Variant: MYBPC3 NM_000256.3.1224-80G>A.

ClinVar: 693982

CAID: CA221697326

gnomAD: The MYBPC3 variant c.1224-80G>A (GRCh38: 11-47343342-C-T) is present at a low frequency in gnomAD v4; Exome allele frequency: 0.00001637; Genome allele frequency: 0.00005261; Total allele frequency: 0.00002005; The highest ancestry-specific allele frequency reported was in the African/African American population, with an allele count (AC) of 8 and allele frequency (AF) of 0.0001113.

AdditionalInfo: The variant was detected by expanding the analysis to include previously characterized intronic splice-altering regions and was subsequently confirmed by Sanger sequencing. Functional evidence for altered splicing was obtained from previously published research rather than new patient-specific RNA experiments.

Case#: 38 year old woman

DiseaseAssertion: Hypertrophic cardiomyopathy (HCM)

FamilyInfo: Father and one brother had HCM, no sudden deaths reported in family

ParentalTesting: Father carried the same heterozygous variant

CasePresentingHPOs: HP:0001962, HP:0031352 “palpitation and chest distress”

CaseHPOFreeText: myocardial fibrosis

CaseNotHPOs: N/A

CaseNotHPOFreeText: Unremarkable

CasePreviousTesting: N/A

Genotyping Method:PCR

FunctionalAnalysis: N/A

Variant: TNNI3 heterozygous c.235C>T (p.Arg79Cys)

ClinVar: Variation ID: 43367

CAID:N/A

gnomAD:N/A

AdditionalInfo: authors describe the variant as novel

Case#:27-year-old woman

DiseaseAssertion: Familial restrictive cardiomyopathy (RCM)

FamilyInfo: Strong family history of cardiac disease and sudden cardiac death

ParentalTesting:N/A

CasePresentingHPOs: “ Fatigue, dyspnoea , palpitations, atrial fibrillation, pulmonary hypertension, and fluid congestion.”

CaseHPOFreeText: age 27, she presented with progressive fatigue and limited exercise capacity, 13-year symptom-free interval, she developed rapid clinical deterioration with dyspnoea and palpitations

CaseNotHPOs: N/A

CaseNotHPOFreeText: N/A

CasePreviousTesting: focused screening of MYH7, TNNT2, and MYBPC3 was unremarkable,

Genotyping Method: NGS

FunctionalAnalysis:N/A

Variant: TNNI3, NP_000354.4: p.Lys193Glu; NC_000019.9: g.55663258T>C. Heterozygous reported as novel

ClinVar: N/A

CAID:N/A

gnomAD: N/A

AdditionalInfo: patient improved with restored sinus rhythm and low-dose diuretic treatment and was symptom-free at the time of reporting. later had an ischaemic stroke

Case#:7 ¾ year old girl

DiseaseAssertion: Hypertrophic cardiomyopathy (HCM), with ventricular tachycardia and Stokes–Adams syndrome.

FamilyInfo: Parents not consanguineous, no family history of cardiovascular disease or sudden cardiac death.

ParentalTesting: Parents not consanguineous, no symptoms

CasePresentingHPOs: HP:0001639, HP:0001695, HP:0001663, HP:0001250, HP:0001678, HP:0001712 “Hypertrophic cardiomyopathy, cardiac arrest, ventricular tachycardia, seizures, atrioventricular block, left ventricular hypertrophy:”

CaseHPOFreeText: Diagnosed with cardiomyopathy in utero. Frequent cardiac arrests from 3-7, multiple seizures a year, cardiac mri showed heart enlargement, hypertrophy, after icd implant no cardiac arrests after 1 year.

CaseNotHPOs: N/A

CaseNotHPOFreeText: N/A

CasePreviousTesting: diagnosed with cardiomyopathy in utero, no other testing stated.

Genotyping Method: Whole exon sequencing

FunctionalAnalysis: protein modeling only, revealed to be “probably damaging” score 1.0

Variant: MYH7 NM_000257.3:c.2723T>C (p.Leu908Pro; p.L908P)

ClinVar: no id but is listed under https://www.ncbi.nlm.nih.gov/clinvar/RCV000158849.1/?cmd=HTOn&utm_source=chatgpt.com

CAID: N/A

gnomAD:N/A

AdditionalInfo: patient underwent ICD implantation and had no further arrests after a years time.

Case#: P2 female; age 2 years; Vietnamese family

DiseaseAssertion: Dilated cardiomyopathy

FamilyInfo: MYL2 variant was reported as de novo in P2

ParentalTesting: Sanger sequencing of family members; variant was absent in the parents

CasePresentingHPOs: HP:0001644 HP:0012664 HP:4000141

CaseHPOFreeText: Dilated cardiomyopathy; reduced ejection fraction of 30%; increased diastolic cardiac diameter of 37 mm

Genotyping Method: Whole-exome sequencing using the SureSelect Human All Exon V7 kit, followed; Sanger sequencing confirmation

Variant: MYL2 c.284C>G, p.Pro95Arg; heterozygous missense variant

Case#: CMP-83-03; sex NR; pediatric age, exact age NR

CasePresentingHPOs: HP:0001644

CaseHPOFreeText: Biopsy proven myocarditis with dilated cardiomyopathy

Genotyping Method: Targeted next-generation sequencing

Variant: TNNT2 c.460C>T (p.Arg154Trp); heterozygous; missense single-nucleotide variant

ClinVar: 132943

CAID: CA004554

gnomAD: 0.000032

Case#: Jenni a 7 year-old female

DiseaseAssertion: Dilated cardiomyopathy with LVNC

FamilyInfo: MYH7 maternally inherited and MYBPC3 paternally inherited

ParentalTesting: Mother has PPCM

CasePresentingHPOs: HP:0004755

heart failure and supraventicular tachycardia

CaseHPOFreeText: Intermittent Wolff-Parkinson- White

CaseNotHPOs:

CaseNotHPOFreeText:

CasePreviousTesting: Echocardiogram and MRI

Genotyping Method: Segregation analysis, targeted next generation sequencing

FunctionalAnalysis: electrophysiology, transplant evaluation, CT angiography

Variant: NM_000256.3(MYBPC3):c.2672G>A (p.Arg891Gln)

ClinVar:177884

CAID: CA012826

gnomAD: 11-47357493-C-T

AdditionalInfo:

Case#: a 2‐year‐old white girl who was found to have RCM following an evaluation precipitated by the sudden death of her older brother.

DiseaseAssertion: Restrictive cardiomyopathy

FamilyInfo: 4‐year‐old brother presented with a sudden cardiac arrest while running on a soccer field and could not be resuscitated. Her father had restrictive physiology and arrhythmias. The TNNT2 variant was found in the father and two children

ParentalTesting: Father carried the variant. For both paternal grandparents tested negative and the proband inherited the variant from her father.

CasePresentingHPOs: HP:0001723, HP:0001639, HP:0031295, HP:0030718, HP:0001695

CaseHPOFreeText: "Her ECG demonstrated criteria for right and left atrial enlargement with marked repolarization abnormality. Echocardiogram revealed mild right atrial and severe left atrial enlargement (left atrial volume, 61.5 cc/m2; normal, 16–28), with mildly increased left ventricular mass and otherwise normal ventricular dimensions. Right and left ventricular systolic function was preserved. "

CaseNotHPOs: not reported

CaseNotHPOFreeText: Ventricular dimensions and biventricular systolic function were reported as normal.

CasePreviousTesting: Cardiomyopathy‐based panel testing (Familion; Transgenomic, Inc., New Haven, CT) was conducted on the proband, demonstrating a single, heterozygous, variant TNNT2‐R94C (c.280C>T).

Genotyping Method: Clinical genetic testing was performed on family members using the same HCM panel test.

FunctionalAnalysis: Biophysical experiments using recombinant human TNNT2‐R94C demonstrated impaired cardiac regulation at the molecular level attributed to reduced calcium‐dependent blocking of myosin's interaction with the thin filament.

Variant: c.835C>T, p.Gln279Ter

ClinVar: 165549

CAID: CA004288

gnomAD: not reported

AdditionalInfo: not reported

Case1: 11 year old female

DiseaseAssertion: hypertrophic cardiomyopathy, restrictive cardiomyopathy

FamilyInfo: twin sister carrier identical mutation, both sisters expired, no family history of cardiomyopathy

ParentalTesting: sanger sequencing

CasePresentingHPOs:HP:0001629, HP:0001640, HP:0034314,HP:0002098, HP:0001723

CaseHPOFreeText:bilateral atrial dilation with mid ventricular hypertrophy, accelerated cardiac sound, liver palpable 2cm below costal margin, cold extremities, cardiac condition was New York Heart Association class (NYHA) III.

CaseNotHPOs: NR

CaseNotHPOFreeText: no gallop rhythm, no murmur, no blood test abnormalities, no metabolic disorders

CasePreviousTesting: cardiac catheterization, cardiac magnetic resonance, echocardiography, Chest Xray, electrocardiography,

Genotyping Method: sanger sequencing

FunctionalAnalysis: NR

Variant: TNNI3 c.574 C>T p.Arg192Cys

ClinVar: 165510

CAID: CA021951

gnomAD:NR

Case 2: 15 year old female

DiseaseAssertion: restrictive cardiomyopathy, hypertrophic cardiomyopathy

FamilyInfo: twin sister carrier identical mutation, both sisters expired, no family history of cardiomyopathy

ParentalTesting: Sanger Sequencing

CasePresentingHPOs:HP:0002875, HP:0001723

CaseHPOFreeText: Abnormal ECG, LV hypertrophy, enlargement of both atria, high LVEF levels, accelerated cardiac sound, liver was palpable 3 cm, extremities were warm, and cardiac condition was NYHA II.

CaseNotHPOs:NR

CaseNotHPOFreeText: no gallop rhythm, no murmur, no blood test abnormalities, no metabolic disorders

CasePreviousTesting: cardiac catheterization, cardiac auscultation, myocardial disarray with myocyte hypertrophy

Genotyping Method: Sanger sequencing

FunctionalAnalysis: NR

Variant: TNNI3 c.574 C>T p.Arg192Cys

ClinVar: 165510

CAID: CA021951

gnomAD:NR

Case#:7-year-old boy

DiseaseAssertion: restrictive cardiomyopathy

FamilyInfo: his parents were in generally good health and other family members denied a history of syncope, sudden death or cardiomyopathy

ParentalTesting: sanger sequencing

CasePresentingHPOs:HP:0007185, HP:0001695, HP:0005943, HP:0002516, HP:0001663

CaseHPOFreeText:electrocardiogram (ECG) revealed Ptf-V 1 < –0.04 mm⋅s and biphasic P waves, indicating left atrial enlargement and the chest X-ray revealed an enlarged heart shadow. The echocardiogram showed biatrial enlargement, especially in the left atrium (LA: 32 mm, RA: 30 mm), decreased diastolic function (E/A = 2.6; E Peak deceleration time 130 ms; Constant volume diastolic time 49 ms), moderate mitral valve regurgitation and left ventricular ejection fraction of 51.8%.

CaseNotHPOs:HP:0000969

CaseNotHPOFreeText:Except for bilateral enlargement, cardiac magnetic resonance (CMR) indicated no abnormalities

CasePreviousTesting: brain MRI, video electrocephalogram, electrocardiogram

Genotyping Method: sanger sequencing

FunctionalAnalysis:

Variant: TNNI3 c.611(exon8)G>A,p.R204H

ClinVar:177679

CAID:CA022060

gnomAD:chr19-55151856-C-T

Case#: A 50-year-old man diagnosed with HCM at age 41, with asymmetric septal hypertrophy and exertional chest discomfort and pain

DiseaseAssertion: Hypertrophic cardiomyopathy

FamilyInfo: His third elder sister, niece and daughter were also clinically affected by HCM. The proband's mother and eldest sister had suffered from sudden cardiac death. The proband's father, second elder sister (HCM1 in Fig. 1A), nephew (HCM5 in Fig. 1A) and son (HCM7 in Fig. 1A) were 87, 65, 35, and 14 years old, respectively, and had normal echocardiographic measurements.

ParentalTesting: The mother had died suddenly; the 87-year-old father had normal echocardiographic measurements. There was no testing reported.

CasePresentingHPOs: HP:0001639, HP:0005144, HP:0100749, HP:0011712

CaseHPOFreeText: chest distress and chest pain during exercise for 9 years. He presented with an asymmetric hypertrophic interventricular septum and a maximum interventricular septum thickness of 18.04 mm

CaseNotHPOs: not reported

CaseNotHPOFreeText: no findings found in the proband

CasePreviousTesting: nothing reported in the article

Genotyping Method: hole exome sequencing (WES) Exons were captured using Sure Select Human All Exon V5 kit. Sequenced by Illumina HiSeq 4000 sequencer.

FunctionalAnalysis: not reported

Variant: rs796925245 (g.1:201359630G>A, c.835C>T, p.Gln279Ter) (c.835C>T, p.Gln279Ter)

ClinVar: 1175037

CAID: CA088451

gnomAD: not reported

AdditionalInfo: n/r

Case#: Male half-brother with mild left ventricular dilation from early childhood and mild left ventricular impairment; latest reported ejection fraction was 51%.

DiseaseAssertion: DCM

FamilyInfo: several of the siblings DCM, two died at age of 12, father had DCM, and heart transplant for an individual who later died

ParentalTesting: father was an obligate carrier

CasePresentingHPOs: HP:0001644

CaseHPOFreeText: Mild left ventricular dilation and mildly increased LV volumes

CaseNotHPOs: not reported

CaseNotHPOFreeText: not reported

CasePreviousTesting: panel-based genetic test was conducted in the index patient.

Genotyping Method: panel-based genetic test was conducted in the index patient. sanger confirmation

FunctionalAnalysis: not reported

Variant: c.659_661del p.(Lys220del)

ClinVar: 43659

CAID: CA006429

gnomAD: not reported

AdditionalInfo: not reported

Case#: A 2-year-old girl with dilated cardiomyopathy (DCM) and left ventricular noncompaction, who received a heart transplant at 26 months.

DiseaseAssertion: DCM with left ventricular noncompaction

FamilyInfo: Family history revealed that three of the father’s (A.II.2) sisters (A.II.4-A.II.6) died in infancy due to cardiac failure.

ParentalTesting: The father and paternal grandmother tested positive for the same TNNT2 variant

CasePresentingHPOs: HP:0001644, HP:0030682, HP:0001635

CaseHPOFreeText: The heart failure signs began at 2 months and got worse. Then got a heart transplant at 26 months.

CaseNotHPOs: not reported

CaseNotHPOFreeText: nothing found

CasePreviousTesting: a genetic panel test, including the genes LDB3, LMNA, MYH7, MYBPC3, and TNNT2, was performed.

Genotyping Method: Variants were confirmed in first-degree family members by Sanger sequencing.

FunctionalAnalysis: not reported

Variant: c.316G>A (p.Glu106Lys)

ClinVar: 224775

CAID: CA088557

gnomAD: not reported

AdditionalInfo: not reported

Case#: 19-year-old boy with coarctation of the aorta who complained of mild exertional dyspnea.

DiseaseAssertion: dilated cardiomyopathy (DCMP). LV systolic dysfunction.

FamilyInfo: The family history was positive to sudden death (his cousin died, aged 3 months) and to DCMP and heart transplant (HT): a maternal aunt and her son who then died at 42 and 24 years old. Cascade family screening revealed a TNNT2 variant in the mother (II.1) and his cousin (IV.3).

ParentalTesting: The 19 year old brother's was tested and carried the TNNT2 variant. She was also affected by DCM. The father’s testing status was not reported.

CasePresentingHPOs: HP:0001680, HP:0002875, HP:0025169, HP:4000141

CaseHPOFreeText: surgically repaired coarctation of the aorta who complained of mild exertional dyspnea repaired at birth (NYHA class II–III). Cardiac MRI showed a moderately dilated, hypokinetic LV with an ejection fraction of 45%; echocardiography showed an ejection fraction around 42%.

CaseNotHPOs: not reported

CaseNotHPOFreeText: Nothing was found that was correlated.

CasePreviousTesting: genetic investigations through next generation sequencing (NGS) with a panel of 325 genes, known to be associated with cardiomyopathies

Genotyping Method: targeted next generation sequencing

FunctionalAnalysis: not reported

Variant: c.518G>A (p. Arg173Gln)

ClinVar: 43649

CAID: CA004727

gnomAD: not reported

AdditionalInfo: not reported

Case#: 1, age 36, age 30 (report)

DiseaseAssertion: Pompe Disease

FamilyInfo: Family 1, no further information

CasePresenting HPOs:HP:0002878, HP:0012378, HP:0003546, HP:0001324 (Respiratory failure, fatigue, exercise intolerance, muscle weakness)

CaseHPOFreeText: myopathy with vacuoles, biopsy with vacuolar myopathy with PAS +

CaseNotHPOs: N/A

CaseNotHPOFreeText: N/A

CasePreviousTesting: single gene sequencing/analysis, 48.47 NaG/AaGIA, 89.61% inhibition of GAA; NaG/AaGIA above 40 and %INH above 87 are in the affected range

Variants:

Variant 1:NM_000152.5(GAA):c.1941C>G (p.Cys647Trp)

ClinVar: 550327

Variant 2: Not reported

ParentalGenotypes: N/A

PreviouslyPublished: N/A

Case#: 2/b, age 11, age 5 (report)

DiseaseAssertion: Pompe Disease

FamilyInfo: Family 2, no further information

CasePresenting HPOs:HP:0002878, HP:0012378, HP:0003546, HP:0001324 (Respiratory failure, fatigue, exercise intolerance, muscle weakness)

CaseHPOFreeText: valvular myopathy, biopsy with vacuolar myopathy with PAS +

CaseNotHPOs: N/A

CaseNotHPOFreeText: N/A

CasePreviousTesting: single gene sequencing/analysis, 105.01 NaG/AaGIA, 90.98% inhibition of GAA; NaG/AaGIA above 40 and %INH above 87 are in the affected range

Variants:

Variant 1:NM_000152.5(GAA):c.377G>A (p.Trp126Ter)

ClinVar: 932904

Variant 2: NM_000152.5(GAA):c.1655T>C (p.Leu552Pro)

ClinVar: 279811

ParentalGenotypes: N/A

PreviouslyPublished: N/A

Case#: 6/b, age 40, age 36 (report)

DiseaseAssertion: gray zone (Pompe Disease)

FamilyInfo: Family 6, no further information

CasePresenting HPOs: HP:0001288 (Impaired gait)

CaseHPOFreeText: Progressive worsening muscle weakness, vacuolar myopathy biopsy PAS +

CaseNotHPOs: N/A

CaseNotHPOFreeText: N/A

CasePreviousTesting: single gene sequencing/analysis, the results of (NaG/AaGIA > 40 and %INH < 87) or (NaG/AaGIA < 40 and %INH > 87) are considered to be within a gray zone.

Variants:

Variant 1: NM_000152.5(GAA):c.1941C>G (p.Cys647Trp)

ClinVar: 550327

Variant 2:NM_000152.5(GAA):c.-32-13T>G

ClinVar: 4027

ParentalGenotypes: N/A

PreviouslyPublished: N/A

Case#: 4, age 24, age 18 (report)

DiseaseAssertion: Pompe Disease

FamilyInfo: Family 4, no further information

CasePresenting HPOs:HP:0012378, HP:0003546, HP:0001324 (fatigue, exercise intolerance, muscle weakness)

CaseHPOFreeText: vacuolar myopathy biopsy PAS +

CaseNotHPOs: N/A

CaseNotHPOFreeText: N/A

CasePreviousTesting: single gene sequencing/analysis, 40.54 NaG/AaGIA, 90.54% inhibition of GAA; NaG/AaGIA above 40 and %INH above 87 are in the affected range

Variants:

Variant 1:NM_000152.5(GAA):c.2608C>T (p.Arg870Ter)

ClinVar: 189009

Variant 2: not reported

ParentalGenotypes: N/A

PreviouslyPublished: N/A

Case#: 2/a, age 20, age 14 (report)

DiseaseAssertion: Pompe Disease

FamilyInfo: Family 2, no further information

CasePresenting HPOs:HP:0002878, HP:0012378, HP:0003546, HP:0001324 (Respiratory failure, fatigue, exercise intolerance, muscle weakness)

CaseHPOFreeText: valvular myopathy, biopsy with vacuolar myopathy with PAS +

CaseNotHPOs: N/A

CaseNotHPOFreeText: N/A

CasePreviousTesting: single gene sequencing/analysis, 64.47 NaG/AaGIA, 92.28% inhibition of GAA; NaG/AaGIA above 40 and %INH above 87 are in the affected range

Variants:

Variant 1:NM_000152.5(GAA):c.377G>A (p.Trp126Ter)

ClinVar: 932904

Variant 2: NM_000152.5(GAA):c.1655T>C (p.Leu552Pro)

ClinVar: 279811

ParentalGenotypes: N/A

PreviouslyPublished: N/A

Case: Patient 6 (Family C), male, 53yo

DiseaseAssertion: VWD2B

FamilyInfo: NA

ParentalGenotype(s): NA

CasePresentingHPOs: HP:0030129, HP:0030130, HP:0011871, HP:0012147

(impaired ristocetin cofactor assay activity, Impaired von Willebrand factor collagen binding activity, Impaired RIPA, Reduced quantity of VWF)

CaseHPOFreeText: blood group O, Platelet count x103/μl = 193, PFA 100 sec = 199, SPA % = 5.2, MADR mg/mL = 0.75, VWF:Ag U/dL = 47.1, VWF:CB U/dL = 44.6, VWF:CB ratio = 0.94, VWF:RCo U/dL = 38.0, VWF:RCo ratio = 0.80, Platelet VWF U/dL = 47.4, FVIII U/dL = 43.8, BAT F/M = 0

CaseNOTHPOs:

CaseNOTHPOFreeText:

CasePreviousTesting: NA

GenotypingMethod: Sequencing of exon 28 of VWF

PreviouslyPublished: NA

Gene: VWF

Variant: c.4135C>T (p.Arg1379Cys)

HGVS: NM_000552.4(VWF):c.4135C>T (p.Arg1379Cys)

ClinVarID: 100333; https://www.ncbi.nlm.nih.gov/clinvar/variation/100333/

CAID: NA

gnomAD: MAF = 0.00003891 (5/128500 alleles) in the European (non-Finnish) population; https://gnomad.broadinstitute.org/variant/12-6128449-G-A?dataset=gnomad_r2_1

Case: Patient 10 (Family D), female, 24yo

DiseaseAssertion: VWD2B

FamilyInfo: NA

ParentalGenotype(s): NA

CasePresentingHPOs: HP:0030129, HP:0030130, HP:0011871, HP:0012147

(impaired ristocetin cofactor assay activity, Impaired von Willebrand factor collagen binding activity, Impaired RIPA, Reduced quantity of VWF)

CaseHPOFreeText: blood group AB, Platelet count x103/μl = 231, PFA 100 sec = 223, SPA % = 6, MADR mg/mL = 0.75, VWF:Ag U/dL = 46.0, VWF:CB U/dL = 49.1, VWF:CB ratio = 1.07, VWF:RCo U/dL = 40.5, VWF:RCo ratio = 0.83, Platelet VWF U/dL = 55.6, FVIII U/dL = 64.8, BAT F/M = NA

CaseNOTHPOs:

CaseNOTHPOFreeText:

CasePreviousTesting: NA

GenotypingMethod: Sequencing of exon 28 of VWF

PreviouslyPublished: NA

Gene: VWF

Variant: c.4135C>T (p.Arg1379Cys)

HGVS: NM_000552.4(VWF):c.4135C>T (p.Arg1379Cys)

ClinVarID: 100333; https://www.ncbi.nlm.nih.gov/clinvar/variation/100333/

CAID: NA

gnomAD: MAF = 0.00003891 (5/128500 alleles) in the European (non-Finnish) population; https://gnomad.broadinstitute.org/variant/12-6128449-G-A?dataset=gnomad_r2_1

Case: Patient 11 (Family E), male, 11yo

DiseaseAssertion: VWD2B

FamilyInfo: NA

ParentalGenotype(s): NA

CasePresentingHPOs: HP:0030129, HP:0030130, HP:0011871, HP:0012147

(impaired ristocetin cofactor assay activity, Impaired von Willebrand factor collagen binding activity, Impaired RIPA, Reduced quantity of VWF)

CaseHPOFreeText: blood group O, Platelet count x103/μl = 220, PFA 100 sec = >300, SPA % = 11.3, MADR mg/mL = 0.3, VWF:Ag U/dL = 28.8, VWF:CB U/dL = 28.4, VWF:CB ratio = 0.98, VWF:RCo U/dL = 23.0, VWF:RCo ratio = 0.80, Platelet VWF U/dL = 54.1, FVIII U/dL = 51.5, BAT F/M = 5

CaseNOTHPOs:

CaseNOTHPOFreeText:

CasePreviousTesting: NA

GenotypingMethod: Sequencing of exon 28 of VWF

PreviouslyPublished: NA

Gene: VWF

Variant: c.4021C>T (p.Arg1341Trp)

HGVS: NM_000552.4(VWF):c.4021C>T (p.Arg1341Trp)

ClinVarID: 100316; https://www.ncbi.nlm.nih.gov/clinvar/variation/100316/

CAID: NA

gnomAD: MAF = 0.00008674 (3/34586 alleles) in the Latino/Admixed American population; https://gnomad.broadinstitute.org/variant/12-6128563-G-A?dataset=gnomad_r2_1

Case: Patient 12 (Family F), female, 50yo

DiseaseAssertion: VWD2B

FamilyInfo: NA

ParentalGenotype(s): NA

CasePresentingHPOs: HP:0030129, HP:0030130, HP:0011871, HP:0012147

(impaired ristocetin cofactor assay activity, Impaired von Willebrand factor collagen binding activity, Impaired RIPA, Reduced quantity of VWF)

CaseHPOFreeText: blood group A, Platelet count x103/μl = 204, PFA 100 sec = 91, SPA % = 10, MADR mg/mL = 0.45, VWF:Ag U/dL = 108, VWF:CB U/dL = 101.9, VWF:CB ratio = 0.94, VWF:RCo U/dL = 97.7, VWF:RCo ratio = 0.91, Platelet VWF U/dL = 136.4, FVIII U/dL = 107.5, BAT F/M = 16

CaseNOTHPOs:

CaseNOTHPOFreeText:

CasePreviousTesting: NA

GenotypingMethod: Sequencing of exon 28 of VWF

PreviouslyPublished: NA

Gene: VWF

Variant: c.3797C>A (p.Pro1266Gln)

HGVS: NM_000552.4(VWF):c.3797C>A (p.Pro1266Gln)

ClinVarID: 100279; https://www.ncbi.nlm.nih.gov/clinvar/variation/100279/

CAID: NA

gnomAD: MAF = 0.001346 (41/30468 alleles; 2 homozygotes) in the South Asian population; https://gnomad.broadinstitute.org/variant/12-6128787-G-T?dataset=gnomad_r2_1

Case: Patient 5 (Family C), female, 50yo

DiseaseAssertion: VWD2B

FamilyInfo: NA

ParentalGenotype(s): NA

CasePresentingHPOs: HP:0030129, HP:0030130, HP:0011871, HP:0012147

(impaired ristocetin cofactor assay activity, Impaired von Willebrand factor collagen binding activity, Impaired RIPA, Reduced quantity of VWF)

CaseHPOFreeText: blood group O, Platelet count x103/μl = 211, PFA 100 sec = 210, SPA % = 10, MADR mg/mL = 0.6, VWF:Ag U/dL = 40.6, VWF:CB U/dL = 42.8, VWF:CB ratio = 1.1, VWF:RCo U/dL = NA, VWF:RCo ratio = NA, Platelet VWF U/dL = 41.2, FVIII U/dL = 62.8, BAT F/M = 8

CaseNOTHPOs:

CaseNOTHPOFreeText:

CasePreviousTesting: NA

GenotypingMethod: Sequencing of exon 28 of VWF

PreviouslyPublished: NA

Gene: VWF

Variant: c.4135C>T (p.Arg1379Cys)

HGVS: NM_000552.4(VWF):c.4135C>T (p.Arg1379Cys)

ClinVarID: 100333; https://www.ncbi.nlm.nih.gov/clinvar/variation/100333/

CAID: NA

gnomAD: MAF = 0.00003891 (5/128500 alleles) in the European (non-Finnish) population; https://gnomad.broadinstitute.org/variant/12-6128449-G-A?dataset=gnomad_r2_1

Case: Patient 7 (Family C), female, 21yo

DiseaseAssertion: VWD2B

FamilyInfo: NA

ParentalGenotype(s): NA

CasePresentingHPOs: HP:0030129, HP:0030130, HP:0011871, HP:0012147

(impaired ristocetin cofactor assay activity, Impaired von Willebrand factor collagen binding activity, Impaired RIPA, Reduced quantity of VWF)

CaseHPOFreeText: blood group O, Platelet count x103/μl = 293, PFA 100 sec = >300, SPA % = 6, MADR mg/mL = 0.6, VWF:Ag U/dL = 26.7, VWF:CB U/dL = 27.2, VWF:CB ratio = 1.01, VWF:RCo U/dL = 25.8, VWF:RCo ratio = 0.92, Platelet VWF U/dL = 59.1, FVIII U/dL = 40.9, BAT F/M = 6

CaseNOTHPOs:

CaseNOTHPOFreeText:

CasePreviousTesting: NA

GenotypingMethod: Sequencing of exon 28 of VWF

PreviouslyPublished: NA

Gene: VWF

Variant: c.4135C>T (p.Arg1379Cys)

HGVS: NM_000552.4(VWF):c.4135C>T (p.Arg1379Cys)

ClinVarID: 100333; https://www.ncbi.nlm.nih.gov/clinvar/variation/100333/

CAID: NA

gnomAD: MAF = 0.00003891 (5/128500 alleles) in the European (non-Finnish) population; https://gnomad.broadinstitute.org/variant/12-6128449-G-A?dataset=gnomad_r2_1

Case: Patient 9 (Family D), male, 18yo

DiseaseAssertion: VWD2B

FamilyInfo: NA

ParentalGenotype(s): NA

CasePresentingHPOs: HP:0030129, HP:0030130, HP:0011871, HP:0012147

(impaired ristocetin cofactor assay activity, Impaired von Willebrand factor collagen binding activity, Impaired RIPA, Reduced quantity of VWF)

CaseHPOFreeText: blood group A, Platelet count x103/μl = 231, PFA 100 sec = 95, SPA % = 0, MADR mg/mL = 0.75, VWF:Ag U/dL = 151.1, VWF:CB U/dL = 175.5, VWF:CB ratio = 1.16, VWF:RCo U/dL = 153.5, VWF:RCo ratio = 0.87, Platelet VWF U/dL = 95.2, FVIII U/dL = 148.2, BAT F/M = NA

CaseNOTHPOs:

CaseNOTHPOFreeText:

CasePreviousTesting: NA

GenotypingMethod: Sequencing of exon 28 of VWF

PreviouslyPublished: NA

Gene: VWF

Variant: c.3797C>A (p.Pro1266Gln)

HGVS: NM_000552.4(VWF):c.3797C>A (p.Pro1266Gln)

ClinVarID: 100279; https://www.ncbi.nlm.nih.gov/clinvar/variation/100279/

CAID: NA

gnomAD: MAF = 0.001346 (41/30468 alleles; 2 homozygotes) in the South Asian population; https://gnomad.broadinstitute.org/variant/12-6128787-G-T?dataset=gnomad_r2_1

Case: Patient 4 (Family C), female, 76yo

DiseaseAssertion: VWD2B

FamilyInfo: NA

ParentalGenotype(s): NA

CasePresentingHPOs: HP:0030129, HP:0030130, HP:0011871, HP:0012147

(impaired ristocetin cofactor assay activity, Impaired von Willebrand factor collagen binding activity, Impaired RIPA, Reduced quantity of VWF)

CaseHPOFreeText: blood group O, Platelet count x103/μl = 236, PFA 100 sec = >300, SPA % = 8.4, MADR mg/mL = 0.75, VWF:Ag U/dL = 35.4, VWF:CB U/dL = 37.3, VWF:CB ratio = 1.05, VWF:RCo U/dL = 25.4, VWF:RCo ratio = 0.72, Platelet VWF U/dL = 66.8, FVIII U/dL = 32.0, BAT F/M = 16

CaseNOTHPOs:

CaseNOTHPOFreeText:

CasePreviousTesting: NA

GenotypingMethod: Sequencing of exon 28 of VWF

PreviouslyPublished: NA

Gene: VWF

Variant: c.4135C>T (p.Arg1379Cys)

HGVS: NM_000552.4(VWF):c.4135C>T (p.Arg1379Cys)

ClinVarID: 100333; https://www.ncbi.nlm.nih.gov/clinvar/variation/100333/

CAID: NA

gnomAD: MAF = 0.00003891 (5/128500 alleles) in the European (non-Finnish) population; https://gnomad.broadinstitute.org/variant/12-6128449-G-A?dataset=gnomad_r2_1

Case: P53, Chinese, female, 70yo

DiseaseAssertion: VWD2A

FamilyInfo: NA

ParentalGenotype(s): NA

CasePresentingHPOs:

CaseHPOFreeText: HP:0012147

(Reduced quantity of VWF)

CaseNOTHPOs: type O blood, bleeding score = 9; VWF:Ag (%) = 32.1, VWF:RCo (%) = 6.0, FVIII:C (%) = 50.8, VWF:CB (%) = 14.7, VWFpp (%) = 56.9, VWF:RCo/VWF:Ag = 0.2, VWF:CB/VWF:Ag = 0.5, VWFpp/VWF:Ag = 1.8, FVIII:C/VWF:Ag = 1.6

CaseNOTHPOFreeText:

CasePreviousTesting: NA

GenotypingMethod: NGS

PreviouslyPublished: NA

Gene: VWF

Variant: c.4790G>T (p.Arg1597Leu)

HGVS: NM_000552.4(VWF):c.4790G>T (p.Arg1597Leu)

ClinVarID: 100392; https://www.ncbi.nlm.nih.gov/clinvar/variation/100392/

CAID: NA

gnomAD: absent

Case: P19, Chinese, male, 35yo

DiseaseAssertion: VWD2M

FamilyInfo: NA

ParentalGenotype(s): NA

CasePresentingHPOs: HP:0012147

(Reduced quantity of VWF)

CaseHPOFreeText: type O blood, bleeding score = 4; VWF:Ag (%) = 17.5, VWF:RCo (%) = 5.1, FVIII:C (%) = 35.9, VWF:CB (%) = 16.9, VWFpp (%) = 45.3, VWF:RCo/VWF:Ag = 0.3, VWF:CB/VWF:Ag = 1.0, VWFpp/VWF:Ag = 2.6, FVIII:C/VWF:Ag = 2.1

CaseNOTHPOs:

CaseNOTHPOFreeText:

CasePreviousTesting: NA

GenotypingMethod: NGS

PreviouslyPublished: NA

Gene: VWF

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

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

ClinVarID: 100329; https://www.ncbi.nlm.nih.gov/clinvar/variation/100329/

CAID: NA

gnomAD: MAF = 0.000008824 (1/113322 alleles) in the European (non-Finnish) population; https://gnomad.broadinstitute.org/variant/12-6128464-G-A?dataset=gnomad_r2_1

Case: P60, Chinese, female, 8yo

DiseaseAssertion: VWD1

FamilyInfo: NA

ParentalGenotype(s): NA

CasePresentingHPOs: HP:0012147

(Reduced quantity of VWF)

CaseHPOFreeText: type O blood, bleeding score = 3; VWF:Ag (%) = 37.8, VWF:RCo (%) = 35.2, FVIII:C (%) = 56.6, VWF:CB (%) = 38.6, VWFpp (%) = 41.1, VWF:RCo/VWF:Ag = 0.9, VWF:CB/VWF:Ag = 1.0, VWFpp/VWF:Ag = 1.1, FVIII:C/VWF:Ag = 1.5

CaseNOTHPOs:

CaseNOTHPOFreeText:

CasePreviousTesting: NA

GenotypingMethod: NGS

PreviouslyPublished: NA

Gene: VWF

Variant: c.5546T>C (p.Ile1849Thr)

HGVS: NM_000038.6(APC):c.5546T>C (p.Ile1849Thr)

ClinVarID: 654506; https://www.ncbi.nlm.nih.gov/clinvar/variation/654506/

CAID: NA

gnomAD: absent

Case: P46, Chinese, female, 55yo

DiseaseAssertion: VWD1

FamilyInfo: NA

ParentalGenotype(s): NA

CasePresentingHPOs: HP:0012147

(Reduced quantity of VWF)

CaseHPOFreeText: type O blood, bleeding score = 14; VWF:Ag (%) = 45.4, VWF:RCo (%) = 44.5, FVIII:C (%) = 76.9, VWF:CB (%) = 54.9, VWFpp (%) = 43.2, VWF:RCo/VWF:Ag = 1.0, VWF:CB/VWF:Ag = 1.2, VWFpp/VWF:Ag = 1.0, FVIII:C/VWF:Ag = 1.7

CaseNOTHPOs:

CaseNOTHPOFreeText:

CasePreviousTesting: NA

GenotypingMethod: NGS

PreviouslyPublished: NA

Gene: VWF

Variant: c.4414G>C (p.Asp1472His)

HGVS: NM_000552.4(VWF):c.4414G>C (p.Asp1472His)

ClinVarID: 256679; https://www.ncbi.nlm.nih.gov/clinvar/variation/256679/

CAID: NA

gnomAD: MAF = 0.5077 (12039/23712 alleles; 3078 homozygotes) in African/African-American population; https://gnomad.broadinstitute.org/variant/12-6128170-C-G?dataset=gnomad_r2_1

Case: P12, Chinese, Male, 11yo

DiseaseAssertion: VWD2A

FamilyInfo: NA

ParentalGenotype(s): NA

CasePresentingHPOs: HP:0012147

(Reduced quantity of VWF)

CaseHPOFreeText: type O blood, bleeding score = 6; VWF:Ag (%) = 15.8, VWF:RCo (%) = 7.3, FVIII:C (%) = 45.5, VWF:CB (%) = 4.1, VWFpp (%) = 50.3, VWF:RCo/VWF:Ag = 0.5, VWF:CB/VWF:Ag = 0.3, VWFpp/VWF:Ag = 3.2, FVIII:C/VWF:Ag = 2.9

CaseNOTHPOs:

CaseNOTHPOFreeText:

CasePreviousTesting: NA

GenotypingMethod: NGS

PreviouslyPublished: NA

Gene: VWF

Variant: c.4517C>T (p.Ser1506Leu)

HGVS: NM_000552.4(VWF):c.4517C>T (p.Ser1506Leu)

ClinVarID: 100369; https://www.ncbi.nlm.nih.gov/clinvar/variation/100369/

CAID: NA

gnomAD: absent

Case: P70, Chinese, female, 51yo

DiseaseAssertion: VWD1

FamilyInfo: NA

ParentalGenotype(s): NA

CasePresentingHPOs: HP:0012147

(Reduced quantity of VWF)

CaseHPOFreeText: type O blood, bleeding score = 2; VWF:Ag (%) = 28.3, VWF:RCo (%) = 36.6, FVIII:C (%) = 61.3, VWF:CB (%) = 34.6, VWFpp (%) = 64.5, VWF:RCo/VWF:Ag = 1.3, VWF:CB/VWF:Ag = 1.2, VWFpp/VWF:Ag = 2.3, FVIII:C/VWF:Ag = 2.2

CaseNOTHPOs:

CaseNOTHPOFreeText:

CasePreviousTesting: NA

GenotypingMethod: NGS

PreviouslyPublished: NA

Gene: VWF

Variant: c.6554G>A (p.Arg2185Gln)

HGVS: NM_000552.4(VWF):c.6554G>A (p.Arg2185Gln)

ClinVarID: 100443; https://www.ncbi.nlm.nih.gov/clinvar/variation/100443/

CAID: NA

gnomAD: MAF = 0.1899 (4712/24810 alleles; 465 homozygotes) in the African/African-American population; https://gnomad.broadinstitute.org/variant/12-6103072-C-T?dataset=gnomad_r2_1

Case: P56, Chinese, male, 6yo

DiseaseAssertion: VWD2A

FamilyInfo: NA

ParentalGenotype(s): NA

CasePresentingHPOs: HP:0012147

(Reduced quantity of VWF)

CaseHPOFreeText: type B blood, bleeding score = 13; VWF:Ag (%) = 15.4, VWF:RCo (%) = 7.1, FVIII:C (%) = 43.3, VWF:CB (%) = 2.4, VWFpp (%) = 43.8, VWF:RCo/VWF:Ag = 0.5, VWF:CB/VWF:Ag = 0.2, VWFpp/VWF:Ag = 2.8, FVIII:C/VWF:Ag = 2.8

CaseNOTHPOs:

CaseNOTHPOFreeText:

CasePreviousTesting: NA

GenotypingMethod: NGS

PreviouslyPublished: NA

Gene: VWF

Variant: c.4517C>T (p.Ser1506Leu)

HGVS: NM_000552.4(VWF):c.4517C>T (p.Ser1506Leu)

ClinVarID: 100369; https://www.ncbi.nlm.nih.gov/clinvar/variation/100369/

CAID: NA

gnomAD: absent

Case: P25, Chinese, female, 23yo

DiseaseAssertion: VWD2M

FamilyInfo: NA

ParentalGenotype(s): NA

CasePresentingHPOs: HP:0012147

(Reduced quantity of VWF)

CaseHPOFreeText: type B blood, bleeding score = 5; VWF:Ag (%) = 15.6, VWF:RCo (%) = 4.4, FVIII:C (%) = 31.0, VWF:CB (%) = 14.2, VWFpp (%) = 47.7, VWF:RCo/VWF:Ag = 0.3, VWF:CB/VWF:Ag = 0.9, VWFpp/VWF:Ag = 3.1, FVIII:C/VWF:Ag = 2.0

CaseNOTHPOs:

CaseNOTHPOFreeText:

CasePreviousTesting: NA

GenotypingMethod: NGS

PreviouslyPublished: NA

Gene: VWF

Variant: c.4121G>A (p.Arg1374His)

HGVS: NM_000552.4(VWF):c.4121G>A (p.Arg1374His)

ClinVarID: 100330; https://www.ncbi.nlm.nih.gov/clinvar/variation/100330/

CAID: NA

gnomAD: absent

Case: P14, Chinese, female, 3yo

DiseaseAssertion: VWD2M

FamilyInfo: NA

ParentalGenotype(s): NA

CasePresentingHPOs: HP:0012147

(Reduced quantity of VWF)

CaseHPOFreeText: type O blood, bleeding score = 5; VWF:Ag (%) = 18.2, VWF:RCo (%) = 7.6, FVIII:C (%) = 29.1, VWF:CB (%) = 17.5, VWFpp (%) = 45.6, VWF:RCo/VWF:Ag = 0.4, VWF:CB/VWF:Ag = 1.0, VWFpp/VWF:Ag = 2.5, FVIII:C/VWF:Ag = 1.6

CaseNOTHPOs:

CaseNOTHPOFreeText:

CasePreviousTesting: NA

GenotypingMethod: NGS

PreviouslyPublished: NA

Gene: VWF

Variant: c.4121G>A (p.Arg1374His)

HGVS: NM_000552.4(VWF):c.4121G>A (p.Arg1374His)

ClinVarID: 100330; https://www.ncbi.nlm.nih.gov/clinvar/variation/100330/

CAID: NA

gnomAD: absent

Case: P84, Chinese, female, 36yo

DiseaseAssertion: VWD2M

FamilyInfo: NA

ParentalGenotype(s): NA

CasePresentingHPOs: HP:0012147

(Reduced quantity of VWF)

CaseHPOFreeText: type B blood, bleeding score = 5; VWF:Ag (%) = 26.6, VWF:RCo (%) = 3.3, FVIII:C (%) = 41.7, VWF:CB (%) = 25.2, VWFpp (%) = 41.1, VWF:RCo/VWF:Ag = 0.1, VWF:CB/VWF:Ag = 1.0, VWFpp/VWF:Ag = 1.6, FVIII:C/VWF:Ag = 1.6

CaseNOTHPOs:

CaseNOTHPOFreeText:

CasePreviousTesting: NA

GenotypingMethod: NGS

PreviouslyPublished: NA

Gene: VWF

Variant: c.4121G>A (p.Arg1374His)

HGVS: NM_000552.4(VWF):c.4121G>A (p.Arg1374His)

ClinVarID: 100330; https://www.ncbi.nlm.nih.gov/clinvar/variation/100330/

CAID: NA

gnomAD: absent

Case: P13, Chinese, male, 8yo

DiseaseAssertion: VWD2A

FamilyInfo: NA

ParentalGenotype(s): NA

CasePresentingHPOs: HP:0012147

(Reduced quantity of VWF)

CaseHPOFreeText: type O blood, bleeding score = 5; VWF:Ag (%) = 21.6, VWF:RCo (%) = 9.4, FVIII:C (%) = 53.2, VWF:CB (%) = 5.7, VWFpp (%) = 56.1, VWF:RCo/VWF:Ag = 0.4, VWF:CB/VWF:Ag = 0.3, VWFpp/VWF:Ag = 2.6, FVIII:C/VWF:Ag = 2.5

CaseNOTHPOs:

CaseNOTHPOFreeText:

CasePreviousTesting: NA

GenotypingMethod: NGS

PreviouslyPublished: NA

Gene: VWF

Variant: c.4517C>T (p.Ser1506Leu)

HGVS: NM_000552.4(VWF):c.4517C>T (p.Ser1506Leu)

ClinVarID: 100369; https://www.ncbi.nlm.nih.gov/clinvar/variation/100369/

CAID: NA

gnomAD: absent

Case: P65, Chinese, female, 24yo

DiseaseAssertion: VWD1

FamilyInfo: NA

ParentalGenotype(s): NA

CasePresentingHPOs: HP:0012147

(Reduced quantity of VWF)

CaseHPOFreeText: type O blood, bleeding score = 3; VWF:Ag (%) = 55.7, VWF:RCo (%) = 51.2, FVIII:C (%) = 72.7, VWF:CB (%) = 57.1, VWFpp (%) = 61.7, VWF:RCo/VWF:Ag = 0.9, VWF:CB/VWF:Ag = 1.0, VWFpp/VWF:Ag = 1.1, FVIII:C/VWF:Ag = 1.3

CaseNOTHPOs:

CaseNOTHPOFreeText:

CasePreviousTesting: NA

GenotypingMethod: NGS

PreviouslyPublished: NA

Gene: VWF

Variant: c.4499C>T (p.Ala1500Val)

HGVS: NM_000552.4(VWF):c.4499C>T (p.Ala1500Val)

ClinVarID: 439339; https://www.ncbi.nlm.nih.gov/clinvar/variation/439339/

CAID: NA

gnomAD: MAF = 0.003511 (70/19938 alleles) in the East Asain population; https://gnomad.broadinstitute.org/variant/12-6128085-G-A?dataset=gnomad_r2_1

Case: P4, Chinese, male, 35yo

DiseaseAssertion: VWD2M

FamilyInfo: NA

ParentalGenotype(s): NA

CasePresentingHPOs: HP:0012147

(Reduced quantity of VWF)

CaseHPOFreeText: type A blood, bleeding score = 6; VWF:Ag (%) = 22.6, VWF:RCo (%) = 5.0, FVIII:C (%) = 36.6, VWF:CB (%) = 25.4, VWFpp (%) = 43.0, VWF:RCo/VWF:Ag = 0.2, VWF:CB/VWF:Ag = 1.1, VWFpp/VWF:Ag = 1.9, FVIII:C/VWF:Ag = 1.6

CaseNOTHPOs:

CaseNOTHPOFreeText:

CasePreviousTesting: NA

GenotypingMethod: NGS

PreviouslyPublished: NA

Gene: VWF

Variant: c.4121G>A (p.Arg1374His)

HGVS: NM_000552.4(VWF):c.4121G>A (p.Arg1374His)

ClinVarID: 100330; https://www.ncbi.nlm.nih.gov/clinvar/variation/100330/

CAID: NA

gnomAD: absent

Case: P45, Chinese, female, 8yo

DiseaseAssertion: VWD2A

FamilyInfo: NA

ParentalGenotype(s): NA

CasePresentingHPOs: HP:0012147

(Reduced quantity of VWF)

CaseHPOFreeText: type A blood, bleeding score = 4; VWF:Ag (%) = 23.5, VWF:RCo (%) = 6.5, FVIII:C (%) = 40.0, VWF:CB (%) = 15.2, VWFpp (%) = 44.5, VWF:RCo/VWF:Ag = 0.3, VWF:CB/VWF:Ag = 0.6, VWFpp/VWF:Ag = 1.9, FVIII:C/VWF:Ag = 1.7

CaseNOTHPOs:

CaseNOTHPOFreeText:

CasePreviousTesting: NA

GenotypingMethod: NGS

PreviouslyPublished: NA

Gene: VWF

Variant: c.4628C>T (p.Ser1543Phe)

HGVS: NM_000552.4(VWF):c.4628C>T (p.Ser1543Phe)

ClinVarID: 100376; https://www.ncbi.nlm.nih.gov/clinvar/variation/100376/

CAID: NA

gnomAD: absent

Case: P52, Chinese, female, 55yo

DiseaseAssertion: VWD2M

FamilyInfo: NA

ParentalGenotype(s): NA

CasePresentingHPOs: HP:0012147

(Reduced quantity of VWF)

CaseHPOFreeText: type O blood, bleeding score = 9; VWF:Ag (%) = 19.1, VWF:RCo (%) = 3.1, FVIII:C (%) = 35.8, VWF:CB (%) = 22.7, VWFpp (%) = 50.3, VWF:RCo/VWF:Ag = 0.2, VWF:CB/VWF:Ag = 1.2, VWFpp/VWF:Ag = 2.6, FVIII:C/VWF:Ag = 1.9

CaseNOTHPOs:

CaseNOTHPOFreeText:

CasePreviousTesting: NA

GenotypingMethod: NGS

PreviouslyPublished: NA

Gene: VWF

Variant: c.4120C>A (p.Arg1374Ser)

HGVS: NM_000552.4(VWF):c.4120C>A (p.Arg1374Ser)

ClinVarID: 100328; https://www.ncbi.nlm.nih.gov/clinvar/variation/100328/

CAID: NA

gnomAD: absent

Case: P26, Chinese, female, 30yo

DiseaseAssertion: VWD2A

FamilyInfo: NA

ParentalGenotype(s): NA

CasePresentingHPOs: HP:0012147

(Reduced quantity of VWF)

CaseHPOFreeText: type AB blood, bleeding score = 14; VWF:Ag (%) = 11.3, VWF:RCo (%) = 3.7, FVIII:C (%) = 45.0, VWF:CB (%) = 4.0, VWFpp (%) = 41.7, VWF:RCo/VWF:Ag = 0.3, VWF:CB/VWF:Ag = 0.4, VWFpp/VWF:Ag = 3.7, FVIII:C/VWF:Ag = 4.0

CaseNOTHPOs:

CaseNOTHPOFreeText:

CasePreviousTesting: NA

GenotypingMethod: NGS

PreviouslyPublished: NA

Gene: VWF

Variant: c.4517C>T (p.Ser1506Leu)

HGVS: NM_000552.4(VWF):c.4517C>T (p.Ser1506Leu)

ClinVarID: 100369; https://www.ncbi.nlm.nih.gov/clinvar/variation/100369/

CAID: NA

gnomAD: absent

Case: P3, Chinese, male, 7yo

DiseaseAssertion: VWD2A

FamilyInfo: NA

ParentalGenotype(s): NA

CasePresentingHPOs: HP:0012147

(Reduced quantity of VWF)

CaseHPOFreeText: type B blood, bleeding score = 5; VWF:Ag (%) = 13.0, VWF:RCo (%) = 5.0, FVIII:C (%) = 39.1, VWF:CB (%) = 3.5, VWFpp (%) = 46.8, VWF:RCo/VWF:Ag = 0.4, VWF:CB/VWF:Ag = 0.3, VWFpp/VWF:Ag = 3.6, FVIII:C/VWF:Ag = 3.0

CaseNOTHPOs:

CaseNOTHPOFreeText:

CasePreviousTesting: NA

GenotypingMethod: NGS

PreviouslyPublished: NA

Gene: VWF

Variant: c.4517C>T (p.Ser1506Leu)

HGVS: NM_000552.4(VWF):c.4517C>T (p.Ser1506Leu)

ClinVarID: 100369; https://www.ncbi.nlm.nih.gov/clinvar/variation/100369/

CAID: NA

gnomAD: absent

Case: P58, Chinese, female, 27yo

DiseaseAssertion: VWD2A

FamilyInfo: NA

ParentalGenotype(s): NA

CasePresentingHPOs:

CaseHPOFreeText: HP:0012147

(Reduced quantity of VWF)

CaseNOTHPOs: type O blood, bleeding score = 8; VWF:Ag (%) = 17.1, VWF:RCo (%) = 2.3, FVIII:C (%) = 44.3, VWF:CB (%) = 3.7, VWFpp (%) = 41.1, VWF:RCo/VWF:Ag = 0.1, VWF:CB/VWF:Ag = 0.2, VWFpp/VWF:Ag = 2.4, FVIII:C/VWF:Ag = 2.6

CaseNOTHPOFreeText:

CasePreviousTesting: NA

GenotypingMethod: NGS

PreviouslyPublished: NA

Gene: VWF

Variant: c.4790G>T (p.Arg1597Leu)

HGVS: NM_000552.4(VWF):c.4790G>T (p.Arg1597Leu)

ClinVarID: 100392; https://www.ncbi.nlm.nih.gov/clinvar/variation/100392/

CAID: NA

gnomAD: absent

Case: P17, Chinese, female, 20yo

DiseaseAssertion: VWD2M

FamilyInfo: NA

ParentalGenotype(s): NA

CasePresentingHPOs: HP:0012147

(Reduced quantity of VWF)

CaseHPOFreeText: type B blood, bleeding score = 10; VWF:Ag (%) = 17.6, VWF:RCo (%) = 7.3, FVIII:C (%) = 35.8, VWF:CB (%) = 15.8, VWFpp (%) = 51.9, VWF:RCo/VWF:Ag = 0.4, VWF:CB/VWF:Ag = 0.9, VWFpp/VWF:Ag = 2.9, FVIII:C/VWF:Ag = 2.0

CaseNOTHPOs:

CaseNOTHPOFreeText:

CasePreviousTesting: NA

GenotypingMethod: NGS

PreviouslyPublished: NA

Gene: VWF

Variant: c.3389G>T (p.Cys1130Phe)

HGVS: NM_000552.4(VWF):c.3389G>T (p.Cys1130Phe)

ClinVarID: 100261; https://www.ncbi.nlm.nih.gov/clinvar/variation/100261/

CAID: NA

gnomAD: absent

Case: P66, Chinese, male, 27yo

DiseaseAssertion: VWD1

FamilyInfo: NA

ParentalGenotype(s): NA

CasePresentingHPOs: HP:0012147

(Reduced quantity of VWF)

CaseHPOFreeText: type O blood, bleeding score = 1; VWF:Ag (%) = 56.0, VWF:RCo (%) = 43.4, FVIII:C (%) = 70.0, VWF:CB (%) = 58.9, VWFpp (%) = 53.6, VWF:RCo/VWF:Ag = 0.8, VWF:CB/VWF:Ag = 1.1, VWFpp/VWF:Ag = 1.0, FVIII:C/VWF:Ag = 1.3

CaseNOTHPOs:

CaseNOTHPOFreeText:

CasePreviousTesting: NA

GenotypingMethod: NGS

PreviouslyPublished: NA

Gene: VWF

Variant: c.4414G>C (p.Asp1472His)

HGVS: NM_000552.4(VWF):c.4414G>C (p.Asp1472His)

ClinVarID: 256679; https://www.ncbi.nlm.nih.gov/clinvar/variation/256679/

CAID: NA

gnomAD: MAF = 0.5077 (12039/23712 alleles; 3078 homozygotes) in African/African-American population; https://gnomad.broadinstitute.org/variant/12-6128170-C-G?dataset=gnomad_r2_1

Case: P32, Chinese, male, 3yo

DiseaseAssertion: VWD1

FamilyInfo: NA

ParentalGenotype(s): NA

CasePresentingHPOs: HP:0012147

(Reduced quantity of VWF)

CaseHPOFreeText: patient coexisting with severe haemophilia A; type O blood, bleeding score = 3; VWF:Ag (%) = 51.3, VWF:RCo (%) = 50.7, FVIII:C (%) = 1.0, VWF:CB (%) = 50.3, VWFpp (%) = 49.2, VWF:RCo/VWF:Ag = 1.0, VWF:CB/VWF:Ag = 1.0, VWFpp/VWF:Ag = 1.0, FVIII:C/VWF:Ag = 0.0

CaseNOTHPOs:

CaseNOTHPOFreeText:

CasePreviousTesting: NA

GenotypingMethod: NGS

PreviouslyPublished: NA

Gene: VWF

Variant: c.646G>A (p.Glu216Lys)

HGVS: NM_000552.5(VWF):c.646G>A (p.Glu216Lys)

ClinVarID: 870925; https://www.ncbi.nlm.nih.gov/clinvar/variation/870925/

CAID: NA

gnomAD: 0.001353 (27/19952 alleles) in East Asian population; https://gnomad.broadinstitute.org/variant/12-6204637-C-T?dataset=gnomad_r2_1

MultipleGeneVariants

Gene: F8

Variant: p.Arg446Ter - Unregistered Variant

Case: IP #28 (also in row 14 table 6)

DiseaseAssertion: type 2A VWF

FamilyInfo:

ParentalGenotype(s):

CasePresentingHPOs: HP:0032370 - blood group A

CaseHPOFreeRext: VWF:Ag [IU/dL] = 39; VWF:Ac [IU/dL] = 7; FVIII:C [IU/dL] = 61

CaseNOTHPOs:

CaseNOTHPOFreeText:

CasePreviousTesting:

GenotypingMethod: Sequence variations in the VWF were identified through direct sequencing, while the detection of large rearrangements utilized the multiplex ligation-dependent probe amplification (MLPA) technique or assessed copy number variations (CNVs). Sanger sequencing methodology was applied for VWF analysis in patients investigated prior to 2016 (approximately 14% of IPs), whereas next-generation sequencing (NGS) was employed for the analysis of VWF in patients inspected from 2017 onwards (about 86% of IPs). In both strategies, the entire VWF, encompassing all 52 exons, intron/exon boundaries, and promoter regions, was analyzed.

PreviouslyPublished:

Gene: VWF

Variant: c.4078G>T p.Val1360Phe

HGVS:

ClinVarID: 1809581

CAID:

gnomAD: absent

Case: IP #9

DiseaseAssertion: type 2A VWD

FamilyInfo:

ParentalGenotype(s):

CasePresentingHPOs:

CaseHPOFreeRext: VWF:Ag [IU/dL] = 17; VWF:Ac [IU/dL] = 10; FVIII:C [IU/dL] = 51

CaseNOTHPOs:

CaseNOTHPOFreeText:

CasePreviousTesting:

GenotypingMethod: Sequence variations in the VWF were identified through direct sequencing, while the detection of large rearrangements utilized the multiplex ligation-dependent probe amplification (MLPA) technique or assessed copy number variations (CNVs). Sanger sequencing methodology was applied for VWF analysis in patients investigated prior to 2016 (approximately 14% of IPs), whereas next-generation sequencing (NGS) was employed for the analysis of VWF in patients inspected from 2017 onwards (about 86% of IPs). In both strategies, the entire VWF, encompassing all 52 exons, intron/exon boundaries, and promoter regions, was analyzed.

PreviouslyPublished:

Gene: VWF

Variant: c.2771G>A p.Arg924Gln

HGVS:

ClinVarID: 100240

CAID:

gnomAD: 0.01774 (28642 in 1614096) 297 homozygotes https://gnomad.broadinstitute.org/variant/12-6031493-C-T?dataset=gnomad_r4

Case: IP #34

DiseaseAssertion: type 2A (IIA) VWD

FamilyInfo:

ParentalGenotype(s):

CasePresentingHPOs:

CaseHPOFreeRext: VWF:Ag [IU/dL] = 55; VWF:Ac [IU/dL] = 35; FVIII:C [IU/dL] = 60

CaseNOTHPOs:

CaseNOTHPOFreeText:

CasePreviousTesting:

GenotypingMethod: Sequence variations in the VWF were identified through direct sequencing, while the detection of large rearrangements utilized the multiplex ligation-dependent probe amplification (MLPA) technique or assessed copy number variations (CNVs). Sanger sequencing methodology was applied for VWF analysis in patients investigated prior to 2016 (approximately 14% of IPs), whereas next-generation sequencing (NGS) was employed for the analysis of VWF in patients inspected from 2017 onwards (about 86% of IPs). In both strategies, the entire VWF, encompassing all 52 exons, intron/exon boundaries, and promoter regions, was analyzed.

PreviouslyPublished:

Gene: VWF

Variant: c.4517C>T p.Ser1506Leu

HGVS:

ClinVarID: 100369

CAID:

gnomAD: absent

Case: IP #26 in type 2N VWD sub-cohort

DiseaseAssertion: type 2N VWD

FamilyInfo:

ParentalGenotype(s):

CasePresentingHPOs: HP:0032442 - blood group O

CaseHPOFreeRext: VWF:Ag [IU/dL] = 50; VWF:Ac [IU/dL] = 29; FVIII:C [IU/dL] = 40

CaseNOTHPOs:

CaseNOTHPOFreeText:

CasePreviousTesting:

GenotypingMethod: Sequence variations in the VWF were identified through direct sequencing, while the detection of large rearrangements utilized the multiplex ligation-dependent probe amplification (MLPA) technique or assessed copy number variations (CNVs). Sanger sequencing methodology was applied for VWF analysis in patients investigated prior to 2016 (approximately 14% of IPs), whereas next-generation sequencing (NGS) was employed for the analysis of VWF in patients inspected from 2017 onwards (about 86% of IPs). In both strategies, the entire VWF, encompassing all 52 exons, intron/exon boundaries, and promoter regions, was analyzed.

PreviouslyPublished:

Gene: VWF

Variant: c.3673T>G p.Cys1225Gly

HGVS:

ClinVarID: 100272

CAID:

gnomAD: absent

Case: IP #15 in type 2M VWD sub-cohort

DiseaseAssertion: type 2M VWD

FamilyInfo:

ParentalGenotype(s):

CasePresentingHPOs:

CaseHPOFreeRext: VWF:Ag [IU/dL] = n/a; VWF:Ac [IU/dL] = 50; FVIII:C [IU/dL] = n/a; VWF:CB (IU/dL) = 25

CaseNOTHPOs:

CaseNOTHPOFreeText:

CasePreviousTesting:

GenotypingMethod: Sequence variations in the VWF were identified through direct sequencing, while the detection of large rearrangements utilized the multiplex ligation-dependent probe amplification (MLPA) technique or assessed copy number variations (CNVs). Sanger sequencing methodology was applied for VWF analysis in patients investigated prior to 2016 (approximately 14% of IPs), whereas next-generation sequencing (NGS) was employed for the analysis of VWF in patients inspected from 2017 onwards (about 86% of IPs). In both strategies, the entire VWF, encompassing all 52 exons, intron/exon boundaries, and promoter regions, was analyzed.

PreviouslyPublished:

Gene: VWF

Variant: c.5191T>A p.Ser1731Thr

HGVS:

ClinVarID: 100420

CAID:

gnomAD: 0.001158 (1870 in 1614210) 5 homozygotes https://gnomad.broadinstitute.org/variant/12-6016636-A-T?dataset=gnomad_r4