CaseHCM16.1: Mixed ancestry

DiseaseAssertion: Hypertrophic Cardiomyopathy

FamilyInfo: N/A

CasePresentingHPOs: HP:0001639 (Hypertrophic cardiomyopathy)

HPOsFreeText: N/A

CaseNotHPOs: HP:0000819 (Diabetes mellitus)

NotHPOsFreeText: N/A

CaseAddInfo: N/A

CasePreviousTesting: See Table 1

CaseGenotypingMethod: DNA was extracted from the probands' blood, followed by sequencing of the coding regions and exon/intron boundaries of 15 genes associated with HCM. From there, they were amplified by PCR ampliication and variants were filtered by pathogenicity. Every unclassified variant was confirmed by Sanger sequencing afterwards.

Variant: NM_000257.3:c.746G>A (p.Arg249Gln)

ClinVar: 14088 https://www.ncbi.nlm.nih.gov/clinvar/variation/14088/

gnomAD: Not in gnomAD

Supplementary Material 4

Case M-3: Japanese

DiseaseAssertion: idiopathic cardiomyopathy (HCM and DCM)

CasePreviousTesting: Clinical evaluation, chest radiography, electrocardiography, echocardiography, cardiac catheterization, full testing (MYBPC3, TNNT2, TNNI3, MYL2, MYL3, TPM1, ACTC1)

CaseGenotypingMethod: 8 sarcomeric genes amplified from genomic DNA via PCR then screened using bidirectional direct sequencing

Variant: NM_000257.3(MYH7):c.746G>A (p.Arg249Gln)

ClinVar: 14088, https://www.ncbi.nlm.nih.gov/clinvar/variation/14088/

gnomAD: n/a

Supplementary Material 4

Case M-4: Japanese

DiseaseAssertion: idiopathic cardiomyopathy (HCM and DCM)

CasePreviousTesting: Clinical evaluation, chest radiography, electrocardiography, echocardiography, cardiac catheterization, full testing (MYBPC3, TNNT2, TNNI3, MYL2, MYL3, TPM1, ACTC1)

CaseGenotypingMethod: 8 sarcomeric genes amplified from genomic DNA via PCR then screened using bidirectional direct sequencing

Variant: NM_000257.3(MYH7):c.746G>A (p.Arg249Gln)

ClinVar: 14088, https://www.ncbi.nlm.nih.gov/clinvar/variation/14088/

gnomAD: n/a

Case1: 16 yo Caucasian female

DiseaseAssertion: MYH7 mediated cardiomyopathy

FamilyInfo: 6 additional family members were found to be at risk for HCM and were advised of recommendations for both

CasePresentingHPOs: HP:0001640, HP:0031319, HP:0031318, HP:0001655, HP:0002181

"Cardiomegaly, Cardiomyocyte hypertrophy, Myofiber disarray, Patent foramen ovale, Cerebral edema"

CaseHPOFreeText: moderately enlarged heart (352 g; expected range, 119–290 g), myocardium exhibited marked myocyte hypertrophy, myocyte disarray, and mild interstitial fibrosis, most notably in the anterolateral left ventricle, fatty streaks present in the thoracic aorta

CaseNotHPOs: n/a

CaseNotHPOFreeText: overt septal hypertrophy

CasePreviousTesting: n/a

GenotypingMethod: Whole Exome Next-Generation DNA Sequencing, gene-specific surveillance of all known LQTS, CPVT, BrS, HCM, DCM, and ACM genes (90 total; Table 1 and Figure 1), pathogenic mutations were confirmed in the SUD case's genomic DNA by using standard polymerase chain reaction and Sanger DNA sequencing methods

Variant: NM_000257.4:c.746G>A

ClinVar: 14088

CAID: CA016781

gnomAD: not found

Case1: deceased 16 year old female

DiseaseAssertion: HCM

FamilyInfo: 6 additional family members were found to be at risk for HCM

CasePresentingHPOs: HP:0001640, HP:0031319, HP:0002181

"Cardiomegaly, Cardiomyocyte hypertrophy, Cerebral edema

CaseHPOFreeText: Moderately enlarged heart, myocyte disarray, mild interstitial fibrosis

CaseNotHPOs: n/a

CaseNotHPOFreeText: n/a

CasePreviousTesting: n/a

GenotypingMethod: Whole Exome Next-Generation DNA Sequencing

Variant: NM_000257.4:c.746G>A

ClinVar: 14088

CAID: CA016781

gnomAD: not found

Case8: Female, 12 yrs onset, Slovakia

DiseaseAssertion: Hypertrophic Cardiomyopathy

FamilyInfo: Affected family members

CasePresentingHPOs: HP:0001639 (Hypertrophic Cardiomyopathy)

HPOsFreeText: N/A

CaseNotHPOs: N/A

NotHPOsFreeText: Normal LV ejection fraction

CasePreviousTesting: ABCC9, ACTC1, ACTN2, ANKRD1, CASQ2, CAV3, CRYAB, CSRP3, CTF1, DES, DSC2, DSG2, DSP, DTNA, EMD, FHL2, GLA, JUP, LAMA4, LAMP2, LDB3, LMNA, MYBPC3, MYH6, MYH7, MYL2, MYL3, MYLK2, MYOZ2, NEXN, PKP2, PLN, PRKAG2, RBM20, RYR2, SGCD, TAZ, TCAP, TMEM43, TNNC1, TNNI3, TNNT2, TPM1, TTN, TTR, VCL were all tested on NGS panel

CaseGenotypingMethod: Genomic DNA was extracted from whole blood and samples were purified and amplified. The coding exons for the above genes were enriched and sequenced. The resulting variants were analyzed and filtered for predicted pathogenicity.

CaseAddInfo: N/A

Variant: NM_000257.4(MYH7):c.746G>A (p.Arg249Gln)

ClinVar: 14088 https://www.ncbi.nlm.nih.gov/clinvar/variation/14088/

gnomAD: Not in gnomAD

MultipleGeneVariants

(1) DSP Variant

Variant: NM_004415.4(DSP):c.484C>G (p.Arg162Gly)

ClinVar: 546994 https://www.ncbi.nlm.nih.gov/clinvar/variation/546994/

gnomAD: 0.00006152, http://gnomad.broadinstitute.org/variant/6-7559520-C-G

CaseMYH7-10: Japan, Asian

CasePresentingHPOs: HP:0001639, HP:0006685 (Hypertrophic cardiomyopathy, Endomyocardial fibrosis)

HPOsFreeText: N/A

CaseNotHPOs: N/A

NotHPOsFreeText: N/A

CaseAddInfo:

CaseGenotypingMethod: Genomic DNA was extracted from whole blood from each proband. They each had 95 genes associated with DCM, HCM, and other inherited cardiovascular diseases sequenced with a panel, including both the exons and spicing regions. Variants were then filtered for rarity and pathogenicity.

Variant: NM_000257.3:c.746G>A (p.Arg249Gln)

ClinVar: 14088 https://www.ncbi.nlm.nih.gov/clinvar/variation/14088/

gnomAD: Not in gnomAD

Case1: 58 year old female

DiseaseAssertion: HCM

FamilyInfo: proband's daughter presents with same disease and mutation

CasePresentingHPOs: HP:0008994, HP:0000218

"Proximal muscle weakness in lower limbs, High palate

CaseHPOFreeText: symmetrical bilateral ptosis without oculomotor palsy, high arched palate but the rest of the cranial examination was normal, weakness of neck flexors, weakness in psoas and tibialis anterior, unable to walk on heels

CaseNotHPOs: n/a

CaseNotHPOFreeText: atrophy of the sternocleidomastoid muscles, hanging big toe

CasePreviousTesting: n/a

GenotypingMethod: "gene sequencing"

Variant: NM_000257.4:c.746G>A

ClinVar: 14088

CAID: CA016781

gnomAD: not found

DiseaseAssertion: Hypertrophic Cardiomyopathy

VariantPreviousTesting: ACTC1, ACTN2, CSRP3, LAMP2, MYBPC3, MYH7, NEXN, PLN, TNNC1, TNNI3, TNNT2, TPM1, CAV3, GLA, MYL2, MYL3, MYOZ2, PRKAG2, and TTR were all tested via NGS.

VariantGenotypingMethod: Next-generation sequencing panels were used to sequence and read the genetic information of the 760 HCM probands for variants within exons and in +/- 10 nucleotides within the flanking introns of the genes provided above. These variants were then interpreted and filtered for pathogenicity.

VariantAddInfo: Seen in one HCM probands

Variant: NM_000257.4(MYH7):c.746G>A (p.Arg249Gln)

ClinVar: 14088 https://www.ncbi.nlm.nih.gov/clinvar/variation/14088/

gnomAD: Not in gnomAD