Case1: A 68-year-old female

DiseaseAssertion: dilated cardiomyopathy (DCM), with the MYH7 p.Lys637Glu (K637E) variant identified in the affected patient. The study investigated sarcomeric gene variants associated with DCM

FamilyInfo: The proband's father (M1, II:3) died suddenly at 36 years of age, 2 years after being diagnosed with DCM.

ParentalTesting: Yes from father

CasePresentingHPOs: HP:0011664, HP:0011713, HP:0003677, HP:0001635,

CaseHPOFreeText: was diagnosed as having left ventricular non-compaction (LVNC) based on echocardiography. A left bundle branch block on her ECG. Echocardiography revealed LVNC and she has had a slow progression towards heart failure and an increasing dilatation of her left ventricle. Age 48 at onset, left ventricular end diastolic diameter 59 mm, left ventricular ejection fraction 25%, ventricular extrasystoles for arrhythmia.

CaseNotHPOs:NR

CaseNotHPOFreeText:NR

CasePreviousTesting:NR

Genotyping Method: genetically screened, also using GenBank with PCR by amplifying the coding region

FunctionalAnalysis:NR

Variant: NM_000257.4:c.1909A>G

ClinVar: 1329402

CAID:CA389049558

gnomAD: NR

AdditionalInfo: NR

Case#: SF No. 29, GC Case No. 22, female, 79

DiseaseAssertion: hypertrophic cardiomyopathy, dilated cardiomyopathy

FamilyInfo: No

CasePresentingHPOs: N/A

CaseHPOFreeText: Colorectal cancer,

CaseNotHPOs: N/A

CaseNotHPOFreeText: N/A

CasePreviousTesting: N/A

CaseGenotypingMethod: WES, Sanger sequencing,

Variant: NM_000257.3(MYH7):c.4522_4524delGAG (p.Glu1508del)

ClinVar: 43023 https://www.ncbi.nlm.nih.gov/clinvar/variation/43023/

CAID: N/A

gnomAD: absent

Case#: SF No. 28, GC Case No. 21, male, 83

DiseaseAssertion: hypertrophic cardiomyopathy, dilated cardiomyopathy

FamilyInfo: No

CasePresentingHPOs: N/A

CaseHPOFreeText: Gastric cancer,

CaseNotHPOs: N/A

CaseNotHPOFreeText: N/A

CasePreviousTesting: N/A

CaseGenotypingMethod: WES, Sanger sequencing,

Variant: NM_000257.3(MYH7):c.4522_4524delGAG (p.Glu1508del)

ClinVar: 43023 https://www.ncbi.nlm.nih.gov/clinvar/variation/43023/

CAID: N/A

gnomAD: absent

Case#: SF No. 30, GC Case No. 23, female, 31

DiseaseAssertion: hypertrophic cardiomyopathy, dilated cardiomyopathy

FamilyInfo: Yes, one affected

CasePresentingHPOs: N/A

CaseHPOFreeText: Gastric cancer,

CaseNotHPOs: N/A

CaseNotHPOFreeText: N/A

CasePreviousTesting: N/A

CaseGenotypingMethod: WES, Sanger sequencing,

Variant: NM_000257.3(MYH7):c.4522_4524delGAG (p.Glu1508del)

ClinVar: 43023 https://www.ncbi.nlm.nih.gov/clinvar/variation/43023/

CAID: N/A

gnomAD: absent

Case12:

DiseaseAssertion: Dilated cardiomyopathy

FamilyInfo: 2 family members tested, 1 family member tested positive for cardiomyopathy

CasePresentingHPOs: HP:0001644

"Dilated cardiomyopathy"

CasePreviousTesting: If there was a normal heart at autopsy, KCNQ1, KCNH2, SCN5A, KCNE1, and KCNE2 were sequenced. If death occurred during exertion, CPVT was considered and RYR2 sequenced. If these tests were negative and there was a family history of sudden death, PKP2, DSC2, DSG and DSP2 were sequenced. If dilated cardiomyopathy (DCM) was suspected, the six sarcomere genes were sequenced along with LMNA and SCN5A.

GenotypingMethod: Sanger sequencing

Variant: NM_000257.4:c.2945T>C

ClinVar: 42941

CAID: CA013227

gnomAD: https://gnomad.broadinstitute.org/variant/14-23892910-A-G

Total Allele Frequency: 0.0008484

European (non-Finnish): 0.001416

Case11:

DiseaseAssertion: Dilated cardiomyopathy

FamilyInfo: 1 family member tested, 0 tested positive for cardiomyopathy

CasePresentingHPOs: HP:0001644

"Dilated cardiomyopathy"

CasePreviousTesting: If there was a normal heart at autopsy, KCNQ1, KCNH2, SCN5A, KCNE1, and KCNE2 were sequenced. If death occurred during exertion, CPVT was considered and RYR2 sequenced. If these tests were negative and there was a family history of sudden death, PKP2, DSC2, DSG and DSP2 were sequenced. If dilated cardiomyopathy (DCM) was suspected, the six sarcomere genes were sequenced along with LMNA and SCN5A.

GenotypingMethod: Sanger sequencing

Variant: NM_000257.4:c.3726G>A

CAID: CA485766621

Case#: DCM-12, female, Yunnan, China

DiseaseAssertion: DCM

FamilyInfo: n/a

CasePresentingHPOs: n/a

CaseHPOFreeText: n/a

CaseNotHPOs: n/a

CaseNotHPOFreeText: n/a

CasePreviousTesting: physical exam, invasive examination, ECG,

CaseGenotypingMethod: NGS and Sanger Sequencing

Variant: NM_000257.4(MYH7):c.77C>T (p.Ala26Val)

ClinVar: 37375 https://www.ncbi.nlm.nih.gov/clinvar/variation/37375/

CAID: n/a

gnomAD: absent

DiseaseAssertion: Dilated Cardiomyopathy

VariantPreviousTesting: ACTC1, ACTN2, CSRP3, LAMP2, MYBPC3, MYH7, NEXN, PLN, TNNC1, TNNI3, TNNT2, TPM1, ABCC9, CTF1, DES, EMD, LDB3, LMNA, MYH6, RBM20, SGCD, TAZ, TCAP, TTN, and VCL were all tested via NGS.

VariantGenotypingMethod: Next-generation sequencing panels were used to sequence and read the genetic information of the 74 DCM 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 two DCM probands and relatives

Variant: NM_000257.4(MYH7):c.5395G>A (p.Glu1799Lys)

ClinVarID: 181278 https://www.ncbi.nlm.nih.gov/clinvar/variation/181278/

gnomAD: Not in gnomAD

DiseaseAssertion: Dilated Cardiomyopathy

VariantPreviousTesting: ACTC1, ACTN2, CSRP3, LAMP2, MYBPC3, MYH7, NEXN, PLN, TNNC1, TNNI3, TNNT2, TPM1, ABCC9, CTF1, DES, EMD, LDB3, LMNA, MYH6, RBM20, SGCD, TAZ, TCAP, TTN, and VCL were all tested via NGS.

VariantGenotypingMethod: Next-generation sequencing panels were used to sequence and read the genetic information of the 74 DCM 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 DCM proband

Variant: NM_000257.4(MYH7):c.2711G>A (p.Arg904His)

ClinVarID: 42926 https://www.ncbi.nlm.nih.gov/clinvar/variation/42926/

gnomAD: Not in gnomAD

Case#: Case III-2, 42 yr, female, early childhood onset

DiseaseAssertion: DCM

FamilyInfo: Family D, her father (II-5) had late-onset foot drop and hand weakness, died at age 51 from a myocardial infarction, her son (IV-3) had a severe generalized weakness since childhood, never walked and had prominent neck extensor contractures, died at age 19 years from a presumed cardiopulmonary event, 2 sisters (III-1 and III-3) had early-onset weakness and foot drop, her nephew (IV-1) had severe weakness and was wheelchair dependent, muscle biopsy, diagnosed with an unspecified muscular dystrophy, her niece (IV-2) had severe congenital weakness and died in her teens, all affected family members had contractures of neck extensors, heel cords, knee flexors, and elbow flexors

CasePresentingHPOs: progressive muscle weakness, lower limb muscle weakness, gait disturbance, slender build, scapular winging, thoracic scoliosis, weakness of facial musculature, generalized limb muscle atrophy, sinus tachycardia (HP:0003323, HP:0007340, HP:0001288, HP:0001533, HP:0003691, HP:0002943, HP:0030319, HP:0009055, HP:0011703)

CaseHPOFreeText: weakness and gait at age 5, nonambulatory by late-teens, wheelchair dependent, neck muscle atrophy, hypoactive myotatic reflexes

CaseNotHPOs: respiratory distress, syncope, chest pain, palpitations (HP:0002098, HP:0001279, HP:0100749, HP:0001962)

CaseNotHPOFreeText: normal sensory

CasePreviousTesting: Analysis included most of the other congenital myopathy genes and was otherwise negative

CaseGenotypingMethod: neurological examination, serum creatine kinase, nerve conduction studies, concentric needle electromyography, electrocardiogram, echocardiogram, forced vital capacity, muscle biopsy, immunohistochemical analyses, strength testing (MRC 3/2), arm flexors (MRC 4/4), arm extensors (MRC 3/3), wrist flexors (MRC 3/3), wrist extensors (MRC 1/3), finger flexors (MRC 2/2), finger extensors (MRC 2/2), finger adductors and abductors (MRC 4/4), hip flexors (MRC 2/2), hip abductors and adductors (MRC 3/3), knee flexors (MRC 2/2), knee extensors (MRC 2/2), foot dorsiflexors (MRC 0/0), plantar flexors (MRC 3/3) toe flexors (MRC 2/2), and toe extensors (MRC 1/1), normal 0.82 L (25%) FVC, 25% left ventricular ejection fraction, normal NCSs, increased small polyphasic MUAP, occasional fibrillation potentials, positive sharp waves, and brief myotonic discharges, facioscapulohumeral muscular dystrophy type 1, Emery–Dreifuss muscular dystrophy (EMD, LMNA, and FHL1 gene disorders), and limb-girdle muscular dystrophy type 1C were negative

Variant: NM_000257.2

ClinVar: absent

CAID: absent

gnomAD: absent

Case3: Male, 53 yrs diagnosis, Portugal

DiseaseAssertion: Dilated Cardiomyopathy

FamilyInfo: No family history

CasePresentingHPOs: HP:0001644, HP:0001635, HP:0011675, HP:0011713, HP:0012665, HP:0031192 (Dilated cardiomyopathy, Congestive heart failure, Arrhythmia, Left bundle branch block, Moderately reduced ejection fraction, Abnormal morphology of left ventricular trabeculae)

HPOsFreeText: Left ventricular end-diastolic diameter of 56 mm

CaseNotHPOs: HP:0002540, HP:0011706, HP:0001709, HP:0006682 (Inability to walk, Second degree atrioventricular block, Third degree atrioventricular block, Premature ventricular contractions)

NotHPOsFreeText: N/A

CasePreviousTesting: Proband has been studied before as part of FATIMA with general clinical assessment and cardiological investigations as well as exclusion criteria subject there. Tested genes include: LMNA, MYH7, MYBPC3, TNNT2, ACTA1, TPM1, CSRP3, TCAP, SGCD, PLN, MYL2, MYL3, TNNI3, TAZ, LDB3.

CaseGenotypingMethod: Blood samples were obtained and NGS was performed on a panel of 15 different genes covering the coding sequence and flanking exon/intron boundaries. Sanger sequencing was used to validate the variants and provide additional coverage. Variants were interpreted and filtered for pathogenicity.

CaseAddInfo: More info in 19044174 but paper is unavailable at this time

Variant: NM_000257.3:c.5623G>T (p.Val1875Phe)

CAID: CA389034842 http://reg.clinicalgenome.org/redmine/projects/registry/genboree_registry/allele?hgvsOrDescriptor=NM_000257.3%3Ac.5623G%3ET

gnomAD: Not in gnomAD

Case#: A 29 years old male DCM patient

DiseaseAssertion:Familial dilated cardiomyopathy (FDCM), associated with the TNNT2 gene missense variant p.Arg144Trp (R144W)

FamilyInfo: Family members had revealed its presence in three other individuals with DCM. A history of sudden cardiac deaths (SCD) in the family. Two individuals in the family, who were diagnosed with DCM, had died due to severe congestive heart failure at the age of 45 and 25 years. However, a 66-year-old individual in the same family with DCM having mild symptoms.

ParentalTesting:NR

CasePresentingHPOs: HP:0001644, HP:4000141, HP:0012664, HP:0004308, HP:0012828, HP:0001653

CaseHPOFreeText: Dilated cardiomyopathy with left ventricular dilation, Age of onset, 25, left ventricular internal diastolic dimension of 72mm, reduced ejection fraction of 26%, has Ventricular arrhythmia, severe mitral regurgitation.

CaseNotHPOs:NR

CaseNotHPOFreeText: NR

CasePreviousTesting:NR

Genotyping Method: Used PCR with GenBank No. NM_000364, then followed by mutation subsequent screening analysis

FunctionalAnalysis:The R144W mutation was found to be within the tropomyosin-binding domain of cTnT and alters the charge of the residue, so it is expected to have a significant impact on the structure and function of the protein. The novel unique/private R144W mutation identified in our present study is associated with FDCM

Variant:NM_001276345.2:c.445C>T

ClinVar:43640

CAID:CA004505

gnomAD: gnomAD v2: 1:201333470 G / A https://gnomad.broadinstitute.org/variant/1-201333470-G-A?dataset=gnomad_r2_1 gnomAD v4: chr1-201364342-G-A https://gnomad.broadinstitute.org/variant/chr1-201364342-G-A?dataset=gnomad_r4

AdditionalInfo:25 bp deletion was found to be associated with both HCM and DCM in India and south Asia

Case1: 42 year old male

DiseaseAssertion:DCM and LVNC

FamilyInfo: Same mutation in proband found in father, sister, and daughter

CasePreviousTesting: Sanger sequencing of coding exons and flanking intronic regions of the main sarcomeric genes, including MYH7, MYBPC3, TNNT2, TNNI3, TPM1, ACTC, MYL2, MYL3 and TNNC.

GenotypingMethod: NGS, Sanger sequencing

Variant: NM_000257.4:c.2146G>A

ClinVar: 14105

CAID: CA011770

gnomAD: not found

Case1: 44 year old male

DiseaseAssertion: Dilated cardiomyopathy

FamilyInfo: Affected brother, two sisters and both parents unaffected, parents denied consanguinity and were assumed to be second cousins

CasePresentingHPOs: HP:0003704, HP:0009060, HP:0000218, HP:0000276, HP:0000819, HP:0003691

"Scapuloperoneal weakness, scapular muscle atrophy, high palate, long face, Diabetes mellitus, Scapular winging

CaseHPOFreeText: decreased cardiac systolic function, clinical symptoms of cardiac failure at 44 years old, left ventricular systolic and diastolic dysfunction, exertional dyspnoea, ymmetrical wasting of the sternocleidomastoideus, trapezius, supraspinatus, pectoralis, deltoid, triceps, biceps, paravertebral and tibialis muscles at age 56

CasePreviousTesting:

GenotypingMethod: genome scan using 730,000 single nucleotide polymorphism (SNP) markers. Sequence analysis for the last four MYH7 exons

Variant: NM_000257.4:c.5458C>T

ClinVar: 181282

CAID: CA016126

gnomAD: https://gnomad.broadinstitute.org/variant/14-23884305-G-A

0.00008011 African

Case: male, 44yo at diagnosis

DiseaseAssertion: Dilated cardiomyopathy (DCM)

CasePresentingHPOs: HP:0012664, HP:0001635, HP:0100598, HP:0001324, HP:0003388 (reduced ejection fraction, heart failure, pulmonary oedema, muscle weakness, fatigability)

HPOsFreeText: reduced LVEF (=29%), diagnosed with congenital myopathy at 36yo, LVIDD = 74mm, creatine kinase < 600 U/l, ICD implantation, waiting heart transplantation at 48yo

CaseNotHPOs:

NotHPOsFreeText: anti-tachycardia pacing or shock therapy from the ICD

CasePreviousTesting: ACTA1, BIN1, DNM2, MYH7, MTM1, NEB, RYR1, SCN4A, SEPN1, TPM2, TPM3

CaseGenotypingMethod: PCR amplification of exons of genes investigated and Sanger sequencing or NGS

Variant: NM_000257.4(MYH7):c.5401G>A (p.Glu1801Lys)

ClinVar: 43069, https://www.ncbi.nlm.nih.gov/clinvar/variation/43069/

gnomAD: absent

CaseCM007

DiseaseAssertion: DCM

GenotypingMethod: NGS

MultipleGeneVariants:

VariantMYH7: NM_000257.4:c.2645A>G

ClinVar: not found

CAID: CA389047904

gnomAD: not found

VariantRBM20: NM_001134363.3:c.2062C>T

ClinVar: 202065

CAID: CA335555

gnomAD: https://gnomad.broadinstitute.org/variant/10-112572217-C-T?dataset=gnomad_r2_1

0.00006494 European (non Finnish)

Case#: 55-year-old man

DiseaseAssertion: single coronary artery (SCA) and presented with dilated cardiomyopathy (DCM)

FamilyInfo: Unremarkable

ParentalTesting: NR

CasePresentingHPOs: HP:0002094, HP:0031352, HP:0001638, HP:0001644, HP:0010741

CaseHPOFreeText: chest tightness and dyspnoea after activity lasting for 2 months. CTCA showed congenital absence of the right coronary artery. TTE revealed enlargement of the left heart and cardiomyopathy. CMR revealed DCM. oedema of both lower limbs. Laboratory data in Table 1.

CaseNotHPOs: NR

CaseNotHPOFreeText: Stenosis

CasePreviousTesting: See NGS results in Supplementary Table 1

Genotyping Method: Genetic screening (NGS results in Supplementary Table 1) with confirmation by Sanger

FunctionalAnalysis: NR

Variant: c.1858C>T (p.Arg620Cys)

ClinVar: 67694

CAID: CA015449

gnomAD: v4.1.0 GrpMax FAF: 0.00002033 (European non-Finnish)

AdditionalInfo: The patient also has APOA5:c.990_993delAACA (p. Asp332Valfs*5) (P/LP in ClinVar with 2 stars)