Case1: 13 year old female

DiseaseAssertion: LVNC

FamilyInfo: No family history of cardiac disease

ParentalTesting:

CasePresentingHPOs: HP:0011664. HP:0002094. HP:0001279. HP:0031295

CaseHPOFreeText:Left Ventricular noncompaction, Dyspnea (shortness of breath), Fainting, Left atrial enlargement

CaseNotHPOs:HP:0000929. HP:0001712

CaseNotHPOFreeText:Abnormal skull morphology, left ventricular hypertrophy

CasePreviousTesting:

Genotyping Method:Next-Generation Sequencing (NGS)

FunctionalAnalysis:

Variant:c.575G>A

ClinVar:12424

CAID:CA021957

gnomAD:

AdditionalInfo:

PMID: 30279906

Gene: TNNI3

HGNC ID: 11947

Case1: 14 month, female

DiseaseAssertion: dilated cardiomyopathy

FamilyInfo:The three siblings' cases were born to consanguineous parents. They had DCM and died from decompensated heart failure around the age of 12 months

ParentalTesting: Sanger Sequencing determined the parents carried the variant at the heterozygous state (Figure 1)

CasePresentingHPOs: HP:0002094, HP:0002099, HP:0002098, HP:0002090,

CaseHPOFreeText:productive cough, and whining, wheezing, mitral and tricuspid regurgitation

CaseNotHPOs: NR

CaseNotHPOFreeText:no sign of ischemia,

CasePreviousTesting: cardiac ultrasound

Genotyping Method: sanger sequencing

FunctionalAnalysis: NR

Variant: TNNI3 (NM_000363.5): c.204delG;p.(Arg69AlafsTer8)

ClinVar: 179447

gnomAD: not found

Case1:eight-year-old female

DiseaseAssertion: Restrictive cardiomyopathy (RCM)

FamilyInfo: The patient's parents denied any positive family history of cardiac attacks, cardiovascular issues, hypertension, or coronary artery diseases. There was also no reported history of diabetes or obesity among family members. No inherited diseases, including cardiomyopathies or metabolic disorders, were identified in the family history

ParentalTesting: NR

CasePresentingHPOs:HP:0012378, HP:0001695, HP:0100539, HP:0031352, HP:0034030

CaseHPOFreeText:bilateral pedal edema, shortness of breath after engaging in physical activity, typically lasting for 15 min, enlarged bilateral atrium, segmental thickening of the interventricular septum, segmentally diffuse reduction and lack of coordination in ventricular wall pulse amplitude, mild mitral regurgitation, and diminished left ventricular systolic and diastolic function identified by increased parameters of early diastolic velocity (E wave) and low late filling velocity (A wave) (Figure 1A) abnormal bilateral atrial signals, premature atrial beats, junctional ectopic beats, first-degree atrioventricular block, complete right bundle branch block, prolonged QT interval, and abnormal Q wave (Figure 1B).

CaseNotHPOs: NR

CaseNotHPOFreeText: NR

CasePreviousTesting: echocardiography, genetic screening, imaging assessments, blood gas analysis, blood cell counts, carbon dioxide retention

Genotyping Method: whole-exome sequencing (WES)

FunctionalAnalysis: computational analysis

Variant:TNNI3 c.509G>A p.R170Q

ClinVar:165516

gnomAD: not found

Case1: 6 year old, female

DiseaseAssertion: Restrictive cardiomyopathy

FamilyInfo:Both parents were heterozygous carriers, supporting an autosomal recessive inheritance pattern, younger brother showed no abnormalities or signs of the same illness

ParentalTesting: Sanger Sequencing

CasePresentingHPOs:HP:0002094, HP:0012378

CaseHPOFreeText: well below normal weight and height range for patients age, preserved systolic function and diastolic dysfunction, revealed preserved left ventricular systolic function with an ejection fraction, impaired diastolic function, enlarged atrium, wall thickening, impaired ventricular filling

CaseNotHPOs: NR

CaseNotHPOFreeText: comprehensive initial workup, including blood tests and urinalysis, revealed no additional abnormalities

CasePreviousTesting: Echocardiogram, blood tests, urinalysis

Genotyping Method: Whole-exome sequencing

FunctionalAnalysis: NR

Variant: TNNI3 c.406 C > T:p.R136

ClinVar: 488853

gnomAD: not found

Case1: 22-day-old newborn boy

**DiseaseAssertion ** neonatal dilated cardiomyopathy

FamilyInfo: no mutation in the parents

ParentalTesting: whole-exome sequencing

CasePresentingHPOs: HP:0002094, HP:0002789, HP:0010741, HP:0001644, HP:0001640, HP:0002202, HP:0000961, HP:0001259

CaseHPOFreeText: weaning difficulties during hospitalization with intractable low EF heart insufficiency, refractory low ejection fraction, cardiac insufficiency, mild pulmonary hypertension, and mitral and tricuspid insufficiency with mild valve regurgitation, difficulty with ventilator withdrawal, irregular fever, pale face, poor feeding after birth

CaseNotHPOs: NR

CaseNotHPOFreeText: no abnormal symptoms or signs from birth to the 10th day

CasePreviousTesting: Whole-exome sequencing (Figure A), Chest CT, Electrocardiography, Doppler echocardiography (Figures 1A,B), arterial blood gas, routine blood examination, liver function, renal function, serum electrolytes, bacterial endotoxin, G-test, blood culture, sputum culture, and stool tests

Genotyping Method: whole-exome sequencing (WES)

FunctionalAnalysis: NR

Variant: homozygous mutation in 554 based on the 7th exon of the TNNI3 gene (chromosomal position CHR 19:55665403, reference genome HG19): c.544G>A(p.Glu182Lys) (Figure 2A)

ClinVar: not found

CAID: CA021848

gnomAD: not reported

Case#: 2 year old, male, from Yunnan Province, China

DiseaseAssertion: Restrictive cardiomyopathy

FamilyInfo:no family history of sudden cardiac death or cardiomyopathy

ParentalTesting: Sanger Sequencing

CasePresentingHPOs:HP:0034031, HP:0001635, HP:0002240,

CaseHPOFreeText:a little puffiness of the eyelids, jugular veins were full but not inflamed, atypical heart beat, edema of the lower limbs, bi-atrial enlargement, left ventricular ejection fraction of 64.6%, normal septal and ventricular wall thickness, trace pericardial effusion atrial and ventricular septum had uneven signals in the localized areas on perfusion imaging under resting state, and gadolinium-delayed enhancement showed a suspicious stripe enhancement of the septum, (Figures 1-3)

CaseNotHPOs:OMIM:145500, HP:0000819, HP:0001698,

CaseNotHPOFreeText: no history of connective tissue disease or radiation therapy, no endocardial thickening

CasePreviousTesting: electrocardiogram, cardiac magnetic resonance,

Genotyping Method: Whole-exome sequencing, sanger sequencing

FunctionalAnalysis: NR

Variant:heterozygous (c.574C > T, p.Arg192Cys) TNNI3

ClinVar: 165510

gnomAD: not found

Case#: Female, 57, Korea

DiseaseAssertion:hypertrophic cardiomyopathy

FamilyInfo: Proband and 13 family members with diverse phenotypic expression of cardiomyopathies, elder sister had expired due to severe RCMP. large family, in which the p.Arg145Trp mutation of TNNI3 causes diverse phenotypes ranging from RCMP to HCMP to near-normal heart. Six relatives with RCMP or near-normal phenotypes also carried this mutation. Family pedigree (Figure 1) and Clinical data of affected individuals in a family with the c.433C>T mutation (table 1)

CasePresentingHPOs:HP:0002094, HP:0001640, HP:0001639, HP:6000313, HP:0031295, HP:0001670, HP:0004757, HP:0005135, HP:0004756, HP:0001962

CaseHPOFreeText:restrictive filling pattern, delayed hyper-enhancement, which reflects fibrotic change in the myocardium in the hypertrophied segments (Fig. 2D).

CaseNotHPOs:HP:0032092

CaseNotHPOFreeText:normal LV systolic function and normal cavity size

CasePreviousTesting:Chromatograms, Echocardiogram

Genotyping Method:Sanger Sequencing (Fig. 5)

FunctionalAnalysis:heterozygous nonsynonymous mutation

Variant:NM_000363.5(TNNI3):c.433C>T (p.Arg145Trp)

CAID:CA021667

gnomAD:chr19-55154146-G-A

PMID: 37646638

Gene: TNNI3

HGNC ID: 11947

Case1: 8 year old female

DiseaseAssertion: Heart failure

FamilyInfo: “ no family history of sudden cardiac death or cardiomyopathy” Pedigree in Figure 5.

ParentalTesting: Variant not found in parents

CasePresentingHPOs:HP:0010741, HP:0001723

CaseHPOFreeText: “biatrial enlargement of ST-segment depression in the inferior and lateral precordial leads” “Echocardiography revealed the enlargement of both atria…slight pericardial effusion, and a LV ejection fraction of 55%”

CaseNotHPOFreeText: “LV global longitudinal strain measured by speckle-tracking echocardiography was normal” “Late gadolinium enhancement was negative.” “no evidence of pulmonary embolism, anomalous pulmonary venous drainage, or pulmonary arteriovenous fistulas” “The following laboratory findings were normal: thyroid function, liver chemistry tests, rheumatic disease antibody levels, troponin I, creatinine, and blood urea nitrogen levels, white blood cell count, urine analysis, D-dimer levels, hepatitis panel screening, and serum free light chain concentrations; HIV antibody levels were negative.”

Genotyping Method: Next generation sequencing

Variant: c.574C > T p.R192C)

ClinVar:CA021951

Case#:24-year-old male

DiseaseAssertion:non-obstructive HCM and long QTc

FamilyInfo:family history of long QT (mother and maternal grandmother), (family pedigree in Fig. 2)

ParentalTesting:Sanger sequencing done in the father confirming the paternal origin of the variant.

CasePresentingHPOs:HP:0001712, HP:0001657, HP:0005135

CaseNotHPOs:HP:0001654, HP:0006673

CasePreviousTesting: Cardiac MRI, Electrocardiogram, echocardiogram

Genotyping Method:Next Generation Sequencing

FunctionalAnalysis:Pathogenic missense variant

Variant:NM_000363.5(TNNI3):c.592C>G (p.Leu198Val)

ClinVar:177694

CAID:CA022019

gnomAD:chr19-55151875-G-C

PMID:33777698

Gene:TNNI3

HGNC ID:11947

PMID:38548731

Gene:TNNI3

HGNC ID:11947

Case#:14 year old Female

DiseaseAssertion:nonobstructive hypertrophic cardiomyopathy

FamilyInfo:lacked any family history of cardiomyopathy, including sudden death or cardiac hypertrophy, Family Pedigree (Figure. 1a)

ParentalTesting:none, healthy parents

CasePresentingHPOs:HP:0001663

CaseHPOFreeText: cardiopulmonary arrest

CaseNotHPOs: HP:4000004

CaseNotHPOFreeText: myocardial edema, ventricular arrhythmias, obstructed outflow tract

CasePreviousTesting: echocardiogram, cardiac MRI

Genotyping Method:Whole genome sequencing

FunctionalAnalysis:heterozygous missense variant

Variant: NM_000363.5:c.583A>T (p.Ile195Phe)

CAID:CA407439576

PMID:28382084

Gene:TNNI3

HGNC ID:11947

PMID:30953456

Gene:TNNI3

HGNC ID:11947

Case#:52 year old female from a small mountainous village in Peloponnese, Greece

DiseaseAssertion:severely manifested phenotype of RCM leading to heart transplant at 41

FamilyInfo:Family Pedigree Figure 1.Parents were third-degree cousins from the same village and were heterozygous for the same variant but displayed no symptoms of the disease. The older sister who was also found heterozygous was asymptomatic. Twin sister and brother were homozygous for the same variant and displayed a restrictive and a hypertrophic phenotype, respectively. Their children are all carriers of the mutation and remain asymptomatic until the age of 21

ParentalTesting:Family of the proband had Electrocardiogram and Doppler Echocardiogram (shown in figure 2) as well as TNNI3 genetic testing by targeted Sanger sequencing for the detection of the variant. Her twin sister and her brother were found homozygous for the variant, while both parents and the older sister were found heterozygous.

CasePresentingHPOs:HP:0002875, HP:0004757

CaseNotHPOFreeText:normal left ventricular dimensions

CasePreviousTesting:electrocardiogram, Right heart catheterization

Genotyping Method:NGS (filtered using VariantStudio Data Analysis Software and the Sophia Genetics DDM platform)

FunctionalAnalysis: a novel homozygous TNNI3 variant

Variant:NM_000363.4:c.586G > C, p.(Asp196His)

ClinVar:2169619

CAID:CA407439559

PMID:40656892

Gene:TNNI3

HGNC ID:11947

Case#:53-year-old female

DiseaseAssertion:non-obstructive HCM with progressive deterioration of right heart function

FamilyInfo:mother has cardiac hypertrophy, the TNNI3 variant was also detected her sister and daughter

CasePresentingHPOs:HP:0005110, HP:0012398, HP:0001962, HP:0031352, HP:0033118, HP:0011675

CaseHPOFreeText:enlarged cardiac silhouette

CaseNotHPOs:HP:0030829, HP:0030148

CaseNotHPOFreeText: jugular venous distention

CasePreviousTesting:Electrocardiogram, Echocardiograms

Genotyping Method:next-generation sequencing (NGS)

FunctionalAnalysis: Hybridized pathogenic variant in exon 7 of TNNI3 (table 1)

Variant:NM_000363.4:c.470C>T(p.Ala157Val)

ClinVar:43388

CAID:CA021720

gnomAD:chr19-55154109-G-A