Comprehensive Analysis of Allelic, Cellular and Molecular Heterogeneity in Human 3-Dimensional Cardiac Microtissues with MYH7 Mutations

具有 MYH7 突变的人三维心脏微组织等位基因、细胞和分子异质性的综合分析

• 批准号: 10210431
• 负责人: John Travis Hinson
• 金额: $ 53.99万
• 依托单位: UNIVERSITY OF CONNECTICUT SCH OF MED/DNT
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-07-01 至 2024-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10210431
• 关键词: 3-Dimensional; ATAC-seq; Actins; Alleles; Architecture; Arginine; Binding; Biological Assay; Biological Markers; Biomimetics; CRISPR/Cas technology; Cardiac; Cardiac Myocytes; Cell Line; Cells; Chromatin; Computing Methodologies; Consumption; Data; Disease; Disease model; Engineering; Epigenetic Process; Future; Gene Expression; Gene Expression Profile; Gene Mutation; Gene Targeting; Genes; Genetic; Genetic Transcription; Genotype-Tissue Expression Project; Glutamine; Goals; Heart failure; Heterogeneity; High-Throughput Nucleotide Sequencing; Human; Human Engineering; Hypertrophic Cardiomyopathy; Image; Immunofluorescence Immunologic; Individual; Inherited; Kinetics; Knock-out; Knowledge; Laboratories; Lead; Link; Location; Mechanics; Metabolic; Methionine; Mitochondria; Mitochondrial Proteins; Modeling; Molecular; Molecular Motors; Mutation; Myocardium; Myofibrils; Myosin Heavy Chains; Nuclear; Pathogenesis; Pathologic; Pathology; Pathway Analysis; Patients; Pattern; Phenotype; Prevalence; Proteins; RNA analysis; Regulatory Element; Research; Resolution; Risk; Role; Sarcomeres; Secondary to; Stress; Structure; Symptoms; Therapeutic; Three-dimensional analysis; Tissues; Transcript; Transposase; Valine; Variant; beta-Myosin; cell type; cohort; disease heterogeneity; genetic variant; genomic tools; heart dimension/size; heart function; in vivo; induced pluripotent stem cell; insight; new therapeutic target; novel; programs; response; single-cell RNA sequencing; stem cell differentiation; stem cell model; sudden cardiac death; therapeutic development; therapeutic target; therapeutically effective; transcriptome; transcriptome sequencing; treatment response

PROJECT SUMMARY/ABSTRACT Hypertrophic cardiomyopathy (HCM) patients are at risk for sudden cardiac death and progressive heart failure (HF), and there are no effective therapeutics, due in part to our limited genetic understanding of HCM pathogenesis. There are critical gaps in our current knowledge of the molecular mechanisms that link specific mutations in the beta myosin heavy chain gene (MYH7) to pathological thickening of the heart muscle that is associated with HCM. Our long-term goals are to utilize genomic tools combined with 3-dimensional cardiac microtissues derived from human induced pluripotent stem cells to interrogate mechanisms of HCM secondary to specific MYH7 variants, and to utilize these insights to identify new disease biomarkers and therapeutic targets for specific HCM patients. Our previous studies identified that two HCM-associated MYH7 variants, arginine 403 to glutamine and valine 606 to methionine that are located in the actin-binding domain of beta myosin heavy chain protein (MHC- β), generate increased microtissue contraction force with associated abnormalities in contraction kinetics. Studies by others have indicated that MYH7 variants located in distinct structural domains of MHC-β cause distinct phenotypes. These results lead to our central hypothesis that HCM is a heterogeneous disorder, in which patient symptoms and therapeutic responses are dependent on the location of the causative MYH7 variant(s) within the gene and on cell-type specific transcriptional and epigenetic programs, which initiate from abnormalities in contractile function. Guided by our comprehensive preliminary data, we propose to pursue three Specific Aims to determine multi-scale insights into HCM pathogenesis: (1) to characterize functional consequences of MYH7 variants localized to the actin-binding, ATP-binding and converter domains of MHC- β, (2) to identify cell type-specific transcriptional and epigenetic mechanisms of HCM in microtissues using paired single-cell RNA-seq and ATAC-seq and (3) to interrogate the function of C1ORF105, a nuclear-encoded mitochondrial protein that is associated with HCM and is specifically expressed in human cardiomyocytes. In summary, the execution of these aims will provide a more precise understanding of the functional role of MYH7 variant localization, generate novel cell-type specific and molecular mechanisms of HCM and identify critical molecular linkages between sarcomere and mitochondrial function that will broadly impact the field of HCM and heart failure.

项目总结/文摘

项目成果

Reading Frame Repair of TTN Truncation Variants Restores Titin Quantity and Functions.

• DOI: 10.1161/circulationaha.120.049997
• 发表时间: 2022-01-18
• 期刊: Circulation
• 影响因子: 37.8
• 作者: Romano R;Ghahremani S;Zimmerman T;Legere N;Thakar K;Ladha FA;Pettinato AM;Hinson JT
• 通讯作者: Hinson JT