Cardiac Myosin Binding Protein-C in Development and Reversal of Heart Failure

心肌肌球蛋白结合蛋白-C 在心力衰竭发生和逆转中的作用

• 批准号: 10544998
• 负责人: Carl Wei-Chan Tong
• 金额: $ 46.18万
• 依托单位: TEXAS A&M UNIVERSITY HEALTH SCIENCE CTR
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-01-15 至 2024-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10544998
• 关键词: Acceleration; Adenoviruses; Adrenergic Agents; Aging; American; Biochemistry; Biological Assay; Blood Vessels; C10; Calcium; Cardiac; Cardiac Myosins; Cardiac Surgery procedures; Companions; Death Rate; Dependovirus; Deterioration; Development; Diagnosis; Disease; EFRAC; Echocardiography; Exclusion; Exhibits; Focal Adhesion Kinase 1; Functional disorder; Heart; Heart Rate; Heart failure; High Fat Diet; Hour; Hypotension; Immunoprecipitation; In Vitro; Knock-in Mouse; Mass Spectrum Analysis; Measurement; Mechanical Stress; Mediating; Mind; Modeling; Mus; Muscle Proteins; Mutagenesis; Mutate; Mutation; Myocardial dysfunction; Myocardium; Myofibrils; NG-Nitroarginine Methyl Ester; Nitric Oxide; Obesity; Operative Surgical Procedures; Pharmaceutical Preparations; Phenotype; Phenylalanine; Phosphorylation; Phosphorylation Site; Phosphotransferases; Physiological; Preparation; Prevalence; Prevention; Protein Dephosphorylation; Pulmonary Edema; Recombinants; Relaxation; Research Project Grants; Running; Sarcomeres; Serine; Signal Transduction; Site; Stress; Structure; Techniques; Thick Filament; Translating; Treatment Failure; Tyrosine; Tyrosine Phosphorylation; Use Effectiveness; age related; aorta constriction; density; effective therapy; efficacy evaluation; experimental study; gene therapy; heart function; heart preservation; improved; in vivo; inhibitor; insight; mimetics; mortality; mouse model; myosin-binding protein C; novel; overexpression; papillary muscle; preservation; pressure; prevent; response; stem

Heart failure (HF) afflicted 6.5 million Americans in 2017, carries 5-year mortality of ~50%, and will likely increase to > 8 million by 2030. The lack of significant improvement in mortality for the last 46 years, the reality of needing to discontinue effective medications due to hypotension in heart failure with reduced ejection fraction (HFrEF), and the paucity of effective treatment for heart failure with preserved ejection fraction (HFpEF) combine to suggest that new treatments are needed. Cardiac myosin binding protein-C (cMyBPC) resides on the thick filament of the heart muscle. Phosphorylation of cMyBPC at its M-domain increases cross-bridge cycling rate. Because increasing cross-bridge cycling rate can improve both contractility and lusitropy, we hypothesize that phosphorylation of cMyBPC provides a novel central mechanism that can prevent and treat HF due to different causes. Using mouse models that mimic de-phosphorylated and phosphorylated cMyBPC at 3 serine (S) sites in the M-domain, we recently discover that cMyBPC phosphorylation mitigates both aged-related development of HFpEF and trans-aortic constriction (TAC) surgery induced HFrEF, as seen by improved survival and better preservation of diastolic function. Thus, cMyBPC phosphorylation holds potential to treat both HFrEF and HFpEF. However, we find evidence that 2 new S phosphorylation sites outside our study of 3 S sites add significant functional effects. Consequently, we have made 2 new knock-in mouse models to include the new sites for elucidating the effects of maximally phosphorylated cMyBPC(5SD) and dephosphorylated cMyBPC(5SA) M-domain. We also find evidence that pressure stress can activate focal adhesion kinase (FAK) to phosphorylate tyrosine (Y) residue(s) in cMyBPC. This discovery leads to a new supporting hypothesis that pressure stress triggers FAK to phosphorylate cMyBPC to increase contractility as a compensatory response. With these discoveries in mind, we intend to determine the efficacy of maximal M-domain phosphorylation and elucidate the novel FAK-cMyBPC mechanism. Aim #1: Determine the efficacy and companion mechanisms of phosphorylated cMyBPC to preserve cardiac function under HF inducing conditions of aging, pressure stress, and obesity. We will challenge mice with 2-yr aging, TAC, or high fat diet. We will use a combination of functional and biochemistry techniques to determine the underlying mechanisms. Ability of cMyBPC(5SD) mouse to resist deterioration and effectiveness of using cMyBPC(5SD) gene therapy to reverse failing WT hearts will quantify the efficacy of cMyBPC phosphorylation for prevention and treatment respectively. Aim #2: Elucidate signaling mechanism and functional results of pressure stress induced tyrosine phosphorylation of cMyBPC. We will identify Y site(s), confirm FAK-cMyBPC interaction, and elucidate effects of FAK phosphorylating cMyBPC on models ranging from intact papillary muscle to new knock-in mouse. Impact: Efficacy results and mechanistic insights stemming from this study provide evidence for translating cMyBPC phosphorylation to new HF treatment.

心力衰竭（HF）在2017年折磨着650万美国人，其5年死亡率约为50%，并且可能会增加

项目成果

Isolation of Adult Mouse Cardiomyocytes Using Langendorff Perfusion Apparatus.

使用 Langendorff 灌注装置分离成年小鼠心肌细胞。

• DOI: 10.1007/978-1-0716-1480-8_16
• 发表时间: 2021
• 期刊: Methods in molecular biology (Clifton, N.J.)
• 作者: Liu,Yang;Dostal,DavidE;Tong,CarlW
• 通讯作者: Tong,CarlW

Preparation and Identification of Cardiac Myofibrils from Whole Heart Samples.

• DOI: 10.1007/978-1-0716-1480-8_2
• 发表时间: 2021-01-01
• 期刊: Methods in molecular biology (Clifton, N.J.)
• 作者: Creed, Heidi A;Tong, Carl W
• 通讯作者: Tong, Carl W