Modulators of Cardiomyopathic Diseases

心肌病调节剂

• 批准号: 9914116
• 负责人: Jose Renato Pinto
• 金额: $ 37.15万
• 依托单位: FLORIDA STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-05-01 至 2022-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9914116
• 关键词: ATP phosphohydrolase; Ablation; Address; Affect; Affinity; Agonist; Arrhythmia; Binding; Biochemical; Biological Assay; Biophysics; Blood; Buffers; Cardiac; Cardiac Muscle Contraction; Cardiac Myocytes; Cardiac Myosins; Cardiac ablation; Cardiomyopathies; Cells; Complex; Coupled; Data; Development; Disease; Echocardiography; Electrocardiogram; Electrons; Elements; Excision; Functional disorder; Genes; Goals; Growth; Health; Heart; Heart Diseases; Heart failure; Histopathology; Human; Hypersensitivity; Image; Knock-in; Knock-in Mouse; Knockout Mice; Lead; Light; Link; Mass Spectrum Analysis; Measurement; Mechanics; Microfilaments; Mitochondria; Molecular; Monitor; Morphology; Movement; Mus; Muscle; Muscle Fibers; Mutant Strains Mice; Mutation; Myocardium; Myosin Light Chain Kinase; N Domain; Organism; Outcome; Pathogenicity; Pathologic; Pathologic Processes; Pathology; Patients; Phenotype; Phosphorylation; Phosphotransferases; Physiological; Post-Translational Protein Processing; Process; Property; Proteins; Relaxation; Role; Skin; Structure; Susceptibility Gene; Tamoxifen; Testing; Therapeutic; Thin Filament; Time; Tropomyosin; Troponin; Troponin C; Troponin I; Troponin T; Trypsin; Work; X ray diffraction analysis; base; biological systems; biophysical techniques; citrate carrier; desensitization; design; disease phenotype; experimental study; genetic approach; improved; in vivo; insight; micrography; mouse model; mutant; new therapeutic target; novel; pressure; public health relevance; reconstitution; response; sensor; targeted treatment; trait

 DESCRIPTION (provided by applicant): The long-term goal of this work is to identify components inside the cardiac cell that are involved with the development of cardiomyopathic diseases. This proposal focuses on cardiac troponin C (cTnC), the on-off switch of the contractile apparatus and a major cardiomyocyte intracellular Ca2+ buffer. Aimed at understanding the regulatory properties of the troponin complex in vivo and its link to abnormal intracellular Ca2+ handling governing heart disease, this proposal is of considerable health relevance. Mutations in the regulatory complex of the thin filament (tropomyosin, troponin T and troponin I) associated with phenotypic outcomes of hypertrophic (HCM) and dilated (DCM) cardiomyopathies are suggested to indirectly disrupt cardiac muscle contraction by altering the Ca2+-binding properties of cTnC. However, effects of cTnC mutants that influence Ca2+-sensitive contractile responses have yet to be tested for their pathogenic capacity in living organisms. The central hypothesis guiding this proposal is that changes in cTnC N-terminus Ca2+-binding affinity, alone, can evoke cardiac remodeling in vivo. We further posit that ablation of a dedicated high-fidelity kinase has the potential to reverse the hypercontractile state imposed by Ca2+-sensitizing HCM-linked cTnC mutants. Aim 1 will evaluate direct changes in cTnC Ca2+-binding affinity in the thin filament as a critical determinant underlying cardiomyopathic development. This Aim tests the hypothesis that cTnC mutants increasing Ca2+-binding affinity in the N-domain (regulatory) can instigate diastolic dysfunction, leading to HCM; while a designed mutant decreasing cTnC Ca2+- binding affinity will recapitulate a DCM-reminiscent phenotype. Cardiac patho-physiological, biophysical and biochemical approaches will be used to dissect the role of cTnC mutations in our newly developed knock-in (KI) mice. The antithetical effects that these cTnC mutants exert on Ca2+-binding dynamics will be investigated early, prior to development of distinctive cardiac remodeling. In addition, this projet will further define TNNC1 (cTnC- encoding gene) as a cardiomyopathy-susceptibility gene. Aim 2 will establish mechanistic and potential therapeutic links regarding normalization of myofilament Ca2+-sensitivity. This Aim examines whether conditional removal of a dedicated sarcomeric kinase will correct the myofilament Ca2+ response, diminish the hypercontractile phenotype and improve cardiac relaxation, thus reversing post-symptomatic HCM disease. The consequences of conditional removal of this kinase in KI cTnC-HCM hearts will be monitored as a function of time. The last aim can serve as a proof-of-concept for the development of targeted therapies aimed at modulating the activity of dedicated sarcomeric kinases. The novel concepts generated here will define the role of cTnC in initiating and modulating one class of cardiomyopathies, thus opening avenues for development of new tailored therapeutic approaches.

 描述（由申请人提供）：这项工作的长期目标是确定心肌细胞内与心肌病发展有关的成分。该提案重点关注心肌肌钙蛋白 C (cTnC)、收缩装置的开关和主要的心肌细胞胞内 Ca2+ 缓冲液。该提议旨在了解体内肌钙蛋白复合物的调节特性及其与控制心脏病的异常细胞内 Ca2+ 处理的联系，具有相当大的健康相关性。与肥厚型 (HCM) 和扩张型 (DCM) 心肌病表型结果相关的细丝调节复合物（原肌球蛋白、肌钙蛋白 T 和肌钙蛋白 I）的突变被认为通过改变 cTnC 的 Ca2+ 结合特性来间接破坏心肌收缩。然而，cTnC 突变体影响 Ca2+ 敏感收缩反应的作用尚未在活生物体中对其致病能力进行测试。指导这一提议的中心假设是，cTnC N 末端 Ca2+ 结合亲和力的变化单独可以引起体内心脏重塑。我们进一步假设，消融专用的高保真激酶有可能逆转 Ca2+ 敏化 HCM 相关的 cTnC 突变体造成的过度收缩状态。目标 1 将评估细丝中 cTnC Ca2+ 结合亲和力的直接变化，作为心肌病发展的关键决定因素。该目的测试了以下假设：cTnC 突变体增加 N 域（调节性）中 Ca2+ 结合亲和力，可引发舒张功能障碍，导致 HCM；而降低 cTnC Ca2+ 结合亲和力的设计突变体将重现 DCM 相关表型。心脏病理生理学、生物物理和生化方法将用于剖析 cTnC 突变在我们新开发的敲入 (KI) 小鼠中的作用。在形成独特的心脏重塑之前，将尽早研究这些 cTnC 突变体对 Ca2+ 结合动力学的相反作用。此外，该项目还将进一步将TNNC1（cTnC编码基因）定义为心肌病易感基因。目标 2 将建立有关肌丝 Ca2+ 敏感性正常化的机制和潜在治疗联系。该目的检查有条件去除专用肌节激酶是否会纠正肌丝 Ca2+ 反应、减少过度收缩表型并改善心脏舒张，从而逆转症状后 HCM 疾病。将监测 KI cTnC-HCM 心脏中有条件去除该激酶的后果，作为时间的函数。最后一个目标可以作为开发旨在调节专用肌节激酶活性的靶向疗法的概念验证。这里产生的新概念将定义 cTnC 在引发和调节一类心肌病中的作用，从而为开发新的定制治疗方法开辟途径。

项目成果

Micro-mechanical response and power-law exponents from the longitudinal fluctuations of F-actin solutions.

• DOI: 10.1039/d2sm01445a
• 发表时间: 2023-05-24
• 期刊: SOFT MATTER
• 影响因子: 3.4
• 作者: Dominguez-Garcia, Pablo;Pinto, Jose R.;Akrap, Ana;Jeney, Sylvia
• 通讯作者: Jeney, Sylvia

Will you still need me (Ca2+ , TnT, and DHPR), will you still cleave me (calpain), when I'm 64?

当我 64 岁时，你还会需要我（Ca2、TnT 和 DHPR）吗？你还会切割我（钙蛋白酶）吗？

• DOI: 10.1111/acel.12560
• 发表时间: 2017
• 期刊: Aging cell
• 影响因子: 7.8
• 作者: Pinto,JoséRenato;Muller-Delp,Judy;Chase,PBryant
• 通讯作者: Chase,PBryant

Hypertrophic Cardiomyopathy Cardiac Troponin C Mutations Differentially Affect Slow Skeletal and Cardiac Muscle Regulation.

肥厚型心肌病心肌肌钙蛋白 C 突变对骨骼和心肌调节缓慢有不同影响。

• DOI: 10.3389/fphys.2017.00221
• 发表时间: 2017
• 期刊: Frontiers in physiology
• 影响因子: 4
• 作者: Veltri,Tiago;Landim-Vieira,Maicon;Parvatiyar,MichelleS;Gonzalez-Martinez,David;DieseldorffJones,KarissaM;Michell,ClaraA;Dweck,David;Landstrom,AndrewP;Chase,PBryant;Pinto,JoseR
• 通讯作者: Pinto,JoseR

Mutation location of HCM-causing troponin T mutations defines the degree of myofilament dysfunction in human cardiomyocytes.

• DOI: 10.1016/j.yjmcc.2020.10.006
• 发表时间: 2021-01
• 期刊: Journal of molecular and cellular cardiology
• 影响因子: 5
• 作者: Schuldt M;Johnston JR;He H;Huurman R;Pei J;Harakalova M;Poggesi C;Michels M;Kuster DWD;Pinto JR;van der Velden J
• 通讯作者: van der Velden J

Amide hydrogens reveal a temperature-dependent structural transition that enhances site-II Ca2+-binding affinity in a C-domain mutant of cardiac troponin C.

酰胺氢揭示了温度依赖性结构转变，可增强心肌肌钙蛋白 C 的 C 结构域突变体中位点 II Ca2 的结合亲和力。

• DOI: 10.1038/s41598-017-00777-6
• 发表时间: 2017
• 期刊: Scientific reports
• 影响因子: 4.6
• 作者: Veltri,Tiago;deOliveira,GuilhermeAP;Bienkiewicz,EwaA;Palhano,FernandoL;Marques,MayradeA;Moraes,AdolfoH;Silva,JersonL;Sorenson,MarthaM;Pinto,JoseR
• 通讯作者: Pinto,JoseR