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Novel Approach to Enhance Myocardial Performance and Improve Heart Failure Outcome

增强心肌性能和改善心力衰竭结果的新方法

基本信息

批准号：
10064633
负责人：
CHARLES C HONG
金额：
$ 38.63万
依托单位：
UNIVERSITY OF MARYLAND BALTIMORE
依托单位国家：
美国
项目类别：
财政年份：
2017
资助国家：
美国
起止时间：
2017-12-15 至 2022-11-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10064633
关键词：
Academic Medical Centers Acute Affect American Animal Model Biochemical Biological Biological Assay Biology CRISPR/Cas technology Calcium Cardiac Cardiac Myocytes Cell Line Cell physiology Cessation of life Chemicals Clinical Data Concentration Camps Congestive Heart Failure Culture Techniques DNA Databases Databases Development Electronic Health Record Evaluation Foundations Genes Genomic approach Geranyltranstransferase Heart Rate Heart failure Human Hypertrophy In Vitro Individual Knock-out Laboratories Life Link Mattresses Measurement Mediating Medicine Methods Modeling Modification Morbidity - disease rate Mus Myocardial Myocardium Natural History Nucleotides Outcome Parents Performance Persons Pharmaceutical Preparations Pharmacology Pharmacotherapy Protein Isoprenylation Regulation Relaxation Risk Role Structure Structure-Activity Relationship Symptoms Therapeutic Time Treatment Failure Validation Variant analog base drug candidate drug development functional genomics gain of function genome editing heart function improved in vivo induced pluripotent stem cell innovation loss of function matrigel mortality new therapeutic target novel novel strategies novel therapeutics prenylation repaired screening side effect small molecule therapeutic target

项目摘要

Heart failure (HF) is a leading cause of morbidity and mortality, contributing to 1 in 9 deaths in the US. Consequently, there is an enormous need for new HF therapies, which can only emerge from discovery of new therapeutic targets. In the past, inotropic drugs that enhance myocardial performance acutely were developed to treat HF, but most of them are now contraindicated because they worsen HF outcomes long-term. Recently, we developed a novel culturing method, termed Matrigel Mattress, which allowed the simultaneous assessment of contractile performance and calcium dynamics in individual human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs). Using the Matrigel Mattress method as a basis for a chemical screening platform, we discovered that the small molecule EGM significantly enhanced both inotropy and lusitropy in hiPSC-CMs and improved cardiac function in vivo. Unlike the traditional inotropes, EGM did not affect calcium (Ca) cycling, cellular cAMP concentrations or increase the beat rate, suggesting it acts by a fundamentally novel mechanism. To unlock the mechanistic underpinnings of EGM's pharmacology, we carried out a biochemical pull-down assay and identified farnesyl diphosphate synthase (FDPS), required for protein prenylation, as a candidate target of EGM. Consistent with prior studies demonstrating that FDPS contributes to hypertrophy and HF in animal models, we found that naturally occurring variants in the FDPS gene were highly associated with HF in Vanderbilt University Medical Center's electronic health record-linked DNA database. The latter result, based on real world clinical data, raises the exciting possibility that modulating the level of FDPS activity over a course of a person's life can significantly alter HF natural history; and that compounds like EGM that inhibit FDPS may improve long-term HF outcomes. Based on these findings, we hypothesize that EGM enhances myocardial performance by inhibiting FDPS, and that FDPS inhibition improves both acute cardiac function and long-term HF outcome. Here, we propose innovative chemical and functional genomic approaches to elucidate the role of FDPS in EGM function. In Aim 1, we will carry out a structure activity relationship (SAR) study of EGM analogs to determine whether FDPS inhibition is essential for EGM function. In Aim 2, we will employ the CRISPR/Cas9-mediated genome editing to determine whether ablating the FDPS gene recapitulates EGM's unique pharmacology in hiPSC-CMs. In Aim 3, we will utilize the CRISPR/Cas9- directed homology directed repair (HDR) to introduce the nucleotide changes corresponding to the CHF- associated FDPS variants, and evaluate their impact on hiPSC-CM performance and FDPS function. The proposed study will delineate the effects of FDPS modulation on myocardial performance, and possibly identify additional targets of EGM. This study leverages the unique pharmacology of EGM to lay the foundation for a new understanding of myocardial regulation and the new therapeutic paradigm of “dual purpose” drugs that acutely relieve HF symptoms as well as improve long-term HF outcomes.

心力衰竭(HF)是发病率和死亡率的主要原因，在美国，每9人中就有1人死于心力衰竭。因此，对新的心衰疗法的巨大需求，只有在新的发现之后才能出现。治疗靶点。在过去，可以显著提高心肌功能的肌力药物被开发出来。用于治疗心力衰竭，但大多数药物现在都是禁忌，因为它们会使心力衰竭的长期结局恶化。最近，我们开发了一种新的培养方法，称为Matrigel床垫，它允许同时评估人诱导多能干细胞来源的个体收缩性能和钙动力学的研究心肌细胞(HiPSC-CMS)。使用Matrigel床垫法作为化学筛选的基础平台上，我们发现小分子EGM显著增强了肌力和肌力 HiPSC-CMS和改善体内心功能。与传统的肌力调节药不同，EGM不影响钙离子 (CA)骑自行车，细胞内cAMP浓度或增加节拍频率，表明它的作用从根本上说是新的机制。为了解开EGM药理学的机制基础，我们进行了一项生化下拉试验和鉴定蛋白质所需的法尼基二磷酸合成酶(FDPS) 异丙烯基化，作为EGM的候选靶点。与先前的研究表明FDPS有助于在动物模型中，我们发现FDPS基因的自然发生的变体高度在范德比尔特大学医学中心的电子健康记录链接的DNA数据库中与HF有关。这个后者的结果，基于真实世界的临床数据，提出了调节FDPS水平的令人兴奋的可能性一个人一生中的活动可以显著改变HF的自然历史；而像EGM这样的化合物抑制FDPS可能会改善长期心力衰竭的结果。基于这些发现，我们假设特别股东大会通过抑制FDPS改善心肌功能，FDPS抑制可改善急性心肌梗死心功能和远期心衰转归。在这里，我们提出了创新的化学和功能基因组阐明FDPS在EGM功能中作用的途径。在目标1中，我们将开展一项结构化活动对EGM类似物进行相关性(SAR)研究，以确定FDPS抑制是否对EGM功能至关重要。在目标2中，我们将使用CRISPR/Cas9介导的基因组编辑来确定是否消融FDPS 基因概括了EGM在HiPSC-CMS中的独特药理作用。在目标3中，我们将利用CRISPR/Cas9- 定向同源定向修复(HDR)，以引入与CHF- 相关的FDPS变体，并评估它们对HiPSC-CM性能和FDPS功能的影响。这个拟议的研究将描绘FDPS调制对心肌功能的影响，并可能确定特别股东大会的其他目标。这项研究利用了EGM独特的药理作用，为对心肌调节的新认识和“两用”药物的治疗新范式显著缓解心力衰竭症状，改善心力衰竭的长期结局。

项目成果

期刊论文数量（15）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Genome Editing and Induced Pluripotent Stem Cell Technologies for Personalized Study of Cardiovascular Diseases.

用于心血管疾病个性化研究的基因组编辑和诱导多能干细胞技术。

DOI：
10.1007/s11886-018-0984-9
发表时间：
2018
期刊：
Current cardiology reports
影响因子：
3.7
作者：
Chun,YoungWook;Durbin,MatthewD;Hong,CharlesC
通讯作者：
Hong,CharlesC

Exosomes mediated fibrogenesis in dilated cardiomyopathy through a MicroRNA pathway.

DOI：
10.1016/j.isci.2023.105963
发表时间：
2023-02-17
期刊：
ISCIENCE
影响因子：
5.8
作者：
Fu, Xuebin;Mishra, Rachana;Chen, Ling;Arfat, Mir Yasir;Sharma, Sudhish;Kingsbury, Tami;Gunasekaran, Muthukumar;Saha, Progyaparamita;Hong, Charles;Yang, Peixin;Li, Deqiang;Kaushal, Sunjay
通讯作者：
Kaushal, Sunjay

Identification of Novel Genetic Variants and Comorbidities Associated With ICD-10-Based Diagnosis of Hypertrophic Cardiomyopathy Using the UK Biobank Cohort.