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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，导致美国九分之一的死亡。因此，对新的心衰疗法的需求巨大，而这种疗法只能通过新的发现来实现。治疗目标。过去，开发了急剧增强心肌功能的正性肌力药物治疗心力衰竭，但现在大多数药物都是禁忌的，因为它们会长期恶化心力衰竭的结果。最近，我们开发了一种新颖的培养方法，称为基质胶床垫，可以同时评估人类诱导多能干细胞来源的收缩性能和钙动力学的影响心肌细胞（hiPSC-CM）。使用基质胶床垫方法作为化学筛选的基础平台上，我们发现小分子 EGM 显着增强了正性肌力和松弛性 hiPSC-CM 并改善体内心脏功能。与传统正性肌力药物不同，EGM 不会影响钙 (Ca) 循环，细胞 cAMP 浓度或增加心跳率，表明它从根本上起作用新颖的机制。为了解开 EGM 药理学的机制基础，我们进行了一项生化下拉分析并鉴定了蛋白质所需的法尼基二磷酸合酶 (FDPS) 异戊二烯化，作为临时股东大会的候选目标。与之前的研究一致，证明 FDPS 有助于在动物模型中，我们发现 FDPS 基因中自然发生的变异与肥大和心力衰竭高度相关。范德比尔特大学医学中心的电子健康记录链接 DNA 数据库中与心力衰竭相关。这后一个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着改变心力衰竭的自然史；以及像 EGM 这样的化合物抑制 FDPS 可能会改善长期心力衰竭结局。基于这些发现，我们假设 EGM 通过抑制 FDPS 来增强心肌性能，并且 FDPS 抑制可改善急性心功能和长期心力衰竭结局。在这里，我们提出创新的化学和功能基因组阐明 FDPS 在 EGM 功能中的作用的方法。在目标 1 中，我们将开展结构活动 EGM 类似物的关系（SAR）研究，以确定 FDPS 抑制是否对 EGM 功能至关重要。在目标 2 中，我们将采用 CRISPR/Cas9 介导的基因组编辑来确定是否消除 FDPS 基因概括了 EGM 在 hiPSC-CM 中的独特药理学。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-相对应的核苷酸变化相关的 FDPS 变体，并评估它们对 hiPSC-CM 性能和 FDPS 功能的影响。这拟议的研究将描述 FDPS 调节对心肌性能的影响，并可能确定临时股东大会的额外目标。本研究利用 EGM 的独特药理学为以下方面奠定了基础：对心肌调节的新认识和“双用”药物的新治疗范式急性缓解心力衰竭症状并改善长期心力衰竭结局。

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.