Project 1 - Endogenous and Exogenous Mechanisms that Promote Myocardial Remuscularization

项目 1 - 促进心肌再肌化的内源性和外源性机制

• 批准号: 10677730
• 负责人: Jianyi Zhang
• 金额: $ 55.33万
• 依托单位: UNIVERSITY OF ALABAMA AT BIRMINGHAM
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-01 至 2027-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10677730
• 关键词: 3-Dimensional; Acute myocardial infarction; Adult; Animals; Apoptotic; Arrhythmia; Back; Biomedical Engineering; Birth; Blood Vessels; CCND2 gene; Cardiac; Cardiac Myocytes; Cell Cycle; Cell Line; Cell Nucleus; Cell Proliferation; Cell Survival; Cell Transplantation; Cells; Cicatrix; Congestive Heart Failure; Cues; Dilatation - action; Dimensions; Electric Stimulation; Endogenous Factors; Endothelial Cells; Engineering; Engraftment; Family suidae; Fibroblasts; Functional disorder; Generations; Heart; Heart failure; Human; Human Engineering; Imaging technology; Infarction; Injury; Left; Left Ventricular Remodeling; Left ventricular structure; Mammals; Maps; Measurement; Modeling; Molecular; Molecular Biology; Mus; Muscle; Muscle Cells; Myocardial; Myocardial tissue; Myocardium; Natural regeneration; Neonatal; Optics; Patients; Periodicity; Pilot Projects; Pluripotent Stem Cells; Proliferating; Protocols documentation; Public Health; RNA; Recovery; Regimen; Regulatory Pathway; Reporting; Rodent Model; Signal Pathway; Site; Smooth Muscle Myocytes; Stretching; Surface; Techniques; Technology; Testing; Therapeutic; Thick; Tissue Engineering; Tissues; Transplantation; Vascularization; Ventricular; Ventricular Arrhythmia; cardiac tissue engineering; cell type; clinically relevant; effectiveness evaluation; endothelial stem cell; functional improvement; heart function; improved; in vivo; injured; ischemic injury; manufacture; mature animal; molecular imaging; myocardial injury; novel; overexpression; porcine model; postnatal; prevent; promoter; regeneration potential; repaired; restoration; success; synergism; transcriptome sequencing

SUMMARY / ABSTRACT (Project 1) Endogenous and Exogenous Mechanisms that Promote Myocardial Remuscularization in Postinfarction LV Remodeling The molecular and cellular basis for progressive heart failure is the result of the inability of damaged and apoptotic myocytes to be replaced. While a number of cell- and tissue-based therapies can limit this dysfunction, the proportion of cells that survive at the site of administration for more than a few weeks after transplantation is extremely low. As such, substantial remuscularization of the infarcted region has rarely been reported; and when limited remuscularization has been reported, it is frequently accompanied by potentially lethal ventricular arrhythmias of unknown mechanism. This proposal aims at remuscularization of the injured ventricle by the definition of key endogenous factors that regulate and promote the cell cycle of the native cardiomyocyte (CM), and from exogenous transplanted bioengineered cardiac muscle patch (hCMP) that overexpresses key regulators of CM cell cycle, and will incorporate a functional vascular network and recapitulate some of the key micro environmental cues of native heart tissue. We recently established a novel hiPSC cell line with MHC-driven overexpression of a key regulator of CMs: CCND2 (hiPSC-MHC-CCND2OE), which can remuscularize injured ventricle in rodent model. The central objective of this proposal is to “turn back the clock” of the myocyte cell cycle, which will facilitate myocardial repair. The Specific Aims (SA) that will examine this objective include: SA1: To identify the key regulators that promote cell-cycle activity in the hearts of early neonatal pigs after myocardial injury. We will: 1) using state-of-the-art fate-mapping molecular biology and imaging technologies, and the single cell/nucleus RNA sequencing (scRNAseq or snRNAseq) technology demonstrate the key regulators/signaling pathways that govern the myocyte cell cycle; and 2) test the remuscularization of the injured ventricle by manipulating the key regulators using either targeted modRNA or AAV9 delivery strategies to selectively modify these regulators in adult pig hearts following ischemic injury. SA2a. To engineer hCMPs containing CMs that are capable of proliferating after transplantation, and characterized by previously unattainable size and thickness that are functionally mature and primed for in-vivo vascularization. SA2b. To evaluate the effectiveness of our hCMP constructs for myocardial recovery and remuscularization in a large-animal (pig) model of myocardial injury. We will use state-of-the-art techniques that includes optical mapping in combination with the 3- dimensional intramural cardiac mapping to delineate the potential arrhythmia mechanisms over the entire transmural and left ventricular surface. These studies will synergize with the other projects and serve as a prelude for therapeutic initiatives focused on remuscularization of the injured human heart.

摘要/摘要 (项目1) 内生和外生机制促进 心肌梗塞后左室重构中的心肌血运重建 进展性心力衰竭的分子和细胞基础是由于受损和 凋亡的肌细胞将被替换。虽然许多基于细胞和组织的疗法可以限制这种功能障碍， 移植后在给药部位存活超过几周的细胞比例为 非常低。因此，梗死区实质上的肌肉化很少被报道； 已有报道称有限度的心肌重建术，它经常伴有潜在的致命的脑室。 机制不明的心律失常。这项建议旨在通过对受伤的脑室进行肌肉化 调节和促进天然心肌细胞(CM)细胞周期的关键内源性因素的定义， 以及来自过量表达Key的外源移植的生物工程心肌补片(HCMP) CM细胞周期的调节，并将结合一个有功能的血管网络，并概括一些关键 天然心脏组织的微环境线索。我们最近建立了一种由MHC驱动的新型HiPSC细胞系 CMS关键调控因子CCND2(hiPSC-MHC-CCND2OE)的过表达 啮齿动物模型的脑室。这项提议的中心目标是让心肌细胞“倒流”。 循环，这将促进心肌修复。将审查这一目标的具体目标(SA)包括：SA1： 确定促进心肌梗死后早期新生仔猪心脏细胞周期活动的关键调控因子 受伤。我们将：1)使用最先进的命运映射分子生物学和成像技术，以及单一的 细胞/核RNA测序(scRNAseq或SnRNAseq)技术展示了关键的调节/信号 控制心肌细胞周期的途径；以及2)通过以下方式测试受损的脑室的肌化 使用靶向modRNA或AAV9递送策略操纵关键调节器以选择性地修改 这些调节剂在缺血损伤后的成年猪心脏中。SA2a。要设计含有CMS的hCMPS， 在移植后能够增殖，其特点是以前无法达到的大小和厚度 它们在功能上是成熟的，并为体内血管形成做好了准备。SA2B。评估我们的计划的有效性 人巨噬细胞集落刺激蛋白在大动物(猪)心肌模型中的心肌恢复和再肌化作用 受伤。我们将使用最先进的技术，包括光学测绘与3- 三维室壁内标测描绘整个心脏的潜在心律失常机制 跨室壁和左心室表面。这些研究将与其他项目产生协同作用，并作为前奏。 用于侧重于受伤心脏肌肉化的治疗举措。