Endogenous and exogenous mechanisms that promote myocardial remuscularization in post infarction LV remodeling

术后促进心肌肌化的内源性和外源性机制 梗死后左室重构 总结/摘要 进行性心力衰竭的分子和细胞基础是受损的心肌细胞不能正常工作的结果。 和凋亡的心肌细胞被替换。虽然一些基于细胞和组织的疗法可以限制这一点， 功能障碍，在施用后在施用部位存活超过几周的细胞的比例 移植率极低。因此，梗死区域的实质性肌肉再生很少 报告;当报告有限的肌肉再生时，经常伴有 潜在致死性室性心律失常的机制不明。这项建议旨在恢复 通过识别细胞周期的关键调节因子， 心肌细胞（CM）重新进入细胞周期，并从“外”通过移植生物工程心肌 贴片（hCMP）与CM细胞周期上调的关键调节因子，并与之结合功能性 血管网络，并概括了一些关键的微环境线索的天然心脏组织。我们 最近建立了一种新的hiPSC细胞系，其具有MHC驱动的CM的关键调节因子CCND 2的过表达 （hiPSC-MHC-CCND 2 OE），其可以在啮齿动物模型中使受损的心室肌化。的中心目标 这一建议是为了心肌修复而将心肌细胞周期的时钟“倒转”。具体目标（SA）是： SA 1：确定促进早期新生猪心脏细胞周期活性的关键调节因子， 心肌损伤我们将：1）使用最先进的分子生物学和成像技术， 细胞/细胞核RNA测序（scRNAseq或snRNAseq）技术，以证明这些关键 控制肌细胞细胞周期的调节因子/信号通路;以及2）测试损伤的 通过使用靶向modRNA或AAV 9操纵关键调节因子来选择性地修饰这些调节因子， 急性心肌梗死成年猪中的调节剂。SA2a。设计以前无法达到的尺寸和厚度的hCMP 其功能成熟并为体内血管化做好准备。SA2b.评估我们的 用于心肌损伤的大动物（猪）模型中的心肌恢复和肌肉重建的hCMP构建体 损伤我们将使用最先进的光学映射技术结合三维 壁内心脏标测以描绘整个左心室的潜在心律失常机制 表面和透壁。

Endogenous and exogenous mechanisms that promote myocardial remuscularization in post infarction LV remodeling Summary / Abstract The molecular and cellular basis for the 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 from “within” by identifying key regulators of the cell cycle and by promoting the native cardiomyocyte (CM) reenter the cell cycle, and from “outside” by transplanting bioengineered cardiac muscle patch (hCMP) with the key regulators of CM cell cycle upregulated, and with that 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 CM: 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 myocyte cell cycle for myocardial repair. The specific Aims ( SA) are: SA1: Identifying 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 molecular biology and imaging technologies, and the single cell/nucleus RNA sequencing (scRNAseq or snRNAseq) technology to demonstrate these key regulators/signaling pathways that control the myocyte cell cycle; and2) test remuscularization of injured ventricle by manipulating the key regulators using either targeted modRNA or AAV9 to selectively modify these regulators in adult pigs with AMI. SA2a. Engineering hCMPs of previously unattainable size and thickness that are functionally mature and primed for in-vivo vascularization. SA2b. Evaluating the effectiveness of our hCMP constructs for myocardial recovery and remuscularization in a large-animal (swine) model of myocardial injury. We will use state-of-the-art techniques of optical mapping in combination with the 3-dimensional intramural cardiac mapping to delineate potential arrhythmia mechanisms over the entire left-ventricular surface and transmurally.