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

促进术后心肌再肌化的内源性和外源性机制 心肌梗死左室重构 摘要/摘要 进展性心力衰竭的分子和细胞基础是受损的不能 和凋亡的肌细胞被取代。虽然许多基于细胞和组织的疗法可以限制这一点 功能障碍，在给药部位存活超过几周的细胞比例 移植的比例极低。因此，梗死区实质上的肌肉化很少发生。 有报道；当有有限的肌肉化时，经常伴随着 机制不明的潜在致命性室性心律失常。这项提议的目的是重组 通过识别细胞周期的关键调节因子和促进天然的 心肌细胞(CM)重新进入细胞周期，并通过移植生物工程心肌从“外部”进入 补丁(HCMP)与CM细胞周期的关键调节因子上调，并与功能 并概括了天然心脏组织的一些关键微环境线索。我们 最近建立了一种由MHC驱动过表达CM关键调控因子CCND2的新的HiPSC细胞系 (hiPSC-MHC-CCND2OE)，可使啮齿动物损伤的脑室重新肌化。的中心目标是 这一建议是为了让心肌细胞周期的时钟倒流，以进行心肌修复。具体目标(SA)包括： SA1：确定促进早期新生猪心脏细胞周期活动的关键调节因子 心肌损伤。我们将：1)使用最先进的分子生物学和成像技术，以及单一的 细胞/核RNA测序(scRNAseq或SnRNAseq)技术来演示这些关键 控制心肌细胞周期的调节/信号通路；和2)测试损伤后的肌肉化 通过使用靶向modRNA或AAV9操纵关键调节器来选择性地修饰这些 急性心肌梗死成年猪的调节剂。SA2a。以前无法达到的尺寸和厚度的工程hcmps 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 人巨噬细胞集落刺激蛋白在大动物(猪)心肌损伤模型中的心肌修复和再肌化作用 受伤。我们将使用最先进的光学测绘技术与三维 室壁心内标测勾画整个左心室潜在的心律失常机制 表面的和跨壁的。

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.