Myocardial remuscularization by cardiac patch delivery of epicardial FSTL1 and CCND2 overexpressing cardiomyocytes

标题： 心外膜FSTL1和CCND2心脏贴片心肌重建术 过表达的心肌细胞 项目摘要 尽管接受了强化治疗方案，严重急性心肌梗死(AMI)患者经常 以终末期充血性心力衰竭(CHF)告终。从分子和细胞的角度来看，心力衰竭 由于左心室收缩单位：心肌细胞(CMS)的丧失而发生。因此，推广 心肌细胞的增殖和了解心肌细胞周期的调节机制可能会产生非常重要的影响 关于心力衰竭的管理。在这项提案中，我们寻求开发3D生物工程心肌 构建了一个功能性血管网络并概括了一些关键的微环境 这是天然心脏组织的线索。我们最近的研究已经确定了主要的生物力学和分子线索， 显著提高成年CMS的细胞周期再入能力。我们证明了心外膜应用心脏 含有卵泡抑素样蛋白-1(FSTL1)的贴片可保护小鼠和猪的心脏免受急性心肌梗死、左心室 扩张和心力衰竭。我们最近报道了细胞周期基因CCND2(细胞周期蛋白D2)的过表达， 诱导移植的人诱导多能干细胞(HiPSC)来源的CMS增殖。这项建议 以我们最新的技术成就为基础，使主要心脏细胞的铸型或生物打印成为可能 在高空间分辨率(20微米)的水凝胶，以制造3D可灌流的血管结构。我们的中央 假说是，携带FSTL1和hiPSC-CCND2 CMS的3D心脏结构可以协同作用 使缺血心肌重新肌化。我们在三个综合的特定目标(SA)中检验这一假说。在SA1中， 我们将利用我们的管型/生物打印的3D心脏组织模型来识别细胞和分子机制 FSTL1处理对体外培养心肌细胞的促增殖作用。在SA2中，我们将评估已识别的 参与FSTL1-CCND2 CM-Patch效应的信号通路促进小鼠模型肌化 心肌梗塞(急性和慢性)。在SA3a中，我们将评估生物工程预临床应用的潜力。 带血管肌片装置治疗猪急性心肌梗死缺血再灌流模型。我们将比较 开胸分娩与新型微创导管心包分娩FSTL1的疗效比较 CCND2-CM载肌片至心外膜。SA3B，我们会评估该计划的成效 工程补片，防止左室扩张，而不会导致心律失常。全景图 光学标测和跨壁电EP标测将评估整个心脏的机电一体化 在肌肉补片结构和受体心肌之间。这些EP研究的结果将指导 设计具有改进的EP特性的新一代细胞系和贴片结构，从而 降低移植物相关性心律失常的风险

Title: Myocardial remuscularization by cardiac patch delivery of epicardial FSTL1 and CCND2 overexpressing cardiomyocytes Project Summary Despite undergoing intensive treatment regimens, patients with severe acute myocardial infarction (AMI) often end up with end stage congestive heart failure (CHF). From the molecular and cellular perspective, heart failure occurs due to the loss of the contractile unit of the left ventricle: cardiomyocytes (CMs). Therefore, promotion of myocyte proliferation and understanding the regulators of myocyte cell cycle could have highly significant impact on the management of heart failure. In this proposal, we seek to develop 3D bioengineered cardiac muscle constructs that incorporate a functional vascular network and recapitulate some of the key microenvironmental cues of native heart tissue. Our recent studies have identified main biomechanical and molecular cues that can significantly enhance cell cycle re-entry of adult CMs. We demonstrated that epicardial application of a cardiac patch, laden with follistatin like-1 (FSTL1) protein, protected the mouse and pig heart against AMI, left ventricle dilatation, and heart failure. We recently reported that overexpression of a cell cycle gene, CCND2 (cyclin D2), induces proliferation of transplanted human induced pluripotent stem cell (hiPSC) derived-CMs. This proposal builds upon our recent technological achievements, enabling cast or bioprinting of major cardiac cells and hydrogels at high spatial resolution (20 µm) to fabricate 3D perfusable vascular constructs. Our central hypothesis is that 3D cardiac constructs, laden with FSTL1 and hiPSC-CCND2 CMs, can synergistically remuscularize ischemic myocardium. We test this hypothesis in three integrated Specific Aims (SAs). In SA1, we will utilize our cast/bioprinted 3D cardiac tissue models to identify the cellular and molecular mechanisms underlying myocyte pro-proliferative effect of FSTL1 treatment in vitro. In SA2, we will assess the identified signaling pathways, involved in FSTL1-CCND2 CM-patch effect, to promote remuscularization in mouse model of MI (both acute and chronic). In SA3a, we will assess the pre-clinical potential of bioengineered pre- vascularized muscle patch device in treating AMI in a pig model of ischemia-reperfusion. We will compare the efficacy of open chest delivery versus a novel minimally invasive, catheter-based, pericardial delivery of FSTL1 and CCND2 CM laden muscle patch to the epicardium. SA3b, we will evaluate the effectiveness of the engineered patch for preventing the LV dilatation without inducing arrhythmogenic complications. The panoramic optical mapping and transmural electrical EP mapping whole heart will assess the electromechanical integration between the muscle patch constructs and recipient myocardium. The findings from these EP studies will guide the design of new generations of cell lines and patch constructs with improved EP characteristics, thereby reducing the risk of graft-associated arrhythmia