Identification and Characterization of Matrikines for Cardiac Differentiation and Regeneration

PI: Black, Lauren d .提案号：1603524对可用于治疗心脏损伤或疾病的材料和技术的改进有很大的需求。研究人员已经证明，来自心脏细胞外基质（cECM）的多种单一蛋白质促进心肌细胞（CM）增殖或干细胞的心脏分化，但很少有工作来鉴定这些蛋白质上的短肽序列，称为基质因子，是导致细胞反应改变的活性成分。基于初步数据表明，与成人心脏相比，心脏细胞对从胎儿心脏中获得的cECM的反应不同，这个为期3年的项目的研究人员将测试一个假设，即在发育中的心脏ECM中存在许多基质因子，通过增强CM的增殖和存活以及干细胞的心脏分化，这些基质因子可能对心脏再生和修复策略有用。这些母因子的鉴定对于理解在心脏生长过程中cECM重塑的重要性以及心脏损伤或疾病治疗方法的潜在发展具有重要意义。通过在课程和实验室中整合技术和成果，将在本科和研究生两级产生教育影响；在高中层面的影响将通过当地高中的研讨会和生物医学工程系的高中生暑期研究项目的一部分来实现。细胞外基质（ECM）的组成是许多细胞过程的强大调节剂。在心脏中，来自心脏ECM （cECM）的多种单一蛋白促进心肌细胞（CM）增殖或干细胞的心脏分化。到目前为止，大多数研究只研究了单一的ECM蛋白作为细胞的结合位点，尽管体内的ECM是一种复杂的蛋白质混合物，具有器官/组织和生命阶段特异性。此外，重点是单一蛋白质，而不是短肽段/基质因子，这可能是导致细胞反应改变的生物活性区域。研究人员的实验室已经证明，与来自成人心脏的ECM相比，来自胎儿心脏的cECM促进心肌细胞增殖，并且心脏祖细胞对来自不同发育阶段的cECM的分化反应发生了改变。因此，该项目的目标是验证这样一个假设，即在发育中的心脏ECM中存在许多基质因子，这些基质因子可以通过增强CM的增殖和存活以及心脏干细胞的分化，在心脏再生和修复策略中发挥作用。该假设将通过实施两个目标来检验：1)鉴定和鉴定来自胎儿和成年cECM（妊娠3-6个月的猪）的母因子，这些母因子促进诱导多能干细胞（iPSC）向心脏细胞分化和/或促进iPSC来源的CMs （iPSC-CMs）的增殖或成熟。2)评估来自cECM的母因子促进人iPSC- cm在健康和病变心肌的3-D工程化心脏组织模型中的存活、增殖和功能。这些母因子的鉴定对于理解在心脏生长过程中cECM重塑的重要性以及心脏损伤或疾病治疗方法的潜在发展具有重要意义。通过在课程和实验室中整合技术和成果，将在本科和研究生两级产生教育影响；在高中层面的影响将通过在当地高中的研讨会和作为BME部门的高中生暑期研究项目的一部分。

PI: Black, Lauren D.Proposal #: 1603524There is great need for improved materials and techniques that can be used for the treatment of cardiac injury or disease. Investigators have shown that a variety of singular proteins derived from the cardiac extracellular matrix (cECM) promote cardiomyocyte (CM) proliferation or cardiac differentiation of stem cells, but little work has been done to identify the short peptide sequences on those proteins, termed matrikines, that are the active ingredients leading to the altered cellular responses. Based on preliminary data that indicates that cardiac cells respond differently to cECM obtained from fetal compared to adult hearts, the investigators of this 3 year project are going to test the hypothesis that there exists a number of matrikines in the developing heart ECM that could be useful in strategies for cardiac regeneration and repair, via enhancement of CM proliferation and survival and the cardiac differentiation of stem cells. Identification of these matrikines would be significant for understanding the importance of cECM remodeling during cardiac growth as well as for the potential development of therapeutics for the treatment of cardiac injury or disease. Educational impact will be made at the undergraduate and graduate levels through integration of techniques and results in courses and laboratories; impact at the high school level will be made through seminars at local high schools and as part of the Biomedical Engineering department's summer research program for high school students. The composition of the extracellular matrix (ECM) is a powerful regulator of a number of cellular processes. In the heart, a variety of singular proteins derived from the cardiac ECM (cECM) promote cardiomyocyte (CM) proliferation or cardiac differentiation of stem cells. Most investigations to date have examined only singular ECM proteins as binding sites for the cells, even though the ECM in vivo is a complex mixture of proteins that is organ/ tissue and life stage specific. Furthermore, the emphasis has been on singular proteins rather than on the short peptide segments/matrkines that are likely to be the bioactive region leading to altered cellular responses. The investigators' lab has shown that cECM derived from fetal hearts promotes cardiomyocyte proliferation as compared to ECM derived from adult hearts and that cardiac progenitor cells have altered differentiation responses to cECM derived from different developmental stages. Thus, the goal of this project is to test the hypothesis that there exists a number of matrikines in the developing heart ECM that could be useful in strategies for cardiac regeneration and repair, via enhancement of CM proliferation and survival and the cardiac differentiation of stem cells. The hypothesis will be tested by carrying out two aims: 1) Identify and characterize matrikines derived from fetal and adult cECM (pigs at 3rd trimester and 3-6 months) that promote induced pluripotent stem cell (iPSC) differentiation to cardiac cells and/or enhance proliferation or maturation of iPSC-derived CMs (iPSC-CMs) and 2) Assess the efficacy of the matrikines derived from cECM in promoting human iPSC-CM survival, proliferation and function in 3-D engineered cardiac tissue models of healthy and diseased myocardium. Identification of these matrikines would be significant for understanding the importance of cECM remodeling during cardiac growth as well as for the potential development of therapeutics for the treatment of cardiac injury or disease. Educational impact will be made at the undergraduate and graduate levels through integration of techniques and results in courses and laboratories; impact at the high school level will be made through seminars at local high schools and as part of the BME department's summer research program for high school students.