Identification and Characterization of Matrikines for Cardiac Differentiation and Regeneration

用于心脏分化和再生的 Matrikines 的鉴定和表征

• 批准号: 1603524
• 负责人: Lauren Black
• 金额: $ 42.46万
• 依托单位: Tufts University
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-07-01 至 2019-06-30
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1603524&HistoricalAwards=false
• 关键词: Identification; Characterization; Matrikines; Cardiac; Differentiation

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.

PI：Black，Lauren D.Proposal#：1603524非常需要改进的材料和技术，可以用于治疗心脏损伤或疾病。研究人员已经证明，来自心脏细胞外基质(CECM)的各种单一蛋白质可以促进心肌细胞(CM)的增殖或干细胞的心脏分化，但很少有人对这些蛋白质上的短肽序列进行鉴定，这些蛋白质被称为基质因子，这些蛋白质是导致细胞反应改变的活性成分。根据初步数据显示，心脏细胞对从胎儿心脏获得的cECM的反应与成人心脏不同，这个为期3年的项目的研究人员将检验这一假设，即在发育中的心脏ECM中存在许多可能有助于心脏再生和修复的策略，通过促进CM的增殖和存活以及干细胞的心脏分化。识别这些基质因子对于理解心脏生长过程中cECM重构的重要性以及开发治疗心脏损伤或疾病的潜在疗法具有重要意义。将通过在课程和实验室中整合技术和结果，在本科和研究生层面产生教育影响；在高中层面，将通过在当地高中举行研讨会，并作为生物医学工程系针对高中生的暑期研究计划的一部分，产生影响。细胞外基质(ECM)的组成是许多细胞过程的强大调节器。在心脏中，来自心脏ECM(CECM)的各种单一蛋白促进心肌细胞(CM)增殖或干细胞向心脏分化。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部位，尽管体内的ECM是器官/组织和生命阶段特有的蛋白质的复杂混合物。此外，重点一直放在单一蛋白质上，而不是很可能是导致细胞反应改变的生物活性区域的短肽片段/基质因子上。研究人员的实验室表明，与来自成人心脏的ECM相比，来自胎儿心脏的CECM促进了心肌细胞的增殖，并且心脏前体细胞改变了对来自不同发育阶段的CECM的分化反应。因此，该项目的目标是验证这一假设，即在发育中的心脏ECM中存在大量的基质因子，这些基质因子可以通过促进CM的增殖和存活以及干细胞的心脏分化，在心脏再生和修复策略中有用。该假说将通过实现两个目标来验证：1)鉴定和鉴定来自胎儿和成人CECM(孕3个月和3-6个月的猪)的基质因子，这些基质因子可促进诱导多能干细胞(IPSC)向心肌细胞分化和/或促进IPSC衍生CMS(iPSC-CMS)的增殖或成熟；2)在健康和患病心肌的三维工程心脏组织模型中，评估CECM衍生的基质因子在促进人IPSC-CM存活、增殖和功能方面的有效性。识别这些基质因子对于理解心脏生长过程中cECM重构的重要性以及开发治疗心脏损伤或疾病的潜在疗法具有重要意义。通过在课程和实验室中整合技术和结果，将在本科生和研究生层面产生教育影响；在高中层面，将通过在当地高中举行研讨会，并作为BME系针对高中生的暑期研究计划的一部分，产生影响。