UNS: Collaborative Research: Stem Cell-inspired Nanotherapeutics for Regenerative Repair of Elastic Matrix

主要研究者：Ramamurthi，Anand/ Rao，Raj R提案编号：1508642 / 1509377将结构受损的软弹性组织恢复到健康状态是困难的，因为成年细胞构建新的弹性纤维的能力很差，这使得组织能够拉伸和回弹。在这个项目中，研究人员提出了识别和表征来自干细胞的因子，以再生和修复弹性纤维组件和结构。此外，这些研究旨在使用可降解聚合物颗粒以持续的方式递送因子，所述可降解聚合物颗粒本身被化学改性以刺激新的弹性纤维形成并防止其分解。然后，研究人员将测试这些颗粒在治疗腹主动脉瘤方面的有效性，腹主动脉瘤是一种以主要弹性血管（主动脉）结构破裂为特征的疾病。在未来，这种平台技术可以扩展到治疗其他非血管弹性组织类型（例如，肺组织）需要结构修复。该提案旨在开发创新的新方法，以实现在蛋白水解损伤的结构受损的软弹性组织中的原位仿生弹性基质再生修复。所提出的方法旨在通过稳定的成体细胞类型来克服破坏的弹性基质的内在不良的自动再生修复。研究人员最近显示，骨髓间充质干细胞（BM-MSC）衍生的平滑肌细胞（BM-SMC），而不是未分化的BM-MSC，比成人血管SMC（健康，患病）显着更具弹性，其分泌物刺激弹性基质再生修复的SMC患病，基质组装受损的表型。由于干细胞的物理递送面临着几个挑战，该项目提出设计和测试一种干细胞启发的，但无细胞的再生方法来原位ECM再生修复。该方法基于BM-SMC分泌组组分的持续局部递送，所述BM-SMC分泌组组分被鉴定为对于来自新型聚合物纳米载体的原弹性蛋白再生刺激是必要和足够的，所述新型聚合物纳米载体本身表现出原弹性蛋白生成和抗蛋白水解性质。通过旨在解决三个具体目标的实验，研究人员将测试假设：a）人类BM-MSC人骨髓间充质干细胞（hBM-MSCs）可以有效地分化为平滑肌细胞（hBM-SMC）表现出不同的弹性决定表型状态; B）hBM-SMC对腹主动脉瘤SMC的促弹性作用由其分泌的营养因子介导（分泌组）; c）hBM-SMC分泌组的关键组分单独或组合对于促弹性蛋白生成和抗蛋白水解作用是必要的和足够的;和d）将关键hBM-SMC分泌组因子的持续递送与纳米载体整合将增加&ECM破坏的3-D组织空间中再生弹性基质修复的数量和质量。该项目的广泛研究影响是基于其潜力，即一种新的纳米方法可能使再生弹性基质修复重现SC分泌物的再生作用。通过该项目，研究人员将a）为不同教育水平的学生开发教育模块，以更好地了解干细胞和组织工程，以及B）为克利夫兰诊所和弗吉尼亚联邦大学的学生提供独特的机构间合作培训机会。通过在这些机构开展完善的暑期实习和推广计划，研究人员将开发教育模块，使高中生、本科生和公众受益。该提案由化学，生物工程，环境和运输系统部门的生物医学工程项目以及材料研究部门的生物材料项目共同资助。

PI: Ramamurthi, Anand/ Rao, Raj R Proposal Number: 1508642 / 1509377 Restoring structurally damaged soft, elastic tissues to a healthy state is difficult since adult cells are poorly capable of building new elastic fibers, which allow tissues to stretch and recoil. In this project, the investigators propose identification and characterization of factors derived from stem cells towards regenerating and repairing elastic fiber assembly and structure. Further, the studies aim to deliver factors in a sustained manner using degradable polymeric particles which are themselves chemically modified to stimulate new elastic fiber formation and prevent its breakdown. The investigators will then test the effectiveness of the particles in treating abdominal aortic aneurysms, a disorder characterized by breakdown of the structure of the major elastic blood vessel (aorta). In the future, this platform technology can be extended to treat other non-vascular elastic tissue types (e.g., lung tissue) in need of structural repair. This proposal aims to develop innovative, new approaches to enable in situ, biomimetic elastic matrix regenerative repair in soft, elastic tissues structurally compromised by proteolytic injury. The proposed approach seeks to overcome intrinsically-poor auto-regenerative repair of disrupted elastic matrix by stable adult cell types. The investigators have recently shown bone marrow mesenchymal stem cell (BM-MSC)-derived smooth muscle cells (BM-SMCs), but not undifferentiated BM-MSCs, to be significantly more elastogenic than adult vascular SMCs (healthy, diseased), and their secretions to stimulate elastic matrix regenerative repair by SMCs of a diseased, matrix assembly-impaired phenotype. As physical delivery of stem cells faces several challenges, this project proposes to design and test a stem cell-inspired, but cell-free regenerative approach to in situ ECM regenerative repair. The approach is based on sustained, local delivery of BM-SMC secretome components identified to be necessary and sufficient for pro-elastin regenerative stimulus from novel polymer nanocarriers that themselves exhibit pro-elastogenic and anti-proteolytic properties. Through experiments designed to address three specific aims, the investigators will test hypotheses that a) human BM-MSCs (hBM-MSCs) can be efficiently differentiated into SMCs (hBM-SMCs) exhibiting distinct, elastogenicity-determining phenotypic states; b) pro-elastogenic effects of hBM-SMCs on abdominal aortic aneurysm SMCs are mediated by their secreted trophic factors (secretome); c) key components of hBM-SMC secretome individually or in combination are necessary and sufficient for pro-elastogenic and anti-proteolytic effect; and d) integrating sustained delivery of key hBM-SMC secretome factor(s) with nanocarriers will augment quantity & quality of regenerative elastic matrix repair in an ECM-disrupted, 3-D tissue space. The broad research impact of this project is based on its potential that a novel nanotherapeutic approach may enable regenerative elastic matrix repair that recapitulates regenerative effects of SC secretions. Through this project, the investigators will a) develop educational modules for students at several educational levels to better understand stem cell- and tissue engineering, and b) provide unique inter-institutional collaborative training opportunities for students at Cleveland Clinic and Virginia Commonwealth University. By working through well-established summer internship and outreach programs at these institutions, the investigators will develop educational modules that will benefit high school students, undergraduate students and the general public. This proposal is co-funded by the Biomedical Engineering Program in the Chemical, Bioengineering, Environmental and Transport Systems Division, and by the Biomaterials Program in the Division of Materials Research.