Role of SLIT-ROBO and MT1-MMP in MSC-Mediated Neovascularization.

项目摘要 这是一份K 08指导临床科学家职业发展奖的申请，以培训博士Ming-Sing Si， 心脏外科助理教授司博士对科学研究有浓厚的兴趣， 他在教育生涯的每一个阶段参与研究的长期记录。司博士的 部门已经保证他受到保护的时间，实验室空间，人员支持和所有其他资源 成功完成本培训计划。拟议的培训计划将为Si博士提供 必要的技能，成为一个成功的独立研究者专注于间充质干细胞（MSC） 和血管周围细胞生物学。培训的关键组成部分将包括高级研究生课程 实验方法，组织再生，负责任的研究行为，先进的 实验设计中的遗传方法和新型转基因小鼠模型，频繁的预定相互作用 与他的指导委员会成员，研讨会和会议参与，手稿写作，R 01赠款 计划书撰写，以及MSC/血管周围细胞在新生血管形成中的功能的基础科学研究- 是的。导师委员会由三位国际知名的MSC相关专家组成， naling和血管生成。这些专家在为年轻临床医生提供指导方面也很有经验， 科学家申请人已经收集了涉及SLIT-ROBO信号传导的初步数据，SLIT-ROBO信号传导是一种用于治疗糖尿病的信号传导轴。 轴突和尖端内皮细胞，在MSC的行为有关的新血管形成和组织再生。基于 根据这些发现和他的一位导师发表的关于MT 1-MMP重要性的相关结果， 蜗牛转录因子在细胞入侵，申请人计划定义如何，并在何种程度上，SLIT-ROBO 信号传导和MT 1-MMP在体外和体内刺激MSC介导的血管生成，并利用这一知识 设计新的治疗策略，有效地刺激新血管形成。具体目标是 研究项目包括：（1）确定SLIT 3-ROBO 1信号传导对MSC运动性、侵袭性、胶原溶解， 以及左心室（LV）压力超负荷情况下的血管生成。(2)确定MT 1-MMP在以下方面的作用： MSC侵袭性MSC介导的血管生成和心力衰竭。(3)描述在体内，天然的作用， 应用条件性基因敲除小鼠研究左室压力超负荷时血管周壁细胞-SLIT 3-ROBO 1轴 模型这项研究将促进对多细胞环境中血管生成反应的理解。 相互作用，并应提供有价值的见解的手段，治疗性新血管形成， 被引出。完成拟议的研究和培训计划将提供知识和经验， 申请人成为MSC生物学、血管生成和心血管领域的独立研究者， 血管再生具体而言，拟议研究的数据将作为R 01的基础 在K 08奖项的第三年提交的申请将探索SLIT-ROBO的功能 在心血管疾病的背景下MSC/周细胞中的信号传导。

PROJECT SUMMARY This is an application for a K08 Mentored Clinical Scientist Career Development Award to train Dr. Ming-Sing Si, an Assistant Professor in Cardiac Surgery. Dr. Si has a strong interest in scientific investigation, as demonstrated by his long track record of participation in research throughout every stage of his educational career. Dr. Si’s Department has guaranteed him protected time, laboratory space, personnel support, and all other resources required to complete this training program successfully. The proposed training program will provide Dr. Si the necessary skills to become a successful independent investigator focused on mesenchymal stem cell (MSC) and perivascular cell biology. Key components of the training will include graduate level courses in advanced experimental methods, tissue regeneration, and responsible conduct of research, incorporation of advanced genetic methods and novel transgenic mouse models in experimental design, frequent scheduled interactions with his mentorship committee members, seminar and conference participation, manuscript writing, R01 grant proposal writing, and basic science investigation into the function of MSC/perivascular cells in neovasculariza- tion. The mentorship committee is comprised of three internationally renowned experts relevant to MSCs, sig- naling, and angiogenesis. These experts are also experienced in providing mentorship to young clinicians and scientists. The applicant has gathered preliminary data that implicate SLIT-ROBO signaling, a signaling axis for axons and tip endothelial cells, in MSC behavior pertinent to neovascularization and tissue regeneration. Based on these findings and related published results from one of his mentors on the importance of MT1-MMP and Snail transcription factors in cell invasion, the applicant plans to define how, and to what extent, SLIT-ROBO signaling and MT1-MMP stimulate MSC-mediated angiogenesis in vitro and in vivo, and to utilize this knowledge to design novel therapeutic strategies that effectively stimulate neovascularization. The specific aims for this project are: (1) Determine the effects of SLIT3-ROBO1 signaling on MSC motility, invasiveness, collagenolysis, and angiogenesis in the setting of left ventricle (LV) pressure overload. (2) Define the role of the MT1-MMP on MSC invasiveness MSC-mediated angiogenesis and heart failure. (3) Characterize the in vivo, native roles of the perivascular mural cell-SLIT3-ROBO1 axis during LV pressure overload using conditional knockout mouse models. This research will advance the understanding of angiogenic responses in the setting of multicellular interactions and should provide valuable insights into the means by which therapeutic neovascularization can be elicited. The completion of the proposed research and training plan will provide the knowledge and experience for the applicant to become an independent investigator in the field of MSC biology, angiogenesis, and cardio- vascular regeneration. Specifically, the data from the proposed study will serve as the foundation for an R01 application to be submitted during the third year of the K08 award that will explore the function of SLIT-ROBO signaling in MSC/pericytes in the setting of cardiovascular disease.