Protein-Engineered Hydrogels for Gene Transplantation for Myocardial Infarction

项目总结/摘要 每年有近一百万美国人经历心肌梗死（MI）。微创，局部 药物、细胞、基因和/或生长因子的递送是有希望的MI再生疗法;然而， 输送到收缩的心脏仍然是一个未解决的挑战。为了成为临床上可行的选择，生物材料必须 是导管可注射的，必须提供快速和牢固的凝胶化以防止从跳动的心肌中挤出， 并且必须允许有效载荷的治疗剂量的受控释放。不幸的是，大多数导管注射 材料是在心肌收缩期间被挤出的弱凝胶。作为替代，我建议形成凝胶 通过生物相容性，动态共价化学（DCC）交联，可逆但强，导致 可注射的凝胶，其具有保持在收缩组织中所必需的机械完整性。具体来说，这些 凝胶由重组的弹性蛋白样蛋白和化学修饰的透明质酸组成。这些 组分通过DCC腙键交联在一起以形成可酶降解的凝胶， 在化学上完全明确，可用于未来的FDA研究。在目标1中，将机械合成一个12种凝胶的家族， 在体外和体内表征和评价，以便于导管注射和保留在收缩的心脏中。 在目标2中，再生有效载荷，编码基质细胞衍生因子-1 α的小环基因，将被拴系到 可注射凝胶通过与肽核酸-肽的DNA杂交来实现持续、长期 release.在目标3中，将制备具有优化的体内保留和持续基因释放的凝胶制剂。 在临床前大鼠MI模型中评估治疗潜力。 这个职业发展奖将使我能够加强我在生物材料方面的强大背景 化学和基因传递与心血管生物学的新专业知识，缺血性心脏病的临床前模型 心肌病和转化生物工程。我的职业发展计划包括（1）正式课程 在再生医学和心血管生物学和疾病，（2）重组技术培训 生物材料、微环基因和临床前MI模型，（3）两位杰出科学家的密切合作 （一名生物工程师和一名外科医生）具有成功培训和合作的良好记录，以及（4）职业生涯 咨询委员会的指导，反映了职业经验和领域的多样性。准备我的 在向独立过渡的过程中，我与我的共同导师协商制定了一项计划，以进一步增加和加强 我的专业网络，成为抛光和自信，在我的口头研究报告，并获得 宝贵的撰写资助申请的技巧该计划充分利用了斯坦福大学现有的优秀资源 心血管研究所和斯坦福大学工程学院与国家和国际活动。期间 在这个奖项的最后一年，我将准备并提交终身教职助理职位的申请材料 生物工程学教授我的长期目标是领导一个转化生物工程实验室， 他还将为心血管疾病提供再生疗法，并成为年轻黑人科学家的榜样。

PROJECT SUMMARY/ABSTRACT Nearly one million Americans experience a myocardial infarction (MI) annually. Minimally invasive, local delivery of drugs, cells, genes, and/or growth factors are promising regenerative therapies for MI; however, delivery to the contracting heart remains an unmet challenge. To be a clinically viable option, biomaterials must be catheter injectable, must provide rapid and robust gelation to prevent extrusion out of the beating myocardium, and must permit controlled release of a therapeutic dosage of payload. Unfortunately, most catheter-injectable materials are weak gels that are extruded during myocardial contraction. As an alternative, I propose gels formed via biocompatible, dynamic covalent chemistry (DCC) crosslinks that are reversible yet strong, resulting in injectable gels with the mechanical integrity necessary to be retained in contractile tissue. Specifically, these gels are composed of recombinant, elastin-like proteins and chemically modified hyaluronic acid. These components are crosslinked together through DCC hydrazone bonds to form enzymatically degradable gels that are fully chemically defined for future FDA studies. In Aim 1, a family of 12 gels will be synthesized, mechanically characterized, and evaluated in vitro and in vivo for easy catheter injection and retention in the contracting heart. In Aim 2, a regenerative payload, minicircle genes encoding stromal cell-derived factor-1α, will be tethered to the injectable gel via DNA hybridization with peptide nucleic acid-peptides to achieve sustained, long-term release. In Aim 3, the gel formulation with optimized in vivo retention and sustained gene release will be evaluated for therapeutic potential in a preclinical rat MI model. This Career Development Award would enable me to enhance my strong background in biomaterials chemistry and gene delivery with new expertise in cardiovascular biology, preclinical models of ischemic cardiomyopathy, and translational bioengineering. My career development plan includes (1) formal coursework in regenerative medicine and cardiovascular biology and disease, (2) technical training in recombinant biomaterials, minicircle genes, and preclinical MI models, (3) close co-mentorship by two outstanding scientists (a bioengineer and a surgeon) with a strong track-record of successful training and collaboration, and (4) career guidance by an Advisory Committee that reflects a diversity of career experiences and fields. To prepare for my transition to independence, I devised a plan in consultation with my co-mentors to further increase and strengthen my professional network, to become polished and confident in my oral research presentations, and to gain invaluable grant-writing skills. This plan leverages the outstanding resources available within the Stanford Cardiovascular Institute and the Stanford School of Engineering with national and international events. During the last year of this award, I will prepare and submit application packets for the position of tenure-track Assistant Professor in Bioengineering. My long-term goal is to lead a translational bioengineering lab that develops regenerative therapies for cardiovascular diseases and to serve as a role model for young black scientists.