Adaptable tissue-specific endothelial cells for organ regeneration

用于器官再生的适应性组织特异性内皮细胞

• 批准号: 9894491
• 负责人: Shahin Rafii
• 金额: $ 103.23万
• 依托单位: WEILL MEDICAL COLL OF CORNELL UNIV
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-04-01 至 2027-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/9894491
• 关键词: Adult; Allogenic; Award; Blood; Blood Vessels; Cardiac; Cells; Cicatrix; Clinic; Clinical Trials; Coculture Techniques; Code; Endothelial Cells; Engineering; FDA approved; Fibrosis; Functional disorder; Genetic Transcription; Goals; Growth Factor; Heart; Hematopoietic; Hepatic; Heterogeneity; Homeostasis; Human; In Vitro; Infusion procedures; Intention; Lung; Macaca nemestrina; Mission; Molecular; Monkeys; Morphogenesis; Mus; National Heart, Lung, and Blood Institute; Natural regeneration; Organ; Organoids; Outcome; Pathway interactions; Production; RNA; Recovery; Regenerative Medicine; Research; Safety; Signal Transduction; Specific qualifier value; Technology; Testing; Therapeutic; Tissue Engineering; Tissue Transplantation; Tissues; Translating; Translations; Transplantation; cellular transduction; experimental study; fetal; follow-up; healing; in vivo; induced pluripotent stem cell; innovation; knock-down; lung repair; nonhuman primate; novel; organ injury; organ regeneration; overexpression; programs; reconstitution; regenerative; repaired; stem cells; tissue repair; tumorigenesis; vascular contributions

PROJECT ABSTRACT The overarching goal of our proposed research program is to develop a discovery pipeline that will enable identification of transcriptional codes for engineering tissue-specific endothelial cells (ECs) for therapeutic organ regeneration of heart, lung and blood. Therapies for organ regeneration promises unlimited access to the replacement tissues. However, despite breakthroughs in uncovering the molecular underpinnings of organ morphogenesis and organoid technology, translation of regenerative medicine to the clinic has confronted with hurdles. These bottlenecks are in part due to the lack of understanding as to how niche cells coordinate organ repair. Specifically, contribution of vascular niche cells that supply regenerative signals has not been realized. This R35 application builds upon the novel proposition that poor healing after organ damage is due to the dysfunction and loss of the tissue-specific ECs. This programmatic proposal examines the hypothesis that reconstitution of stem cells in injured organs is dependent on the pro-regenerative angiocrine signals supplied by tissue-specific vascular niche ECs. We have shown that organotypic ECs by deploying defined angiocrine factors support lung, cardiac, hepatic and hematopoietic regeneration. Thus, ECs perform actively as dynamic, tissue-specified niche cells critical for tissue homeostasis and repair. To test this and to set up the stage for therapies, we have engineered adaptable mouse, nonhuman primate and human ECs by transducing the transduction factor (TF) ETV2 into adult mature ECs (R-VECs) and differentiating human induced pluripotent stem cells (iPSCs) into generic fetal-like ECs (iVECs) that could inform on the pathways that induce organotypic TFs. These adaptive iVECs and R-VECs will be cocultured with heart, lung, and blood organoids in vitro or infused in vivo in mice undergoing organ repair to identify the induction of organotypic TFs in these cells. The educated iVECs and R-VECs will be recovered and subjected to RNA profiling and de novo motif discovery to identify induced tissue-specific TF(s). The identified TFs will be overexpressed or knocked down in ECs, to validate their function in sustaining organotypic and angiocrine profile for organ repair. We anticipate that transplantation of organotypic ECs will promote long-lasting tissue repair without provoking tumorigenesis or fibrosis. We have initiated FDA-approved human clinical trials to examine the safety and efficacy of allogeneic generic EC infusion for hematopoietic recovery. As a follow up, we intend to assess the contribution of R-VECs or iVECs-derived from nonhuman primates to regeneration in the pigtail macaque monkeys with the intention of translating the potential of organotypic ECs to clinic. The expected outcomes of the proposed research are identification of molecular signals and transcriptional determinants of tissue-specific vascular and angiocrine heterogeneity. Goals of this proposal fit with the mission of NHLBI R35 award to develop innovative regenerative discovery pipeline to promote safe and efficacious treatments for cardiac, pulmonary and blood maladies.

项目摘要 我们提出的研究计划的总体目标是开发一个发现管道，使 鉴定用于工程化组织特异性内皮细胞 (EC) 的转录密码以用于治疗 心脏、肺和血液的器官再生。器官再生疗法有望无限制地获得 替换组织。然而，尽管在揭示器官的分子基础方面取得了突破 形态发生和类器官技术，再生医学向临床的转化面临着 障碍。这些瓶颈部分是由于缺乏对生态位细胞如何协调器官的了解 维修。具体而言，血管生态位细胞提供再生信号的贡献尚未被认识到。 该 R35 应用建立在一个新的命题之上，即器官损伤后愈合不良是由于 组织特异性 EC 功能障碍和丧失。该纲领性提案检验了以下假设： 受损器官中干细胞的重建取决于所提供的促再生血管分泌信号 通过组织特异性血管生态位 EC。我们已经证明，通过部署定义的血管分泌素，器官型 EC 支持肺、心脏、肝脏和造血再生的因素。因此，EC 表现活跃，充满活力， 组织特异的生态位细胞对于组织稳态和修复至关重要。为了测试这一点并为 疗法，我们通过转导 转导因子 (TF) ETV2 转化为成人成熟 EC (R-VEC) 并分化人类诱导多能性 干细胞（iPSC）转化为通用的胎儿样 EC（iVEC），可以了解诱导的途径 器官型 TF。这些适应性 iVEC 和 R-VEC 将与心脏、肺和血液类器官共培养 体外或体内输注到正在进行器官修复的小鼠中，以鉴定这些器官中器官型转录因子的诱导 细胞。经过训练的 iVEC 和 R-VEC 将被回收并进行 RNA 分析和 de novo 基序 识别诱导组织特异性转录因子的发现。已识别的 TF 将被过度表达或被敲除 在 EC 中，验证其在维持器官修复的器官型和血管分泌谱方面的功能。我们预计 器官型 EC 移植将促进持久的组织修复，而不会引发肿瘤发生 或纤维化。我们已启动 FDA 批准的人体临床试验，以检查其安全性和有效性 用于造血恢复的同种异体通用 EC 输注。作为后续行动，我们打算评估贡献 源自非人灵长类动物的 R-VEC 或 iVEC 在猪尾猕猴中的再生 旨在将器官型 EC 的潜力转化为临床。拟议的预期成果 研究方向是识别组织特异性血管和细胞的分子信号和转录决定因素。 血管分泌异质性。该提案的目标符合 NHLBI R35 奖项开发创新型产品的使命 再生发现管道，以促进心脏、肺和血液的安全有效的治疗 疾病。