Unlocking Regenerative Potential through in Vivo Genetic Reprogramming

通过体内基因重编程释放再生潜力

• 批准号: 8358718
• 负责人: P. Duc Si Dong
• 金额: $ 288.23万
• 依托单位: SANFORD BURNHAM PREBYS MEDICAL DISCOVERY INSTITUTE
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-09-30 至 2017-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8358718
• 关键词: Adult; Animals; Biology; Cardiac Myocytes; Cells; Cultured Cells; Development; Diabetes Mellitus; Disease; Effectiveness; Embryo; Endoderm; Endoderm Cell; Engineering; Epigenetic Process; Genetic; Goals; Heart; Heart Diseases; Immune system; In Vitro; Injury; Limb structure; Liver; Liver diseases; Mutation; Natural regeneration; Oncogenic; Organ; Pancreas; Primitive foregut structure; Process; Research Proposals; Somatic Cell; Stem cells; Therapeutic; Tissues; Transplantation; Vertebrates; Work; abstracting; cancer risk; cell type; efficacy testing; embryonic stem cell; gene induction; genetic manipulation; immunogenic; in vivo; induced pluripotent stem cell; insight; mature animal; new technology; progenitor; public health relevance; regenerative; repaired; tissue culture; transgene expression

DESCRIPTION (Provided by the applicant) Abstract: We propose to genetically reprogram cells in vivo to induce new tissue development in post-embryonic vertebrate animals. Our ultimate goal is to grow replacement organs in an adult vertebrate animal using genetic reprogramming. The ability to grow replacement tissues will be critical for treating and curing diseases such as diabetes, liver disease, and heart diseas and for repairing/replacing damaged or lost organs and limbs sustained from traumatic injury. Our primary strategy is test for the efficacy of different combinations of factors implicated in th process of development, regeneration, and of making iPSCs (induced pluripotent stem cells) to induce new tissue development in post-embryonic animals. Our research proposal will focus on growing ectopic pancreas and liver tissues in the foregut endoderm, and ectopic ss-cells in different various places in the body. Currently, the most popular approach to engineering potential therapeutic cells or tissues is primarily via an in vitro strategy: through directed differentiation of embryonic stem cells (ESCs) or from iPSCs. However, there are several potential fundamental obstacles with this approach. Cultured cells may acquire extensive deleterious genetic and epigenetic changes (oncogenic or immunogenic mutations) that would normally be rejected and eradicated from the body, but would continue to thrive in culture. Also, although much progress has been made in differentiating various cell types in the dish, it remains challenging to grow whole organs in 3D. Further, it is still unclear how tissues cultured in a dish such as cardiomyocytes derived stem cells would integrate and become a functional part of the host heart following transplantation. Therefore, many complications have arisen with in vitro approaches to generating therapeutic cells and organs. Much less emphasis has been invested in generating new tissues in vivo, which may circumvent these obstacles. We propose to apply our unique insight on liver and pancreas progenitor biology and combine it with our new technology to precisely induce transgene expression to reprogram endoderm cells to grow ectopic pancreas, liver, and ss-cells. Many fundamental questions regarding the feasibility and effectiveness of in vivo cell reprogramming is largely unexplored. Our studies will allow us to evaluate these issues: Is it possible to grow whole new organs in an adult animal? How amendable are different somatic cells (particularly in the gut endoderm for this proposal) to in vivo genetic reprogramming? Are certain adult cells, maybe progenitors, more poised for genetic reprogramming? Will induced organs integrate effectively to adjacent tissues and be functional? Are they less likely to accumulate mutations or be rejected by the immune system? Will cancer risks increase with our genetic manipulations? Can expression of iPSC induction genes make most somatic cells in vivo more reprogrammable? Our proposed work will allow us to begin to ask these fundamental questions. Public Health Relevance: We propose to genetically reprogram cells in vivo to induce new tissue development in post-embryonic vertebrate animals. Our ultimate goal is to grow replacement organs in an adult vertebrate animal using genetic reprogramming. The ability to grow replacement tissues will be critical for treating and curing diseases such as diabetes, liver disease, and heart disease and for repairing/replacing damaged or lost organs and limbs sustained from traumatic injury.

描述（由申请人提供） 翻译后摘要：我们建议在体内基因重编程细胞诱导新的组织发育后胚胎脊椎动物。我们的最终目标是利用基因重编程在成年脊椎动物体内培育替代器官。生长替代组织的能力对于治疗和治愈诸如糖尿病、肝病和心脏病的疾病以及对于修复/替换因创伤性损伤而受损或丢失的器官和肢体将是至关重要的。我们的主要策略是测试在发育，再生和制造iPSC（诱导多能干细胞）的过程中涉及的不同因子组合的功效，以诱导胚胎后动物的新组织发育。我们的研究计划将集中在前肠内胚层异位胰腺和肝脏组织的生长，以及体内不同部位异位ss细胞的生长。 目前，工程化潜在治疗细胞或组织的最流行方法主要是通过体外策略：通过胚胎干细胞（ESC）或iPSC的定向分化。然而，这种方法存在几个潜在的基本障碍。培养的细胞可能会获得广泛的有害遗传和表观遗传变化（致癌或免疫原性突变），这些变化通常会被人体排斥和根除，但会在培养中继续茁壮成长。此外，尽管在培养皿中分化各种细胞类型方面取得了很大进展，但在3D中培养整个器官仍然具有挑战性。此外，尚不清楚培养皿中培养的组织如心肌细胞衍生的干细胞如何整合并在移植后成为宿主心脏的功能部分。因此，在体外产生治疗性细胞和器官的方法中出现了许多并发症。 在体内产生新的组织，这可能会绕过这些障碍，已经投入的重点少得多。我们建议应用我们对肝脏和胰腺祖细胞生物学的独特见解，并将其与我们的新技术联合收割机相结合，以精确诱导转基因表达，重新编程内胚层细胞，使其生长异位胰腺、肝脏和ss细胞。关于体内细胞重编程的可行性和有效性的许多基本问题在很大程度上尚未探索。我们的研究将使我们能够评估这些问题：是否有可能在成年动物体内培养出全新的器官？不同的体细胞（特别是在本建议的肠道内胚层）对体内遗传重编程的适应性如何？某些成年细胞，也许是祖细胞，更适合基因重编程吗？诱导器官是否能有效地与邻近组织整合并发挥功能？它们是否不太可能积累突变或被免疫系统排斥？癌症风险会随着我们的基因操作而增加吗？iPSC诱导基因的表达能否使体内大多数体细胞更具可重编程性？我们提议的工作将使我们能够开始提出这些根本问题。 公共卫生相关性：我们建议在体内对细胞进行基因重编程，以诱导胚胎后脊椎动物的新组织发育。我们的最终目标是利用基因重编程在成年脊椎动物体内培育替代器官。生长替代组织的能力对于治疗和治愈诸如糖尿病、肝病和心脏病的疾病以及对于修复/替换因创伤性损伤而受损或丢失的器官和肢体将是至关重要的。

项目成果

Identification of Annexin A4 as a hepatopancreas factor involved in liver cell survival.

• DOI: 10.1016/j.ydbio.2014.08.025
• 发表时间: 2014-11-01
• 期刊: DEVELOPMENTAL BIOLOGY
• 影响因子: 2.7
• 作者: Zhang, Danhua;Golubkov, Vladislav S.;Han, Wenlong;Correa, Ricardo G.;Zhou, Ying;Lee, Sunyoung;Strongin, Alex Y.;Dong, P. Duc Si
• 通讯作者: Dong, P. Duc Si