Innate activation and death signals in health and disease

描述（由申请人提供）：这一变革性的研究提案源于我的发现，即胱天蛋白酶8（Casp 8）缺陷小鼠的胚胎致死是释放的RIP 3坏死的结果。Casp 8-/-Rip 3-/-小鼠发育成存活的、可生育的和免疫活性的成年小鼠，其通过NF-κ B和其他因子保留阳性先天信号传导，但缺乏与炎性疾病、自身免疫和肿瘤发生相关的有害凋亡和坏死途径，这引发了一个问题，“在缺乏外源性细胞死亡途径的情况下，生命有什么益处，以及如何在临床中利用这一点”？发育失调表明，在组织移植和核重编程等不同领域可能会获得益处。令人惊讶的是，Casp 8-/-Rip 3-/-小鼠是免疫活性的，但耐受与错配骨髓的移植。Casp 8-/-Rip 3-/-细胞以显著提高的效率支持核重编程。此外，Casp 8-/-Rip 3-/-小鼠表现出对炎性癌症的抗性。挑战是通过遗传操作保留先天信号传导的积极机制并消除失调的有害后果，然后将这些发现转化为改善组织修复和再生的治疗干预措施。我们建议研究遗传缺陷与Casp 8蛋白酶和RIP 3激酶的小分子抑制剂一起，以改善组织移植和核重编程。创新已经开始出现。炎症性疾病和缺血-再灌注损伤减少，同种异体移植在缺乏Casp 8和RIP 3激酶的情况下显著改善。Casp 8-/-Rip 3-/-小鼠不能对足以杀死WT小鼠的炎性损伤作出反应。来自这些小鼠的成纤维细胞重新编程为诱导多能干细胞（iPSC），效率显著提高。基因敲除小鼠以及小分子抑制剂将出现额外的高影响力发现，改善小鼠，恒河猴和人类成纤维细胞向iPSC和/或治疗上重要的体细胞祖细胞的重编程。影响将来自现有和预期的创新，从基因消除小鼠中有害的细胞死亡，到对恒河猴和人类细胞使用强大的遗传和治疗策略。通过平行的遗传和小分子治疗干预，这一知识将走向实际的临床应用，在这些医学相关领域的专家顾问的重要参与。从小鼠模型开始，我将：（1）改善组织再生，（2）优化同种异体移植，（3）增强核重编程，通过消除外源性细胞凋亡建立一个新的范例，而不会引发坏死。通过这些研究获得的知识将在当前的组织修复和移植方法中具有变革性，并将彻底改变未来的再生医学策略。

DESCRIPTION (provided by applicant): This transformative research proposal extends from my discovery that embryonic lethality in caspase 8 (Casp8)-deficient mice the result of unleashed RIP3 necrosis. Casp8-/-Rip3-/- mice develop into viable, fertile and immunocompetent adults that retain positive innate signaling through NF-kappaB and other factors, but lack detrimental apoptosis and necrosis pathways associated with inflammatory disease, autoimmunity and oncogenesis, prompting the question, "What benefit(s) comes from life in the absence of extrinsic cell death pathways and how can this be harnessed in the clinic"? The developmental dysregulation suggested benefits might be gained in areas as diverse as tissue engraftment and nuclear reprogramming. Surprisingly, Casp8-/- Rip3-/- mice are immunocompetent but tolerate engraftment with mismatched bone marrow. Casp8-/-Rip3-/- cells support nuclear reprogramming with dramatically improved efficiency. In addition, Casp8-/-Rip3-/- mice exhibit resistance to inflammatory cancer. The Challenge is to retain positive mechanisms of innate signaling and eliminate deleterious consequences of dysregulation through genetic manipulation, and then to translate these findings into therapeutic interventions that improve tissue repair and regeneration. We propose to investigate genetic deficiency together with small molecule inhibitors of Casp8 protease and RIP3 kinase to improve tissue transplantation and nuclear reprogramming. The Innovation has begun to emerge. Inflammatory disease and ischemia-reperfusion damage is reduced and allograft transplantation is dramatically improved in the absence of Casp8 and RIP3 kinase. Casp8-/-Rip3-/- mice fail to respond to inflammatory insult sufficient to kill WT mice. Fibroblasts from these mice reprogram into induced pluripotent stem cells (iPSCs) with dramatically improved efficiencies. Additional high impact findings will emerge with genetic knock-out mice as well as with small molecule inhibitor, improving reprogramming of mouse, rhesus and human fibroblasts to iPSCs and/or therapeutically important somatic cell progenitors. The Impact will come from existing and expected innovations, ranging from genetic elimination detrimental cell death in mice, to the use of powerful genetic and therapeutic strategies on rhesus and human cells. Through parallel genetic and small molecule therapeutic intervention, this knowledge will move towards practical clinical applications, with the important involvement of expert consultants in these medically relevant areas. Starting with mouse models, I will: (1) improve tissue regeneration, (2) optimize allogeneic engraftment, and (3) enhance nuclear reprogramming, establishing a new paradigm by eliminating extrinsic apoptosis without the disadvantage of triggering necrosis. The knowledge gained through these studies will be transformative in current tissue repair and transplantation approaches, and will revolutionize future regenerative medicine strategies.