Benefits of Eliminating Cell Death Pathways in Health and Disease

消除细胞死亡途径对健康和疾病的好处

基本信息

批准号：
8766753
负责人：
EDWARD S. Edward S Mocarski
金额：
$ 29.64万
依托单位：
EMORY UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2014
资助国家：
美国
起止时间：
2014-09-01 至 2015-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8766753
关键词：
Acute Acute Graft Versus Host Disease Adult Allogenic Antigens Apoptosis Apoptotic Area Autoimmune Diseases Blood Vessels Bone Marrow C57BL/6 Mouse Cardiovascular system Caspase Inhibitor Cell Death Cell membrane Cells Cessation of life Clinic Complex Defect Development Disadvantaged Disease Embryo Engraftment Ensure Extravasation Family Fibroblasts Future Genetic Health Hematopoietic Homeostasis Homologous Transplantation Host Defense Human Immune Immunocompetent Infection Inflammation Inflammatory Interleukin-1 Intervention Invaded Knock-out Knowledge Left Life Light Macaca mulatta Malignant Neoplasms Modeling Mouse Strains Mus Natural regeneration Necrosis Pathway interactions Pattern recognition receptor Peptide Hydrolases Pharmaceutical Preparations Phosphotransferases Play Polyubiquitin Predisposition Process Publishing RIPK3 gene Regenerative Medicine Regulation Relative (related person)Reperfusion Injury Research Risk Role Signal Transduction Stress Symptoms System T-Cell Receptor T-Lymphocyte TLR3 gene TNF gene Therapeutic Therapeutic Intervention Tissue Transplantation Tissues Translating Transplantation Ubiquitination Virus Virus Diseases Work Wound Healing caspase-8 genetic manipulation immune function improved inhibitor/antagonist mimetics nuclear reprogramming pathogen prevent programs public health relevance receptor regenerative repaired research study sensor small molecule stem cell therapy success tissue regeneration tissue repair transdifferentiation ubiquitin isopeptidase ubiquitin-protein ligase

项目摘要

DESCRIPTION (provided by applicant): Benefits of Eliminating Cell Death Pathways in Health and Disease the underlying processes influencing vascular health remain largely undefined, leaving current therapeutic interventions to treat symptoms without a precise understanding the underlying, physiologically relevant disease and regenerative pathways. We propose to restrict inflammation and orchestrate repair and regeneration of hematopoietic and cardiovascular tissues by eliminating nonvital cell death and associated inflammatory processes. Our discovery that embryonic lethality of caspase 8 (Casp8)-deficiency in mice results from unleashed RIP3 necrosis opens the path to harnessing benefits of eliminating these pathways. Casp8-/- Rip3-/- (DKO) mice develop into viable and fertile adults that lack a susceptibility to inflammatory insult but retain sufficient immune function to control viral infection, prompting pilot efforts to harnes this setting to improve tissue repair, allogeneic engraftment and tissue regeneration. The Casp8 and RIP3 pathways evolved to counteract virus-encoded cell death suppressors, but become unleashed to cause inflammatory disease and undermine tissue repair, homeostasis and regeneration. This proposal employs genetic as well as small molecule approaches to eliminate cell death pathways, all modeled by DKO mice, as well as a additional mouse strains we have created. We have already shown that DKO mice resist acute inflammation, inflammatory cancer and ischemia reperfusion injury even though overall wound repair rates appear normal, prompting the very important question, "How can benefit(s) from life in the absence of extrinsic cell death pathways be harnessed in the clinic"? DKO cells engraft and differentiate with higher efficiencies than WT cells. Remarkably, Casp8-/-Rip3-/- mice survive allogeneic (MHC mismatched) bone marrow engraftment that causes acute graft versus host disease (GvHD) in WT C57BL/6 mice. Surprisingly, DKO fibroblasts support nuclear reprogramming with improved efficiency over WT cells. We propose to eliminate deleterious consequences of cell death through genetic manipulation, moving this area closer to a therapeutic intervention strategy to improve hematopoietic and cardiovascular tissue repair and regeneration. We propose to investigate genetic deficiency in mice and mouse cells, and develop knock-out strategies on rhesus macaque and human fibroblasts to combine with newly described small molecule inhibitors of RIP3 kinase and commercially available caspase inhibitors, to improve tissue repair, allogeneic transplantation and nuclear reprogramming. Our Aims are to: (1) optimize allogeneic engraftment, and (2) enhance nuclear reprogramming by eliminating extrinsic apoptosis without the disadvantage of triggering necrosis. The knowledge gained through these studies will have broad impact in current tissue repair and transplantation approaches, as well as future regenerative medicine strategies.

描述（由申请人提供）：消除细胞死亡途径对健康和疾病的益处影响血管健康的潜在过程在很大程度上仍然不明确，使得当前的治疗干预措施无法准确理解潜在的、生理相关的疾病和再生途径来治疗症状。我们建议通过消除非生命细胞死亡和相关的炎症过程来限制炎症并协调造血和心血管组织的修复和再生。我们发现小鼠体内 caspase 8 (Casp8) 缺陷的胚胎致死性是由释放的 RIP3 坏死引起的，这为利用消除这些途径的益处开辟了道路。 Casp8-/- Rip3-/- (DKO) 小鼠发育成可存活且具有生育能力的成年小鼠，且对炎症损伤缺乏敏感性但保留足够的免疫功能来控制病毒感染，促使试点努力利用这种环境来改善组织修复、同种异体移植和组织再生。 Casp8 和 RIP3 通路的进化是为了对抗病毒编码的细胞死亡抑制因子，但它们被释放后会引起炎症性疾病并破坏组织修复、体内平衡和再生。该提案采用遗传和小分子方法来消除细胞死亡途径，所有模型均以 DKO 小鼠以及我们创建的其他小鼠品系为模型。我们已经证明，即使总体伤口修复率看起来正常，DKO 小鼠也能抵抗急性炎症、炎性癌症和缺血再灌注损伤，这提出了一个非常重要的问题：“在没有外在细胞死亡途径的情况下，如何从生命中获益？”在诊所里利用”？ DKO 细胞的移植和分化效率高于 WT 细胞。值得注意的是，Casp8-/-Rip3-/- 小鼠在同种异体（MHC 不匹配）骨髓移植中存活下来，这种移植导致 WT C57BL/6 小鼠发生急性移植物抗宿主病 (GvHD)。令人惊讶的是，DKO 成纤维细胞支持核重编程，其效率比 WT 细胞更高。我们建议通过基因操作消除细胞死亡的有害后果，使该领域更接近于改善造血和心血管组织修复和再生的治疗干预策略。我们建议研究小鼠和小鼠细胞的遗传缺陷，并开发恒河猴和人类成纤维细胞的敲除策略，与新描述的 RIP3 激酶小分子抑制剂和市售 caspase 抑制剂相结合，以改善组织修复、同种异体移植和核重新编程。我们的目标是：(1) 优化同种异体植入，(2) 通过消除外源性细胞凋亡来增强核重编程，同时避免引发坏死的缺点。通过这些研究获得的知识将对当前的组织修复和移植方法以及未来的再生医学策略产生广泛影响。