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

• 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
• 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

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）：消除健康和疾病中细胞死亡途径的好处。影响血管健康的基本过程仍然在很大程度上不确定，从而使当前的治疗干预措施治疗症状，而无需精确理解潜在的，生理上相关的疾病和再生途径。我们建议通过消除非重要细胞死亡以及相关的炎症过程来限制造血和心血管组织的修复和再生。我们发现，小鼠的caspase 8（CASP8）缺陷的胚胎致死性是由释放的RIP3坏死引起的，这为消除这些途径的利用带来了途径。 casp8 - / - rip3 - / - （dko）小鼠成长为缺乏炎症性侮辱的可行而肥沃的成年人 但是，保留足够的免疫功能来控制病毒感染，促使试点努力为哈恩斯的设置改善组织修复，同种异体植入和组织再生。 CASP8和RIP3途径演变为抵消病毒编码的细胞死亡抑制子，但被释放以引起炎症性疾病并破坏组织修复，稳态和再生。该提案采用遗传和小分子方法来消除均由DKO小鼠建模的细胞死亡途径以及我们创建的其他小鼠菌株。我们已经表明，尽管总体伤口修复率看起来正常，但DKO小鼠抵抗了急性炎症，炎症癌和缺血再灌注损伤，这促使一个非常重要的问题：“在没有外部细胞死亡途径的情况下，在临床中利用外部细胞死亡途径，如何从生命中受益”？与WT细胞相比，DKO细胞植入并以更高的效率分化。值得注意的是，CASP8 - / - RIP3 - / - 小鼠在WT C57BL/6小鼠中导致急性移植与宿主疾病（GVHD）的同种异体（MHC不匹配）骨髓植入。令人惊讶的是，DKO成纤维细胞支持核重编程，并提高了WT细胞的效率。我们建议通过遗传操纵消除细胞死亡的有害后果，使该区域更接近治疗性干预策略，以改善造血和心血管组织修复和再生。我们建议研究小鼠和小鼠细胞中的遗传缺乏，并在恒河猴和人成纤维细胞上开发敲除策略，以与新描述的RIP3激酶的小分子抑制剂结合使用，并提供商业上可用的CASPase抑制剂，以改善组织修复，同种异体移植和核重编程。我们的目的是：（1）优化同种异体植入，（2）通过消除外在凋亡而没有触发坏死的不利条件来增强核重编程。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的影响。