Anastasis: A Novel Cell Survival Mechanism

转移：一种新的细胞生存机制 项目摘要 ANASTASIS是一种新发现的细胞恢复机制，可以将凋亡细胞从死亡的边缘拯救出来。 挑战经典的不可逆转的细胞凋亡观点，我们发现了细胞凋亡的强大可逆性 三种类型的小鼠/大鼠原代细胞，12个人类癌细胞系，以及果蝇的卵室。何谓 血管转移症的生理功能、病理作用和治疗潜力？阿纳斯塔西可能 是一种意想不到的细胞保护机制，用于保存终末分化的细胞和组织 难以替代，如心肌细胞和神经元。如果是真的，增强Anastsis可能会对 治疗心力衰竭和脑损伤。此外，Anastsis可能是一种未被认识的逃避策略，使癌症成为可能 细胞在癌症治疗中存活，从而有助于疾病复发。如果得到证实，则抑制Anastasis 癌细胞死亡可能会促进癌细胞死亡，降低复发的机会。阿纳斯塔西也可能 在胚胎发育和正常动态平衡过程中发挥着重要的作用。如果被确认， 了解其调控机制可以为生理学中控制细胞死亡和存活提供新的见解。 条件。然而，检验这些假说有几个挑战。很难追踪到阿纳斯塔西， 特别是在活体内，因为逆转了细胞凋亡的细胞在形态上与健康的细胞没有区别 细胞。目前还没有发现ANASTASIS特有的特征，而且ANASTASIS的调节者仍然没有被发现。 在这里，我们将通过开发一种新颖且高度具体的跟踪系统来克服其中的许多挑战 标记用于哺乳动物研究的再生细胞，并识别再生细胞的关键调节因子。标记为Anastatic 细胞，我们将创造一种ANASTASIS生物传感器，可以标记永久表达的ANASTASIS 荧光蛋白只有在它们从线粒体外膜通透性恢复后才能 (MOMP)和caspase-3激活，这两个最广为人知的凋亡事件，使这个生物传感器系统 高度特异的阿纳斯塔西斯病。我们将建立ANASTASIS生物传感器稳定的细胞系以确定其可逆性 体外细胞凋亡，并将使用生物传感器异种移植来询问体内转移使用临床相关 老鼠模型。为了阐明Anastasis的机制，我们将通过蛋白质组学确定其关键调控因子， 遗传学和药理学方法。我们将确定哪些基因表现出上调或下调表达 (潜在的血管吻合调节剂和治疗靶点)在血管吻合的不同阶段，决定是否 特定的翻译后修饰区分再生细胞，确定细胞是否从 不同的细胞死亡诱导具有相似的分子特征，并研究如何干扰转移 候选调节器可以调节细胞凋亡的可逆性。我们将识别目标为 通过生物信息学，测试它们在体外促进或抑制转移的效果。 该项目的成功完成将为研究AnastASIS提供必要的工具和知识，从而 通过控制复发，为开发革命性的新治疗方法奠定坚实的基础。

Anastasis: A Novel Cell Survival Mechanism Project Summary Anastasis is a newly discovered cell recovery mechanism that rescues apoptotic cells from the brink of death. Challenging the classic view of irreversible apoptosis, we have discovered robust reversibility of apoptosis in three types of mouse/rat primary cells, twelve human cancer cell lines, and egg chambers in fruit flies. What are the physiological functions, pathological roles, and therapeutic potentials of anastasis? Anastasis could be an unexpected cytoprotective mechanism for preserving terminally differentiated cells and tissues that are difficult to replace, such as cardiomyocytes and neurons. If true, enhancing anastasis may be beneficial for treating heart failure and brain injury. Besides, anastasis could be an unrecognized escape tactic enabling cancer cells to survive cancer therapy, thereby contributing to disease recurrence. If confirmed, suppressing anastasis in dying cancer cells may promote cancer cell death and reduce the chances of recurrence. Anastasis may also play important roles in limiting apoptosis during embryonic development and normal homeostasis. If identified, understanding its regulation can provide new insights into the control of cell death and survival in physiological conditions. However, there are several challenges of testing these hypotheses. It is difficult to track anastasis, especially in vivo, because cells that have reversed apoptosis are morphologically indistinguishable from healthy cells. There are no anastasis-specific hallmarks identified, and the regulators of anastasis remain undiscovered. Here, we will overcome many of these challenges by developing a novel and highly specific tracking system to label anastatic cells for mammalian studies, and to identify the key regulators of anastasis. To mark anastatic cells, we will create an anastasis biosensor that can tag anastatic cells with permanent expression of a fluorescent protein only after they have recovered from both mitochondrial outer membrane permeabilization (MOMP) and caspase-3 activation, the two most recognized apoptotic events, making this biosensor system highly specific to anastasis. We will establish anastasis biosensor stable cell lines to determine reversibility of apoptosis in vitro, and will employ biosensor xenografts to interrogate anastasis in vivo using clinically relevant mouse models. To elucidate the mechanism of anastasis, we will identify its key regulators, through proteomics, genetics, and pharmacological approaches. We will identify which genes exhibit up- or down-expression (potential anastasis regulators, and therapeutic targets) during different stages of anastasis, determine whether specific post-translational modifications distinguish anastatic cells, establish whether cells that recover from different cell death inductions share similar molecular features, and investigate how interfering with anastasis regulator candidates could modulate the reversibility of apoptosis. We will identify small molecules that target the candidates by bioinformatics, and test their efficacy in promoting or suppressing anastasis in vitro. Successful completion of this project will generate essential tools and knowledge for studying anastasis, thereby laying a strong foundation for developing revolutionary new therapeutic approaches by controlling anastasis.