Anastasis: A Novel Cell Survival Mechanism

Anastasis：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，可将凋亡细胞从死亡边缘拯救出来。 支持不可逆细胞凋亡的经典观点，我们已经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，十二种人类癌细胞系，以及果蝇的卵室。是什么 吻合的生理功能、病理作用和治疗潜力？Anastasis可以 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， 难以替代的细胞，如心肌细胞和神经元。如果是真的，增强吻合可能对 治疗心力衰竭和脑损伤此外，anastasis可能是一种未被识别的逃避策略， 细胞在癌症治疗中存活，从而有助于疾病复发。如果证实，抑制愈合 在垂死的癌细胞中，可能会促进癌细胞死亡，减少复发的机会。Anastasis也可以 在胚胎发育和正常体内平衡过程中限制细胞凋亡中起重要作用。如果被发现， 了解它的调节可以提供新的见解，控制细胞死亡和生存的生理 条件然而，测试这些假设存在一些挑战。很难追踪anastasis， 特别是在体内，因为逆转凋亡的细胞在形态上与健康细胞无法区分， 细胞目前还没有发现anastasis的特异性标志，anastasis的调节因子也尚未发现。 在这里，我们将通过开发一种新颖且高度特异性的跟踪系统来克服其中的许多挑战， 标记哺乳动物研究的anastatic细胞，并确定anastasis的关键调节因子。为了马克·阿纳斯塔西娅 细胞，我们将创建一个anastasis生物传感器，可以标记anastasis细胞与永久表达 荧光蛋白只有在它们从线粒体外膜透化和线粒体外膜透化中恢复后才能表达。 （MOMP）和半胱天冬酶-3激活，这两个最被认可的凋亡事件，使得该生物传感器系统 对anastasis有很强的特异性。我们将建立anastasis生物传感器稳定的细胞系，以确定可逆性， 细胞凋亡，并将采用生物传感器异种移植物，使用临床相关的 小鼠模型。为了阐明anastasis的机制，我们将通过蛋白质组学鉴定其关键调控因子， 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， （潜在的促渗调节剂和治疗靶点）在促渗的不同阶段， 特定的翻译后修饰可以区分无主枝细胞，确定从无主枝细胞恢复的细胞是否 不同的细胞死亡诱导具有相似的分子特征，并研究了如何干扰细胞凋亡。 调节剂候选物可以调节凋亡的可逆性。我们将识别出小分子， 通过生物信息学方法筛选候选物，并在体外测试其促进或抑制愈合的功效。 该项目的成功完成将为研究anastasis提供必要的工具和知识， 为通过控制回输来开发革命性的新治疗方法奠定了坚实的基础。

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.