Anastasis: A Novel Cell Survival Mechanism

Anastasis：一种新的细胞生存机制

• 批准号: 10713750
• 负责人: Ho Lam Tang
• 金额: $ 40.94万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-09-22 至 2028-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10713750
• 关键词: Animals; Apoptosis; Apoptotic; Bioinformatics; Biosensor; Brain Injuries; CASP3 gene; Cancer cell line; Cardiac Myocytes; Cell Death; Cell Death Induction; Cell Death Process; Cell Differentiation process; Cell Membrane Permeability; Cell Survival; Cells; Cessation of life; Cytoprotection; Drosophila genus; Embryonic Development; Event; Exhibits; Foundations; Genes; Genetic; Heart failure; Homeostasis; Human; In Vitro; Knowledge; Label; Mammalian Cell; Molecular; Morphology; Mus; Neurons; Outer Mitochondrial Membrane; Pathologic; Physiological; Play; Post-Translational Protein Processing; Proteins; Proteomics; Rattus; Recovery; Recurrence; Recurrent disease; Regulation; Role; System; Testing; Therapeutic; Tissues; Xenograft procedure; cancer cell; cancer survival; cancer therapy; clinically relevant; egg; in vivo; insight; mouse model; novel; novel therapeutic intervention; pharmacologic; preservation; small molecule; stable cell line; therapeutic target; tool

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.

Anastasis：一种新型的细胞存活机制 项目摘要 Anastasis是一种新发现的细胞回收机制，可从死亡的边缘挽救凋亡细胞。 挑战不可逆凋亡的经典观点，我们发现了凋亡的强大可逆性 三种类型的小鼠/大鼠原代细胞，十二个人类癌细胞系和果蝇中的鸡蛋室。什么是 Anastasis的生理功能，病理作用和治疗潜力？ anastasis可以 成为一种意外的细胞保护机制，用于保存终末分化的细胞和组织 难以替代，例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，阿纳斯塔症可能是一种无法识别的逃生策略，使癌症能够 细胞生存于癌症治疗，从而导致疾病复发。如果确认，请抑制anastasis 在垂死的癌细胞中可能会促进癌细胞死亡并减少复发的机会。 anastasis也可能 在限制胚胎发育和正常稳态期间凋亡中起重要作用。如果确定， 了解其调节可以为控制细胞死亡和生理生存的新见解提供新的见解 状况。但是，测试这些假设存在一些挑战。很难跟踪anastasis， 特别是体内，因为逆转凋亡的细胞在形态上与健康无法区分 细胞。没有确定的特定标志，阿纳斯塔斯的调节剂仍未发现。 在这里，我们将通过开发一个新颖且高度特定的跟踪系统来克服许多这些挑战 标记用于哺乳动物研究的肛门细胞，并确定厌食的关键调节剂。标记肛门 细胞，我们将创建一个吻合的生物传感器，该生物传感器可以标记具有永久表达的肛门细胞 荧光蛋白仅在两种线粒体外膜通透性中回收后才回收 （MOMP）和caspase-3激活，这是两个最知名的凋亡事件，使该生物传感器系统 高度特定于anastasis。我们将建立Anastasis生物传感器稳定的细胞系，以确定 体外凋亡，并将使用生物传感器异种移植物在体内询问Anastasis，并使用临床上的相关 鼠标模型。为了阐明Anastasis的机制，我们将通过蛋白质组学确定其关键调节剂 遗传学和药理方法。我们将确定哪些基因表现出上表达或下表达 （潜在的Anastasis调节剂和治疗靶标）在不同阶段，在Anastasis的不同阶段，确定是否确定 特定的翻译后修饰区分肛门细胞，确定是否从 不同的细胞死亡归因具有相似的分子特征，并研究了如何干扰厌氧菌 候选者可以调节凋亡的可逆性。我们将确定针对目标的小分子 通过生物信息学的候选者，并测试其在体外促进或抑制吻合方面的功效。 该项目的成功完成将生成研究阿纳斯塔症的基本工具和知识，从而 通过控制anastasis来发展革命性的新治疗方法，为开发革命性的新治疗方法奠定了基础。