Anastasis: A Novel Cell Survival Mechanism

Anastasis：一种新的细胞生存机制

基本信息

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
项目类别：
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

项目摘要

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：一种新的细胞生存机制项目概要 Anastasis是一种新发现的细胞恢复机制，可以将凋亡细胞从死亡边缘拯救出来。挑战不可逆细胞凋亡的经典观点，我们发现细胞凋亡具有强大的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和果蝇的卵室。什么是 anastasis 的生理功能、病理作用和治疗潜力？阿纳斯塔西斯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，如心肌细胞和神经元。如果属实，增强吻合可能有益于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，使癌症细胞在癌症治疗中存活下来，从而导致疾病复发。如果得到证实，则抑制阿斯塔西斯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。阿纳斯塔西斯也可能在胚胎发育和正常稳态过程中限制细胞凋亡中发挥重要作用。如果确定的话，了解其调节可以为生理学中细胞死亡和存活的控制提供新的见解。状况。然而，检验这些假设存在一些挑战。很难追踪阿纳斯塔西斯，尤其是在体内，因为逆转细胞凋亡的细胞在形态上与健康细胞没有区别细胞。没有发现任何阿斯塔西斯特定的标志，并且阿斯塔西斯的调节剂仍未被发现。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许多这些挑战标记用于哺乳动物研究的anastasis细胞，并确定anastasis的关键调节因子。标记 anastatic 细胞，我们将创建一个 anastasis 生物传感器，可以用永久表达的 astasis 细胞来标记仅在线粒体外膜透化恢复后才出现荧光蛋白 (MOMP) 和 caspase-3 激活，这两个最受认可的细胞凋亡事件，使得该生物传感器系统对 anastasis 具有高度特异性。我们将建立 anastasis 生物传感器稳定细胞系来确定体外细胞凋亡，并将利用生物传感器异种移植物利用临床相关的方法来询问体内的吻合情况鼠标模型。为了阐明 anastasis 的机制，我们将通过蛋白质组学来确定其关键调节因子，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（潜在的吻合调节剂和治疗靶点）在吻合的不同阶段，确定是否特定的翻译后修饰区分变形细胞，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，并研究如何干扰细胞分裂候选调节因子可以调节细胞凋亡的可逆性。我们将识别靶向的小分子通过生物信息学分析候选药物，并在体外测试其促进或抑制 anastasis 的功效。该项目的成功完成将为研究解剖学提供必要的工具和知识，从而通过控制吻合，为开发革命性的新治疗方法奠定了坚实的基础。