Necrotic survivors and plasma membrane integrity signaling

坏死幸存者和质膜完整性信号传导

• 批准号: 10241019
• 负责人: Yi-Nan Gong
• 金额: $ 135.81万
• 依托单位: UNIVERSITY OF PITTSBURGH AT PITTSBURGH
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-22 至 2024-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10241019
• 关键词: Animal Model; Caspase; Cell Culture System; Cell Death; Cell membrane; Cells; Data; Electron Transport Complex III; Family member; Goals; Grant; Granzyme; Human; Ion Channel; Lead; Lytic; Malignant Neoplasms; Membrane; Molecular; Names; Necrosis; Neoplasm Metastasis; Process; RIPK3 gene; Research; Role; Signal Pathway; Signal Transduction; Signal Transduction Pathway; Small Interfering RNA; Specimen; Survivors; Testing; Time; Transplantation; cancer therapy; chemokine; cytokine; genome-wide; improved; innovation; paracrine; repaired; sensor; tumorigenesis; ubiquitin-protein ligase

PROJECT SUMMARY/ABSTRACT After decades of cell death study, it is now well accepted that cell death can be highly programmed, including some lytic forms. For example, the plasma membrane (PM) pore-forming executor for necroptosis is MLKL, when phosphorylated by RIPK3. For pyroptosis, gasdermin family members are the PM damaging executors, once processed by caspases or granzymes. Intriguingly, the PM pore-forming is not the “point of no return” for necroptosis or pyroptosis. This is because ESCRT- III complex can repair the damaged PM by membrane remodeling. The broken PM fractions can be shed off. Therefore, for both necroptotic and pyroptotic cells, they can tolerate a limited level of PM damage and survive for a long time if ESCRT-III repairing capacity is not overwhelmed. In this grant, we presented our UNPUBLISHED data showing if a necrotic cell can manage to stay alive, the sub-lethal PM damage is sufficient to initiate a sophisticated signal transduction network. We named this signaling transduction pathway as Plasma Membrane Integrity signaling (PMI signaling). PMI signals are efficient in promoting pro-tumor chemokines/cytokines secretions. We hypothesize that the necrotic “survivors” can stimulate oncogenesis via the PMI signaling pathway and the consequent chemokines/cytokines paracrine. Our ultimate goal is to mechanistically understand and further target these “survivors” to improve cancer treatment. To achieve this end, we will firstly hunt for the sensors for PM integrity loss. We proposed to test a promising ion channel candidate that is very likely to be one of the sensors for PM damage. We have also planned an elegant cell culture system to perform both siRNA and sgRNA screen to search for the PM damage sensors genome-wide. Next, we will use both animal models and human specimens to probe the necrotic “survivors.” We will further test the roles of the chemokines/cytokines secreted by these “survivors” in tumorigenesis and metastasis. Third, we will try to eliminate the necrotic “survivors” directly. Our preliminary data lead us to a candidate RING E3 ligase. We will test whether targeting this E3 ligase can help us suppress the ESCRT- III repair action and eradicate necrotic “survivors.” As the necrotic “survivors” were only discovered not long ago (by us and others), the massive impacts of cell death “survivors” in contributing to cancers, transplantation, and other illnesses are still fresh and needed to be fully defined. Innovative research, like this one, will promote the revelation.

项目总结/摘要 经过几十年的细胞死亡研究，现在人们普遍认为细胞死亡可以高度编程， 包括一些裂解形式。例如，质膜（PM）成孔执行器， 坏死性凋亡是MLKL，当被RIPK 3磷酸化时。对于焦亡，gasdermin家族成员 是PM破坏执行者，一旦被半胱天冬酶或颗粒酶处理。有趣的是，首相 孔形成不是坏死性凋亡或焦亡的“不归点”。这是因为ESCRT- III复合物可通过膜重塑修复受损的PM。破碎的PM级分可以 被甩掉。因此，对于坏死性和热变性细胞，它们可以耐受有限的水平， 如果ESCRT-Ⅲ的修复能力不被压倒，则PM损坏的可能性很小，并且可以存活很长时间。在 在这项资助中，我们展示了我们未公布的数据，显示坏死细胞是否可以设法留在 在存活的情况下，亚致死PM损伤足以启动复杂的信号转导网络。 我们将这一信号转导途径命名为质膜完整性信号转导（Plasma Membrane Integrity Signaling，PMI 信令）。PMI信号在促进促肿瘤趋化因子/细胞因子分泌方面是有效的。我们 假设坏死的“幸存者”可以通过PMI信号通路刺激肿瘤发生 以及随之产生的趋化因子/细胞因子旁分泌。我们的最终目标是 了解并进一步针对这些“幸存者”，以改善癌症治疗。为了达到这个目的， 我们将首先寻找PM完整性损失的传感器。我们提议测试一种有希望的离子 很可能是PM损伤的传感器之一的通道候选者。我们还 计划了一个优雅的细胞培养系统来进行siRNA和sgRNA筛选， PM会破坏全基因组的传感器接下来，我们将使用动物模型和人类模型。 标本来探测坏死的“幸存者”我们将进一步测试 在肿瘤发生和转移中由这些“幸存者”分泌的趋化因子/细胞因子。三是 会试图直接消灭坏死的“幸存者”我们的初步数据显示 RING E3连接酶。我们将测试靶向这种E3连接酶是否可以帮助我们抑制ESCRT。 修复行动，根除坏死“幸存者”。因为坏死的“幸存者” 不久前（由我们和其他人）发现，细胞死亡“幸存者”对人类的巨大影响， 导致癌症，移植和其他疾病仍然是新鲜的，需要充分考虑。 定义了像这样的创新研究将促进启示。