Necrotic survivors and plasma membrane integrity signaling

项目摘要/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程序化， 包括一些抒情形式。例如，质膜(PM)成孔执行器 当被RIPK3磷酸化时，坏死性下垂是MLKL。针对下睑下垂，Gasdermin家族成员 一旦被半胱氨酸酶或颗粒酶处理，PM会破坏执行者吗？耐人寻味的是，首相 毛孔形成不是坏死性上睑下垂或下垂的“不归路”。这是因为ESCRT- III复合体可通过膜重塑修复受损的PM。破碎的PM分数可以 被甩掉。因此，对于嗜坏死性和嗜焦性细胞来说，它们都可以耐受有限的水平。 如果ESCRT-III的修复能力没有被压垮，PM损坏的可能性和生存时间很长。在……里面 在这项资助中，我们展示了我们未发表的数据，表明坏死细胞是否能够存活 活着，亚致命的PM损伤足以启动一个复杂的信号转导网络。 我们将这个信号转导途径命名为质膜完整性信号(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 假设坏死性“幸存者”可以通过PMI信号通路刺激肿瘤发生 以及随之而来的趋化因子/细胞因子旁分泌。我们的最终目标是机械地 了解并进一步瞄准这些“幸存者”，以改善癌症治疗。为了实现这一目标， 我们将首先寻找PM完整性损失的传感器。我们提议测试一种很有前途的离子 通道候选，很可能是PM损坏的传感器之一。我们还有 设计了一个精巧的细胞培养系统，同时进行siRNA和sgRNA筛选，以寻找 PM破坏了全基因组的传感器。接下来，我们将同时使用动物模型和人类 标本探查坏死的“幸存者”。我们将进一步测试 这些“幸存者”在肿瘤发生和转移中分泌的趋化因子/细胞因子。第三，我们 将设法直接消灭坏死性“幸存者”。我们的初步数据让我们找到了一个候选人 环状E3连接酶。我们将测试靶向E3连接酶是否能帮助我们抑制ESCRT- 修复行动和根除坏死性“幸存者”。因为腐烂的“幸存者”只是 不久前(由我们和其他人发现)，细胞死亡的巨大影响 导致癌症、移植和其他疾病仍然是新鲜的，需要充分 已定义。像这样的创新研究将促进这一发现。

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 In this grant, we presented our UNPUBLISHED data showing if a necrotic cell can manage to stay alive, the sub-lethal PM damage is sufficient to initiate a sophisticated signal transduction network.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.