Gasdermin E and pyroptosis in cancer

Gasdermin E 和癌症焦亡

基本信息

批准号：
10583571
负责人：
Judy Lieberman
金额：
$ 49.4万
依托单位：
BOSTON CHILDREN'S HOSPITAL
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-04-03 至 2025-03-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10583571
关键词：
Affect Apoptosis Apoptotic Back Brain C-terminal CASP3 gene CD8-Positive T-Lymphocytes Cancer cell line Caspase Cell Death Cell Line Cell Survival Cell membrane Cells Cellular Stress Cessation of life Chemotherapy and/or radiation Cochlea Colorectal Colorectal Cancer Cytotoxic T-Lymphocytes Data Decitabine Drug Targeting Drug resistance Effectiveness Epidermal Growth Factor Receptor Epigenetic Process Etoposide Family member Genes Goals Granzyme Human Immune Immunity Immunocompetent Immunologic Deficiency Syndromes Immunotherapy In Vitro Infection Infiltration Inflammation Mediators Inflammatory Invertebrates KRAS2 gene Kidney Killer Cells Knock-out Large Intestine Lipid Binding Lung Lymphocyte Malignant Neoplasms Mediating Melanoma Cell Metastatic Neoplasm to Lymph Nodes Methylation Mitochondria Modeling Mucous Membrane Mus Mutate Mutation N-terminal Natural Killer Cells Normal tissue morphology Peptide Hydrolases Permeability Phagocytosis Pharmaceutical Preparations Placenta Predisposition Protein Family Radiation Resistance Role Small Intestines Stimulus Stomach Swelling Syndrome T-Cell Depletion Testing Therapeutic Transplantation Tumor Immunity Tumor Promotion Tumor Suppressor Proteins Tumor-infiltrating immune cells VDAC1 gene Western Blotting cancer cell cancer vaccination chemotherapy cytokine cytotoxic deafness drug sensitivity flexibility immune clearance immunogenic improved in vivo inhibitor insight loss of function mutation lung cancer cell malignant breast neoplasm malignant stomach neoplasm melanoma neoplastic cell overexpression perforin recruit response treatment response tumor tumor growth tumorigenesis

项目摘要

Caspase-3 activation during apoptosis can trigger caspase-3 cleavage of gasdermin E (GSDME). The gasdermins (GSDM) are a family of proteins, whose cleavage activates inflammatory death, called pyroptosis. N-terminal GSDM fragments form pores in the cell membrane that cause rapid cell death in which the cell swells, activates and releases inflammatory cytokines and other inflammatory mediators, and eventually bursts. GSDME cleavage converts noninflammatory apoptotic death to more rapid inflammatory pyroptotic death. GSDME is not expressed in most cancer cell lines, is epigenetically inactivated in gastric, colorectal and breast cancer, relative to normal tissue, and mutated in some others. GSDME expression suppresses colony formation in gastric and colorectal cancer and invasivity of breast cancer. Worse 5-year survival and an increase in lymph node metastases are associated with reduced GSDME in breast cancer. Moreover, lack of GSDME promotes drug resistance in melanoma and lung cancer cell lines. We hypothesize that GSDME acts like a tumor suppressor, that some tumor cells avoid pyroptosis by downregulating or mutating GSDME and that the switch from apoptosis to pyroptosis profoundly affects tumor cell survival, anti-tumor immunity and response to chemotherapy and radiation. In preliminary data, cancer-associated GSDME mutations are shown to be primarily loss of function mutations. Gsdme knockout in cancer lines promoted tumor growth while ectopic GSDME expression strongly inhibited tumor growth in immune competent mice. The tumor suppressive role of GSDME was lost in immunodeficient NOD.scid.Il2rg-/- (NSG) and Prf1-/- mice deficient in perforin and strongly inhibited by NK or CD8 T cell depletion. GSMDE expression by the tumor increased infiltrating, functional CD8 T cells and NK cells. Based on these data, we hypothesize that GSDME suppresses tumor growth by recruiting and activating anti-tumor killer lymphocytes. We also found that granzymes B and M, death- inducing proteases of killer lymphocytes, directly cleave GSDME during killer cell attack to activate pyroptosis in a caspase-independent manner, which is amplified by caspase activation. We also hypothesize that direct granzyme cleavage of GSDME in cancer cells triggers pyroptosis and enhances their anti-tumor immunity. Killer lymphocyte mediated death has previously been thought to be non-inflammatory. Our goal is to test these hypotheses and develop mechanistic insights to understand whether and how suppression of GSDME activation of pyroptosis in cancer cells impacts oncogenesis, drug sensitivity and protective immunity. Our specific aims are to investigate the effect of GSDME mutation and expression on GSDME activation, lipid binding, oligomerization and pore formation, on whether cell death by apoptotic stimuli or killer cells is inflammatory and immunogenic, and whether and how GSDME affects tumor growth, immunity and responses to therapy in vitro and in transplanted mouse tumor models.

细胞凋亡过程中 Caspase-3 的激活可触发 Caspase-3 对 Gasdermin E (GSDME) 的裂解。 Gasdermin (GSDM) 是一个蛋白质家族，其裂解会激活炎症性死亡，称为细胞焦亡。 N 端 GSDM 片段在细胞膜上形成孔，导致细胞快速死亡，细胞肿胀，激活并释放炎症细胞因子和其他炎症介质，并最终爆发。裂解将非炎性细胞凋亡转变为更快速的炎性焦亡，GSDME 则不然。在大多数癌细胞系中表达，在胃癌、结直肠癌和乳腺癌中表观遗传失活，相对正常组织中的 GSDME 表达突变会抑制胃和胃中的集落形成。结直肠癌和乳腺癌的侵袭性更差，并且淋巴结增加。乳腺癌转移与 GSDME 减少有关，此外，缺乏 GSDME 会促进药物治疗。我们假设 GSDME 具有肿瘤抑制因子的作用，一些肿瘤细胞通过下调或突变 GSDME 来避免细胞焦亡，并且从细胞凋亡至焦亡深刻影响肿瘤细胞的存活、抗肿瘤免疫和对肿瘤细胞的反应初步数据显示，癌症相关的 GSDME 突变与化疗和放疗有关。主要是癌细胞系中的功能缺失突变促进了肿瘤生长，而异位。 GSDME 表达强烈抑制免疫活性小鼠的肿瘤生长。 GSDME 在免疫缺陷的 NOD.scid 中强烈丢失。Il2rg-/- (NSG) 和 Prf1-/- 小鼠穿孔素和 NK 或 CD8 T 细胞耗竭所抑制。肿瘤浸润、功能性 CD8 的 GSMDE 表达增加 T 细胞和 NK 细胞。根据这些数据，我们发现 GSDME 通过以下方式抑制肿瘤生长：我们还发现，颗粒酶 B 和 M 可以招募和激活抗肿瘤杀伤淋巴细胞。诱导杀伤淋巴细胞的蛋白酶，在杀伤细胞攻击期间直接裂解 GSDME，从而激活细胞焦亡一种不依赖于半胱天冬酶的方式，通过半胱天冬酶激活来放大。癌细胞中 GSDME 的颗粒酶裂解引发细胞焦亡并增强其抗肿瘤作用杀伤淋巴细胞介导的死亡以前被认为是非炎症性的。测试这些假设并发展机制见解，以了解是否以及如何抑制 GSDME 激活癌细胞焦亡会影响肿瘤发生、药物敏感性和保护性免疫。我们的具体目标是研究 GSDME 突变和表达对 GSDME 激活、脂质结合、寡聚化和孔形成，与细胞凋亡刺激或杀伤细胞导致的细胞死亡有关炎症和免疫原性，以及 GSDME 是否以及如何影响肿瘤生长、免疫和反应用于体外治疗和移植小鼠肿瘤模型。