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Gasdermin E and pyroptosis in cancer

Gasdermin E 和癌症焦亡

基本信息

批准号：
10375415
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/10375415
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 Immunotherapy In Vitro Infection 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 Phagocytosis Pharmaceutical Preparations Placenta Protein Family Radiation Resistance Role Small Intestines Stimulus Stomach Syndrome T-Cell Depletion Testing Therapeutic Transplantation Tumor Immunity Tumor Suppressor Proteins Tumor-infiltrating immune cells VDAC1 gene base cancer cell cancer invasiveness 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.

细胞凋亡过程中半胱天冬酶-β 3的激活可以触发半胱天冬酶-β 3切割gasdermin E（GSDME）。 gasdermins（GSDM）是蛋白质家族，其裂解激活炎性死亡，称为焦亡。 N-末端GSDM片段在细胞膜中形成孔，引起细胞膨胀的快速细胞死亡，激活并释放炎性细胞因子和其他炎症介质，并最终爆发。裂解将非炎性凋亡性死亡转化为更快速的炎性热凋亡性死亡。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学失活，相对于 GSDME的表达抑制了胃粘膜上皮细胞集落的形成，结直肠癌和乳腺癌的侵袭性。更差的5年生存率和淋巴结转移的增加，转移与乳腺癌GSDME减少有关。此外，缺乏GSDME促进药物治疗，我们假设GSDME的作用类似于肿瘤抑制因子，一些肿瘤细胞通过下调或突变GSDME来避免焦亡，细胞凋亡对细胞凋亡的影响深刻地影响了肿瘤细胞的存活、抗肿瘤免疫和对在初步的数据中，癌症相关的GSDME突变被证明是与化疗和放疗相关的。 Gsdme基因敲除可促进肿瘤生长，而肿瘤细胞的异位生长则可促进肿瘤细胞的生长。 GSDME表达强烈抑制免疫活性小鼠中的肿瘤生长。 GSDME在免疫缺陷型NOD. scid. Il 2 rg-Rg/-Rg（NSG）和Prf 1-Rg/-Rg小鼠中缺失，而在免疫缺陷型NOD. scid. Il 2 rg-Rg/-Rg（NSG）小鼠中缺失，肿瘤的GSMDE表达增加了浸润性的功能性CD 8 + T细胞，基于这些数据，我们假设GSDME通过以下途径抑制肿瘤生长：我们还发现，颗粒酶B和M，死亡相关的诱导杀伤淋巴细胞的蛋白酶，在杀伤细胞攻击过程中直接切割GSDME，以激活一种不依赖于半胱天冬酶的方式，这种方式会被半胱天冬酶激活放大。我们还假设直接癌细胞中GSDME的颗粒酶切割触发细胞凋亡并增强其抗肿瘤活性杀伤淋巴细胞介导的死亡以前被认为是非炎症性的，我们的目标是为了验证这些假设，并发展机械的见解，以了解是否以及如何抑制 GSDME激活癌细胞中的焦亡影响肿瘤发生、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、脂质过氧化作用和细胞凋亡的影响。结合，寡聚化和孔形成，对细胞死亡是否由凋亡刺激或杀伤细胞，炎症和免疫原性，以及GSDME是否以及如何影响肿瘤生长，免疫和反应涉及体外和移植小鼠肿瘤模型中的治疗。