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.

凋亡过程中的caspase-3激活会触发caspase-3 Gasdermin E（GSDME）的裂解。这加油动物（GSDM）是一个蛋白质家族，其裂解激活了炎症死亡，称为凋亡。 N末端GSDM片段在细胞膜中形成毛孔，导致细胞肿胀的快速细胞死亡，其中，激活并释放炎症细胞因子和其他炎症介质，有时会破裂。 GSDME 切割将非炎性凋亡死亡转化为更快的炎症性凋亡死亡。 GSDME不是在大多数癌细胞系中表达，在胃，结直肠癌和乳腺癌中表观遗传灭活，相对到正常的组织，并在其他一些组织中突变。 GSDME表达抑制胃中的菌落形成和结直肠癌和乳腺癌的侵入性。更糟糕的5年生存率和淋巴结的增加转移与乳腺癌中的GSDME降低有关。此外，缺乏GSDME会促进药物黑色素瘤和肺癌细胞系的抗性。我们假设GSDME的作用像肿瘤抑制剂，某些肿瘤细胞通过下调或突变GSDME避免了凋亡，并且从凋亡的凋亡深刻影响肿瘤细胞的存活，抗肿瘤免疫和对化学疗法和放射线。在初步数据中，与癌症相关的GSDME突变显示为功能突变的主要丧失。 GSDME敲除癌品系促进了肿瘤的生长，而Ecopic GSDME表达强烈抑制免疫胜任小鼠的肿瘤生长。肿瘤抑制作用的作用 GSDME在免疫缺陷nod.scid.il2rg - / - （nsg）和prf1 - / - 小鼠中丢失了被NK或CD8 T细胞耗竭抑制。肿瘤的GSMDE表达增加了浸润，功能CD8 T细胞和NK细胞。基于这些数据，我们假设GSDME抑制了肿瘤的生长募集和激活抗肿瘤杀伤型淋巴细胞。我们还发现颗粒状B和M，死亡 - 诱导的杀伤型淋巴细胞的蛋白酶，在杀伤细胞攻击过程中直接清除GSDME，以激活凋亡 caspase独立的方式，通过caspase激活扩大。我们还假设这是直接的癌细胞中GSDME的颗粒状切割会触发凋亡并增强其抗肿瘤免疫。杀手型淋巴细胞介导的死亡以前一直认为是非炎症的。我们的目标是测试这些假设并开发机械见解，以了解是否以及如何抑制 GSDME激活癌细胞中的凋亡会影响肿瘤生成，药物敏感性和保护性免疫。我们的具体目的是研究GSDME突变和表达对GSDME激活，脂质的影响结合，寡聚和孔形成，凋亡刺激或杀伤细胞的细胞死亡是炎症性和免疫原性，以及GSDME是否影响肿瘤生长，免疫组织切和反应在体外和移植小鼠肿瘤模型中进行治疗。