A Pathway for Necrotic Cell Death

坏死细胞死亡的途径

基本信息

批准号：
10680459
负责人：
DAVID J SHAPIRO
金额：
$ 37.89万
依托单位：
UNIVERSITY OF ILLINOIS AT URBANA-CHAMPAIGN
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-08-09 至 2027-07-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10680459
关键词：
Antineoplastic Agents Apoptosis Apoptosis Promoter Autophagocytosis Brain Breast Cancer Cell Breast cancer metastasis CRISPR screen Calcium Cell Culture Techniques Cell Death Cell Death Induction Cell Death Inhibition Cell Line Cell membrane Cells Cessation of life Clinical Clinical Trials Colon Colon Carcinoma Data Development Diapause Disseminated Malignant Neoplasm Drug Targeting Electroporation Embryo Endometrial Carcinoma Estrogen Receptor alpha Estrogens Goals Immune Immune checkpoint inhibitor Immunocompromised Host Immunotherapeutic agent Immunotherapy Knock-out Knowledge Link MYC Family Protein Malignant Neoplasms Malignant neoplasm of ovary Mammary Neoplasms Mediating Mediator Membrane Metastatic Neoplasm to the Bone Metastatic Neoplasm to the Liver Metastatic Neoplasm to the Lung Metastatic malignant neoplasm to brain Mitochondria Mus Necrosis Necrosis Induction Neoplasm Metastasis Oncolytic Osmosis Pathway interactions Patients Peptides Pharmaceutical Preparations Protein Synthesis Inhibition Proteins Recurrence Regulatory Pathway Resistance Retreatment Rupture Sodium Channel Stress Swelling Testing Therapeutic Tumor Markers Uterus Water Work Xenograft Model Xenograft procedure cancer cell cancer immunotherapy cancer therapy cancer type drug development endoplasmic reticulum stress flexibility genome-wide immunogenic immunogenic cell death malignant ascites malignant breast neoplasm mouse model neoantigens novel anticancer drug novel therapeutics protein biomarkers protein folding proteostasis receptor resistance mechanism response sensor success targeted agent therapeutic target therapy resistant transcriptome sequencing triple-negative invasive breast carcinoma tumor virtual

项目摘要

Unlike other death pathways, protein mediators of drug-induced necrotic cell death were poorly defined. Necrosis activates immune cells, inducing immunogenic cell death. Therefore, understanding necrosis provides new avenues for enhancing drug development and cancer immunotherapy. Our anticancer drugs BHPI and ErSO act via estrogen receptor α (ERα) to induce lethal necrosis-inducing hyperactivation of the anticipatory Unfolded Protein Response (a-UPR). In orthotopic xenografts and a PDX, ErSO eradicates primary and metastatic therapy-resistant ERα+ breast cancer, induces near complete regression of lethal breast cancer in brain, and of endometrial cancer and ovarian cancer, and kills most ovarian cancer cells in patient malignant ascites. From CRISPR screens against BHPI and ErSO, we identified the Ca2+ activated, plasma membrane Na+ channel TRPM4 as the executioner protein that BHPI and ErSO use to induce necrosis and the likely membrane flexibility modulator FGD3. BHPI and ErSO-induced elevated Ca2+ opens the TRPM4 channel, eliciting a rapid influx of external Na+, Cl- and accompanying water. This swells the cells, causing osmotic stress, which hyperactivates the UPR, leading to ATP depletion, FGD3 enhanced membrane rupture and necrotic cell death. TRPM4 knockout abolished ATP depletion, sustained UPR hyperactivation, cell swelling and death. Notably, TRPM4 knockout also inhibited necrosis induced by unrelated anticancer therapies, the mitochondrial targeting oncolytic peptide, LTX-315, the Ca2+ channel targeting agent, Englerin A and Ca2+ electroporation (CaEP). Aim 1. Identify and functionally characterize known and additional shared components of the TRPM4 pathway. We will combine data from completed CRISPR screens, new screens using LTX-315, Englerin A, and CaEP and RNA-seq data from our recently developed ErSO resistant cell lines. Aim 2. Using cell and tumor studies, test the hypothesis that diverse necrosis-inducing anticancer therapies, in which Ca2+ levels are increased by transient a-UPR activation or other mechanisms, share a common pathway that converges on the UPR-TRPM4-FGD3 pathway. To extend UPR activation therapies to ERα- cancers, test the idea that the clinically promising, mechanistically obscure, necrosis-inducing therapy, Ca2+ electroporation, works in part through the UPR-TRPM4-FGD3 necrosis pathway. Aim 3. Using syngeneic mouse models establish whether necrosis-inducing agents extend the reach of immunotherapy to rapidly lethal breast cancer that has metastasized to brain and does not express neoantigens. Aim 4. Mechanisms of resistance to necrosis inducing cancer drugs are largely unexplored. Using our Myc down-regulated reversibly quiescent cells, we will identify ErSO resistance mechanisms and test whether loss of Myc in the quiescent cells is due to a-UPR mediated ATP depletion activating AMPK, thereby inhibiting protein synthesis via eEF2. These studies will establish a new pathway of immunogenic anticancer therapy-induced necrotic cell death through the UPR.

与其他死亡途径不同，药物诱导的坏死细胞死亡的蛋白质介质的定义很差。坏死激活免疫原性细胞死亡。因此，了解坏死为增强药物发育和癌症免疫疗法提供了新的途径。我们的抗癌药 BHPI和ERSO ACT通过雌激素受体α（ERα）诱导致命坏死诱导的过度激活预期展开的蛋白质反应（A-UPR）。在原位异种移植物和PDX中，erso radiotications 原发性和转移性治疗的ERα+乳腺癌，诱导致死的完全消退大脑中的乳腺癌，子宫内膜癌和卵巢癌，并杀死大多数卵巢癌细胞患者恶性腹水。从针对BHPI和ERSO的CRISPR屏幕中，我们确定了CA2+激活的，质膜Na+通道TRPM4作为BHPI和ERSO诱导的execution子蛋白坏死和可能性膜柔韧性调节剂FGD3。 BHPI和ERSO诱导的高架Ca2+打开 TRPM4通道引起了外部Na+，Cl-和参与水的快速影响。这膨胀了导致渗透应激的细胞，使UPR过度激活，导致ATP耗竭，FGD3增强了膜破裂和坏死细胞死亡。 TRPM4淘汰赛废除了ATP部署，持续了UPR 过度活化，细胞肿胀和死亡。值得注意的是，TRPM4敲除也抑制了由无关的抗药疗法，线粒体靶向溶瘤肽，LTX-315，Ca2+通道靶向剂，Englerin A和Ca2+电穿孔（CAEP）。目标1。识别并在功能上表征 TRPM4途径的已知和其他共享组件。我们将结合完成的数据 CRISPR屏幕，使用LTX-315，Englerin A以及CAEP和RNA-Seq数据的新屏幕，我们最近开发了ERSO抗性细胞系。 AIM 2。使用细胞和肿瘤研究，检验潜水员的假设坏死引起的抗癌疗法，其中瞬态A-UPR激活或其他机制，共享一种在UPR-TRPM4-FGD3途径上收敛的通用途径。到将UPR激活疗法扩展到ERα-Canters，测试临床上承诺的，机械机械的想法晦涩的，坏死诱导的治疗，Ca2+电穿孔，部分通过UPR-TRPM4-FGD3起作用坏死途径。 AIM 3。使用合成小鼠模型确定坏死诱导的药物是否将免疫疗法的范围扩展到已转移到大脑的快速致命的乳腺癌不是表达新抗原。目标4。对坏死诱发癌症药物的抗性机制在很大程度上是意外。使用我们的MYC下调的可逆静止单元，我们将识别ERSO抗性机理和测试是否静止单元中的MYC是否是由于UPR介导的ATP部署激活AMPK，从而通过EEF2抑制蛋白质合成。这些研究将建立一个新的免疫原性抗癌治疗诱导的坏死细胞死亡的途径。