Development of irreversible electroporation-based rational combinations to potentiate the activity of cancer immunotherapy against pancreatic ductal adenocarcinoma

开发基于不可逆电穿孔的合理组合以增强癌症免疫疗法对胰腺导管腺癌的活性

• 批准号: 10363932
• 负责人: CHUN LI
• 金额: $ 59.68万
• 依托单位: UNIVERSITY OF TX MD ANDERSON CAN CTR
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-02-01 至 2027-01-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10363932
• 关键词: Anabolism; Antibodies; Antidiabetic Drugs; Antigens; Attenuated; Bioenergetics; C57BL/6 Mouse; CD8-Positive T-Lymphocytes; CTLA4 gene; Cell Maturation; Cell membrane; Cells; Clinic; Clinical; Clinical Trials; Coagulative necrosis; Cross Presentation; Cytometry; Cytotoxic T-Lymphocytes; Data; Dendritic Cells; Development; Disease; Electroporation; Flow Cytometry; Glutamates; Glutaminase; Glutamine; Glycolysis; Human; IL8RB gene; ITGAM gene; Immune; Immunocompetent; Immunologics; Immunosuppression; Immunotherapy; Infiltration; Inflammatory; Inflammatory Response; Ionizing radiation; KRASG12D; Knowledge; Lead; Malignant Neoplasms; Metabolic; Metabolism; Metastatic Neoplasm to the Liver; Metformin; Mitochondria; Modeling; Mus; Mutation; Myelogenous; Myeloid-derived suppressor cells; Operative Surgical Procedures; Oxidative Phosphorylation; PTPRC gene; Pancreatic Ductal Adenocarcinoma; Pathway Analysis; Pathway interactions; Patients; Penetration; Pharmacology; Phase; Phenformin; Physiologic pulse; Production; Prognosis; Protein Array Analysis; Quality of life; Recurrence; Resistance; Respiration; Role; Signal Transduction; Survival Rate; T-Lymphocyte; Techniques; Testing; Time; Tumor Antigens; Tumor Immunity; Tumor-infiltrating immune cells; Up-Regulation; advanced disease; analog; anti-PD-1; attenuation; base; cancer immunotherapy; cell injury; chemoradiation; clinically relevant; cytokine; effective therapy; immune checkpoint blockade; immunogenic cell death; improved; inhibitor; innovation; mouse model; neoplastic cell; novel; novel therapeutics; pancreatic ductal adenocarcinoma cell; pancreatic ductal adenocarcinoma model; programmed cell death protein 1; programs; recruit; response; single-cell RNA sequencing; small hairpin RNA; success; theranostics; transcriptome sequencing; tumor; tumor ablation; tumor eradication; tumor microenvironment; tumor-immune system interactions; voltage

PROJECT SUMMARY Pancreatic ductal adenocarcinoma (PDAC) is one of the most lethal human cancers, with a 5-year overall survival (OS) rate of 7% for metastatic disease and less than 20% for locally advanced disease. Benefit from current therapies including chemoradiation and surgery is often modest and transient. The significant challenge in the field is how to turn immunologically cold PDAC into hot tumors that respond to immune checkpoint blockade (ICB) therapy. We recently showed that irreversible electroporation (IRE), a tumor ablative technique currently used in the clinics, significantly sensitized PDAC to anti-PD-1 ICB, leading to long-term survival in ~40% mice in an aggressive orthotopic PDAC model. The remarkable anti-PDAC activity was attributed to efficient induction of immunogenic cell death and stromal perturbation in favor of tumor infiltration of CTLs. As part of an effort to define approaches to further enhance the efficacy of IRE + anti-PD-1 combination against PDAC, we uncovered novel immune suppressive mechanism through time-of-flight mass cytometry (CyTOF) immune profiling and single cell RNAseq of PDACs, which showed significant infiltration of CXCR2-expressing myeloid suppressive cells (MDSCs). Furthermore, we found that IRE collapsed glycolysis and oxidative phosphorylation (OxPhos) while upregulated glutaminase and glutamate, suggesting glutaminolysis as a compensatory mechanism to satisfy energy and biosynthesis needs of IRE-treated cells. These data, taken together with the known critical role of MDSCs and heightened glutamine metabolism in immune suppression, the findings by others that the anti-diabetic drugs metformin and phenformin fundamentally change the tumor metabolic program to sensitize tumors to ICB therapy, and our preliminary findings that both IRE and Re-Phen, a newly developed analogue of phenformin, downregulated the OxPhos pathway while displaying an opposite effect on glutamate production, lead us to hypothesize that attenuation of the immunosuppressive TME by depletion of MDSCs or suppression of glutaminolysis by Re-Phen potentiates IRE + ICB to further prolong overall survival and increase the rate of durable response. To test our hypothesis, we will pursue the following specific aims: 1) To identify immunosuppressive factors associated with long-term versus short-term response to IRE + ICB. We will use CyTOF immune profiling, scRNAseq, and cytokine array analyses to fully characterize the impact of IRE in the presence and absence of anti-PD-1 on the immunosuppressive TME. 2) To determine the extent to which therapies directed at MDSCs potentiate IRE + ICB. 3) To determine the extent to which disruption of the metabolic program by theranostic agent Re-Phen potentiates IRE + ICB. The findings from this project are expected to reveal previously undefined roles of MDSCs and deregulated metabolic programming in immune suppression in the context of combined IRE + ICB therapy. Success of this project will have exceptional impact because it will offer a potentially effective therapy for PDAC.

项目概要 胰腺导管腺癌 (PDAC) 是最致命的人类癌症之一，总体生存期为 5 年 转移性疾病的 (OS) 率为 7%，局部晚期疾病的 (OS) 率低于 20%。受益于当前 包括放化疗和手术在内的治疗通常是温和且短暂的。中的重大挑战 研究领域是如何将免疫学上冷的 PDAC 转变为对免疫检查点封锁做出反应的热肿瘤 （ICB）治疗。我们最近展示了不可逆电穿孔（IRE），这是目前的一种肿瘤消融技术 在临床中使用，使 PDAC 对抗 PD-1 ICB 显着敏感，导致约 40% 的小鼠长期存活 积极的原位 PDAC 模型。显着的抗 PDAC 活性归因于有效诱导 免疫原性细胞死亡和基质扰动有利于 CTL 的肿瘤浸润。作为努力的一部分 我们发现，确定进一步增强 IRE + 抗 PD-1 组合对抗 PDAC 功效的方法 通过飞行时间质谱细胞术 (CyTOF) 免疫分析和 PDAC 的单细胞 RNAseq，显示表达 CXCR2 的骨髓抑制显着浸润 细胞（MDSC）。此外，我们发现 IRE 破坏了糖酵解和氧化磷酸化 (OxPhos) 同时上调谷氨酰胺酶和谷氨酸，表明谷氨酰胺分解是一种补偿机制 满足 IRE 处理细胞的能量和生物合成需求。这些数据与已知的关键数据一起 MDSC 和增强的谷氨酰胺代谢在免疫抑制中的作用，其他人的发现表明 抗糖尿病药物二甲双胍和苯乙双胍从根本上改变肿瘤代谢程序致敏 肿瘤对 ICB 治疗的影响，我们的初步发现 IRE 和 Re-Phen（一种新开发的类似物） 苯乙双胍，下调 OxPhos 途径，同时对谷氨酸产生产生相反的影响， 导致我们假设通过消耗 MDSC 或抑制来减弱免疫抑制性 TME Re-Phen 的谷氨酰胺分解作用增强 IRE + ICB，进一步延长总生存期并提高死亡率 持久的反应。为了检验我们的假设，我们将追求以下具体目标：1）确定 与 IRE + ICB 的长期与短期反应相关的免疫抑制因素。我们将使用 CyTOF 免疫分析、scRNAseq 和细胞因子阵列分析，以全面表征 IRE 对 免疫抑制 TME 上是否存在抗 PD-1。 2) 确定程度 针对 MDSC 的疗法可增强 IRE + ICB。 3) 确定中断的程度 治疗诊断剂 Re-Phen 的代谢程序可增强 IRE + ICB。该项目的调查结果是 有望揭示 MDSC 先前未定义的作用以及免疫中失调的代谢编程 IRE + ICB 联合治疗中的抑制。该项目的成功将产生非凡的影响 因为它将为 PDAC 提供潜在有效的治疗方法。