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.

项目摘要 胰腺导管腺癌（PDAC）是最致命的人类癌症之一，总生存期为5年 （OS）转移性疾病的率为7％，局部晚期疾病的率少于20％。从当前受益 包括化学放疗和手术在内的疗法通常是适度和短暂的。在 场是如何将免疫学冷PDAC变成对免疫检查点封锁反应的热肿瘤 （ICB）治疗。我们最近表明，不可逆的电穿孔（IRE），这是一种目前的肿瘤消融技术 在诊所使用，对抗PD-1 ICB显着敏感，导致长期生存率在〜40％的小鼠中 一种侵略性的原位PDAC模型。显着的抗PDAC活性归因于有效诱导 免疫原性的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 定义方法以进一步提高IRE +抗PD-1组合对PDAC的功效，我们发现了 新型免疫抑制机制通过飞行时间质量细胞仪（Cytof）免疫分析和 PDAC的单细胞RNASEQ，显示出表达CXCR2的明显浸润 细胞（MDSC）。此外，我们发现IRE崩溃了糖酵解和氧化磷酸化（Oxphos） 虽然上调谷氨酰胺酶和谷氨酸，这表明谷氨酰胺分解是一种补偿机制 满足IRE处理的细胞的能量和生物合成需求。这些数据，以及已知的关键 MDSC的作用和增强谷氨酰胺代谢在免疫抑制中的作用，他人的发现 抗糖尿病药物二甲双胍和苯甲呈蛋白从根本上改变肿瘤代谢程序以使其敏感 ICB治疗的肿瘤，以及我们的初步发现，即愤怒和重新培训，这是一个新开发的类似物 苯甲酸蛋白，在对谷氨酸产生的相反作用时下调Oxphos途径， 导致我们假设通过耗尽MDSC或抑制来衰减免疫抑制TME 通过重新获得的谷氨酰胺溶解增强IRE + ICB，以进一步延长总体生存率并提高 持久的响应。为了检验我们的假设，我们将追求以下具体目的：1）确定 与长期对IRE + ICB的长期反应相关的免疫抑制因素。我们将使用 细胞的免疫分析，SCRNASEQ和细胞因子阵列分析，以完全表征IRE的影响 免疫抑制性TME上存在抗PD-1。 2）确定在多大程度上 针对MDSCS的疗法增强IRE + ICB。 3）确定破坏的程度 Theranoticatic的代谢程序重新启动增强IRE + ICB。这个项目的发现是 预计将揭示MDSC的先前不确定的作用和免疫管制的免疫程序 在IRE + ICB治疗的背景下进行抑制。该项目的成功将产生特别的影响 因为它将为PDAC提供潜在的有效疗法。