Develop Conditionally Armored CAR Macrophage Therapy for Pancreatic Cancer

开发针对胰腺癌的条件装甲 CAR 巨噬细胞疗法

• 批准号: 10710883
• 负责人: Pingping Hou
• 金额: $ 60.25万
• 依托单位: RUTGERS BIOMEDICAL AND HEALTH SCIENCES
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-09-05 至 2028-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10710883
• 关键词: Address; Adjuvant Therapy; Adverse effects; Antigens; Ascites; Biological Assay; Biology; CD147 antigen; CD47 gene; Cancer Etiology; Cancer Model; Cell Culture Techniques; Cell Therapy; Cells; Cessation of life; Chemicals; Clinical Trials; Collaborations; Colon Carcinoma; Combined Modality Therapy; Cyclic AMP; Cyclic AMP-Dependent Protein Kinases; Data; Engineering; Enhancers; Ensure; Gene Expression; Genes; Genetic; Genetic Engineering; Genetic Transcription; Goals; Growth; Hematopoietic stem cells; Homing; Homologous Transplantation; Human; Hypoxia; Immune; Immune response; Immunity; Immunologist; Immunophenotyping; Impairment; In Vitro; Inflammatory; Intravenous; KRAS2 gene; Knowledge; Liver; Lung; MSLN gene; Macrophage; Maintenance; Malignant Neoplasms; Malignant neoplasm of lung; Malignant neoplasm of pancreas; Mediating; Methods; Mus; Neoplasm Metastasis; Oncogenic; Organ; Outcome; Oxygen; PTPNS1 gene; Pancreatic Ductal Adenocarcinoma; Pathway interactions; Patients; Phagocytosis; Physiological; Pre-Clinical Model; Primary Neoplasm; Proliferating; RXR; Receptor Activation; Recurrence; Recurrent tumor; Research; Resistance; Risk; Role; SIRT1 gene; Safety; Signal Transduction; Solid Neoplasm; Spleen; System; Tertiary Protein Structure; Therapeutic; Tissues; Toxic effect; Transplantation; Tumor-associated macrophages; United States; allotransplant; anti-tumor immune response; cellular engineering; chimeric antigen receptor; clinical application; clinical risk; cytokine; cytokine therapy; cytotoxic; cytotoxicity; design; disorder control; experience; human disease; in vivo; inhibitor; mouse model; mutant; neoantigens; novel; pancreatic ductal adenocarcinoma model; pancreatic tumorigenesis; permissiveness; pre-clinical; programs; promoter; screening; self-renewal; single cell sequencing; single-cell RNA sequencing; suicide gene; synergism; systemic toxicity; targeted treatment; therapeutically effective; transgene expression; tumor; tumor growth; tumor microenvironment

SUMMARY/ABSTRACT Pancreatic ductal adenocarcinoma (PDAC) is one of the deadliest cancers with limited therapeutic options. About 95% PDAC patients harbor oncogenic mutant KRAS (KRAS*) that promotes pancreatic carcinogenesis and is required for PDAC maintenance. Given that KRAS* can be druggable now, therapy resistance occurs in several pre-clinical models and clinical trials, suggesting that therapy combination is required to achieve durable disease control. Targeting KRAS* results in enrichment of tumor associated macrophages (TAMs) in pancreatic, colon and lung cancer models, offering opportunities for chimeric antigen receptor (CAR) macrophage (Mφ) therapy and for Mφ-mediated cancer-specific delivery of pro-inflammatory cytokines. However, transplanted Mφs were prone to accumulate in liver, lung, and spleen despite that most of the remaining Mφs were localized in tumors, rising safety concerns that might be addressed by conditional gene expression systems. The hypothesis is that tumor-conditionally expressed cytokine armored CAR Mφs (ca-CAR-Ms) may synergize with KRAS* inhibitors (KRASi) to effectively suppress primary tumor growth, metastasis, and tumor recurrence and to prolong survival in PDAC spontaneous metastasis mouse models. A successful outcome of proposed study will generate the first ca-CAR-M therapy for PDAC treatment, pave the way for clinical trials in KRAS* PDAC patients and provide applicable methods to develop ca-CAR-Ms for other Mφ-enriched cancers. Aim 1. Optimize cell culture and engineering platforms for adoptive macrophage therapy. Primary Mφs are non-proliferative in vitro, and accumulation of donor Mφs in healthy organs is a latent safety risk. Aim 1a will enable in vitro expansion of Mφs by genetic or chemical screening of Mφ self-renewal regulators. Aim 1b will limit off-tumor activity of donor Mφs by employing dual oxygen-sensing switch and identifying TAM-specific genes versus tissue-resident Mφs, whose promoter/enhancers will be used for tumor-specific gene expression. Aim 2. Design CAR Mφs to target PDAC. The comprehensive CAR optimization for effective antigen-specific activation of phagocytosis and cytotoxicity in Mφs is lacking. Aim 2a will design and optimize constituent protein domains of CAR to direct Mφ activities against PDAC. Aim 2b will determine the tumoricidal effect of CAR-Ms as an adjuvant therapy of KRASi in PDAC models. Aim 2c will dissect mechanisms how CAR activation reprograms Mφ transcription, secretion, and pathway transduction to elicit tumoricidal effect. Aim 3. Engineer conditionally armored Mφs to trigger tumoricidal immunity. Mφs are tumor-homing, so delivery of pro-inflammatory cytokines by Mφs may reduce systemic toxicity while elicit strong anti-tumor immune response. Aim 3a will develop safe and effective cytokine armored Mφ therapy by cytokine screening and employing conditional transgene expression system. Aim 3b will assess tumoricidal activity of Mφs engineered by the optimal CAR construct, the top cytokine candidate or both as an adjuvant therapy of KRASi in various PDAC models. Aim 3c will delineate the role of cytokine armored Mφs in the tumor microenvironment remodeling by single cell RNA sequencing and immunophenotyping.

摘要/摘要 胰腺导管腺癌（PDAC）是最致命的癌症之一，治疗选择有限。关于 95％的PDAC患者具有促进胰腺癌的致癌突变体KRAS（KRAS*） PDAC维护所需。鉴于Kras*现在可以吸毒，因此在几个 临床前模型和临床试验，表明需要治疗组合才能达到持久性疾病 控制。靶向kras*会导致胰腺相关巨噬细胞（TAM）富含胰腺，颜色 和肺癌模型，为嵌合抗原受体（CAR）巨噬细胞（Mφ）治疗提供机会 以及用于Mφ介导的癌细胞因子的癌症特异性递送。但是，移植的Mφ是 容易积聚在肝，肺和脾脏目的地，其余的Mφ位于肿瘤中， 有条件基因表达系统可能解决的安全问题上升。假设是 肿瘤条件表达的细胞因子装甲车MφS（CA-CAR-MS）可能与KRAS*抑制剂协同 （KRASI）有效抑制原发性肿瘤的生长，转移和肿瘤复发，并延长生存率 在PDAC自发转移小鼠模型中。拟议研究的成功结果将产生第一个 用于PDAC治疗的CA-CAR-M治疗，为KRAS* PDAC患者的临床试验铺平了道路 适用的方法，用于开发其他Mφ-富含癌症的CA-CAR-MS。目标1。优化细胞培养和 适应性巨噬细胞疗法的工程平台。一级Mφ在体外不相差，并且 供体Mφ在健康器官中的积累是一种潜在的安全风险。 AIM 1A将在体外扩展Mφ 通过Mφ自我更新调节剂的遗传或化学筛选。 AIM 1B将限制供体MφS的肿瘤活动 通过采用双氧感应开关并识别TAM特异性基因与组织居民​​MφS（谁） 启动子/增强子将用于肿瘤特异性基因表达。 AIM 2。设计汽车MφS靶向PDAC。 用于有效抗原特异性吞噬作用和细胞毒性激活的综合汽车优化 缺乏Mφ。 AIM 2A将设计和优化汽车的组成蛋白域，以指导Mφ活动 PDAC。 AIM 2B将确定CAR-MS作为PDAC模型中KRASI的可调节疗法的结核效应。 AIM 2C将剖析汽车激活如何重新编程Mφ转录，分泌和途径的机制 转导向结核效应。目标3。有条件装甲的工程师以触发结核 免疫。 MφS是肿瘤的，因此MφS递送促炎细胞因子可能会降低全身性 毒性，同时引起强烈的抗肿瘤免疫反应。 AIM 3A将发展安全有效的细胞因子装甲 通过细胞因子筛查和采用条件转化表达系统的Mφ治疗。 AIM 3B将评估 由最佳汽车构建体设计的MφS的肿瘤活性，顶部细胞因子候选者或两者兼而有之 各种PDAC模型中Krasi的辅助治疗。 AIM 3C将描述细胞因子装甲Mφ在 单细胞RNA测序和免疫表型通过肿瘤微环境重塑。