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患者存在致癌突变KRAS(KRAS*)，该突变可促进胰腺癌的发生，并 PDAC维护所需。鉴于KRAS*现在可以用药，在几种情况下出现了治疗耐药性 临床前模型和临床试验，表明需要联合治疗才能获得持久的疾病 控制力。靶向KRAS*导致肿瘤相关巨噬细胞(TAMs)在胰腺和结肠中的聚集 和肺癌模型，为嵌合抗原受体(CAR)巨噬细胞(Mφ)治疗提供了机会 以及M-φ介导的致炎细胞因子的癌症特异性递送。然而，移植的MφS被 易于在肝、肺、脾中蓄积，尽管剩余的M-φS大部分定位于肿瘤， 越来越多的安全担忧可能通过条件基因表达系统来解决。假设是这样的 肿瘤条件性表达细胞因子装甲CAR MφS(ca-CAR-ms)与KRAS*抑制剂的协同作用 (Krasi)有效抑制原发肿瘤生长、转移和肿瘤复发，延长生存期 在PDAC自发转移小鼠模型上。拟议研究的成功结果将产生第一个 CA-CAR-M治疗PDAC，为KRAS*PDAC患者的临床试验铺平道路，并提供 为其他M-φ富集型癌症开发CA-CAM的适用方法。目标1.优化细胞培养和 过继巨噬细胞治疗的工程平台。原代MφS体外无增殖性， 供者M-φS在健康器官内蓄积存在安全隐患。AIM 1a将使MφS的体外扩增成为可能 由遗传或化学筛选的M-φ自我更新调节因子。目标1b将限制供者MφS的肿瘤外活动 通过使用双重氧气感应开关并识别特有的基因和组织驻留的MφS，其 启动子/增强子将用于肿瘤特异性基因的表达。目的2.针对PDAC设计MφS轿车。 有效抗原特异性激活小鼠巨噬细胞吞噬功能和细胞毒作用的CAR综合优化 MφS缺阵。Aim 2a将设计和优化CAR的组成蛋白结构域，以指导Mφ针对 PDAC。目的2b将确定CAR-MS作为Krasi在PDAC模型中的辅助治疗的杀瘤作用。 Aim 2c将剖析CAR激活如何重新编程Mφ转录、分泌和途径的机制 转导以诱导杀瘤作用。目的3.工程师有条件装甲MφS触发杀瘤 豁免权。MφS是肿瘤归巢者，因此MφS释放促炎细胞因子可能会降低系统性红斑狼疮 毒副作用大，同时可引起较强的抗肿瘤免疫反应。Aim 3a将开发安全有效的细胞因子装甲 应用细胞因子筛选和条件转基因表达系统治疗M-φ。目标3b将评估 最佳CAR结构构建的M-φS的杀瘤活性 Krasi在各种PDAC模型中的辅助治疗。AIM 3c将阐明细胞因子装甲Mφ在S中的作用 单细胞RNA测序和免疫表型对肿瘤微环境重塑的影响