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% PDAC 患者携带致癌突变体 KRAS (KRAS*)，可促进胰腺癌发生，并且 PDAC 维护所需。鉴于 KRAS* 现在可以药物化，几种药物中都会出现治疗耐药性 临床前模型和临床试验表明需要联合治疗才能实现持久疾病 控制。靶向 KRAS* 可导致胰腺、结肠中肿瘤相关巨噬细胞 (TAM) 的富集 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 Mφ 介导的癌症特异性促炎细胞因子的传递。然而，移植的Mφs是 尽管大多数剩余的 Mφ 位于肿瘤中，但仍容易在肝脏、肺和脾中积累， 条件基因表达系统可能会解决日益严重的安全问题。假设是 肿瘤条件表达的细胞因子装甲 CAR Mφs (ca-CAR-Ms) 可能与 KRAS* 抑制剂产生协同作用 （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 疗法，为 KRAS* PDAC 患者的临床试验铺平道路，并提供 开发用于其他富含 Mφ 的癌症的 ca-CAR-M 的适用方法。目标 1. 优化细胞培养和 过继性巨噬细胞治疗的工程平台。原发性 Mφ 在体外不增殖，并且 供体Mφ在健康器官中的积累是一个潜在的安全风险。目标 1a 将实现 Mφ 的体外扩增 通过遗传或化学筛选 Mφ 自我更新调节剂。目标 1b 将限制供体 Mφ 的肿瘤外活性 通过采用双氧传感开关并识别 TAM 特异性基因与组织驻留 Mφ，其 启动子/增强子将用于肿瘤特异性基因表达。目标 2. 设计 CAR Mφ 以针对 PDAC。 用于有效抗原特异性激活吞噬作用和细胞毒性的全面 CAR 优化 缺乏Mφs。目标 2a 将设计和优化 CAR 的组成蛋白结构域，以指导 Mφ 活性 PDAC。目标 2b 将确定 CAR-M 在 PDAC 模型中作为 KRASi 辅助疗法的杀肿瘤作用。 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 转导以引发杀肿瘤作用。目标 3. 设计有条件装甲的 Mφ 来触发杀肿瘤作用 免疫。 Mφs 是肿瘤归巢性的，因此通过 Mφs 传递促炎细胞因子可能会减少全身性 毒性同时引发强烈的抗肿瘤免疫反应。目标3a将开发安全有效的细胞因子装甲 通过细胞因子筛选和采用条件转基因表达系统进行 Mφ 治疗。目标 3b 将评估 由最佳 CAR 构建体、顶级候选细胞因子或两者共同设计的 Mφ 的肿瘤活性 KRASi 在各种 PDAC 模型中的辅助治疗。目标 3c 将描述细胞因子装甲 Mφs 在 通过单细胞RNA测序和免疫表型重塑肿瘤微环境。