Engineering In Vivo Chimeric Antigen Receptor Macrophages (CARMs) using mRNA-exosomes for Cancer Immunotherapy

使用 mRNA-外泌体工程体内嵌合抗原受体巨噬细胞 (CARM) 用于癌症免疫治疗

基本信息

批准号：
10740743
负责人：
Wen Jiang
金额：
$ 56.78万
依托单位：
UNIVERSITY OF TX MD ANDERSON CAN CTR
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-08-01 至 2027-07-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10740743
关键词：
Animals Antibodies Antigen Presentation Antitumor Response Autologous Breast Breast Cancer Cell Breast Cancer Model CAR T cell therapy CD8B1 gene Cell Therapy Cells Clinical Complex Consumption Dose Engineering Epidermal Growth Factor Receptor Exhibits Face Flow Cytometry Gene Expression Generations Genes Hematologic Neoplasms Human ITGAM gene Immune Immunocompetent Immunofluorescence Immunologic Immunologic Stimulation In Vitro Infusion procedures Macrophage Malignant Neoplasms Maps Mediating Membrane Messenger RNA Metastatic breast cancer Mus Myeloid-derived suppressor cells Nature Pathway interactions Phagocytes Phagocytosis Phenotype Population Pre-Clinical Model Preparation Production Receptor Cell Regimen Research Solid Solid Neoplasm Spatial Distribution Specificity T cell infiltration T-Lymphocyte Testing Therapeutic Time Toxic effect Viral Vector antigen-specific T cells antitumor effect cancer immunotherapy chimeric antigen receptor chimeric antigen receptor T cells cytokine design draining lymph node engineered exosomes exosome experimental study extracellular immune activation in vivo innovation insight interest lymph nodes mRNA delivery malignant breast neoplasm multiphoton microscopy novel strategies patient derived xenograft model programs protein expression receptor reconstitution transcriptome sequencing transcriptomics tumor tumor growth tumor heterogeneity tumor microenvironment

项目摘要

PROJECT SUMMARY Chimeric antigen receptor (CAR) T cell therapy has revolutionized the treatment of hematologic cancers. However, for solid tumors, CAR T cells face challenges including intratumor heterogeneity, dynamic expression of target receptors, and often the inability for T cells to traffic to tumors to mediate the desired antitumor effect. In contrast to the lack of T cell infiltrates, many solid tumors are abundant in immune suppressive myeloid cells including macrophages. Therefore, converting these immune suppressive cells into tumoricidal phenotype represents a promising strategy for cell-based therapy. There are now strong interest in generating CAR macrophages in which autologous macrophages are transduced with CAR delivered by viral vectors ex vivo to enhance their phagocytosis, antigen presentation and cytokine producing capabilities following re-infusion. However, ex vivo preparation of CAR macrophages is complex, time consuming, and due to the non-dividing nature of macrophages, is often inefficient. With the recent advances in mRNA-based therapeutics, it is now possible to reprogram specific immune cell populations in vivo, thus eliminating the complex ex vivo production of autologous CAR cells. Our present proposal aims to propose an innovative strategy of generating CAR macrophages in vivo using mRNA-loaded exosomes to treat HER2 receptor positive breast cancer. This will be the first study to evaluate the feasibility of producing CAR macrophages in vivo using mRNA delivery platforms and assessing the antitumor efficacy of CAR macrophages for cancer immunotherapy. We hypothesize that our strategy represents a revolutionary way to produce CAR macrophages in vivo using CAR mRNA-loaded exosome and offers a promising new approach for cell therapy against solid tumors. Our previous study showed that we can efficiently produce mRNA-loaded exosomes to restore protein expression in solid tumors. Furthermore, our preliminary experiments showed that the exosomes loaded with HER2 CAR mRNA can produce CAR macrophages in vivo with enhanced effector functions. Our current study will test our overall hypothesis by using the following specific aims. In Aim 1, we will evaluate the dynamics and toxicity of CAR macrophage production in vivo using CAR mRNA exosomes. In Aim 2, we will evaluate transcriptomic and functional profiles of in vivo generated CAR macrophages, Finally, in Aim 3, we will assess the antitumor effect of in vivo generated CAR macrophages against both murine and human HER2 expressing breast cancer. If successful, our proposed research can overcome a major technical hurdle that is currently facing cell therapy. The mRNA exosome platform could potentially be expanded to other CAR constructs and greatly expand the potential utility of cell therapy for breast and other solid cancers.

项目概要嵌合抗原受体 (CAR) T 细胞疗法彻底改变了血液病的治疗癌症。然而，对于实体瘤，CAR T细胞面临着瘤内异质性、动态性等挑战。靶受体的表达，以及 T 细胞通常无法运输到肿瘤以介导所需的作用抗肿瘤作用。与缺乏 T 细胞浸润相反，许多实体瘤具有丰富的免疫细胞浸润。抑制性骨髓细胞，包括巨噬细胞。因此，将这些免疫抑制细胞转化为杀肿瘤表型代表了基于细胞的治疗的一种有前途的策略。现在大家有浓厚的兴趣产生 CAR 巨噬细胞，其中自体巨噬细胞被病毒传递的 CAR 转导载体离体增强其吞噬作用、抗原呈递和细胞因子产生能力重新输注后。然而，CAR巨噬细胞的离体制备过程复杂、耗时，并且由于由于巨噬细胞的不可分裂性质，这种方法通常效率低下。随着基于 mRNA 的最新进展疗法，现在可以在体内重新编程特定的免疫细胞群，从而消除自体 CAR 细胞的复杂离体生产。我们目前的提案旨在提出一种创新的使用装载mRNA的外泌体在体内产生CAR巨噬细胞来治疗HER2受体的策略阳性乳腺癌。这将是第一个评估在中生产 CAR 巨噬细胞的可行性的研究。使用 mRNA 递送平台进行体内实验并评估 CAR 巨噬细胞对癌症的抗肿瘤功效免疫疗法。我们假设我们的策略代表了一种革命性的 CAR 生产方式使用负载 CAR mRNA 的外泌体对体内巨噬细胞进行研究，为细胞治疗提供了一种有前景的新方法对抗实体瘤。我们之前的研究表明，我们可以有效地生产装载 mRNA 的外泌体恢复实体瘤中的蛋白质表达。此外，我们的初步实验表明装载HER2 CAR mRNA的外泌体可以在体内产生具有增强效应的CAR巨噬细胞功能。我们当前的研究将通过以下具体目标来检验我们的总体假设。在目标 1 中，我们将使用 CAR mRNA 外泌体评估 CAR 巨噬细胞在体内产生的动力学和毒性。在目标 2，我们将评估体内生成的 CAR 巨噬细胞的转录组和功能图谱，最后，在目标 3 中，我们将评估体内生成的 CAR 巨噬细胞对小鼠和小鼠的抗肿瘤作用。表达人类 HER2 的乳腺癌。如果成功，我们提出的研究可以克服重大技术问题细胞治疗目前面临的障碍。 mRNA 外泌体平台有可能扩展到其他领域 CAR 构建并极大地扩展了细胞疗法对乳腺癌和其他实体癌的潜在效用。