Immunotherapy Modeling in Organoids Co-preserving Tumor and Infiltrating Immune Compartments

共保存肿瘤和浸润免疫区室的类器官的免疫治疗模型

基本信息

批准号：
10586140
负责人：
CALVIN J KUO
金额：
$ 64.39万
依托单位：
STANFORD UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-03-17 至 2026-02-28
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10586140
关键词：
3-Dimensional Acute Address Adoptive Cell Transfers Adoptive Transfer Aftercare Air Antigens Autologous B-Lymphocytes Biopsy Bioreactors Cells Cellular immunotherapy Clinical Coculture Techniques Complex Cutaneous Data Detection Development Epithelium Equilibrium Exhibits Human Immune Immune Evasion Immune checkpoint inhibitor Immune response Immune system Immunologic Surveillance Immunological Models Immunotherapeutic agent Immunotherapy In Vitro Interleukin-2 Invaded Ligands Liquid substance Macrophage Malignant Neoplasms Methods Modeling Mus Natural Killer Cells Nature Nodal Organoids Outcome Patients Peptides Phosphoric Monoester Hydrolases Population Primary Neoplasm Progressive Disease Protein Dephosphorylation Receptor Inhibition Resistance Resolution Sampling System T cell receptor repertoire sequencing T-Cell Receptor T-Lymphocyte T-Lymphocyte Subsets T-cell receptor repertoire TNFRSF5 gene Techniques Testing Therapeutic Therapeutic antibodies Time Tumor Antigens Tumor Cell Line Tumor Escape Tumor-Derived Tumor-Infiltrating Lymphocytes air treatment anti-PD-1 anti-PD-1/PD-L1 anti-PD-L1 anti-PD1 therapy anti-tumor immune response cancer immunotherapy cell killing checkpoint inhibition chimeric antigen receptor T cells clinical decision-making clinical efficacy cohort cytotoxicity exhaust follow-up human model immune cell infiltrate immune checkpoint blockade immunoregulation improved in vitro Model in vitro activity in vivo inhibitor therapy melanoma mouse model neoplastic cell network models next generation novel patient response patient subsets peripheral blood predictive marker preservation programmed cell death ligand 1 programmed cell death protein 1 prospective receptor reconstitution recruit response restraint single-cell RNA sequencing skin squamous cell carcinoma success treatment trial trial comparing tumor tumor immunology tumor progression tumor specificity tumorigenesis

项目摘要

PROJECT SUMMARY The immune system remarkably distinguishes between self and non-self/self-aberrant antigens, affording exquisite anti-tumor specificity and inhibition of tumorigenesis. However, tumor immunosurveillance is unfortunately opposed by tumor cell evasion of the immune response. Immune checkpoint blockade (ICB) targeting PD-1, PD-L1, CD40 and others, as well as adoptive cell transfer (CAR-T, bulk TILs) favorably modulate this equilibrium for therapeutic benefit. However, response rates are often incomplete, progressive disease is common, and predictive biomarkers are suboptimal. The development of next-generation immunotherapies has been hindered by a lack of in vitro models that functionally recapitulate syngeneic interactions between tumor and infiltrating immune cells. In response, we have developed organoid methods that culture primary human tumor biopsies together with their infiltrating immune components as a cohesive unit without reconstitution. These “patient-derived tumor organoids” (PDO) preserve tumor cells alongside endogenous T, B, NK cells and macrophages, robustly recapitulate the T cell receptor clonotype repertoire of the original tumor, and crucially, manifest tumor-infiltrating lymphocyte (TIL) expansion, activation and tumor cell killing in response to anti-PD-1/PD-L1 therapeutic antibodies (Cell, 2018). The PDO system thus represents a holistic organoid model of human tumor-immune interactions. Here, we leverage the PDO technique to investigate immunotherapeutic mechanisms and treatments in PD-1-responsive cutaneous squamous cell carcinoma (cSCC) and melanoma, exploiting pre- and post-treatment human biopsies and mouse models. Aim 1 hypothesizes that checkpoint inhibition induces a complex and sequential network response involving immune-tumor and immune-immune crosstalk. Thus, Aim 1 employs the ability to perform serial time-course sampling of PDOs to define a single cell RNA-seq network cellular crosstalk model of the early anti-PD-1-stimulated anti-tumor immune response over multiple acute time points typically inaccessible to clinical biopsies performed after months. Importantly, comparison of this immune propagation in responding versus non-responding mouse and human organoids will define nodal points conferring resistance. Aim 2 improves bulk TIL adoptive transfer immunotherapy by using PDOs as living bioreactors to enrich tumor-reactive mouse and human melanoma TILs by anti-PD-1 checkpoint inhibition, followed by testing of enhanced anti-tumor activity in vitro and in vivo. Lastly, Aim 3 performs a co-treatment trial comparing anti-PD-1 responses of pre-treatment biopsy cSCC PDOs to clinical outcomes. Further, post- treatment biopsy PDOs are re-challenged with anti-PD-1 and a novel agent inactivating PD-1 by dephosphorylation. We thus utilize the holistic PDO model preserving endogenous tumor epithelial and immune components en bloc to investigate and improve cancer immunotherapy via our team of Calvin Kuo (organoids), Mark Davis and Chris Garcia (tumor immunology) and Anne Chang and Dimitri Colevas (cSCC clinicians).

项目概要免疫系统显着区分自身和非自身/自身异常抗原，从而提供精致的抗肿瘤特异性和抑制肿瘤发生的作用。不幸的是，肿瘤细胞逃避免疫反应（ICB）。针对 PD-1、PD-L1、CD40 等以及过继细胞转移（CAR-T、批量 TIL）进行有利调节然而，缓解率通常是不完全的，疾病进展是有可能的。常见的生物标志物和预测性生物标志物均不理想。缺乏体外模型阻碍了下一代免疫疗法的发展在功能上概括了肿瘤和浸润免疫细胞之间的同基因相互作用。我们开发了类器官方法，可培养原发性人类肿瘤活检及其浸润这些“患者来源的肿瘤类器官”（PDO）作为一个凝聚单元的免疫成分。将肿瘤细胞与内源性 T、B、NK 细胞和巨噬细胞一起保存，有力地再现 T 细胞原发肿瘤的受体克隆型库，最重要的是，表现出肿瘤浸润淋巴细胞（TIL）对抗 PD-1/PD-L1 治疗性抗体的反应导致肿瘤细胞的扩增、激活和杀伤（Cell，2018）。因此，PDO 系统代表了人类肿瘤-免疫相互作用的整体类器官模型。在这里，我们利用 PDO 技术来研究免疫治疗机制并 PD-1 反应性皮肤鳞状细胞癌 (cSCC) 和黑色素瘤的治疗，利用治疗前和治疗后的人体活检和小鼠模型。诱导涉及免疫-肿瘤和免疫-免疫串扰的复杂且连续的网络反应。因此，目标 1 利用对 PDO 进行连续时间过程采样的能力来定义单细胞 RNA-seq 早期抗PD-1刺激的抗肿瘤免疫反应的网络细胞串扰模型重要的是，几个月后进行的临床活检通常无法获得急性时间点。这种在有反应与无反应的小鼠和人类类器官中的免疫传播将定义节点目标 2 通过使用 PDO 作为活体来改善批量 TIL 过继转移免疫疗法。通过抗 PD-1 检查点抑制来丰富肿瘤反应性小鼠和人类黑色素瘤 TIL 的生物反应器，接下来是体外和体内增强的抗肿瘤活性测试，最后，Aim 3 进行联合治疗。比较治疗前活检 cSCC PDO 的抗 PD-1 反应与临床结果的试验。治疗活检 PDO 再次受到抗 PD-1 和一种新型灭活 PD-1 药物的攻击因此，我们利用整体 PDO 模型来保护内源性肿瘤上皮和免疫。通过我们的 Calvin Kuo（类器官）团队研究和改进癌症免疫疗法， Mark Davis 和 Chris Garcia（肿瘤免疫学）以及 Anne Chang 和 Dimitri Colevas（cSCC 赞助人）。