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：
10374163
负责人：
CALVIN J KUO
金额：
$ 64.36万
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/10374163
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 Epithelial 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 MHC antigen 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 T-Lymphocyte T-Lymphocyte Subsets T-cell receptor repertoire TNFRSF5 gene Techniques Testing Therapeutic Therapeutic antibodies Time Transgenic Mice Tumor Antigens Tumor Cell Line Tumor-Derived Tumor-Infiltrating Lymphocytes Tumor-infiltrating immune cells air treatment anti-PD-1 anti-PD-1/PD-L1 anti-PD-L1 anti-PD1 therapy anti-tumor immune response base cancer immunotherapy cell killing checkpoint inhibition clinical decision-making clinical efficacy cohesion cohort cytotoxicity exhaust follow-up human model immune checkpoint blockade improved in vitro Model in vitro activity in vivo inhibitor therapy macrophage 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).

项目摘要免疫系统明显区分自我和非自我/自我灭绝的抗原，可提供精致的抗肿瘤特异性和抑制肿瘤发生。但是，肿瘤免疫监视是不幸的是，免疫反应的肿瘤细胞演变反对。免疫检查点封锁（ICB）靶向PD-1，PD-L1，CD40等这对治疗益处的同等效果。但是，回应率通常不完整，进行性疾病是常见和预测生物标志物是次优的。缺乏体外模型阻碍了下一代免疫疗法的发展这在功能上概括了肿瘤与浸润的免疫细胞之间的合成性相互作用。作为回应，我们开发了培养原发性人类肿瘤活检及其浸润的器官方法免疫组件作为无凝结单元而无需重新构成的单位。这些“患者衍生的肿瘤器官”（PDO）与内源性T，B，NK细胞和巨噬细胞一起保存肿瘤细胞，可稳固地概括T细胞原始肿瘤的受体克隆型曲目，并完全表现出肿瘤浸润淋巴细胞（TIL）响应抗PD-1/PD-L1治疗抗体的膨胀，激活和肿瘤细胞杀死（Cell，2018）。因此，PDO系统代表了人类肿瘤免疫相互作用的整体器官模型。在这里，我们利用PDO技术来研究免疫治疗机制和 PD-1响应性皮肤鳞状细胞癌（CSCC）和黑色素瘤的治疗治疗前后的人类活检和小鼠模型。目标1假设检查点抑制诱导涉及免疫肿瘤和免疫免疫串扰的复杂和顺序的网络响应。这是AIM 1具有执行PDO的串行时间顺序采样的能力来定义单个细胞RNA-Seq 早期抗PD-1刺激的抗肿瘤免疫激素的网络蜂窝串扰模型超过多个几个月后进行的临床活检通常无法访问急性时间点。重要的是，比较反应与非反应小鼠和人类器官的这种免疫磷脂将定义淋巴结积分会议抵抗。 AIM 2通过使用PDO作为生活通过抗PD-1检查点抑制来富含肿瘤反应性小鼠和人黑色素瘤的生物反应器，然后在体外和体内测试增强的抗肿瘤活性。最后，AIM 3执行共同处理试验比较了治疗活检CSCC PDOS与临床结果的抗PD-1反应。此外，用抗PD-1重新挑战治疗活检PDO，并通过新颖的药物使PD-1灭活PD-1 去磷酸化。因此，我们利用保留内源性肿瘤上皮和免疫的整体PDO模型组成部分通过我们的Calvin Kuo团队（Organoids），调查和改善癌症免疫疗法，马克·戴维斯（Mark Davis）和克里斯·加西亚（Chris Garcia）（肿瘤免疫学）以及安妮·张（Anne Chang）和迪米特里·科尔瓦斯（Dimitri Colevas）（CSCC临床医生）。