Tumor Organoid-Mediated Drug Testing and Clonality Analysis in Peritoneal Surface Disease of Intestinal Origin

肠源性腹膜表面疾病中肿瘤类器官介导的药物检测和克隆分析

基本信息

批准号：
10578822
负责人：
Lance David Miller
金额：
$ 49.43万
依托单位：
WAKE FOREST UNIVERSITY HEALTH SCIENCES
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：
2021-03-01 至 2026-02-28
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10578822
关键词：
3-Dimensional Abdomen Adjuvant Chemotherapy Anatomy Antigens Architecture Biomedical Engineering Biopsy CT26 Cancer Patient Candidate Disease Gene Cell Death Cell Survival Cell physiology Cells Characteristics Chemoresistance Chemosensitivity Assay Clinical Clinical Management Clinical Treatment Clinical Trials Design Clonality Colon Carcinoma Colorectal Cytotoxic T-Lymphocytes DNA Sequence Alteration Devices Disease Resistance Drug Combinations Drug Interactions Drug Screening Drug resistance Effector Cell Evaluation Exhibits FDA approved Foundations Gene Mutation Genes Genomics Genotype Goals Heterogeneity Homeostasis Human Hyperthermia Immune Immune checkpoint inhibitor Immunocompetent Immunological Models Immunooncology In Situ Intestinal Diseases Intestines Lesion Lung Neoplasms MC38 Malignant - descriptor Malignant Neoplasms Malignant neoplasm of appendix Malignant neoplasm of gastrointestinal tract Measures Mediating Mesothelioma Methods Microfluidic Microchips Microsatellite Instability Modality Modeling Mus Mutate Mutation Neoplasm Metastasis Non-Malignant Operative Surgical Procedures Organoids Pathway interactions Patients Performance Peritoneal Pharmaceutical Preparations Phenotype Postoperative Period Prediction of Response to Therapy Relapse Reporting Research Resistance Resolution Role Sampling Selection for Treatments Site Surface System Testing Tumor Debulking Validation Variant Work Xenograft Model cancer cell cancer type carcinogenesis cell killing chemotherapeutic agent chemotherapy clinical efficacy clinical predictive model clinical predictors clinically actionable colon cancer cell line design driver mutation drug efficacy drug response prediction drug sensitivity drug testing gene function guided inquiry immunoregulation insight intraperitoneal therapy melanoma metastatic colorectal mouse model neoplastic cell novel parity patient response preservation response sarcoma targeted exome sequencing treatment response tumor

项目摘要

Project Summary The overarching goal of this project is to develop a tumor organoid-based drug screening platform that quantifies the differential drug responsiveness of tumor clonal heterogeneity to predict a patient’s therapeutic response prior to treatment selection. Our research is directed at metastatic intestinal peritoneal surface malignancies (IPSM), with a focus on colon and appendiceal cancers. These cancers are similarly treated and present clinically with numerous spatially-distinct peritoneal metastases associated with different anatomic structures. The clinical course of these cancers typically follows a path of chemoresistance, thus, ex vivo models that can identify drug combinations and sequencing strategies able to mitigate chemoresistance would be of significant clinical value. Innate and acquired chemoresistance results from genetic alterations that manifest within the tumor’s clonal architecture and its changing clonal composition. In IPSM, we have observed distinct clonal variants among spatially distinct lesions within the same patient indicating the potential for differential drug responses among intrapatient lesions. Our central hypothesis is that a patient’s clinical response to chemotherapy is proportional to the magnitude of the clonal response exhibited by the patient’s predominating cancer clones, which can be assessed by chemosensitivity testing and clonality analysis using patient-derived tumor organoids (PTOs) constructed from intra-patient lesions that reflect the patient’s metastatic clonal diversity. In Aim 1, we will construct multi-lesion PTOs from a number of IPSM patients to investigate interactions between tumor clonal fraction and PTO chemosensitivity. We will test the hypothesis that summative measures of chemo resistant and sensitive clonal fractions will predict patient clinical treatment responses better than simple measures of organoid cell viability. Furthermore, we will seek to identify genes in IPSM that are frequently mutated in chemo sensitive and/or resistant clonal fractions across IPSM patients, and characterize candidate gene functions in colon cancer cell line organoids and mouse xenograft models of colon cancer. In Aim 2, we will seek to extend the reach of the PTO platform to include evaluation of immune checkpoint inhibitors (ICIs) which cannot yet be faithfully modeled in human systems, and that have emerging roles in gastro-intestinal cancers, yet remain unexplored in IPSM management. We will test the hypothesis that immune-enhanced PTOs (iPTOs) that preserve the patient- specific homeostatic relationship between cancer cells and cytotoxic T cells (CTLs) can be used to model ICI- induced cancer cell killing by tumor-reactive CTLs as well as to gain insight into tumor clonal alterations that influence ICI responsiveness. We will use established murine immune-oncology models (CT26 and MC38) to study tumor-CTL homeostasis in iPTOs, and to confirm ICI-response parity between ex vivo iPTOs and in situ mouse tumors. We will confirm the involvement of antigen specific CTL-mediated cancer cell killing in ICI-treated iPTOs. Finally, parallel to Aim 1, we will identify genes that are frequently mutated in ICI sensitive and/or resistant clonal fractions of IPSM, and characterize their ICI-modulatory functions in immunocompetent mouse models.

项目摘要该项目的总体目标是开发一个基于肿瘤的基于器官的药物筛查平台，以量化肿瘤克隆异质性的差异药物反应性预测患者的治疗反应在选择之前。我们的研究针对转移性肠腹膜表面恶性肿瘤（IPSM），重点是结肠和阑尾癌。这些癌症也类似地治疗并在临床上存在与不同的解剖结构相关的许多空间呈腹膜转移。临床这些癌症的过程通常遵循化学耐药性的途径，因此可以识别药物的模型可以采用结合和测序策略来减轻化学耐药性将具有显着的临床价值。先天和获得的化学耐药性来自遗传变化，在肿瘤的克隆中表现出来建筑及其不断变化的克隆组成。在IPSM中，我们观察到了不同的克隆变体同一患者内的空间不同病变，表明在肠内病变。我们的中心假设是患者对化学疗法的临床反应是比例的对于患者占主导的癌症克隆表现出的克隆反应的大小，这可以是通过使用患者衍生的肿瘤器官（PTO）评估化学敏度测试和克隆性分析由反映患者转移性克隆多样性的患者内病变建造。在AIM 1中，我们将从许多IPSM患者中构建多层PTO，以研究肿瘤克隆之间的相互作用分数和PTO化学敏感性。我们将检验以下假设，即化学抗性和敏感的克隆分数将比类带器官的简单测量更好地预测患者临床治疗反应细胞活力。此外，我们将寻求鉴定IPSM中经常在化学敏感中突变的基因 IPSM患者的克隆分数和/或抗性克隆分数，并表征结肠癌的候选基因功能结肠癌的细胞系器官和小鼠异种移植模型。在AIM 2中，我们将寻求扩展 PTO平台包括评估免疫检查点抑制剂（ICI），该抑制剂尚不忠实以人类系统建立模型，并且在胃肠道癌中具有新兴角色，但在 IPSM管理。我们将检验以下假设：保留患者的免疫增强PTO（IPTO）癌细胞与细胞毒性T细胞（CTL）之间的特定稳态关系可用于模拟ICI- 肿瘤反应性CTL诱导癌细胞杀死，并深入了解肿瘤克隆改变影响ICI的响应能力。我们将使用已建立的鼠免疫肿瘤模型（CT26和MC38）研究IPTOS中的肿瘤-CTL稳态，并确认离体IPTO和原位之间的ICI反应均衡小鼠肿瘤。我们将确认抗原特异性CTL介导的癌细胞杀死ICI治疗 iptos。最后，平行于目标1，我们将鉴定出在ICI敏感和/或抗性中经常突变的基因 IPSM的克隆分数，并在免疫功能的小鼠模型中表征其ICI调节功能。