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.

项目概要该项目的总体目标是开发一个基于肿瘤类器官的药物筛选平台，该平台可量化肿瘤克隆异质性的不同药物反应性可预测患者的治疗反应在选择治疗方法之前，我们的研究针对转移性肠腹膜表面恶性肿瘤。 (IPSM)，重点关注结肠癌和阑尾癌，这些癌症的治疗方法和临床表现类似。具有许多与不同解剖结构相关的空间上不同的腹膜转移瘤。这些癌症的病程通常遵循化学耐药性的路径，因此，可以识别药物的离体模型能够减轻化疗耐药性的组合和测序策略将具有重要的临床价值。先天性和获得性化疗耐药性是由肿瘤克隆内的基因改变引起的在 IPSM 中，我们观察到了不同的克隆变异。同一患者体内空间上不同的病变表明不同药物反应的可能性我们的中心假设是患者对化疗的临床反应是成比例的。患者的主要癌症克隆所表现出的克隆反应的程度，这可以是使用患者来源的肿瘤类器官 (PTO) 通过化学敏感性测试和克隆性分析进行评估在目标 1 中，我们将根据反映患者转移克隆多样性的患者内病变构建。从许多 IPSM 患者构建多病灶 PTO，以研究肿瘤克隆之间的相互作用我们将检验化疗耐药性和 PTO 的总结性测量的假设。敏感的克隆分数比简单的类器官测量更好地预测患者的临床治疗反应此外，我们将寻求鉴定 IPSM 中在化学敏感性中经常突变的基因。和/或 IPSM 患者的耐药克隆部分，并表征结肠癌中候选基因的功能在目标 2 中，我们将寻求扩大结肠癌的细胞系类器官和小鼠异种移植模型。 PTO 平台将包括免疫检查点抑制剂 (ICIs) 的评估，但目前尚不能如实评估以人体系统为模型，在胃肠道癌症中发挥着新的作用，但尚未得到探索我们将检验免疫增强 PTO（iPTO）可以保护患者的假设。癌细胞和细胞毒性 T 细胞 (CTL) 之间的特定稳态关系可用于模拟 ICI- 通过肿瘤反应性 CTL 诱导癌细胞杀伤，并深入了解肿瘤克隆改变我们将使用已建立的小鼠免疫肿瘤学模型（CT26 和 MC38）来影响 ICI 反应性。研究 iPTO 中的肿瘤-CTL 稳态，并确认离体 iPTO 和原位 iPTO 之间的 ICI 反应同等性我们将确认 ICI 治疗中抗原特异性 CTL 介导的癌细胞杀伤作用。最后，与目标 1 平行，我们将识别 ICI 敏感和/或耐药中经常突变的基因。 IPSM 的克隆部分，并表征其在免疫活性小鼠模型中的 ICI 调节功能。