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

• 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
• 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

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类器官的药物筛选平台， 肿瘤克隆异质性的不同药物反应性预测患者的治疗反应 在治疗选择之前。我们的研究是针对转移性肠腹膜表面恶性肿瘤 （IPSM），重点是结肠癌和阑尾癌。这些癌症的治疗和临床表现相似 具有与不同解剖结构相关的许多空间上不同的腹膜转移。临床 这些癌症的过程通常遵循化学抗性的路径，因此，可以鉴定药物的离体模型 能够减轻化学抗性的组合和测序策略将具有显著的临床价值。 先天性和获得性化疗耐药性是由肿瘤克隆内的遗传改变引起的。 结构和不断变化的克隆组成。在IPSM中，我们观察到不同的克隆变体， 同一患者内的空间上不同的病变，表明不同患者之间可能存在不同的药物反应。 患者体内病变。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表现出的克隆应答的大小，这可以是 使用患者源性肿瘤类器官（PTO）通过化学敏感性测试和克隆性分析进行评估 由反映患者转移性克隆多样性的患者内病变构建。在目标1中，我们 从许多IPSM患者中构建多病灶PTO，以研究肿瘤克隆之间的相互作用， 分数和PTO化学敏感性。我们将检验这一假设，即化疗耐药和 敏感的克隆分数将比简单的类器官指标更好地预测患者的临床治疗反应 细胞活力此外，我们将寻求鉴定IPSM中在化疗敏感性中经常突变的基因， 和/或耐药克隆部分，并表征结肠癌中的候选基因功能 细胞系类器官和结肠癌的小鼠异种移植模型。在目标2中，我们将努力扩大 PTO平台包括对免疫检查点抑制剂（ICI）的评估， 在人类系统中建模，并且在胃肠道癌症中具有新兴作用，但在 IPSM管理。我们将检验这样一个假设，即免疫增强型PTO（iPTO）可以保护患者- 癌细胞和细胞毒性T细胞（CTL）之间的特异性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，以及深入了解肿瘤克隆改变， 影响ICI反应性。我们将使用已建立的鼠免疫肿瘤学模型（CT 26和MC 38）， 研究iPTO中肿瘤-CTL体内平衡，并确认离体iPTO和原位iPTO之间的ICI反应等同性 小鼠肿瘤。我们将证实在ICI治疗的肿瘤细胞中，抗原特异性CTL介导的癌细胞杀伤参与其中。 iPTO。最后，与目标1平行，我们将鉴定在ICI敏感和/或抗性中频繁突变的基因。 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。