Ex vivo analysis of human brain tumor cells in a microvascular niche model

微血管生态位模型中人脑肿瘤细胞的离体分析

• 批准号: 10339325
• 负责人: Rong Fan
• 金额: $ 52.67万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-02-04 至 2025-01-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10339325
• 关键词: 3-Dimensional; Animal Model; Animals; Biocompatible Materials; Biological; Biological Assay; Biology; Biomedical Engineering; Bone Marrow; Brain; Brain Neoplasms; Cancer Biology; Cancer Center; Cell Culture Techniques; Cells; Cerebrovascular system; Clinic; Clinical Data; Coculture Techniques; Data; Development; Disease; Disease Progression; Distant; Drug Delivery Systems; Environment; Gel; Genetic Transcription; Glioblastoma; Glioma; Heterogeneity; Human; In Vitro; Individual; Infiltration; Laboratories; Libraries; Link; Maintenance; Malignant Neoplasms; Malignant neoplasm of brain; Measures; Microfluidics; Modeling; Molecular; Molecular Biology; Motivation; Oncology; Pathologic; Pathology; Patients; Pericytes; Perivascular Neoplasm; Pharmaceutical Preparations; Phenotype; Pilot Projects; Primary Brain Neoplasms; Primary Neoplasm; Prognosis; Proliferating; Property; Proteomics; Relapse; Reproducibility; Resistance development; Resolution; Sampling; Specimen; Structure; System; Technology; Testing; Tissue Engineering; Training; Tumor Cell Migration; Tumor Stem Cells; Tumor Subtype; Universities; angiogenesis; cell behavior; cell type; chemotherapy; clinical application; cohort; cost; drug testing; experience; genetic signature; improved; in vitro Model; in vivo Model; interest; leukemic stem cell; medical schools; microsystems; mid-career faculty; migration; mind control; mouse model; multidisciplinary; neoplastic cell; neuropathology; neurosurgery; novel; patient derived xenograft model; patient prognosis; precision drugs; precision medicine; predict clinical outcome; real time monitoring; response; self-renewal; single cell analysis; single-cell RNA sequencing; small molecule; stem cells; stem-like cell; success; synergism; targeted treatment; therapy resistant; transcriptome; transcriptome sequencing; transcriptomics; treatment stratification; tumor; tumor heterogeneity

PROJECT SUMMARY The region near the brain vasculature in human brain tumors, called the perivascular niche (PVN), is an important microenvironment for the maintenance of brain tumor stem-like cells (BTSCs), the development of resistance to chemo or targeted therapies, and the path for tumor infiltration to distant regions in the whole brain, leading to incurable diseases. Current in vitro models such as 2D cell cultures or 3D tumor spheroids do not contain this niche environment. Mouse models of brain tumors can recapitulate some aspects of the PVN, but have challenges in terms of costly assays, low throughput, and lack of the ability for high-resolution live cell tracking of BTSC dynamics. Herein, we propose to develop a tissue-engineered 3D microvascular niche-on-a- chip model that can incorporate primary brain tumor cells from patients in order to bridge this gap between in vitro and in vivo models. Our pilot study has demonstrated the success in co-culture of patient-derived glioblastoma cells and microvasculature in a microfluidic gel system and observed preferential localization of BTSCs in the PVN. Comparing ex vivo dynamics of individual tumor cells on-chip to single-cell transcriptomes across 10 patients further revealed a correlation between perivascular localization and transcriptional subtypes. In this project, we propose to further examine tumor cell migration and localization using a larger cohort of patient specimens and compare the results to pathological and clinical data, aiming to develop it into an ex vivo functional assay for patient prognosis and subclassification (Aim 1). We will apply scRNA-seq to the same samples to generate correlative data to identify subtypes associated with distinct ex vivo dynamics in the tissue-engineered PVN model, which can help elucidate the molecular mechanisms of PVN in tumor cell fate and invasion (Aim 2). Finally, we will investigate the response of tumor cells in PVN to chemo and targeted therapies administered through the perfusable microvascular network to assess the potential to perform personalized drug test and therapeutic stratification (Aim 3). This project will lead to a novel tissue-engineered microsystem to not only study the biology of PVN in human brain tumor development but also develop new assays for ex vivo test of human tumor cells for precision medicine.

项目概要 人脑肿瘤中靠近脑血管系统的区域称为血管周围生态位（PVN），是一个 维持脑肿瘤干样细胞（BTSC）的重要微环境， 对化疗或靶向治疗的耐药性，以及肿瘤整体向远处区域浸润的路径 脑，导致不治之症。当前的体外模型（例如 2D 细胞培养物或 3D 肿瘤球体）确实可以 不包含这个利基环境。小鼠脑肿瘤模型可以概括 PVN 的某些方面， 但面临检测成本高、通量低以及缺乏高分辨率活细胞能力等挑战 跟踪 BTSC 动态。在此，我们建议开发一种组织工程 3D 微血管利基- 芯片模型可以整合来自患者的原发性脑肿瘤细胞，以弥补之间的差距 体外和体内模型。我们的试点研究证明了患者来源的共培养的成功 微流体凝胶系统中的胶质母细胞瘤细胞和微血管，并观察到 PVN 中的 BTSC。比较芯片上单个肿瘤细胞与单细胞转录组的离体动力学 对 10 名患者的研究进一步揭示了血管周围定位与转录亚型之间的相关性。 在这个项目中，我们建议使用更大的队列进一步检查肿瘤细胞迁移和定位 患者标本并将结果与​​病理和临床数据进行比较，旨在将其开发为离体 用于患者预后和亚分类的功能测定（目标 1）。我们将 scRNA-seq 应用于相同的 样本生成相关数据，以识别与不同的离体动力学相关的亚型 组织工程PVN模型，有助于阐明PVN在肿瘤细胞命运中的分子机制 和入侵（目标 2）。最后，我们将研究 PVN 中肿瘤细胞对化疗和靶向治疗的反应。 通过可灌注微血管网络进行治疗，以评估执行的潜力 个性化药物测试和治疗分层（目标 3）。该项目将产生一种新型的组织工程 微系统不仅可以研究 PVN 在人脑肿瘤发展中的生物学，还可以开发新的 用于精密医学的人体肿瘤细胞离体测试的测定。