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: 10599100
• 负责人: Rong Fan
• 金额: $ 51.72万
• 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/10599100
• 关键词: 3-Dimensional; Animal Model; Animals; Biocompatible Materials; Biological; Biological Assay; Biology; Biomedical Engineering; Blood Vessels; Bone Marrow; Brain; Brain Neoplasms; Cancer Biology; Cancer Center; Cell Culture Techniques; Cells; Cerebrovascular system; Classification; Clinic; Clinical Data; Coculture Techniques; Cultured Tumor Cells; Data; Development; Disease; Disease Progression; Distant; Drug Delivery Systems; Environment; Gel; Genetic Transcription; Glioblastoma; Glioma; Heterogeneity; Human; In Vitro; Individual; Infiltration; Invaded; Laboratories; Libraries; Link; Maintenance; Malignant Neoplasms; Malignant neoplasm of brain; Measures; Microfluidics; Modeling; Molecular; Molecular Biology; Motivation; Oncology; Pathologic; Pathology; Patients; Perfusion; 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; technology validation; therapy resistant; transcriptome; transcriptome sequencing; transcriptomic profiling; 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.

项目摘要 人脑肿瘤中脑脉管系统附近的区域称为周围的壁re（PVN），是一种 维持脑肿瘤样细胞（BTSC）的重要微环境，发展 对化学疗法或靶向疗法的抗性，以及肿瘤浸润到整个远处的路径 大脑，导致无法治愈的疾病。当前的体外模型，例如2D细胞培养或3D肿瘤球体DO 不包含这个利基环境。脑肿瘤的小鼠模型可以概括PVN的某些方面 但是在昂贵的测定，低通量和缺乏高分辨率活细胞的能力方面面临挑战 跟踪BTSC动力学。本文中，我们建议开发组织工程的3D微血管壁rike-a-a- 芯片模型可以结合患者的原发性脑肿瘤细胞，以弥合IN之间的差距 体外模型。我们的试点研究表明，在患者衍生的共同文化中取得了成功 微流体凝胶系统中的胶质母细胞瘤细胞和微脉管系统，并观察到优先定位 PVN中的BTSC。比较片上个体肿瘤细胞的离体动力学与单细胞转录组 在10名患者中，进一步显示血管周定位与转录亚型之间存在相关性。 在这个项目中，我们建议使用较大的队列进一步检查肿瘤细胞的迁移和定位 患者标本并将结果与​​病理和临床数据进行比较，旨在将其发展为离体 患者预后和亚分类的功能分析（AIM 1）。我们将把scrna-seq应用于同一 样品生成相关数据以识别与与不同的离体动力学相关的亚型 组织工程的PVN模型，可以帮助阐明肿瘤细胞命运中PVN的分子机制 和入侵（目标2）。最后，我们将研究PVN中肿瘤细胞对化学疗法的反应，并靶向 通过灌注微血管网络进行的疗法，以评估执行的潜力 个性化药物测试和治疗分层（AIM 3）。该项目将导致新型组织工程 微系统不仅要研究人类脑肿瘤发育中PVN的生物学，而且还发展了新的 对精确医学的人类肿瘤细胞的离体检测测定。