Elucidating the Role of Perivascular Niche in Glioblastoma Invasion and Therapeutic Resistance at Single Cell Resolution using Biomimetic Tumor Microenvironment Models

使用仿生肿瘤微环境模型以单细胞分辨率阐明血管周围微环境在胶质母细胞瘤侵袭和治疗耐药中的作用

• 批准号: 10279283
• 负责人: Mehdi Nikkhah
• 金额: $ 39.15万
• 依托单位: ARIZONA STATE UNIVERSITY-TEMPE CAMPUS
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-15 至 2026-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10279283
• 关键词: 3-Dimensional; Address; Affect; Architecture; Astrocytes; Behavior; Bioinformatics; Biological; Biological Assay; Biological Models; Biology; Biomedical Engineering; Biomimetics; Blood capillaries; CXCL12 gene; CXCR; CXCR4 gene; Cell Communication; Cell Culture Techniques; Cell Maintenance; Cells; Coculture Techniques; Complex; Data; Development; Disease; Disease Progression; Dose; Endothelial Cells; Excision; Exposure to; Foundations; Genetic; Genomics; Glioblastoma; Glioma; Goals; Homing; Human; Hypoxia; Immune; In Vitro; Individual; Interruption; Knowledge; Ligands; Mediating; Methods; Microfluidic Microchips; Microfluidics; Microglia; Modeling; Molecular; Oncology; Operative Surgical Procedures; Organoids; Pathway interactions; Patients; Pericytes; Pharmacology; Physiological; Positioning Attribute; Prediction of Response to Therapy; Property; Radiation; Radiation therapy; Recurrence; Research Design; Resistance; Resolution; Role; Sampling; Shelter facility; Site; Slice; Specificity; Time; Tissue Engineering; Tissues; Tumor Tissue; Tumorigenicity; Validation; base; cell behavior; cell type; chemotherapeutic agent; conventional therapy; endothelial stem cell; genotoxicity; in vivo; in vivo Model; insight; interdisciplinary approach; irradiation; mouse model; multidisciplinary; neoplastic cell; novel; radiation resistance; receptor; response; self-renewal; single-cell RNA sequencing; small hairpin RNA; stem; stem cell biology; stem cell proliferation; stem cells; stemness; targeted treatment; therapy resistant; three dimensional cell culture; three-dimensional modeling; transcriptome sequencing; transcriptomics; treatment response; tumor; tumor microenvironment

Summary One of the critical challenges in the treatment of Glioblastoma (GBM) is the presence of highly resistant cells with stem-like properties, called glioma stem cells (GSCs), that evade surgical resection, resist conventional treatments and are primarily responsible for tumor recurrence. The perivascular niche within the GBM tumor microenvironment (TME) has been well recognized as a critical site that shelters GSCs and promotes their stemness, invasion, and therapeutic resistance. Extensive studies from others and our lab, using in vitro and in vivo models, have demonstrated that the crosstalk between the endothelial cells (ECs) and GSCs regulates GSC proliferation, tumorigenicity and self- renewal capacity. However, the perivascular niche is a complex microenvironment comprised not only of ECs but multiple other cell types including astrocytes, pericytes, and immune cells. How the cell-cell interactions between the various cellular components of the perivascular niche modulate GSC behavior (proliferation vs. quiescence and invasion vs. homing) and therapy resistance is poorly understood. To address these unmet biological knowledge gaps, there is a critical need for sophisticated and more realistic ex vivo tumor models that better recapitulate the physiological complexities of the GBM perivascular niche to advance our fundamental understanding of the biology of the disease and predict therapeutic responses. Recently, we have established and validated an on-chip microfluidic tumor model of GBM, with a unique 3D organotypic architecture, to study the influence of the perivascular niche on GSC invasion. We have shown that co-culturing of astrocytes enhances EC-induced invasion of GSCs, where RNA-seq analysis of mono-culture vs. tri-culture provided a mechanistic insight into the receptor-ligand pairs that mediate the interactions between cells. Based on these foundational developments, in this study our goal is to develop an ex vivo tumor model of GBM, bioinspired from the native perivascular niche, with patient-derived cells to dissect the role of cellular components within the niche on GSC biology and response to treatment at single cell resolution. In Aim 1, our objective is to determine the influence of the key cell types within the perivascular niche on GSC-EC interactions. In Aim 2, we plan to mechanistically unveil the impact of radiation treatment on GSCs- perivascular niche interactions, while in Aim 3, we will blunt invasion and sensitize GSCs through disruption of niche-tumor cell interactions. Our study design uniquely employs an interdisciplinary approach including microengineering of a bioinspired ex vivo tumor model, single-cell level resolution analysis, molecular-level transcriptomics, and validation using ex vivo patient tumor samples. Successful completion of these studies will not only further our understanding of the interactions of GSCs with the perivascular niche but will also facilitate identification of novel targets to block disease progression.

概括 胶质母细胞瘤 (GBM) 治疗的关键挑战之一是高度耐药细胞的存在 具有类似干细胞的特性，称为神经胶质瘤干细胞（GSC），可以逃避手术切除，抵抗传统疗法 治疗和肿瘤复发的主要原因。 GBM 肿瘤内的血管周围生态位 微环境（TME）已被公认为是庇护 GSC 并促进其发展的关键场所 干性、侵袭性和治疗耐药性。 其他人和我们的实验室使用体外和体内模型进行的广泛研究表明， 内皮细胞 (EC) 和 GSC 之间的串扰调节 GSC 增殖、致瘤性和自我修复 更新能力。然而，血管周围生态位是一个复杂的微环境，不仅由 EC 组成 但多种其他细胞类型，包括星形胶质细胞、周细胞和免疫细胞。细胞与细胞如何相互作用 血管周围生态位的各种细胞成分之间调节 GSC 行为（增殖与增殖） 静止和入侵与归巢）和治疗耐药性知之甚少。为了解决这些未满足的问题 由于生物学知识差距，迫切需要复杂且更现实的离体肿瘤模型 更好地概括 GBM 血管周围生态位的生理复杂性，以推进我们的基础研究 了解疾病的生物学并预测治疗反应。 最近，我们建立并验证了 GBM 的片上微流控肿瘤模型，具有独特的 3D 器官型结构，研究血管周围生态位对 GSC 侵袭的影响。我们已经证明 星形胶质细胞的共培养增强了 EC 诱导的 GSC 侵袭，其中单一培养与单独培养的 RNA-seq 分析 三元培养提供了对介导受体-配体之间相互作用的受体-配体对的机制洞察。 细胞。基于这些基础进展，在本研究中，我们的目标是开发一种离体肿瘤模型 GBM，受天然血管周围生态位的生物启发，使用患者来源的细胞来剖析细胞的作用 GSC 生物学领域内的组成部分以及单细胞分辨率下的治疗反应。 在目标 1 中，我们的目标是确定血管周围生态位内的关键细胞类型对 GSC-EC 相互作用。在目标 2 中，我们计划从机制上揭示放射治疗对 GSC 的影响—— 血管周围生态位相互作用，而在目标 3 中，我们将通过破坏 生态位-肿瘤细胞相互作用。我们的研究设计独特地采用了跨学科方法，包括 仿生离体肿瘤模型的微工程、单细胞水平分辨率分析、分子水平 转录组学，并使用离体患者肿瘤样本进行验证。成功完成这些研究将 不仅进一步加深了我们对 GSC 与血管周围生态位相互作用的理解，而且还将促进 确定阻止疾病进展的新靶点。