Biomimetic Materials to Elucidate the Role of Microenvironment in Glioblastoma Stem Cell Maintenance In Vitro

该项目致力于开发和表征基于透明质酸的水凝胶3D基质，作为研究胶质母细胞瘤干细胞（GSCs或脑肿瘤起始细胞）的体外试验平台，胶质母细胞瘤干细胞在多形性胶质母细胞瘤（GBM）的肿瘤形成和肿瘤复发中起着重要作用，GBM是人类最致命的癌症之一。这些基质将用于研究微环境影响的作用，特别是机械和化学信号对GSC表型的作用，并检查其在测量治疗反应方面的效用。该系统有可能提供一个可控和可调的环境，密切模仿体内的大脑。该系统可以为GBM患者（超过22,000/年）提供更好的治疗选择。该系统可广泛用于理解微环境信号在多种类型癌症中的作用，特别是那些转移到大脑的癌症，以及神经发育的基础研究。通过为高中学生设立“工程日”，为代表性不足的女性和非裔美国学生提供研究机会，将研究相关原则纳入现有课程，并为高中学生和教师提供暑期生物工程讲习班，实现了教育影响。该项目将结合生物材料工程策略和当前对胶质母细胞瘤干细胞（GSC，也称为脑肿瘤起始细胞）生物学的理解，设计一种模拟天然GSC微环境的新型3D培养系统，通过研究GSC与基质的相互作用，提高体外治疗反应预测。GSCs细胞模型优于已建立的多形性胶质母细胞瘤（GBM）细胞系，后者通常用于药物开发管道。新近从GBM患者身上提取的GSCs被认为能更好地概括GBM生物学。虽然众所周知，当暴露在3D微环境中时，细胞行为会发生巨大变化，但目前GSCs主要在人工、2D和无基质环境（即硬组织培养聚苯乙烯）中培养，这些环境不能适当地捕获大脑微环境。此外，柔软的类组织生物材料提供了提供最佳线索组合的能力，以提供更多生理相关的环境，这可以诱导通常在体内观察到的细胞表型。为了提高对微环境对GSC表型影响的基本理解，必须使用模仿大脑微环境中发现的体内机械和化学线索的生物材料。在计划的研究中，研究人员将使用仿生透明质酸（HA）水凝胶来确定机械和化学信号在维持GSC表型中的作用。此外，他们将利用他们的培养系统来量化体外治疗反应。因此，该项目的主要目标是创造一种新的使能技术，以推进GSC生物学的研究，并帮助未来GBM药物的开发管道。GBM是最致命的人类癌症之一，平均存活时间约为一年。这项工作旨在提供一种新的治疗评估系统，可以更好地预测临床结果，从而可以改善治疗选择，每年影响超过22,000人的生活。该系统可广泛用于理解微环境信号在多种类型癌症中的作用，特别是那些转移到大脑的癌症，以及神经发育的基础研究。外展活动包括：1)在阿拉巴马州农村地区的高中发展“工程日”，这些地区的学生主要是非洲裔美国人；2)为传统上代表性不足的女性和非洲裔美国学生提供工程研究机会；3)将各种生物学、材料科学和工程原理整合到新的和现有的本科和研究生水平的化学工程课程中；4)每年为高中学生和教师举办暑期生物工程研讨会。

#1604677-KimThis project focuses on developing and characterizing hyaluronic acid-based hydrogel 3D matrices as an in vitro test bed for studying glioblastoma stem cells (GSCs or brain tumor initiating cells), which play an important role in tumor formation and tumor recurrence in glioblastoma multiforme (GBM), one of the deadliest forms of human cancer. The matrices will be used to investigate the role of microenvironmental influences, particularly the role of mechanical and chemical signals on GSC phenotype and examine its utility in measuring therapy response. The system has the potential to provide a controllable and tunable environment closely mimicking the in vivo brain. The proposed system could lead to improved therapeutic options for patients (over 22,000/year) suffering from GBM. The system developed could be broadly utilized in understanding the role of microenvironmental signals in multiple types of cancer, particularly those that metastasize to the brain as well as in fundamental studies of neural development. Educational impact is achieved through development of an "Engineering Day" for high school students, providing research opportunities for under-represented female and African-American Students, integration of research related principles into existing courses and providing summer bioengineering workshops for high school students and teachers.This project will combine biomaterials engineering strategies with current understanding of glioblastoma stem cell (GSC; also known as brain tumor initiating cell) biology to design a novel 3D culture system mimicking the native GSC microenvironment that would improve in vitro prediction of therapeutic response by allowing investigation of GSC-matrix interactions. GSCs cell models are superior to established glioblastoma multiforme (GBM) cell lines that are routinely employed in the drug development pipeline. GSCs freshly-derived from GBM patients are known to better recapitulate GBM biology. Though it is well-known that cell behavior is drastically altered when exposed to 3D microenvironments, GSCs are currently mainly cultured in artificial, 2D, and matrix-free environments (i.e., stiff tissue culture polystyrene) that do not appropriately capture the brain microenvironment. Furthermore, softer tissue-like biomaterials afford the ability to present the optimal combination of cues to provide more physiologically relevant environments, which can induce cellular phenotypes typically observed in vivo. For an improved fundamental understanding of microenvironmental influences on GSC phenotype, biomaterials mimicking the in vivo mechanical and chemical cues found in the brain microenvironment must be employed. In the planned studies, the investigators will use biomimetic hyaluronic acid (HA) hydrogels to establish the role of mechanical and chemical signals in maintenance of the GSC phenotype. Furthermore, they will utilize their culture system to quantify therapy response in vitro. Thus the main goal of the project is to create a novel enabling technology that will advance studies of GSC biology and aid future GBM drug development pipelines. GBM is among the deadliest forms of human cancer with an average survival of approximately one year. The proposed work is poised to provide a novel therapeutic evaluation system that can better predict clinical outcomes, thus could lead to improved therapeutic options affecting the lives of over 22,000 individuals per year. The system developed could be broadly utilized in understanding the role of microenvironmental signals in multiple types of cancer, particularly those that metastasize to the brain as well as in fundamental studies of neural development. Outreach activities include: 1) developing "Engineering Day" in high schools of rural Alabama's socioeconomically disadvantaged locality where the students are predominantly African-Americans, 2) providing engineering research opportunities to traditionally under-represented female and African-American students, 3) integrating various biology, materials science, and engineering principles into new and existing undergraduate and graduate level chemical engineering courses, and 4) providing annual summer bioengineering workshops for high school students and teachers.