Biomimetic hydrogel niches to study the malignant phenotype of glioblastoma multiforme

 描述：多形性胶质母细胞瘤(GBM)是最常见、最具侵袭性和致命性的脑癌，GBM通过不同的侵袭过程迅速扩散，因此有必要研究发生在肿瘤边缘的现象。考虑到与GBM相关的独立基因组突变的数量，开发仿生组织工程方法来直接研究患者来源的生物干细胞而不是通用细胞系是至关重要的。该领域的一个主要瓶颈是目前尚不清楚跨越GBM肿瘤的生物物理和生物分子信号的组合如何影响恶性表型和治疗反应。能够复制天然肿瘤微环境的调控元素的空间图案化生物材料是必不可少的。我们开发了一种微流体形成技术，以创建光学半透明工程胶质瘤生物材料库，其中包含受GBM边缘启发的细胞、基质和生物分子线索的重叠图案。我们能够通过转录、分泌和成像指标对EG离散亚区的细胞进行多路分析，将细胞反应映射为局部微环境的函数。虽然利用永生化细胞系成功地解决了与临床相关的现象，但临床上迫切需要能够从患者来源的生物标本中收集类似信息的护理点工具。这项应用的主要目的是展示一种仿生组织工程方法，利用患者体内来源的生物旋毛虫研究潜在的表型机制。目标1将剖析肿瘤边缘激发信号的重叠模式如何塑造恶性表型。目的2将确定血管周围信号对侵袭性表型的贡献。AIM 3将使用工程化胶质瘤通过与临床表型的定量基准来解决原位和异位异种移植瘤之间的不一致性。基因工程的胶质瘤生物材料提供了洞察大脑皮层微环境的空间和时间方面的潜力，而目前的实验方法是不可能的。我们使用可扩展的微流控平台，以及通过传统和新一代分子分析工具对GBM细胞进行多路评估，大大减少了所需患者生物样品的大小，加快了分析速度，同时保留了询问稀有细胞亚群(如胶质瘤癌症干细胞)的能力。工程化的胶质瘤生物材料有可能成为

 DESCRIPTION: Glioblastoma multiforme (GBM) is the most common, aggressive, and deadly form of brain cancer GBM spreads rapidly and diffusely via distinct invasive processes, making it essential to investigate phenomena occurring at the tumor margins. And given the number of independent genomic mutations associated with GBM, it is critical to develop biomimetic tissue engineering approaches to directly study patient-derived biospecimens rather than generic cell lines. A major bottleneck in the field is that it remains unclear how combinations of biophysical and biomolecular signals that exist in close spatial and temporal order across the GBM tumor affect malignant phenotype and response to therapy. Spatially-patterned biomaterials capable of replicating regulatory elements of the native tumor microenvironment such as the margins are essential. We have developed a microfluidic forming technique to create libraries of optically-translucent engineered glioma biomaterials containing overlapping patterns of cell, matrix, and biomolecular cues inspired by the GBM margins. We are able to map cell response as a function of local microenvironment via multiplexed analyses of cells from discrete sub-regions of the EG via transcriptomic, secretomic, and imaging metrics. While successful for resolving clinically-relevant phenomena using immortalized cell lines, there is an acute clinical need for point-of-care tools able to gather similar information from patient-derived biospecimens. The primary objective of this application is to demonstrate a biomimetic tissue engineering approach to investigate mechanisms underlying phenotype using patient-derived biospecimens ex vivo. Aim 1 will dissect how overlapping patterns of tumor margin-inspired signals shape malignant phenotype. Aim 2 will define the contribution of perivascular signals on invasive phenotype. Aim 3 will employ engineered gliomas to resolve discordances between orthotopic and heterotopic xenograft tumors via quantitative benchmarking against clinical phenotype. Engineered glioma biomaterials offer the potential for insight regarding spatial and temporal aspects of the GBM microenvironment in ways not possible with current experimental approaches. Our use of a scalable microfluidic platform as well as multiplexed assessment of GBM cells via conventional and next generation molecular analysis tools greatly reduces the size of the required patient biospecimen, accelerates the speed of analysis, and yet preserves the capability of interrogating rare cell subpopulations such as glioma cancer stem cells. Engineered glioma biomaterials have the potential to b