Perivascular tissue models to overcome MGMT-mediated temozolomide resistance in glioblastoma

摘要 本申请是根据PA-21-071的要求提交的。胶质母细胞瘤(GBM)是最常见和 致命的脑癌。护理的标准是手术切除，然后用烷化处理 替莫唑胺(TMZ)。切除切除肿瘤块，TMZ为许多人提供了一些好处 病人。亲代肿瘤组织工程合作项目(R01 CA256481)正在开发组织 加速评价克服TMZ的新型抗癌化合物的工程方法 抵抗。该项目正在开发过程，以创建血管周围壁龛的工程模型 (PVN)从肿瘤延伸到周围实质并被认为起主导作用的 在侵袭、复发、抗TMZ和低存活率方面起作用。传统的块状水凝胶，甚至 小型化的变体不提供定制或跟踪当地微环境演变的途径 围绕着独特的细胞亚群。本NCI多元化行政副刊的目标是 支持一项新的倡议，创造能够模拟多细胞肿瘤的颗粒状水凝胶组件 微环境仍然适用于常规用于检查的高通量筛查方法 使用二维培养的药物反应。我们已经生成了创建颗粒状产品的技术基础 水凝胶研究GBM的治疗效果。颗粒状水凝胶是一种大尺度结构，由 堵塞的微尺度水凝胶颗粒集合。到目前为止，它们主要用于脱细胞。 水凝胶颗粒与细胞在颗粒之间的空隙中培养。作为最近一次行政管理的一部分 作为补充，我们开发了将GBM细胞包裹在不同纳升体积的水凝胶中的能力 可以快速形成的微滴具有为离散细胞群体量身定做的基质成分， 而且是无毒降解的。现在，我们试图用颗粒状水凝胶系统扩大努力，以检查 胶质母细胞瘤细胞的高通量反应数据。要做到这一点，这个项目将首先测量治疗 颗粒水凝胶中的GBM细胞对模拟脑的HA和血管周围分泌体的反应(目标S1)。 我们随后将研究多细胞聚集在基底膜细胞侵袭和治疗中的作用。 使用宏观水凝胶模型和颗粒水凝胶模型的有效性(目标S2)。这项拟议的补编将支持 来自生物医学研究历史上代表性不足的群体的研究生，以发展层级结构 胶质母细胞瘤肿瘤微环境模型。这种颗粒状水凝胶方法为 询问胶质母细胞瘤聚集大小和相对间距对胶质母细胞瘤干细胞活性的影响， 基底膜的侵袭，以及对一线治疗的抵抗。我们将展示颗粒水凝胶可以集成到 高通量筛选方法加速靶向创建的新型TMZ衍生物的评估 弥漫性的GBM细胞，与MGMT状态无关。

ABSTRACT This application is being submitted in response to PA-21-071. Glioblastoma (GBM) is the most common and lethal form of brain cancer. Standard of care is surgical resection followed by treatment with the alkylating agent temozolomide (TMZ). Resection removes the tumor bulk, and TMZ provides some benefit to many patients. The parent Cancer Tissue Engineering Collaborative project (R01 CA256481) is developing tissue engineering approach to accelerate the evaluation of new anticancer compounds that overcome TMZ resistance. This project is developing processes to create engineered models of the perivascular niches (PVNs) that extend from the tumor into the surrounding parenchyma and which are believed to play a dominant role in invasion, recurrence, TMZ resistance, and poor survival. Conventional bulk hydrogels, even miniaturized variants, do not provide an avenue to tailor, or trace the evolution of, the local microenvironment surrounding unique cell subpopulations. The objective of this NCI Diversity Administrative supplement is to support a novel initiative to create granular hydrogel assemblies that can mimic the multicellular tumor microenvironment yet are amenable to high-throughput screening approaches conventionally used to examine drug responses using two-dimensional culture. We have generated the technical foundation to create granular hydrogel to study GBM therapeutic response. Granular hydrogels are macroscale structures generated as jammed assemblies of microscale hydrogel particles. To date they have been predominantly used as acellular hydrogel particles with cells cultured in the voids between particles. As part of a recent administrative supplement, we developed capacity to encapsulate GBM cells in distinct nanoliter-volume hydrogel microdroplets that can be rapidly formed, have their matrix composition tailored for discrete cell populations, and be non-toxically degraded. Now, we seek to expand efforts with granular hydrogel systems to examine high-throughput response data for glioblastoma cells. To do this, this project will first measure therapeutic responses of GBM cells to brain-mimetic HA and the perivascular secretome in granular hydrogels (Aim S1). We will subsequently examine the role of multicellular aggregations on GBM cell invasion and therapeutic efficacy using both macroscale and granular hydrogel models (Aim S2). This proposed supplement will support a graduate student from a historically underrepresented group in biomedical research to develop hierarchical models of the glioblastoma tumor microenvironment. This granular hydrogel approach provides the basis to interrogate the role of glioblastoma aggregation size and relative spacing on glioblastoma stem cell activity, GBM invasion, and resistance to frontline therapies. We will show granular hydrogels can be integrated into high-throughput screening approaches to accelerate the evaluation of novel TMZ derivatives created to target diffuse GBM cells regardless of MGMT status.