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

摘要 本申请是根据PA-21-071提交的。胶质母细胞瘤（GBM）是最常见的， 致命的脑癌标准治疗是手术切除，然后用烷化剂治疗。 替莫唑胺（TMZ）。切除肿瘤块，TMZ提供了一些好处，许多 患者母公司癌症组织工程合作项目（R 01 CA 256481）正在开发组织 加速评价克服TMZ的新抗癌化合物的工程方法 阻力该项目正在开发过程，以创建血管周围壁龛的工程模型 （PVN）从肿瘤延伸到周围的实质，并被认为是发挥主导作用， 在侵袭、复发、TMZ耐药和生存率差中的作用。传统的散装水凝胶，甚至 小型化的变体，不提供一个途径来定制或跟踪局部微环境的演变， 围绕着独特的细胞亚群。本NCI多样性管理补充文件的目的是 支持一项新的倡议，创造颗粒水凝胶组装，可以模拟多细胞肿瘤， 微环境中，但适合于高通量筛选方法，传统上用于检查 使用二维培养的药物反应。我们已经建立了技术基础， 水凝胶以研究GBM治疗反应。颗粒水凝胶是宏观结构， 微型水凝胶颗粒的堵塞组件。迄今为止，它们主要用作脱细胞的 水凝胶颗粒与在颗粒之间的空隙中培养的细胞。作为最近一次行政 作为补充，我们开发了将GBM细胞封装在不同纳升体积水凝胶中的能力 可以快速形成的微滴，具有为离散细胞群定制的基质组成， 并且无毒降解。现在，我们寻求扩大颗粒水凝胶系统的努力， 胶质母细胞瘤细胞的高通量应答数据。为了做到这一点，该项目将首先衡量治疗 GBM细胞对颗粒水凝胶中的脑模拟HA和血管周围分泌蛋白的反应（Aim S1）。 我们随后将研究多细胞聚集体在GBM细胞侵袭和治疗中的作用。 使用宏观尺度和颗粒水凝胶模型来评估疗效（Aim S2）。本补充建议将支持 一个来自生物医学研究领域历史上代表性不足的群体的研究生， 胶质母细胞瘤肿瘤微环境的模型。这种颗粒状水凝胶方法提供了基础， 询问胶质母细胞瘤聚集体大小和相对间距对胶质母细胞瘤干细胞活性的作用， GBM侵袭和对一线治疗的耐药性。我们将展示颗粒状水凝胶可以整合到 高通量筛选方法，以加速评估新的TMZ衍生物， 弥漫性GBM细胞，无论MGMT状态如何。

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.