Synthetic manipulation of engineered perivascular niches

摘要 本申请是为了响应被标识为NOT-CA的特别利益通知（NOSI）而提交的- 23-045.胶质母细胞瘤（GBM）是最常见和致命的脑癌形式。标准治疗是外科手术 切除后用烷化剂替莫唑胺（TMZ）和放疗治疗。切除 TMZ可以去除肿瘤块，并且TMZ为许多患者提供了一些益处。母体癌组织 工程合作项目（R 01 CA 256481）正在开发一种组织工程方法，以加速 新的抗癌化合物，克服TMZ耐药性的评价。我们正在发育组织 从肿瘤延伸到周围的血管周围龛（PVN）的工程模型 这些细胞被认为在侵袭、复发、TMZ抗性和不良反应中起主导作用。 生存我们的努力集中在开发一种工程PVN生物材料，研究病理生理学， 驱动GBM侵袭和TMZ抗性的过程，并加速新型TMZ衍生物的评价 针对GBM而不考虑MGMT状态。本NOT-CA-23-045 NOSI管理的目的 补充是为了支持一个新的合作倡议，将合成基因电路纳入我们的 工程PVN模型。目前的组织工程脑血管模型缺乏正交的、可调控的控制 在血管周围生态位的生长和成熟上。制定独立的定量控制的能力 超过PVN的生长和成熟将代表一个重大的进步，并将使我们能够深入研究 检查PVN内可能产生新的治疗靶点以改善结局的相互作用。 为了实现这一目标，我们建议与Ahmad Khalil博士（波士顿大学）开展新的合作， 应用他实验室的基因组正交合成锌指转录调节因子（synZiFTR）技术 在血管周围生态位模型中，能够对PVN生长与成熟进行药物调节的正交控制 在这个项目的开发中。为了做到这一点，我们将调节合成工程的生长和成熟， 血管周围龛（Aim S1）。我们随后将对TMZ耐药和侵袭的模式进行基准测试， 对合成血管系统的反应（目标S2）。这一拟议补充将支持一个协作小组， 开发新的互补能力，将先进的组织工程和合成生物学相结合 工具集，以提供对脑血管周围生态位模型的可调节控制。通过合作，我们将建立 人类合成组织构建物作为研究GBM-PVN相互信号传导的重要新工具， 脑肿瘤微环境这种能力对于研究GBM细胞药物的模式至关重要。 抵抗、侵袭和血管重塑是改善患者预后所必需的。

ABSTRACT This application is being submitted in response to the Notice of Special Interest (NOSI) identified as NOT-CA- 23-045. 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) and radiotherapy. Resection removes the tumor bulk, and TMZ provides some benefit to many patients. The parent Cancer Tissue Engineering Collaborative project (R01 CA256481) is developing a tissue engineering approach to accelerate the evaluation of new anticancer compounds that overcome TMZ resistance. We are developing tissue 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. Our efforts focus on developing an engineered PVN biomaterial, investigating pathophysiological processes driving GBM invasion and TMZ resistance, and accelerating evaluation of novel TMZ derivatives that target GBM regardless of MGMT status. The objective of this NOT-CA-23-045 NOSI Administrative Supplement is to support a new collaborative initiative to incorporate synthetic gene circuits into our engineered PVN models. Current tissue engineering brain vascular models lack orthogonal, regulatable control over the growth and maturation of the perivascular niche. The ability to enact independent, quantitative control over PVN growth and maturation would represent a significant advance and would enable us to deeply examine reciprocal interactions within the PVN that may yield novel therapeutic targets to improve outcomes. To realize this objective, we propose a new collaborative effort to with Dr. Ahmad Khalil (Boston University) to apply his laboratory’s genome-orthogonal synthetic zinc finger transcriptional regulator (synZiFTR) technology to enable drug-regulated, orthogonal control over PVN growth vs. maturation in the perivascular niche models under development by this project. To do this, we will regulate growth and maturation of a synthetic engineered perivascular niche (Aim S1). We will subsequently benchmark patterns of TMZ resistance and invasion in response to synthetic vasculature (Aim S2). This proposed supplement will support a collaborative team to develop a new yet complementary capability to integrate advanced tissue engineering and synthetic biology toolsets to provide regulatable control over brain perivascular niche models. Collaboratively, we will establish human synthetic tissue constructs as an important new tool to investigate reciprocal GBM-PVN signaling within the brain tumor microenvironment. Such capabilities are essential for investigating patterns of GBM cell drug resistance, invasion, and vascular remodeling necessary for improving patient outcomes.