Synthetic manipulation of engineered perivascular niches

摘要 本申请是为了回应被确认为非CA-CA的特殊利益通知(NOSI)而提交的 23-045.胶质母细胞瘤(GBM)是最常见和致命的脑癌。标准的护理是外科手术 切除，然后用烷化剂替莫唑胺(TMZ)治疗和放射治疗。切除 去除肿瘤体积，TMZ为许多患者提供了一些好处。亲代癌组织 工程协作项目(R01 CA256481)正在开发一种组织工程方法，以加速 克服TMZ耐药性的新型抗癌化合物的评价。我们正在开发组织 从肿瘤延伸到周围的血管周围壁龛(PVN)的工程模型 实质组织被认为在侵袭、复发、TMZ抵抗和不良反应中起主导作用 生死存亡。我们致力于开发一种工程化的PVN生物材料，研究病理生理 驱动GBM侵袭和TMZ抗性的过程，并加速新TMZ衍生物的评估 无论MGMT状态如何，目标GBM。NOT-CA-23-045 NOT-SI管理的目标 补充内容是支持一项新的合作计划，将合成基因电路整合到我们的 工程设计的PVN模型。目前的组织工程脑血管模型缺乏正交性、可调性控制 血管周围壁龛的生长和成熟。制定独立的量化控制的能力 PVN的生长和成熟将是一个重大的进步，并将使我们能够深入 检查PVN内可能产生新的治疗靶点以改善结果的相互作用。 为了实现这一目标，我们提议与艾哈迈德·哈利勒博士(波士顿大学)进行新的合作努力 应用他的实验室的基因组正交合成锌指转录调控因子(SynZiFTR)技术 为了在血管周围壁龛模型中实现对PVN生长和成熟的药物调节、正交控制 该项目正在开发中。为了做到这一点，我们将调节合成工程菌的生长和成熟 血管周围生态位(目标S1)。我们随后将对TMZ的抵抗和入侵模式进行基准测试 对合成血管系统的反应(目标2)。这一提议的补充将支持一个协作团队 开发一种新的但互补的能力，以整合先进的组织工程和合成生物学 提供对脑血管周围生态位模型的可调节控制的工具集。通过协作，我们将建立 人合成组织结构作为研究相互作用的GBM-PVN信号的重要新工具 脑瘤的微环境。这种能力对于研究GBM细胞药物的模式是必不可少的 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.