Engineered Enteric Nervous System-Peri Neural Invasion platform to improve predictive preclinical screens in early-stage colorectal adenocarcinomas

项目摘要 本提案的目的是设计和利用肠神经丛作为工具，以提高预测性 早期II-III期结直肠癌（CRC）临床前筛查的准确性。神经周围浸润（PNI）或CRC 通过神经（尤其是肠神经系统的神经; ENS）传播是 预后不良。CRC肿瘤不仅积极招募神经，ENS神经然后具有控制神经的能力， 上皮细胞增殖和上皮干细胞分化。这是值得注意的，因为这两个 过程可以改变CRC动力学并调节其对辅助化疗的反应。用于目前的临床 在实践中，PNI是通过病理分级确定的，但没有考虑PNI进入ENS的具体情况。我们 我认为ENS-PNI是临床决策的重要考虑因素，因为它可以从根本上 改变肿瘤动力学和对辅助化疗的反应。尽管有明确的证据表明，PNI是一个积极的 由于ENS-PNI是CRC的一个重要贡献者，目前还没有ENS-PNI在CRC中的临床前模型，这严重限制了我们的研究。 研究和靶向ENS-PNI的能力。我们建议通过发展和 一种新型3D自体患者源性ENS-PNI平台的验证，该平台源自患者源性肿瘤 上皮细胞和来自成体肠神经干细胞（ENSC）的工程化神经。我们假设 工程ENS-PNI平台可通过ENS高保真预测患者对辅助化疗的反应 调节CRC动力学和癌症干细胞自我更新。该提案独特地利用了PI的 （Raghavan）发表了先进的生物材料和生物工程平台的专业知识， 肿瘤和结肠微环境，结合Co-I（Esnaola）的临床专业知识， 病人的生物样本通过我们的方法，我们将研究以下目标：（1）开发和验证一个在 （2）利用ENS-PNI体外平台筛选患者源性结直肠癌早期肿瘤细胞， 用于辅助化疗的分期CRC肿瘤。我们将使用以下组织病理学来验证ENS-PNI平台： 它们来源于患者的肿瘤，并将对化疗的反应与临床反应进行比较。这 将是CRC中基于ENS的PNI的体外3D模型的第一个实例。拟议的工作非常适合 探索/发展生物工程研究资助计划，由于其组织的发展- 基于工程的技术，以提高临床前癌症筛查的能力。这项新技术 将用于开发具有PNI成分的II/III期CRC的高通量可接受筛选， 目的是确定靶向辅助治疗的新途径。

PROJECT SUMMARY The objective of this proposal is to engineer and utilize enteric nerve plexi as a tool to improve predictive accuracy of preclinical screens in early stage II-III colorectal cancer (CRC). Peri-neural invasion (PNI) or CRC spread through nerves (especially nerves of the enteric nervous system; ENS) is an independent predictor of poor prognosis. Not only do CRC tumors actively recruit nerves, the ENS-nerves then have the ability to control epithelial cell proliferation and epithelial stem cell differentiation. This is noteworthy because both of these processes can alter CRC dynamics and modulate its response to adjuvant chemotherapy. In current clinical practice, PNI is established by pathological grading, but PNI into the ENS specifically is not considered. We believe that ENS-PNI is an important consideration in clinical decision making, because it could fundamentally alter tumor dynamics and response to adjuvant chemotherapy. Despite clear evidence that PNI is an active contributor to CRC, there are currently no preclinical models of ENS-PNI in CRC which severely limits our ability to investigate and target ENS-PNI. We propose to address this limitation through the development and validation of a novel 3D autologous patient-derived ENS-PNI platform, derived from patient-derived tumor epithelial cells and engineered nerves from adult enteric neural stem cells (ENSCs). We hypothesize that the engineered ENS-PNI platform can predict patient response to adjuvant chemotherapy with high fidelity via ENS regulation of CRC dynamics and cancer stem cell self-renewal. This proposal uniquely leverages the PI’s (Raghavan) published expertise with advanced biomaterial and bioengineered based platforms for engineering the tumor and colon microenvironment, combined with the Co-I’s (Esnaola) clinical expertise and access to patient biospecimen. Through our approach, we will examine the following aims: (1) Develop and validate an in vitro platform of ENS-PNI in CRC; (2) Utilize the engineered ENS-PNI platform to screen patient-derived early stage CRC tumors for adjuvant chemotherapy. We will validate the ENS-PNI platform using histopathology of the patient tumors they are derived from, and compare responses to chemotherapy to clinical responses. This will be the first instance of an in vitro 3D model of ENS-based PNI in CRC. The proposed work fits well into the Exploratory/Developmental Bioengineering Research Grant program, owing to its development of tissue- engineering based technology to improve the capability of preclinical cancer screening. This novel technology will be leveraged to develop a high-throughput amenable screen of stage II/III CRCs with a PNI component, for the purpose of identifying new avenues of targeted adjuvant therapy.