3D Engineered Model of Microscopic Colorectal Cancer Liver Metastasis for Adjuvant Chemotherapy Screens

项目总结 50%的结直肠癌患者发生肝转移(CRLM)。尽管有佐剂 化疗和手术后，75%的CRLM复发是由于微小的残留病变(~1-3 mm) 逃脱了射线检测。这项提议目的是设计和利用一种佐剂 针对微观CRLM的化疗筛查平台，以识别可以 根除微小残留病。临床前模型，如球体和患者来源的异种移植 模型缺乏肝转移瘤的结构和细胞外基质(ECM)组成 微环境。我们假设，通过设计肝脏转移的微环境，我们 可以生产可用于治疗性筛查的可重现的显微镜CRLM实例。 工程化微观CRLM模型将包括：i)脱细胞猪肝生物基质支架 包含肝脏细胞外基质成分和3D结构；ii)来自细胞的微观球体 从IV期CRLM患者来源的异种移植物中建立。我们的方法提供了比 其他脱细胞模型：i)使用患者来源的细胞；以及ii)使用光学成像 定量基准建立显微CRLM；iii)使用正交验证 包括从工程模型到治疗学的临床患者反应基准；iv) 中、高吞吐量兼容。该项目将建立在PI(Raghavan)在 癌症组织工程学，但代表着研究方向的重大变化：PI的专业知识 正在研究卵巢癌的启动和化疗耐药，这项建议侧重于结直肠癌 转移和CRLM。为了支持这些研究重点，PI组建了一个调查小组 在结直肠癌(Kopetz)的临床治疗和管理方面拥有专业知识，高通量药物 筛查(斯蒂芬)和显微镜(沃尔什)。建议的具体目标包括：(1)建立和 描述来自人类IV期结直肠癌的微观CRLM的生物工程模型；(2)治疗性 使用NCI批准的文库筛选微观CRLM；(3)治疗靶向的验证 显微CRLM在原位转移模型中的作用。这将是第一个 显微CRLM体外生物工程模型的建立。使用设计好的 微观CRLM模型包括：i)筛选NCI批准的化合物库；ii)识别 治疗显微镜下CRLM的有效策略。预期的长期结果包括：i)预测 微小残留病的侵袭性，对高危患者进行分层 复发；ii)测试可转化为I期临床试验的化合物 用循环肿瘤DNA生物标记物鉴定的微观CRLM。

PROJECT SUMMARY 50% of colorectal cancer (CRC) patients develop liver metastasis (CRLM). Despite adjuvant chemotherapy and surgery, 75% of CRLM recurs due to microscopic residual disease (~1-3mm) that escapes radiographic detection. The objective of this proposal is to engineer and utilize an adjuvant chemotherapy screening platform specific to microscopic CRLM, to identify therapeutics that can eradicate microscopic residual disease. Preclinical models like spheroids and patient-derived xenograft models lack the architecture and extracellular matrix (ECM) composition of the liver metastatic microenvironment. We hypothesize that by engineering the liver metastatic microenvironment, we can produce reproducible instances of microscopic CRLM that can be used for therapeutic screening. Engineered microscopic CRLM models will include: i) decellularized porcine liver biomatrix scaffolds that contain both liver ECM composition and 3D architecture; ii) microscopic spheroids from cells established from patient-derived xenografts of stage IV CRLM. Our methods offer advantages over other decellularized models by: i) using patient-derived cells; and ii) using optical imaging to quantitatively benchmark the establishment of microscopic CRLM; iii) using orthogonal validation including clinical patient-response benchmarking of engineered models to therapeutics; iv) being medium- and high-throughput amenable. The project will build on the PI's (Raghavan) expertise in cancer tissue engineering, but represent significant changes in research directions: The PI's expertise is in studying ovarian cancer initiation and chemoresistance, and this proposal focuses on CRC metastasis and CRLM. To support these research pivots, the PI has assembled an investigative team with expertise in clinical treatment and management of CRC (Kopetz), high-throughput drug screening (Stephan) and microscopy (Walsh). Specific Aims proposed include: (1) : Establish and characterize a bioengineered model of microscopic CRLM from human stage IV CRC; (2) Therapeutic screening of microscopic CRLM using an NCI-approved library; (3) Validation of therapeutic targeting of microscopic CRLM in orthotopic metastasis models. This will be the first instance of a bioengineered in vitro model of microscopic CRLM. Immediate outcomes using the engineered microscopic CRLM model include: i) screening a library of NCI-approved compounds; ii) identifying efficacious strategies to treat microscopic CRLM. Long-term outcomes envisioned include: i) predict the aggressiveness of microscopic residual disease, stratifying patients who are at high-risk of recurrence; ii) testing compounds that can translate into phase I clinical trials in patients with microscopic CRLM identified with circulating tumor DNA biomarkers.