Collaborative Research: Microengineered Tumor-Mimetic Collagen Landscapes to Test the Role of Prognostic Structural Cues on Cell Migration Through the Extracellular Matrix

癌症从原发肿瘤扩散到身体其他部位被称为转移，是世界范围内癌症相关死亡的主要原因。这个合作研究项目的首要目标是确定预测转移的胶原蛋白特性如何协同作用，引导癌细胞向血管运动。该项目团队将创建一个三维（3D）水凝胶库，其中包含胶原蛋白特性的不同组合，并确定引导癌细胞通过肿瘤微环境的特性。该项目开发的新知识将帮助研究人员了解不同的癌细胞亚群如何与环境相互作用，并确定未来抗转移治疗的可能靶点。该项目还将通过建立一个有指导的暑期项目来支持罗切斯特理工学院和罗切斯特大学校园的本科生研究，从而加强罗切斯特地区的科学、技术、工程和数学（STEM）管道。肿瘤转移是肿瘤细胞呈现迁移表型，侵入周围细胞外基质（ECM），并浸润淋巴和血管的过程。然后这些循环肿瘤细胞可以在继发部位形成种子。在基质侵袭过程中，富含胶原的肿瘤ECM提供了促进细胞迁移的结构指导线索。利用肿瘤胶原蛋白的二次谐波成像（SHG），已经确定垂直于肿瘤-宿主界面排列的胶原纤维可以预测患者的转移。该项目团队最近表明，SHG正向/向后（F/B）成像对构成胶原纤维的胶原原纤维的空间组织敏感，是乳腺癌患者转移的独立预测因子。这些由SHG F/B测量的原纤维水平的特性被称为胶原纤维内部结构（FIS）。虽然排列的胶原纤维和SHG F/B都能预测人类患者的转移，但尚不清楚这些多尺度特性如何联合影响基质侵袭过程中肿瘤细胞的运动。本项目旨在验证FIS和纤维排列特性以协同和分层方式结合在一起，通过肿瘤ECM影响细胞迁移的假设。第一个目标是使用已知转移结果的人类肿瘤样品的F/B测量和纤维排列特性作为微工程3D胶原蛋白支架的指导，以复制多尺度肿瘤模拟胶原蛋白特征。第二个目标是系统地研究肿瘤模拟F/B和纤维排列的组合如何影响具有不同转移潜力的肿瘤细胞的迁移特性，然后评估盘状蛋白结构域受体作为胶原FIS受体的作用。该项目代表了首个结合和独立调整两种临床相关结构线索（F/B和纤维排列）的3D微工程努力，并系统地评估了它们对肿瘤细胞运动的影响。这项工作可以帮助推进转移预测算法，支持未来的治疗设计工作，并提供对调节细胞- ecm相互作用的受体的见解。该奖项反映了美国国家科学基金会的法定使命，并通过使用基金会的知识价值和更广泛的影响审查标准进行评估，被认为值得支持。

The spread of cancer from the primary tumor to other locations in the body is called metastasis and is the leading cause of cancer-related deaths worldwide. The overarching goal of this collaborative research project is to determine how collagen properties that predict metastasis work together to guide cancer cell movement toward blood vessels. The project team will create a library of three-dimensional (3D) hydrogels containing different combinations of collagen properties and identify the properties that guide cancer cells through the tumor microenvironment. The new knowledge developed in this project will help researchers understand how different cancer cell sub-populations interact with their environment and identify possible targets for future anti-metastatic treatments. This project will also enhance the Rochester-area science, technology, engineering, and mathematics (STEM) pipeline by establishing a mentored summer program to support undergraduate student research at the Rochester Institute of Technology and University of Rochester campuses. Cancer metastasis is a process wherein tumor cells take on a migratory phenotype, invade the surrounding extracellular matrix (ECM), and infiltrate lymph and blood vessels. These circulating tumor cells can then seed secondary sites. The collagen-rich tumor ECM provides structural guidance cues that promote cell migration during the matrix invasion process. Using second harmonic generation (SHG) imaging of tumor collagen, it has been established that collagen fibers aligned perpendicular to the tumor-host interface are predictive of patient metastasis. The project team has recently shown that SHG forward/backward (F/B) imaging, sensitive to the spatial organization of collagen fibrils that comprise collagen fibers, is an independent predictor of metastasis in breast cancer patients. These fibril-level properties measured by SHG F/B are referred to as the collagen fiber internal structure (FIS). Although aligned collagen fibers and SHG F/B are both predictive of metastasis in human patients, it is unclear how these multiscale properties combine to influence the motility of tumor cells during matrix invasion. This project aims to test the hypothesis that FIS and fiber alignment properties combine in a synergistic and hierarchal manner to influence cell migration through the tumor ECM. The first objective is to use F/B measurements and fiber alignment properties from human tumor samples with known metastatic outcomes as a guide to microengineer 3D collagen scaffolds that replicate the multiscale tumor-mimetic collagen characteristics. The second objective is to systematically investigate how combinations of tumor-mimetic F/B and fiber alignment influence migratory characteristics of tumor cells with different metastatic potentials and then evaluate the role of Discoidin domain receptors as collagen FIS receptors. This project represents the first 3D microengineering efforts to combine and independently tune two clinically relevant structural cues, F/B and fiber alignment, and systematically evaluate their effects on tumor cell motility. This work could help advance metastatic prediction algorithms, support future therapeutic design efforts, and provide insight into the receptors that modulate cell-ECM interactions.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.