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称为转移，是全球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共同作用，以引导癌细胞向血管移动。该项目团队将创建一个包含不同胶原特性组合的三维（3D）水凝胶库，并确定引导癌细胞通过肿瘤微环境的特性。该项目中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其环境相互作用，并确定未来抗转移治疗的可能靶点。该项目还将加强罗切斯特地区的科学，技术，工程和数学（STEM）管道，建立一个指导暑期计划，以支持本科生在罗切斯特理工学院和罗切斯特大学校园的研究。癌症转移是其中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接种次要部位。富含胶原蛋白的肿瘤ECM提供了在基质侵入过程中促进细胞迁移的结构引导线索。使用肿瘤胶原的二次谐波产生（SHG）成像，已经确定垂直于肿瘤-宿主界面排列的胶原纤维预测患者转移。该项目团队最近表明，SHG前向/后向（F/B）成像对构成胶原纤维的胶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。通过SHG F/B测量的这些原纤维水平性质被称为胶原纤维内部结构（FIS）。尽管排列的胶原纤维和SHG F/B都是人类患者转移的预测因子，但尚不清楚这些多尺度特性如何联合收割机影响肿瘤细胞在基质侵袭期间的运动性。该项目旨在测试FIS和纤维排列特性联合收割机以协同和分级方式结合以影响细胞迁移通过肿瘤ECM的假设。第一个目标是使用F/B测量和来自具有已知转移结果的人类肿瘤样品的纤维排列特性作为指导，以复制多尺度肿瘤模拟胶原特性的微工程3D胶原支架。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能的肿瘤细胞的迁移特性，然后评估盘状结构域受体作为胶原FIS受体的作用。该项目代表了第一个3D微工程的努力，联合收割机和独立调整两个临床相关的结构线索，F/B和纤维对齐，并系统地评估其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，并提供深入了解调节细胞-ECM相互作用的受体。该奖项反映了NSF的法定使命，并已被认为值得通过使用基金会的智力价值和更广泛的影响审查标准进行评估。

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.