CAREER: Probing the Physiochemical Regulators of Tumor Stroma Interactions Using Microfluidic Biomimicry

实体瘤由癌细胞和癌细胞周围的组织组成，称为间质。间质包括血管细胞、炎症/免疫细胞和结缔组织，已成为肿瘤行为的重要决定因素。因此，迫切需要鉴定基质的促癌特性。该项目将使用微尺度工程技术来构建肿瘤间质的微流体模型。该模型将包括肿瘤间质的细胞、细胞外基质和生物分子成分。这个新的模型平台将被用来精确地检查肿瘤间质的细胞成分促进癌症进展的机制。这项研究的成功结果将消除研究肿瘤微环境的技术障碍，并将提供对肿瘤间质在癌症中作用的更深层次的理解。该研究计划将与PI的教育使命紧密结合在一起，即通过促进工程学学生对工程学与癌症交界处这条令人兴奋的学术职业道路的机会的认识，增加为跨学科癌症研究做出贡献的工程学学生的渠道。该项目将通过一项综合的教育和外联计划来支持这一愿景，该计划被组织为一系列举措，从初中和高中开始，一直持续到高年级，同时也吸引当地社区的参与。总而言之，外展工作将利用癌症研究作为最优先的社会需求的强大吸引力，强调吸引、留住和提高工程学中未被充分代表的女学生的韧性。该项目旨在促进对基质成纤维细胞通过自分泌、旁分泌和细胞周围基质通讯重新编程肿瘤微环境(TME)的物理化学机制的理解，特别关注基质激活，该激活受趋化因子CXCL12(或SDF-1)的不同亚型控制，趋化因子CXCL12(或SDF-1)是一种信号分子，推动超过23种不同癌症的癌症转移。一种创新的方法，结合了a)精确模拟TME内基质细胞-内皮细胞相互作用的微尺度工程技术，b)ECM的定量微结构分析，以及c)检测局部生物分子微环境变化的分子成像探针，将被用来系统地研究激活的基质微环境的发病机制，并具有高度的保真度和简单的实验控制。本研究旨在验证两个假说：1)量化CXCL12亚型对成纤维细胞激活的影响，利用微流控肿瘤间质模型结合ECM重塑的定量评估，检验间质成纤维细胞对ECM含量和微观结构的改变是区分不同CXCL12亚型潜力的重要决定因素的假设；2)利用PI的芯片上血管生成(AOC)平台，区分特定CXCL12亚型对血管生成和血管通透性促进特性的影响，以检验不同CXCL12亚型对血管生成和血管通透性的影响是由ECM含量和微结构的变化决定的假说。这项研究将在乳腺癌的背景下进行，选择性地分泌三种不同CXCL12亚型的乳腺成纤维细胞将被用作基质成纤维细胞的模型。完成这些目标将大大提高我们对由CXCL12调节电路介导的反应性基质反应的定量理解。该奖项反映了NSF的法定使命，并通过使用基金会的智力优势和更广泛的影响审查标准进行评估，被认为值得支持。

Solid tumors are composed of cancer cells and the tissue surrounding the cancer cells, called the stroma. The stroma, which includes vascular cells, inflammatory/immune cells, and connective tissues, has emerged as an important determinant of tumor behavior. Therefore, a critical need exists for identifying the tumor promoting properties of the stroma. This project will use micro-scale engineering technology to construct a microfluidic model of the tumor stroma. The model will include the cellular, extracellular matrix, and biomolecular constituents of the tumor stroma. This novel model platform will be used to precisely examine the mechanisms by which the cellular components of the tumor stroma promote cancer progression. The successful outcome of this research will remove technical barriers to studying the tumor microenvironment, and it will provide a deeper understanding of the role of tumor stroma in cancer. The research program will be closely coupled with the PI's educational mission of increasing the pipeline of engineering students contributing to interdisciplinary cancer research by promoting awareness of the opportunities for engineering students towards this exciting academic career path at the interface of engineering and cancer. This project will support this vision through a combined education and outreach plan that is organized as a series of initiatives that start at the middle and high school level and proceed to the senior undergraduate level while also engaging the local community. Collectively, the outreach efforts will leverage the strong appeal that cancer research has as a top priority societal need with an emphasis on attracting, retaining, and promoting resilience of female students, who are underrepresented in engineering.This project seeks to advance understanding of the physiochemical mechanisms by which stromal fibroblasts reprogram the tumor microenvironment (TME) through autocrine, paracrine, and pericellular matrix communication, with a particular focus on stroma activation that is controlled by different isoforms of the chemokine CXCL12 (or SDF-1), a signaling molecule that drives cancer metastasis in more than 23 different cancers. An innovative approach, combining a) microscale engineering technologies that precisely model the stromal cell-endothelial cell interactions within the TME, b) quantitative microstructural analysis of the ECM, and c) molecular imaging probes that detect changes in the local biomolecular microenvironment, will be used to systematically investigate, with high degrees of fidelity and facile experimental control, the pathogenesis of an activated stroma microenvironment. The research aims are designed to test two hypotheses: 1)To quantify the effects on fibroblast activation that are CXCL12 isoform specific, using the microfluidic tumor stroma model combined with quantitative assessment of ECM remodeling, to test the hypothesis that alterations to ECM content and microstructure by stromal fibroblasts are important determinants in distinguishing the potencies of different CXCL12 isoforms and 2) To distinguish specific isoforms of CXCL12 on their angiogenesis and vascular permeability promoting properties, using the PI's angiogenesis-on-a-chip (AoC) platform, to test the hypothesis that the potency of different CXCL12 isoforms on angiogenesis and vascular permeability is determined by changes to the ECM content and microstructure. The studies will be performed in the context of breast cancer, and mammary fibroblasts that selectively secrete three distinct CXCL12-isoforms will be used as the model for stromal fibroblasts. Completion of these aims will result in a significant advancement of our quantitative understanding of reactive stroma responses mediated by CXCL12 regulatory circuits.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.