Directed Cell Motility Along Gradients in Extracellular Matrix Fiber Alignment

项目摘要 在天然组织中，细胞的招募和定位是由定向迁移控制的，细胞感觉到 在他们的微环境中有多个引导线索，并优先朝向或远离特定的 信号。尽管已经确定了几种类型的指导或“出租车”提示，但 在定向迁移过程中同时呈现的信号仍然知之甚少。解剖这些 关系将有助于增强我们对复杂环境中细胞运动性的理解 对促进或抑制迁移活动的治疗发展的影响。 为了扩大我们对促进细胞定向迁移的线索的了解，我们提出了一项新的实验 将微工程纤维取向梯度与受控可溶生化相结合的平台 在复杂的3D环境中，内皮细胞、免疫细胞和癌细胞迁移的梯度。我们的平台 将允许我们以独特的方式将细胞群体暴露在包含比对的受控多线索3D环境中 梯度和生化梯度，以剖析细胞如何区分优先顺序和对同时指导做出反应 暗示。我们假设生物物理和生物化学之间存在协同(相加)和层次(竞争)关系。 影响细胞定向迁移的生化引导信号。为了检验这一假设，我们将进行 以下目标：1)在微工程3D胶原胶环境中使用可调节的 对齐景观，2)在3D凝胶中创建生化梯度并量化趋化反应 环境，以及3)在组合的生物物理和生物化学梯度内量化运动响应 环境。 该项目的成功将建立一个独特的平台，研究细胞如何感知、优先排序和迁移到 对多种指导线索(这里是生物物理和生化梯度)的反应，与早期 发育、免疫细胞运输、血管形成和癌症侵袭。

Project Summary In native tissue, cell recruitment and positioning are controlled by directed migration, where cells sense multiple guidance cues in their microenvironment and migrate preferentially toward or away from particular signals. Although several types of guidance or "taxis" cues have been identified, the hierarchy between simultaneously presented signals during directed migration remains poorly understood. Dissecting these relationships will help enhance our understanding of cell motility in complex environments with broad implications in therapeutic development to promote or inhibit migratory activity. To expand our knowledge about the cues that promote directed cell migration, we propose a new experimental platform that combines microengineered fiber alignment gradients with controlled soluble biochemical gradients to endothelial, immune, and cancer cell migration within a complex 3D environment. Our platform will allow us to uniquely expose cell populations to a controlled multi-cue 3D environment containing alignment gradients and biochemical gradients to dissect how cells prioritize and respond to simultaneous guidance cues. We hypothesize synergetic (additive) and hierarchal (competitive) relationships between biophysical and biochemical guidance cues that influence directed cell migration. To test this hypothesis, we will carry out the following aims: 1) Quantify migration within a microengineered 3D collagen gel environment with tunable alignment landscapes, 2) Create biochemical gradients and quantify chemotactic responses within 3D gel environments, and 3) Quantify motility responses within combined biophysical and biochemical gradient environments. Success in this project will establish a platform to uniquely study how cells sense, prioritize, and migrate in response to multiple guidance cues (here, biophysical and biochemical gradients), with relevance to early development, immune cell trafficking, vascularization, and cancer invasion.