Directed Cell Motility Along Gradients in Extracellular Matrix Fiber Alignment

项目摘要 在天然组织中，细胞募集和定位由定向迁移控制，其中细胞感知 在他们的微环境中有多种引导线索，并优先朝着或远离特定的方向迁移。 信号.虽然已经识别了几种类型的引导或“出租车”线索，但是 在定向迁移期间同时呈现的信号仍然知之甚少。解剖这些 关系将有助于提高我们对细胞运动的理解，在复杂的环境中， 在促进或抑制迁移活性的治疗开发中的意义。 为了扩大我们对促进定向细胞迁移的线索的了解，我们提出了一个新的实验方法。 该平台将微工程纤维排列梯度与可控可溶性生化 在复杂的3D环境中，内皮细胞、免疫细胞和癌细胞的迁移梯度。我们的平台 将使我们能够独特地将细胞群暴露于包含对齐的受控多线索3D环境中， 梯度和生化梯度来剖析细胞如何优先考虑和响应同步指导 线索我们假设生物物理和生物化学之间的协同（加性）和层次（竞争）关系， 影响定向细胞迁移的生化引导线索。为了验证这一假设，我们将进行 1）用可调的微工程化3D胶原凝胶环境量化迁移 对齐景观，2）创建生化梯度并量化3D凝胶内的趋化反应 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. 这个项目的成功将建立一个平台，独特地研究细胞如何感知，优先考虑和迁移到 对多种指导线索的反应（这里，生物物理和生物化学梯度），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.