Combinatorial analysis of migration stimuli for enhanced wound healing

描述（由申请人提供）：在体内和体外环境中，细胞暴露于许多调节其功能的细胞外刺激。例如，皮肤伤口愈合中的细胞事件受到可溶性和固定化生物分子线索以及机械环境的强烈影响。细胞如何解释和响应这些单独和组合传递的有影响的信号的特征不仅对告知环境的构建具有重要意义，这些环境可以更好地控制细胞行为和伤口愈合，而且对理解天然生理现象也很重要。该研究的目标是描述细胞如何解释、整合和响应简单和复杂的微环境信号组合，特别关注角质细胞迁移和皮肤伤口愈合事件。促进加速和定向细胞迁移的刺激组合可用于告知伤口敷料的设计，使伤口愈合更快，更有效。这项工作不仅旨在应用于伤口愈合的临床问题，而且还有助于科学家和工程师更好地理解和预测细胞决策过程，以便更好地控制细胞功能。以下目的将使我们能够描述和预测角化细胞如何整合可溶性、固定化和机械线索，以决定增殖、迁移和凋亡。特异性目标1：表征可溶性EGF、固定化EGF和底物机制对角质形成细胞信号传导和功能的组合效应。在这个目标中，我们将量化可溶性表皮生长因子（EGF）、固定化EGF和底物力学对细胞行为的单独和联合影响，以确定角化细胞如何整合和响应来自这些不同刺激的信号，特别是在皮肤伤口愈合事件的背景下。具体目标2：表征加法和对立的趋化、触致和重致梯度刺激在角化细胞信号传导和功能调节方面的贡献和整合。本目的目的是了解空间定向可溶性、固定化和机械刺激在控制角化细胞功能方面的作用。具体来说，趋化、触致和重致梯度刺激将以叠加和对立的方式结合起来，以检查细胞如何解释和响应这些不同的线索，以及确定哪些线索主导细胞行为及其原因。具体目标3：开发一个数据驱动的网络模型来解释和预测角质形成细胞信号传导和功能微环境调节中的多元连接。将开发一个偏最小二乘回归模型来分析目标1和目标2中获得的数据集，使我们不仅能够描述而且能够预测角质形成细胞如何整合微环境线索的众多组合。

DESCRIPTION (provided by applicant): In both in vivo and in vitro environments, cells are exposed to numerous extracellular stimuli that regulate their function. For instance, cellular events in dermal wound healing are strongly influenced by soluble and immobilized biomolecule cues as well as the mechanical environment. The characterization of how cells interpret and respond to these influential cues delivered both individually and in combination is important in not only informing the construction of environments that allow greater control over cell behavior and wound healing, but also in understanding native physiological phenomena. The goal of the proposed research is to characterize how cells interpret, integrate, and respond to simple and complex combinations of microenvironmental cues, with a specific focus on keratinocyte migration and dermal wound healing events. The combination of stimuli that promote accelerated and directed cell migration can be used to inform the design of wound dressing materials that enable faster and more efficient wound closure. This work is intended to not only apply to the clinical problem of wound healing, but also to help scientists and engineers better understand and predict cellular decision-making processes in order to achieve greater control over cell function. The following aims will allow us to characterize and predict how keratinocytes integrate soluble, immobilized, and mechanical cues to make decisions about proliferation, migration, and apoptosis. Specific Aim 1: Characterize the combinatorial effects of soluble EGF, immobilized EGF, and substrate mechanics on keratinocyte signaling and function. In this aim we will quantify the individual and combined effects of soluble epidermal growth factor (EGF), immobilized EGF, and substrate mechanics on cellular behavior to determine how keratinocytes integrate and respond to signals received from these different stimuli, particularly in the context of dermal wound healing events. Specific Aim 2: Characterize the contributions and integration of additive and opposing chemotactic, haptotactic, and durotactic gradient stimuli with respect to regulation of keratinocyte signaling and function. The purpose of this aim is to understand the contributions of spatially-directed soluble, immobilized, and mechanical stimuli with respect to controlling the function of keratinocytes. Specifically, chemotactic, haptotactic, and durotactic gradient stimuli will be combined in both additive and opposing manners in order to examine how cells interpret and respond to these different cues, as well as to determine which cues dominate cell behavior and why. Specific Aim 3: Develop a data-driven network model to interpret and predict multivariate connections in the microenvironmental regulation of keratinocyte signaling and function. A partial least squares regression model will be developed to analyze the data set acquired in Aims 1 and 2, enabling us to not only describe but also predict how keratinocytes integrate numerous combinations of microenvironmental cues. PUBLIC HEALTH RELEVANCE: Cells in the body receive signals from their environment that instruct them on how to function. In this proposal, we aim to better understand how cells integrate and respond to these signals, particularly in the context of dermal wound healing. By studying how these signals stimulate different wound healing events, we will gain information that will help us to design improved wound dressings and treatments.