Combinatorial analysis of migration stimuli for enhanced wound healing

描述(申请人提供)：在体内和体外环境中，细胞暴露在许多调节其功能的细胞外刺激中。例如，皮肤创伤愈合中的细胞事件受到可溶和固定的生物分子线索以及机械环境的强烈影响。表征细胞如何解释和响应这些单独和联合传递的有影响力的信号，不仅对于构建允许更好地控制细胞行为和伤口愈合的环境是重要的，而且对于理解自然的生理现象也很重要。这项拟议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线索组合，并特别关注角质形成细胞迁移和真皮伤口愈合事件。促进细胞加速和定向迁移的刺激组合可用于指导伤口敷料材料的设计，使伤口更快、更有效地闭合。这项工作不仅旨在应用于创伤愈合的临床问题，还旨在帮助科学家和工程师更好地了解和预测细胞决策过程，以便实现对细胞功能的更大控制。以下目标将使我们能够描述和预测角质形成细胞如何结合可溶的、固定的和机械的信号来做出关于增殖、迁移和凋亡的决定。具体目标1：研究可溶性EGF、固定化EGF和底物力学对角质形成细胞信号和功能的联合作用。在这一目标中，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，以确定角质形成细胞如何整合和响应来自这些不同刺激的信号，特别是在皮肤创伤愈合事件的背景下。具体目标2：确定相加和相反的趋化、趋化和趋化梯度刺激在角质形成细胞信号和功能调节中的作用和整合。这个目的是为了了解空间定向的可溶的、固定化的和机械的刺激在控制角质形成细胞功能方面的作用。具体地说，趋化、趋化和趋杜性梯度刺激将以相加和相反的方式组合，以检查细胞如何解释和响应这些不同的提示，以及确定哪些提示主导细胞行为以及为什么。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和功能的微环境调节中的多变量联系。将开发偏最小二乘回归模型来分析在AIMS 1和2中获得的数据集，使我们不仅能够描述并预测角质形成细胞如何整合多种微环境线索组合。 与公共健康相关：体内的细胞接收来自环境的信号，指示它们如何发挥作用。在这项建议中，我们的目标是更好地了解细胞如何整合和响应这些信号，特别是在皮肤伤口愈合的背景下。通过研究这些信号如何刺激不同的伤口愈合事件，我们将获得有助于我们设计改进的伤口敷料和治疗方法的信息。

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.