Hydrogels to Study Synergistic Effects of Signaling Factors and Matrix Mechanics on Valve Disease Progression

水凝胶研究信号因子和基质力学对瓣膜疾病进展的协同作用

• 批准号: 9247569
• 负责人: KRISTI S. ANSETH
• 金额: $ 35.01万
• 依托单位: UNIVERSITY OF COLORADO
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-12-15 至 2020-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9247569
• 关键词: Address; Aftercare; Aortic Valve Stenosis; Attention; Back; Biochemical; Cell Culture Techniques; Cells; Cues; Cytokine Signaling; Disease; Disease Progression; Encapsulated; Epigenetic Process; Excision; Experimental Designs; Exposure to; Fibroblasts; Fibrosis; Gel; Goals; Growth Factor; Heart Valve Diseases; Heart Valves; Histologic; Homeostasis; Hydrogels; IL6 gene; In Situ; In Vitro; Inflammation; Inflammatory; Interleukin-1 beta; Knowledge; Light; Mechanics; Mediating; Medicine; Methods; Molecular; Molecular Target; Morphology; Myofibroblast; Operative Surgical Procedures; Organ; PI3K/AKT; Pathogenicity; Pathologic Processes; Pathway Analysis; Pathway interactions; Phenotype; Phosphorylation; Play; Population; Process; Role; Signal Pathway; Signal Transduction; Signaling Protein; Stenosis; Stimulus; System; TNF gene; Testing; Therapeutic; Time; Tissues; Transforming Growth Factor beta; Work; Wound Healing; aortic valve; aortic valve replacement; clinically relevant; combinatorial; culture plates; cytokine; efficacy testing; ethylene glycol; inhibitor/antagonist; innovation; insight; interest; interstitial cell; mechanical properties; mechanotransduction; molecular drug target; molecular targeted therapies; mouse model; receptor; small molecule inhibitor; stem; targeted treatment; transcriptome; transcriptome sequencing; valvular stenosis

Myofibroblast activation of Valvular Interstitial Cells (VICs) is considered to be a primary driver of valvular fibrosis and stenosis. For this reason, the external cues that act to control the myofibroblast phenotype of VICs have been topics of considerable attention in the field. Increasing evidence suggests that beyond receptor-mediated activation of VICs by soluble growth factors, physical cues from the matrix play a critical role in this process. Unfortunately, traditional methods used to culture VICs inherently leads to their myofibroblast activation, such that it becomes difficult to determine the effects of environmental stiffness on activation and especially de-activation. To address this issue, our group has demonstrated that unique hydrogel materials can be used to create soft, non-activating substrates for VIC culture that allow VICs to maintain a phenotype that more closely resembles that of freshly isolated cells. Now, we aim to examine how matrix stiffness in combination with pro-inflammatory cytokines influence the VIC fibroblast-to- myofibroblast transition, the epigenetic changes that may occur to these cells over time, and the pathways in matrix signaling that might be useful in reversing the pathogenic myofibroblast phenotype. Specifically, we propose to: 1) Use a combinatorial approach to study the effect of pro-inflammatory cytokines on VIC phenotypes as a function of microenvironmental stiffness 2) Identify the effects of mechanical and inflammatory cues on the fibroblast-to-myofibroblast transition and its reversal using hydrogels with dynamically tunable mechanical properties, and 3) Discover new molecular targets for therapeutics to temper pathogenic VIC myofibroblast activation under inflammatory conditions. Together, work completed within each of these Aims will provide unique insight into the progression of fibrotic aortic valvular stenosis. The creation of tunable cell culture platforms will allow us to answer questions about differences between reversible (transient, wound healing state) and irreversible (persistent, pathogenic state) VIC myofibroblasts that cannot be adequately addressed with traditional methods. Subsequent analysis of the signaling pathways and genes will be used to identify new targets with therapeutic potential to reverse VIC activation and treat valve disease. Moreover, successful completion of these Aims should be of general interest to the field of medicine, as mechanisms of fibrosis are likely shared among most fibrosis-related diseases.

瓣膜间质细胞 (VIC) 的肌成纤维细胞激活被认为是 瓣膜纤维化和狭窄。出于这个原因，控制行为的外部线索 VIC 的肌成纤维细胞表型一直是该领域备受关注的话题。 越来越多的证据表明，除了受体介导的可溶性生长激活 VIC 之外， 因素，来自矩阵的物理线索在此过程中发挥着关键作用。不幸的是，传统 用于培养 VIC 的方法本质上会导致其肌成纤维细胞激活，因此 很难确定环境刚度对激活的影响，尤其是 停用。为了解决这个问题，我们的团队证明了独特的水凝胶材料 可用于创建用于 VIC 培养的柔软、非活化基质，使 VIC 能够保持 表型更接近于新鲜分离的细胞。现在，我们的目标是研究如何 基质硬度与促炎细胞因子相结合影响 VIC 成纤维细胞 肌成纤维细胞转变，这些细胞随着时间的推移可能发生的表观遗传变化，以及 基质信号通路可能有助于逆转致病性肌成纤维细胞 表型。具体来说，我们建议：1）使用组合方法来研究 促炎细胞因子对 VIC 表型的影响作为微环境硬度的函数 2) 确定机械和炎症信号对成纤维细胞到肌成纤维细胞的影响 使用具有动态可调机械性能的水凝胶进行转变及其逆转，以及3） 发现治疗致病性 VIC 肌成纤维细胞的新分子靶点 炎症条件下的激活。共同完成这些目标中的工作 将为纤维化主动脉瓣狭窄的进展提供独特的见解。的创建 可调谐细胞培养平台将使我们能够回答有关细胞之间差异的问题 可逆（短暂、伤口愈合状态）和不可逆（持续、致病状态）VIC 传统方法无法充分解决肌成纤维细胞问题。随后的 对信号通路和基因的分析将用于识别新的治疗靶标 逆转 VIC 激活和治疗瓣膜疾病的潜力。此外，还顺利完成了 这些目标应该引起医学领域的普遍兴趣，因为纤维化的机制是 可能是大多数纤维化相关疾病共有的。