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

基本信息

批准号：
9397567
负责人：
KRISTI S. ANSETH
金额：
$ 36.28万
依托单位：
UNIVERSITY OF COLORADO
依托单位国家：
美国
项目类别：
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/9397567
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

项目摘要

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治疗的新靶点逆转 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。