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.

瓣膜间隙细胞（VIC）的肌纤维细胞激活被认为是瓣膜纤维化和狭窄。因此，为控制该行动的外部提示 VIC的肌纤维细胞表型一直是该领域引起的大量关注的主题。越来越多的证据表明，超出受体介导的可溶性生长激活的VIC激活因素，矩阵中的物理线索在此过程中起着至关重要的作用。不幸的是，传统用于培养VIC的方法固有地导致其肌纤维细胞激活，以便它难以确定环境僵硬对激活的影响，尤其是去激活。为了解决这个问题，我们的小组证明了独特的水凝胶材料可用于为VIC文化创建柔软的，无激活的基质，使Vics保持表型更类似于新鲜分离的细胞。现在，我们旨在研究如何基质刚度与促炎性细胞因子结合影响VIC成纤维细胞至肌纤维细胞转变，随着时间的推移可能发生的表观遗传变化，以及基质信号传导中的途径可能有助于逆转致病性成肌纤维细胞表型。具体而言，我们建议：1）使用组合方法研究 VIC表型的促炎性细胞因子与微环境刚度的关系2）确定机械和炎症性线索对成纤维细胞至肌纤维细胞的影响过渡及其使用具有动态可调机械性能的水凝胶的逆转，3）发现治疗剂的新分子靶标，以使致病性VIC肌纤维细胞调节在炎症条件下激活。共同完成的每个目标都完成了将为纤维化主动脉瓣狭窄的进展提供独特的见解。创建可调的细胞培养平台将使我们回答有关差异的问题可逆（瞬态，伤口愈合状态）和不可逆（持续性，致病状态）VIC 传统方法无法充分解决的肌纤维细胞。随后的信号通路和基因的分析将用于识别具有治疗性的新靶逆转VIC激活和治疗瓣膜疾病的潜力。此外，成功完成这些目标应该对医学领域具有普遍的兴趣，因为纤维化的机制是可能在大多数与纤维化有关的疾病中共享。