Investigating sex differences in persistent valvular myofibroblast activation using hydrogel culture substrates

使用水凝胶培养基质研究持续瓣膜肌成纤维细胞活化的性别差异

• 批准号: 10631477
• 负责人: Brian Alberto Aguado
• 金额: $ 8.25万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN DIEGO
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-05 至 2024-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10631477
• 关键词: Address; Algorithms; Aortic Valve Stenosis; Biochemical; Biological Assay; Cells; Chromatin; Cues; Disease; Drug Combinations; Evolution; Female; Fibroblasts; Genes; Heart failure; Human; Hydrogels; Lead; Link; Molecular; Myofibroblast; Operative Surgical Procedures; Pathologic; Patients; Pharmaceutical Preparations; Pharmacotherapy; Progressive Disease; Research; Serum; Sex Differences; Sum; Tissues; Transposase; X Inactivation; aortic valve; computational platform; epigenome; interstitial cell; male; mechanical signal; precision medicine; response; sex; small molecule; valve replacement

PROJECT SUMMARY Aortic valve stenosis (AVS) is a progressive disease where fibroblast-like valvular interstitial cells (VICs) become persistently activated myofibroblasts, which contribute to pathologic aortic valve leaflet stiffening. AVS is treated with valve replacement surgeries, which may be avoided if small molecule drug combinations could be identified to inhibit persistent myofibroblast activation. However, the molecular mechanisms regulating persistent myofibroblast activation are unknown and may vary from patient-to-patient and/or by sex. My proposed R00 research seeks to continue characterizing the sex- and patient-specific differences that lead to persistent myofibroblast activation during AVS and optimize drug combinations to inhibit myofibroblast activation as a function of patient-specific cues. We hypothesize (i) sex-linked differences in how male and female VICs respond to mechanical cues and (ii) patient-specific biochemical cues found in AVS patient sera contribute to persistent activation and subsequent myofibroblast response to small molecule drugs. In Aim 1 (completed), we have characterized genes that escape X-chromosome inactivation that regulate myofibroblast activation uniquely in female VICs seeded on PEG hydrogels. In Aim 2 (in progress), we will generate persistently activated myofibroblasts in human AVS patient sera and determine alterations (e.g. open chromatin regions) in the VIC epigenome due to patient-specific serum factors using Assay for Transposase-Accessible Chromatin with sequencing (ATAC-seq). In Aim 3 (proposed), we will identify optimal combinations of small molecule drugs to inhibit persistent myofibroblast activation in the presence of AVS patient serum using a differential evolution algorithm. In sum, the proposed research will address an urgent, unmet need for sex-specific and precision medicine approaches for identifying molecular mechanisms of myofibroblast persistence, which may provide a bridge toward non-surgical AVS therapies.

项目摘要 主动脉瓣狭窄（AVS）是一种进行性疾病，成纤维细胞样瓣膜间质细胞（VIC） 持续激活的肌纤维细胞，这有助于病理主动脉瓣叶片僵硬。 AV被治疗 使用瓣膜置换手术，如果可以鉴定出小分子药物组合，则可以避免这种手术 抑制持续的肌纤维细胞激活。但是，调节持续的分子机制 肌纤维细胞的激活尚不清楚，可能从患者到患者和/或性别变化。我建议的R00 研究旨在继续表征性别和患者特定的差异，导致持久性 AVS期间的肌纤维细胞激活，并优化药物组合以抑制肌纤维细胞激活为一种 患者特异性提示的功能。我们假设（i）男性和女性维克斯的反应方式（i）性别联系的差异 在AVS患者血清中发现的机械提示和（ii）患者特异性生化线索有助于持续 激活和随后对小分子药物的肌纤维细胞反应。在AIM 1（完成）中，我们有 逃脱X染色体灭活的特征基因，这些基因在肌成纤维细胞激活中独特地调节肌纤维细胞 雌性维克斯播种在钉凝胶上。在AIM 2（正在进行中）中，我们将持续激活 人类AVS患者血清中的肌纤维细胞，并确定VIC中的改变（例如开放染色质区域） 由于患者特异性的血清因子而使用转座酶可访问的染色质而引起的表观基因组 测序（ATAC-SEQ）。在AIM 3（提议）中，我们将确定小分子药物的最佳组合 使用差异进化，在存在AVS患者血清的情况下抑制持续的肌纤维细胞激活 算法。总而言之，拟议的研究将解决对性别特定和精确的紧急，未满足的需求 鉴定肌纤维细胞持久性的分子机制的医学方法，这可能会提供 迈向非手术AV疗法的桥梁。