Cyclic stretch of bicuspid aortic valves: elucidating its implications for cell signaling and tissue mechanics.

二叶式主动脉瓣的循环拉伸：阐明其对细胞信号传导和组织力学的影响。

• 批准号: 10607130
• 负责人: Toni Mcclish West
• 金额: $ 7.38万
• 依托单位: UNIVERSITY OF TEXAS AT AUSTIN
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-04-01 至 2026-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10607130
• 关键词: 3-Dimensional; Actins; Address; Affect; Age; Anatomy; Aortic Valve Stenosis; Architecture; Basal Cell; Biochemical; Cell Shape; Cell physiology; Cells; Cellular Morphology; Clinical; Commissure; Congenital Abnormality; Cytoskeleton; Data; Development; Disease; Disease Progression; Early Diagnosis; Early identification; Echocardiography; Environment; Ethylenes; Event; Exposure to; Extracellular Matrix; Fellowship; Gel; Glycols; Goals; Heart Valves; Human; Image; In Situ; Investigation; Lead; Legal patent; Length; Link; Measures; Mechanics; Mediating; Microscopy; Modeling; Morbidity - disease rate; Morphology; Myofibroblast; Nuclear; Organ; Pathway interactions; Patients; Pattern; Periodicity; Persons; Pharmacological Treatment; Phenotype; Procedures; Process; Proteins; Proteomics; Quality of life; Research; Research Personnel; Risk; Role; Running; Sampling; Series; Sex Differences; Signal Transduction; Smooth Muscle; Stimulus; Stretching; System; Texas; Tissues; Training; Transforming Growth Factor beta; Travel; Universities; Work; aortic valve; aortic valve disorder; aortic valve replacement; bicuspid aortic valve; biobank; biochemical model; biomechanical model; body system; cell type; conditioning; effective therapy; experience; experimental study; interstitial cell; malformation; mathematical model; mechanical signal; metaplastic cell transformation; mortality; negative affect; novel; pharmacologic; response; therapeutic target; transcription factor; treatment strategy; two photon microscopy; valve replacement

PROJECT SUMMARY: While 1.4% of people have the congenital defect of bicuspid aortic valve (BAV), BAV patients make up 50% of the patients that receive aortic valve replacements. Moreover, patients with BAVs develop aortic stenosis (AS) earlier, and thus require replacements at younger ages than patients with normal tricuspid aortic valves (TAV). Given the limited durability of replacements, BAV patients have a high procedure burden that negatively affects their length and quality of life. Therefore, development of a pharmacological therapy will reduce morbidity and mortality from AS. Our long-term goal is to understand the BAV disease process at the cellular level to develop effective treatments that mitigate AS. Since no pharmacological treatment has been forthcoming, our hypothesis is that abnormal valve interstitial cell (VIC) deformation patterns present in BAVs have a crucial role in the biochemical signaling events in AS. Since AS is associated with VIC activation into myofibroblasts and transforming growth factor-beta (TGFB) signaling, this study addresses the relationship between mechanically-conditioned cellular morphology and TGFB signaling in two specific aims: 1. Determine what parameters of mechanical conditioning experienced in varying aortic valve anatomies affect VIC morphology. The level of myofibroblast activation will first be determined in native human valve leaflet explants from BAV and TAV. The morphologies of cells from native tissues will be compared to that of cells mechanically conditioned in a novel 3D high-throughput biaxial oscillatory stretch screen (3D HT-BOSS) to determine what cyclical biaxial stretch, matrix stiffness, and VIC basal contractility is required to produce morphologies seen in BAVs and TAVs. 2. Ascertain how altering mechanical pattern will modify VIC response to TGFB. Quantitative proteomics will be employed to develop steady-state models of VIC TGFB signaling from VICs freshly isolated from native human valve leaflets. Microscopy of 3D HT-BOSS samples will then be employed to analyze shifts in: 1) EC50 of αSMA protein and 2) nuclear localization of TGFB- mediated transcription factors upon exogenous TGFB stimulation. Through this investigation, underlying drivers of AS and novel target pathways for pharmacological treatment will therefore be uncovered. Furthermore, the training that the fellowship applicant, Dr. Toni West, will receive will enable her to make the leap to becoming an independent investigator. Dr. West will be conducting research in the lab of her sponsor, Dr. Michal Sacks, and in the lab of her collaborator, Dr. Aaron Baker, at the University of Texas. As part of her training, Dr. West will travel to Columbia University, where her co- sponsor Dr. Giovanni Ferrari runs the biobank she will be collecting tissues and cells from.

项目摘要：虽然1.4％的人患有双质主动脉瓣（BAV）的先天性缺陷，但 BAV患者占接受主动脉瓣置换术的患者的50％。而且，患者 随着BAV的发展主动脉狭窄（AS），因此需要年轻时的替换 正常三尖瓣主动脉瓣（TAV）的患者。鉴于更换的耐用性有限，BAV 患者的手术伯恩（Burnen）具有很高的过程，会对他们的生活长度和生活质量产生负面影响。所以， 药物疗法的开发将降低AS的发病率和死亡率。 我们的长期目标是了解细胞水平的BAV疾病过程以发展有效 减轻AS的治疗。由于没有任何药物治疗，我们的假设 是，BAV中存在的异常瓣膜间隙（VIC）变形模式在 AS中的生化信号事件。因为与VIC激活成肌纤维细胞有关 并改变生长因子-Beta（TGFB）信号传导，本研究解决了 机械条件的细胞形态和TGFB信号传导两个具体目的： 1。确定在不同主动脉瓣中经历的机械调节参数 解剖影响VIC形态。肌纤维细胞激活的水平将首先确定 来自BAV和TAV的本地人瓣膜外植体。来自天然细胞的形态 将组织与在新型3D高通量中机械条件的细胞进行比较 双轴振荡拉伸屏（3D HT-BOSS）确定哪种周期性双轴矩阵矩阵 在BAV和TAV中产生形态需要刚度和VIC基本收缩力。 2。确定改变机械模式将如何改变对TGFB的VIC响应。定量 将采用蛋白质组学来开发VIC的VIC TGFB信号的稳态模型 从本地人类瓣膜传单中新鲜分离。然后，3D HT-Boss样品的显微镜将是 用于分析αSMA蛋白的EC50和2）TGFB-的核定位置。 外源TGFB刺激后介导的转录因子。 通过这项调查，AS和新型目标途径的基础驱动因素 因此，将发现治疗。 此外，奖学金申请人托尼·韦斯特（Toni West）的培训将使她能够 飞跃成为一名独立调查员。韦斯特博士将在 她的赞助商Michal Sacks博士的实验室，以及她的合作者Aaron Baker博士的实验室 德克萨斯大学。作为培训的一部分，韦斯特博士将前往哥伦比亚大学，她的共同 赞助商Giovanni Ferrari博士经营着她将收集组织和细胞的生物库。