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：虽然有1.4%的人患有先天性二尖瓣缺陷症， 在接受主动脉瓣置换术的患者中，BAV患者占50%。此外，患者 患有BAVs的患者更早发展为主动脉瓣狭窄(AS)，因此需要在以下年龄进行置换 正常三尖瓣主动脉瓣(TAV)患者。考虑到更换件的耐用性有限，BAV 患者有很高的手术负担，这对他们的长度和生活质量产生了负面影响。因此， 药物治疗的发展将降低AS的发病率和死亡率。 我们的长期目标是在细胞水平上了解BAV疾病的过程，以开发有效的 缓解AS的治疗方法。由于没有即将到来的药物治疗，我们的假设 BAVs中存在的异常瓣膜间质细胞(VIC)变形模式在 AS中的生化信号事件。因为AS与VIC激活成肌成纤维细胞有关 和转化生长因子-β(TGFb)信号，本研究探讨了 机械调节的细胞形态和TGFb信号在两个特定的目的： 1.确定不同的主动脉瓣经历了哪些机械调节参数 解剖结构影响VIC的形态。肌成纤维细胞的激活水平将首先在 来自BAV和TAV的本地人瓣叶组织块。天然细胞的形态特征 在一种新的3D高通量技术中，组织将与机械条件下的细胞进行比较 双轴振动拉伸筛(3D HT-BOSS)确定哪些循环双轴拉伸，矩阵 硬度和VIC基础收缩能力是产生BAVs和TAVs中所见形态所必需的。 2.确定改变机械模式将如何改变VIC对TGFb的响应。量化 蛋白质组学将被用来开发VICTGFb信号的稳态模型 新鲜从本地人的瓣膜叶中分离出来。然后将对3D HT-BOSS样本进行显微镜检查 用来分析：1)αSMA蛋白的EC50值和2)TGFb的核定位 转录因子介导的外源TGFb刺激。 通过这项研究，AS的潜在驱动因素和新的药理靶向途径 因此，治疗将被揭开。 此外，奖学金申请者托尼·韦斯特博士将接受的培训将使她能够 飞跃成为一名独立的调查员。韦斯特博士将在 她的赞助者米哈尔·萨克斯博士的实验室，以及她的合作者亚伦·贝克博士的实验室 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，在那里她的同事 赞助商乔瓦尼·费拉里博士经营着一个生物库，她将从中收集组织和细胞。