Vascular cell sexual dimorphism in complex mechanical microenvironments

复杂机械微环境中的血管细胞性别二态性

基本信息

批准号：
9760337
负责人：
Bryan Daniel James
金额：
$ 3.92万
依托单位：
UNIVERSITY OF FLORIDA
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-09-16 至 2022-09-15
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9760337
关键词：
4D MRI Address Architecture Area Arterial Fatty Streak Atherosclerosis Atomic Force Microscopy Behavior Biological Biological Assay Bioreactors Blood Vessels Blood flow Cardiovascular Diseases Cardiovascular system Cell physiology Cells Cellular Morphology Clinical Complex Computational Molecular Biology Development Devices Diagnosis Disease Disease susceptibility Early Diagnosis Endothelial Cells Estradiol Exhibits Fellowship Female Florida Foundations Functional disorder Future Gene Expression Gene Expression Regulation Genes Germ Cells Glycocalyx Health Human Immunofluorescence Microscopy In Vitro Incidence Knowledge Learning Life Link Liquid substance Location MRI Scans Measurement Measures Mechanics Medicine Mentors Methods Mission Molecular Morphology Nature Organism Outcome Output Patients Pattern Periodicity Peripheral arterial disease Physical Stimulation Physical environment Physiological Play Polymerase Chain Reaction Probability Property Regulation Reporting Research Reverse Transcription Role Scientist Sex Differences Site Smooth Muscle Myocytes Statistical Data Interpretation Statistical Methods Statistical Models Stretching Structure Symptoms Techniques Testing Training Umbilical vein United States National Institutes of Health Universities Vascular Endothelial Cell Vascular Endothelium Vascular Smooth Muscle Vision Woman Work base cell behavior design disability experimental study genetic analysis hemodynamics human female imaging modality improved male mechanical force mechanical properties medical attention men polyacrylamide gels pre-doctoral response sex sex disparity sexual dimorphism shear stress stem cell differentiation

项目摘要

The sex disparity in the incidence and progression of cardiovascular disease (CVD), including peripheral arterial disease (PAD), is a troubling clinical observation. It is well known that CVD manifests differently in men and women, with more women than men suffering from the disease. Further, within the vascular network, there are mechanical microenvironments that favor CVD development, for example, atherosclerotic plaques preferentially form at regions of disturbed flow and low fluid shear stresses such as, vasculature bifurcations. Yet, no reports have correlated these two separate observations at the cellular level, specifically, the sex differences in CVD and local vascular mechanics. This project aims to 1) assess the functional changes in male and female human umbilical vein endothelial cells in response to combined physiological fluid shear stresses and substrate stiffness by morphometric, secretory, and genetic analyses, 2) assess the functional changes in male and female human aortic smooth muscle cells in response to combined physiological cyclic stretching and substrate stiffness by morphometric, mechanical, and genetic analyses, and 3) correlate the disparate functional response of male and female vascular cells using statistical methods. Upon completion of these aims, a foundational understanding of vascular cell functionality in vitro as it pertains to sexual dimorphism in complex mechanical microenvironments will be gained. A factorial design of experiments will be used to systematically assess the influence of cell sex, laminar fluid shear stress or cyclic stretch, and the underlying substrate stiffness. The effects of the mechanical microenvironment will be assessed using well-defined flow and stretch bioreactors with RGD-conjugated polyacrylamide gel substrates. Cell morphology will be quantified by immunofluorescence microscopy methods. Glycocalyx, vasoregulatory, and estradiol secretory products will be quantified by commercial assays. Cellular mechanical properties will be measured by atomic force microscopy. The regulation of a large set of vascular cell genes including a subset related to CVD will be measured by quantitative reverse transcription polymerase chain reaction. Statistical analyses will be performed to relate gene expression results to morphological, secretory, and mechanical responses and then to correlate them to a given sex and mechanical force combination. This work follows the 2015 NIH mandate to consider sex as a biological variable by investigating the clinical observations of the sex disparity in cardiovascular disease at the cellular level. The significance for this work is that future imaging modalities that quantify vascular forces such as with 4D MRI, will be able to indicate areas within the vasculature that are susceptible to localized cellular dysfunction using correlative statistical models and ultimately predict a patient's location-specific vulnerability for disease. This proposal is part of a predoctoral training fellowship at the University of Florida. The trainee will learn to conduct the experiments proposed and to grow professionally from the guidance of a comprehensive mentoring team of clinicians, cell and molecular biologists, computational theorists, and material scientists.

心血管疾病（CVD）（包括外周动脉疾病）的发病率和进展存在性别差异疾病（PAD）是一个令人不安的临床观察结果。众所周知，CVD 在男性和女性中的表现不同。女性，患有这种疾病的女性多于男性。此外，在血管网络内，还有有利于CVD发展的机械微环境，例如优先选择动脉粥样硬化斑块形成于流动受干扰和低流体剪切应力的区域，例如脉管系统分叉处。然而，还没有任何报道在细胞水平上将这两个单独的观察结果关联起来，特别是性别差异 CVD 和局部血管力学。该项目旨在 1) 评估男性和女性的功能变化人脐静脉内皮细胞对生理流体剪切应力和通过形态测量、分泌和遗传分析来确定基质硬度，2) 评估功能变化男性和女性人主动脉平滑肌细胞对联合生理循环拉伸的反应通过形态测量、机械和遗传分析确定基底刚度，3) 将不同的使用统计方法测量男性和女性血管细胞的功能反应。完成这些后目标是对体外血管细胞功能的基本了解，因为它与性别二态性有关将获得复杂的机械微环境。实验的因子设计将用于系统地评估细胞性别、层流剪切应力或循环拉伸的影响以及潜在的影响基材刚度。机械微环境的影响将使用明确的流动和具有 RGD 共轭聚丙烯酰胺凝胶基质的拉伸生物反应器。细胞形态将通过以下方式量化免疫荧光显微镜方法。 Glycocalyx、血管调节和雌二醇分泌产品将通过商业化验进行量化。细胞机械特性将通过原子力显微镜测量。一大组血管细胞基因（包括与 CVD 相关的子集）的调节将通过以下方法测量定量逆转录聚合酶链反应。将进行统计分析以关联基因表达结果与形态、分泌和机械反应，然后将它们与给定的相关联性与机械力的结合。这项工作遵循 2015 年 NIH 的指令，将性视为一种生物学行为通过研究细胞中心血管疾病性别差异的临床观察结果来确定变量等级。这项工作的意义在于，未来量化血管力的成像方式，例如 4D MRI 将能够指示脉管系统内易受局部细胞功能障碍影响的区域使用相关统计模型并最终预测患者特定位置的疾病易感性。该提案是佛罗里达大学博士前培训奖学金的一部分。受训者将学习进行建议的实验，并在全面的指导下专业成长由临床医生、细胞和分子生物学家、计算理论家和材料科学家组成的指导团队。