Nanomechanical effects of aldosterone on vaso-occulation in sickle cell disease

描述(申请人提供)：过去十年的心血管研究明确表明，激素醛固酮具有促炎血管作用，包括内皮细胞的生物物理变化，如：细胞僵硬，细胞体积增加，表面黏附分子表达增加，一氧化氮产生减少等。然而，尚未研究的是这些由醛固酮引起的内皮细胞力学性质的改变如何影响镰状细胞血管闭塞，这也是一种生物物理现象，以及醛固酮如何改变血细胞的生物物理性质。我们假设醛固酮介导的细胞机械效应影响镰状细胞血管闭塞，并建议使用我们先前开发的结合常规检测的单细胞机械技术来探索醛固酮如何影响微血管闭塞。具体地说，我们将在微血管的三维体外模型中确定和研究醛固酮对内皮与镰状红细胞和白细胞的黏附(目标1)、镰状红细胞和白细胞的变形性(目标2)以及细胞聚集和微循环流动的潜在机制(目标3)。应聘者选择了一条新颖的培训和职业道路，最近完成了儿科血液学/肿瘤学的奖学金和生物工程博士学位的结合。这笔K08拨款将让林博士接受所有必要的培训，以发展成为学术医学领域的新一代独立研究员：一名应用生物工程和纳米科学技术开发新的生物医学设备的医生-科学家-工程师，用于诊断和研究儿科血液疾病。具体地说，这一奖项将用于进一步培训他在血管生物学、黏附细胞生物学以及先进的微制造和微流体技术方面的培训，以开发用于镰状细胞疾病的复杂的芯片实验室设备。这项研究将在加州大学伯克利分校一个独特的生物工程实验室进行，该实验室专注于应用和开发新的光学显微镜、力显微镜和微流体技术来研究健康和疾病中的细胞力学，与加州大学旧金山分校有着密切的临床联系。相关性(参见说明)：通过调查激素醛固酮是否在镰状细胞性贫血中发挥作用，这项工作将提高我们对这种复杂且不治之症的理解和新的治疗方法。此外，我们为这个项目开发的设备将作为新的诊断和药物发现工具的原型，用于治疗这种每天影响数万美国人生活的虚弱和潜在致命的疾病。

DESCRIPTION (provided by applicant): Cardiovascular research in the last decade has definitively shown that the hormone aldosterone has pro- inflammatory vascular effects, including biophysical changes in endothelial cells such as: increased cell stiffness, increased cell volume, increased surface adhesion molecule expression, and decreased nitric oxide production, among others. What has not been studied, however, is how these aldosterone-induced alterations in endothelial cell mechanical properties affect sickle cell vaso-occlusion, which is also a biophysical phenomenon, and how aldosterone may also alter the biophysical properties of blood cells. We hypothesize that aldosterone-mediated cell mechanical effects influence sickle cell vaso-occlusion and propose to use single-cell mechanical techniques that we have previously developed in combination with conventional assays to explore how aldosterone affects microvascular occlusion. Specifically, we will determine and investigate the underlying mechanisms of aldosterone's effect on endothelial adhesion to sickle red cells and leukocytes (Aim 1), sickle red cell and leukocyte deformability (Aim 2), and cell aggregation and microcirculatory flow in a three-dimensional in vitro model of the microvasculature (Aim 3). The candidate has chosen a novel training and career path and has recently completed a fellowship in pediatric hematology/oncology combined with a PhD in bioengineering. This K08 grant will allow Dr. Lam to receive all the necessary training to develop into a new breed of independent researcher in academic medicine: a physician-scientist-engineer who applies bioengineering and nanoscience techniques to develop new biomedical devices for diagnosis and study of pediatric blood diseases. Specifically, this award will be used to further his training in vascular biology, the cell biology of adhesion, and advanced microfabrication and microfluidics to develop complex lab-on-chip devices for sickle cell disease. This research will be conducted in a unique bioengineering lab at UC Berkeley that focuses on applying and developing new optical microscopy, force microscopy, and microfluidic techniques to study cell mechanics in health and disease, with close clinical connections at UC San Francisco. RELEVANCE (See instructions): By investigating whether the hormone aldosterone plays a role in sickle cell anemia, this work will improve our understanding of- and new treatments for-this complex and incurable disease. In addition, the devices we develop for this project will serve as prototypes for new diagnostic and drug discovery tools for this debilitating and potentially fatal disease that affects the lives of tens of thousands of Americans every day.