Regulation of vascular metabolism in acute lung injury

项目摘要/摘要 该提案概述了医学博士David Wu的综合研究和职业发展计划，以- 在Gokhan Mutlu医学博士和Yun Fang博士的实验室进行全面的博士后培训，并过渡到 独立的学术地位--与美国国立卫生研究院建立血管病理生理学研究项目 职业指导奖(K99/R00)。PI最近完成了NIH F32奖学金(F32 HL134288)和 在新陈代谢、分子生物学、生物物理学、光学、生物工程和血管生物等领域接受过培训。 GY。在为期两年的辅导期(K99)内，少年辅导团将从以下男子那里获得额外的学术指导- Tors和芝加哥大学的咨询委员会。职业发展计划旨在 让私人助理具备生物医学研究方面的必要知识和技能，以便成功过渡为 独立院士，随着工作的R00阶段的进展而导致R01。整体研究 目的是确定内皮代谢在调节内皮表型中的作用。 急性肺损伤的发生和扩散。内皮代谢在促炎性血液循环中的作用 人们对典型的水流类型了解甚少。PI在他的博士后F32期间进行的初步研究 时相显示，低切应力或血流动力学紊乱导致内皮细胞进入血管内皮细胞。 与单向流动相比，增加糖酵解代谢。增加这种糖酵解对体内- 内皮细胞炎症加剧(eLife，2017)。PI进一步表明，在急性肺损伤时，内皮细胞-血管紧张素转换酶活性增加。 AL细胞不能感知血液动力学流动，其表型使人联想到紊乱的血流。 血流动力学(AJRCCM，2017)。这项K99/R00研究提案测试了内皮细胞的总体假设 细胞功能障碍导致内皮细胞代谢的改变，这对血管病变的发生和发展是至关重要的 急性肺损伤。目标1将验证细胞骨架对启动局部糖酵解至关重要的假设 它通过RhoA介导的醛缩酶A的释放来推动血管屏障的破坏，醛缩酶A是甘氨酸-甘氨酸脱氢酶中的一个关键酶。 结肠裂解途径。目标2将验证氧化磷酸化下调作为一种结果- 内皮细胞功能障碍导致线粒体反向电子传递，这是血管内皮细胞功能障碍发生的关键。 糖酵解过程中活性氧的产生和上调。目标3将检验这样的假设，即调制 内皮代谢可减轻小鼠模型肺损伤。该目标将通过集成单个 体外和体内系统中的细胞显微镜和分子分析，导致对 新陈代谢如何影响损伤的传播，或许还能发现治疗靶点。实现这一目标 拟议的职业奖项将加速吴彦祖向独立内科医生-科学家的转变 导致获得具有竞争力的R01资金。

Project Summary/Abstract The proposal outlines an integrated research and career development plan for David Wu, MD PhD, to com- plete postdoctoral training in the laboratories of Gokhan Mutlu, MD, and Yun Fang, PhD and transition to an independent academic position by establishing a research program in vascular pathophysiology with a NIH mentored career award (K99/R00). The PI has recently completed an NIH F32 fellowship (F32 HL134288) and is trained in the fields of metabolism, molecular biology, biophysics, optics, bioengineering, and vascular biolo- gy. During the 2-year mentored period (K99), the PI will receive additional academic guidance from the men- tors and the advisory committee at the University of Chicago. The career development plan is designed to equip the PI with the necessary knowledge and skills in biomedical research for a successful transition as an independent academician, leading to an R01 as the R00 phase of the work progresses. The overall research goal is to determine the role of endothelial metabolism in mediating endothelial phenotypes in relation to the initiation and propagation of acute lung injury. The role of endothelial metabolism in pro-inflammatory hemody- namical flow types is poorly understood. Preliminary studies conducted by the PI during his post-doctoral F32 phase demonstrated that low shear stress or disturbed flow hemodynamics causes endothelial cells to in- crease glycolytic metabolism, compared to unidirectional flow. Increasing this glycolysis is necessary for in- creased endothelial inflammation (eLife, 2017). The PI furthermore showed that in acute lung injury, endotheli- al cells are unable to sense hemodynamic flow, and have a phenotype that is reminiscent of disturbed flow hemodynamics (AJRCCM, 2017). This K99/R00 research proposal tests the overall hypothesis that endothelial cell dysfunction causes a change in endothelial metabolism that is critical to the initiation and progression of acute lung injury. Aim 1 will test the hypothesis that the cytoskeleton is critical for initiating localized glycolysis that powers vascular barrier breakdown by RhoA mediated release of aldolase A, a critical enzyme in the gly- colytic pathway. Aim 2 will test the hypothesis that down-regulation of oxidative phosphorylation as a conse- quence of endothelial cell dysfunction causes mitochondrial reverse electron transport, which is critical for re- active oxygen species production and upregulation of glycolysis. Aim 3 will test the hypothesis that modulating endothelial metabolism can reduce lung injury in mouse models. The goal will be achieved by integrating single cell microscopy and molecular analysis in vitro and in vivo systems, leading to a mechanistic understanding of how metabolism influences injury propagation, and perhaps uncover therapeutic targets. Attainment of this proposed career award will accelerate the transition for David Wu to an independent physician-scientist and lead to acquisition of competitive R01 funding.