Perioperative high-density lipoproteins and postoperative AKI

项目摘要 该奖项将支持罗兰·E·史密斯医学博士的一项前瞻性临床研究和职业发展。Dr。 史密斯与肖恩·戴维森博士一起完成了高密度脂蛋白(HDL)分子生物学的博士学位 辛辛那提大学与范德比尔特大学医学中心麻醉学住院医师培训 (VUmc)，她于2017年被任命为教员。VUMC是一家以其高度的 协作研究环境、卓越的职业发展计划和广泛的核心设施。 在她的T32奖学金期间，史密斯博士和她的导师林顿、比林斯和维克斯博士发现了一种 心脏手术患者术前高密度脂蛋白升高与AKI降低的关系在她的建议中 史密斯博士将对围手术期较高的载脂蛋白(Apo)A-I浓度和 特定的高密度脂蛋白-微RNA(MiRNAs)与术后较少的AKI(目标1)有关，而较高的高密度脂蛋白 抑制内皮细胞间黏附分子-1(ICAM-1)和肾近端功能的功能 肾小管细胞Toll样受体(TLR)2和TLR4的表达与较少的AKI(目标2)相关，而外源性 小鼠肾缺血再灌注(IR)前给予高密度脂蛋白降低肾脏ICAM-1、TLR2和TLR4的表达 Expression和AKI(目标3)。AIMS 1和AIMS 2将在一项150名受试者的前瞻性研究中进行测试，这些患者正在接受 心脏或血管外科手术。血液、尿液和临床数据将在整个 围手术期。载脂蛋白A-I浓度将用ELISA法测定；高密度脂蛋白-miRNAs将被分离， 用RT-PCR进行测序和定量；高密度脂蛋白抑制ICAM-1、TLR2和TLR4的功能能力将 用肿瘤坏死因子-α和低氧刺激的人血管内皮细胞和肾近端小管细胞进行体外测定 分别进行了分析。研究对象的高密度脂蛋白特征将与他们肾小管压力的严重程度和 损伤，通过尿液中金属蛋白酶组织抑制物2，胰岛素样生长因子的浓度来量化- 结合蛋白7和肾损伤分子1及其AKI严重程度与48小时血清定量 使用潜变量回归模型比较基线内生肌酐的变化。在AIM 3研究期间，有三个不同的 在肾脏IR损伤或假手术前给小鼠注射剂量的高密度脂蛋白颗粒，以及高密度脂蛋白的影响 肾脏损害的剂量将被量化。在第二个实验中，包含不同miRNA的高密度脂蛋白将被 在肾脏IR损伤前给予，以确定特定的高密度脂蛋白-miRNAs对AKI的影响。 通过完成这项研究，史密斯博士将获得翻译研究团队管理方面的培训，并 来自林顿博士的活体实验设计；围手术期临床研究执行和AKI表型。 Vickers博士的高密度脂蛋白相关微RNA(MiRNA)测序和生物信息学分析。在……里面 以这种方式，在这个职业发展奖的支持下，她的导师团队和她的机构 环境，史密斯博士将获得建立独立翻译研究所需的技能 该计划的重点是开发基于脂蛋白的新疗法，以减少术后AKI。

Project Abstract This award will support a prospective clinical study and the career development of Loren E. Smith, MD, PhD. Dr. Smith completed a PhD in high-density lipoprotein (HDL) molecular biology with Dr. Sean Davidson at the University of Cincinnati and residency training in Anesthesiology at Vanderbilt University Medical Center (VUMC), where she was appointed faculty in 2017. VUMC is a tertiary academic hospital renowned for its highly collaborative research environment, exceptional career development programs, and extensive core facilities. During her T32 fellowship Dr. Smith and her mentors, Drs. Linton, Billings, and Vickers, discovered an association between higher preoperative HDL and reduced AKI in cardiac surgery patients. In her proposed study, Dr. Smith will test the hypotheses that higher perioperative concentrations of apolipoprotein (apo) A-I and specific HDL-microRNAs (miRNAs) are associated with less postoperative AKI (Aim 1), that a higher HDL functional capacity to suppress endothelial cell intercellular adhesion molecule 1 (ICAM-1) and renal proximal tubule cell toll-like receptor (TLR) 2 and TLR4 expression is associated with less AKI (Aim 2), and that exogenous HDL administration to mice before renal ischemia and reperfusion (IR) reduces renal ICAM-1, TLR2, and TLR4 expression and AKI (Aim 3). Aims 1 and 2 will be tested in a 150-subject prospective study of patients undergoing cardiac or vascular surgery. Blood, urine, and clinical data will be collected longitudinally throughout the perioperative period. ApoA-I concentration will be determined with an ELISA; HDL-miRNAs will be isolated, sequenced and quantified with RT-PCR; and HDL functional capacity to suppress ICAM-1, TLR2, and TLR4 will be measured in vitro using human endothelial and renal proximal tubule cells stimulated with TNF-α and hypoxia respectively. Study subjects’ HDL characteristics will be compared to their severity of renal tubule stress and damage, quantified by urinary concentrations of tissue inhibitor of metalloproteinase 2, insulin-like growth factor- binding protein 7, and kidney injury molecule 1 and to their severity of AKI, quantified with 48-hour serum creatinine change from baseline using latent variable regression modeling. During Aim 3 studies, threedifferent doses of HDL particles will be administered to mice before renal IR injury or sham surgery, and the effects of HDL dose on renal damage will be quantified. In a second experiment, HDL containing different miRNAs will be administered before renal IR injury to determine the effect of specific HDL-miRNAs on AKI. By completing this study, Dr. Smith will obtain training in the management of a translational research team and in vivo experimental design from Dr. Linton; perioperative clinical study execution and AKI phenotyping from Dr. Billings; and HDL-associated microRNA (miRNA) sequencing and bioinformatics analysis from Dr. Vickers. In this manner, with the support of this career development award, her mentorship team, and her institutional environment, Dr. Smith will obtain the necessary skills to establish an independent translational research program focused on developing novel lipoprotein-based therapies to reduce postoperative AKI.