Cellular and Molecular Response to Gentamicin-Induced Injury in Underdeveloped Kidneys

项目摘要 在孕龄34-36周之前出生的人肾单位禀赋低，风险增加2-3倍。 慢性肾脏病（CKD）早产儿患急性肾损伤（阿基）的风险也会增加， 一种独特的致病过程，因为损伤发生在出生后肾脏生长和成熟的关键窗口期间。 然而，由于有限的动物模型和很少的早产人类肾脏活检， 对早产肾的阿基和CKD的认识不足，治疗是支持性的，我们缺乏策略， 预防阿基和CKD在该人群中的发展。我们已经建立了一种新的小鼠模型， 先天性低肾单位数，模拟人类早产肾，并提出5年的职业生涯 旨在满足未满足需求的发展项目。我用庆大霉素诱导新生儿阿基， 经常给早产儿开肾毒性药物，发现肾单位数量少的小鼠 与同窝对照组相比，阿基更严重，炎症持续，CKD进展更快 肾单位数目正常我还发现了庆大霉素诱导的 低肾单位数小鼠的阿基。我假设近端小管中庆大霉素摄取增加， 肾单位数量少的小鼠导致更多的细胞损伤和死亡，引起严重和持续的炎症， 损害肾脏生长，加速CKD的发展。目标1将确定细胞基础， 通过评估庆大霉素摄取增加，发现庆大霉素诱导的损伤和炎症更严重， 积累，然后通过确定庆大霉素积累是否导致溶酶体和线粒体 损伤和由此产生的炎症。我们将确定庆大霉素后哪些细胞死亡途径被激活- 在低肾单位数的小鼠中诱导损伤，这一发现可以为未来的靶向治疗提供信息。 目的2研究庆大霉素诱导的肾脏炎症对肾脏生长发育的影响， 慢性肾脏病在小鼠与低肾单位数。我们将采用单细胞RNA测序的无偏方法 并分析调节细胞代谢、生长、炎症和细胞死亡的途径， 急性损伤后肾单位数量减少，以及出生后肾脏生长和成熟的关键时期。的 以发现为基础的研究有可能指导未来的分子靶向方法， 在早产儿中发生CKD。拟议的职业发展战略将使古德博士能够 获得必要的培训，成为一个成功的医生科学家研究细胞和分子 在不断增长的早产儿人群中，肾脏健康和疾病的基础。古德博士是一位精神病学家 和哥伦比亚大学儿科学助理教授，在那里她完成了临床培训， 研究奖学金在丰富的制度环境和导师的奉献精神的支持下， 林芳明和乔纳森·巴拉施，古德博士将很好地过渡到独立。

PROJECT SUMMARY Humans born before 34-36 weeks of gestation age have low nephron endowment and a 2-3-fold increased risk of chronic kidney disease (CKD). Preterm infants are also at increased risk of acute kidney injury (AKI), which is a unique pathogenic process as injury occurs during a critical window of postnatal renal growth and maturation. However, due to limited animal models and few preterm human kidney biopsies, the cellular and molecular basis of AKI and CKD in premature kidneys is poorly understood, treatment is supportive, and we lack strategies to prevent AKI and the development of CKD in this population. We have generated a novel mouse model of congenitally low nephron number that simulates human premature kidneys and propose a 5-year career development project that is designed to fill the unmet needs. I induced neonatal AKI with gentamicin, a nephrotoxic medication often prescribed to preterm infants, and discovered that mice with low nephron number have more severe AKI, sustained inflammation and rapid progression to CKD compared to littermate controls with normal nephron number. I have also identified a unique inflammatory profile following gentamicin-induced AKI in mice with low nephron number. I hypothesize that increased gentamicin uptake in proximal tubules of mice with low nephron number leads to more cell injury and death, causing profound and sustained inflammation, impairing renal growth, and accelerating the development of CKD. Aim 1 will determine the cellular basis for more severe gentamicin-induced injury and inflammation by assessing for increased gentamicin uptake and accumulation, and then by determining whether gentamicin accumulation causes lysosomal and mitochondrial damage and resultant inflammation. We will determine which cell death pathways are activated after gentamicin- induced injury in mice with low nephron number, the discovery of which could inform future targeted therapies. In Aim 2 we will study the effect of gentamicin-induced renal inflammation on renal growth and the development of CKD in mice with low nephron number. We will take the unbiased approach of single cell RNA sequencing and analyze pathways regulating cell metabolism, growth, inflammation, and cell death in mice with normal and low nephron number after acute injury, and during a critical period of postnatal renal growth and maturation. The discovery-based studies have the potential to guide future molecular targeting approaches to abrogate the development of CKD in humans born preterm. The career development strategy proposed will allow Dr. Good to acquire the training necessary to become a successful physician scientist studying the cellular and molecular basis of renal health and disease in a growing population of humans born preterm. Dr. Good is a neonatologist and Assistant Professor of Pediatrics at Columbia University, where she completed her clinical training and research fellowship. With the support of a rich institutional environment and the dedication of her mentors, Drs. Fangming Lin and Jonathan Barasch, Dr. Good will be well positioned to transition to independence.