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

项目总结 在34-36周孕龄之前出生的人肾单位发育程度低，风险增加2-3倍。 慢性肾脏疾病(CKD)。早产儿患急性肾损伤(AKI)的风险也会增加 损伤发生在出生后肾脏生长和成熟的关键时期，是一种独特的致病过程。 然而，由于有限的动物模型和很少的早产人类肾脏活检，细胞和分子基础 AKI和CKD在早产儿肾脏中的作用知之甚少，治疗是支持性的，我们缺乏策略来 预防AKI和CKD在这一人群中的发展。我们已经产生了一种新的小鼠模型 与生俱来的低肾单位数，模拟人类早产儿肾脏，并提出5年的职业生涯 旨在满足未得到满足的需求的开发项目。我用庆大霉素诱导新生儿AKI，a 经常给早产儿开肾毒性药物，发现肾单位数低的小鼠 与产仔对照组相比，有更严重的AKI、持续的炎症和快速进展为CKD 肾单位数正常。我还发现了庆大霉素诱导的一种独特的炎症状态 肾单位数低的小鼠的Aki。我推测庆大霉素在肾小管近端的摄取增加 肾单位数低的小鼠会导致更多的细胞损伤和死亡，导致严重和持续的炎症， 损害肾脏生长发育，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和炎症来评估庆大霉素引起的更严重的损伤和炎症 积累，然后通过确定庆大霉素积累是否会导致溶酶体和线粒体 损害和由此产生的炎症。我们将确定庆大霉素后哪些细胞死亡途径被激活- 在低肾单位数小鼠中诱导损伤，这一发现可能为未来的靶向治疗提供参考。 在目标2中，我们将研究庆大霉素引起的肾炎对肾脏生长发育的影响。 对低肾单位数小鼠CKD的影响。我们将采取单细胞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 急性肾损伤后和出生后肾脏发育成熟的关键时期，肾单位数量较少。这个 基于发现的研究有可能指导未来的分子靶向方法，以消除 人类早产儿慢性肾脏病的发生发展。提出的职业发展战略将使古德博士能够 获得必要的培训，成为一名研究细胞和分子的成功的内科科学家 在越来越多的早产人口中肾脏健康和疾病的基础。古德医生是一位新生儿科医生 和哥伦比亚大学儿科学助理教授，在那里她完成了她的临床培训和 研究奖学金。在丰富的机构环境和导师们的奉献精神的支持下，杜兰特博士。 林方明和乔纳森·巴拉施，Good博士将处于向独立过渡的有利地位。

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.