Defining the Pathogenesis and Prognosis of Human Acute Interstitial Nephritis

急性间质性肾炎(Ain)是由药物暴露、感染或自身免疫性疾病引起的。 在接受肾活检的患者中，高达20%的患者发生了急性肾损伤(AKI)。尽管我们目前 有2种临床干预措施可用于治疗AIN患者(停用违规药物和 皮质类固醇)，40%-60%的AIN患者继续发展为慢性肾脏疾病(CKD)，即使当 得到了适当的对待。AIN的肾脏损害被认为是免疫介导的肾小管损伤所致 最终会导致纤维化和永久性的肾脏损伤。 最近对多种自身免疫性疾病和癌症的免疫基础的否认导致了 靶向治疗的开发与治疗相比，显示出更好的疗效和更少的毒性 皮质类固醇。因此，免疫渗入和由此产生的驻留细胞(管状和血管)的分析 提供对启动和传播AIN的特定免疫事件的病原学理解的响应 应导致开发和/或改变靶向疗法的用途，以更有效地解决AIN 并防止进展为CKD，以及潜在的毒性较低。 来自几个小组的数据，包括我们自己的，表明CD4+T辅助细胞(特别是TH2/TH9 子集)是AIN的潜在驱动因素。我们发现TH2/TH9细胞因子IL-5和IL-9以及某些类型的细胞 2 AIN患者尿液或肾脏中的肥大细胞和嗜酸性粒细胞免疫水平较高。基于这些 数据显示，我们的假设是TH2/TH9辅助T细胞在肾脏本身起重要的致病作用。 促进AIN的肾小管或血管损伤。我们将通过执行定量评估来检验这一假设 AIN患者肾脏免疫浸润及伴随的肾小管和血管反应。我们会 使用现有的肾脏活检，由3名肾脏病理学家判定为AIN，来自两所大学的健康状况 中心(耶鲁大学和约翰霍普金斯大学)，作为这项研究的发现和验证队列。要执行量化 分析我们将使用一种名为成像质量细胞术(IMC)的成像技术，该技术支持同时、 在单个组织切片上对多达42个抗体进行空间保存的量化。我们有一个现有的图书馆 27个有效的肾脏和免疫抗体，并开发了一种机器学习协议来快速和 准确地量化和定位使用IMC鉴定的人类肾脏中的所有细胞。 我们将首先增加我们的有效抗体小组，并优化我们的IMC方案，以用于AIN的研究 (SA 1)。然后，我们将使用IMC来识别、量化和定位30例AIN的免疫和驻留细胞反应 来自耶鲁大学的病例和60例非AIN对照活检(发现队列)，其次是30例AIN病例和60例非AIN病例 来自JHU的对照(验证队列，SA 2)。最后，我们将定义细胞决定因素之间的关系 AIN和肾功能恢复以及对类固醇的反应(SA3)。我们的发现不仅会导致 在AIN中发现新的可用药靶点，也有助于改善AIN的临床组织学诊断。

Acute interstitial nephritis (AIN), resulting from drug exposure, infection or autoimmune disease, is the cause of acute kidney injury (AKI) in up to 20% of patients who undergo a kidney biopsy. Even though we currently have 2 clinical interventions available to treat patients with AIN (withdrawal of the offending drug and corticosteroids), 40-60% of patients with AIN go on to develop chronic kidney disease (CKD) even when appropriately treated. Kidney damage in AIN is believed to result from immune-mediated tubular injury that eventually leads to fibrosis and permanent kidney damage. The recent dileniation of the immune underpinnings of multiple autoimmune diseases and cancers has led to the development of targeted therapies that exhibit improved efficacy and less toxicity compared to corticosteroids. Therefore, an analysis of the immune infiltrate and resulting resident cell (tubular and vascular) responses that provides pathogenic understanding of the specific immune events that initiate and propogate AIN should lead to development and/or repurposing of targeted therapies that are more effective at resolving AIN and preventing the progression to CKD, as well as potentially less toxic. Data from several groups, including our own, suggest that CD4+ T-helper cells (particularly the TH2/TH9 subsets) are potential drivers of AIN. We have found that TH2/TH9 cytokines IL-5 and IL-9 and some cells of type 2 immunity, mast cells and eosinophils, are higher in the urine or kidneys of patients with AIN. Based on these data, it is our hypothesis that TH2/TH9 T-helper cells in the kidney itself play an important pathogenic role in promoting tubular or vascular injury in AIN. We will test this hypothesis by performing a quantitative evaluation of the kidney immune infiltrate and accompanying tubular and vascular response in humans with AIN. We will use existing kidney biopsies, adjudicated by 3 nephropathologists as exhibiting AIN, from two university health centers (Yale and Johns Hopkins), as discovery and validation cohorts for this study. To perform the quantitative analysis we will use an imaging technique called Imaging Mass Cytometry (IMC) that supports the simultaneous, spatially-preserved quantification of up to 42 antibodies on a single tissue section. We have an existing library of 27 validated kidney and immune antibodies and have developed a machine learning protocol to rapidly and accurately quantify and localize all cells in the human kidney identified using IMC. We will first increase our validated antibody panel and optimize our IMC protocol for use in the study of AIN (SA 1). We will then use IMC to identify, quantify and localize the immune and resident cell responses in 30 AIN cases and 60 non-AIN control biopsies from Yale (discovery cohort), followed by 30 AIN cases and 60 non-AIN controls from JHU (validation cohort, SA 2). Finally, we will define the relationship between cellular determinants of AIN and recovery of kidney function as well as response to steroids (SA3). Our findings will not only lead to identification of novel druggable targets in AIN, but also lead to improving clinical histological diagnosis of AIN.