Iron homeostasis and monocyte differentiation in patients with chronic kidney disease

慢性肾病患者的铁稳态和单核细胞分化

• 批准号: 10648527
• 负责人: Oleh Akchurin
• 金额: $ 12.71万
• 依托单位: WEILL MEDICAL COLL OF CORNELL UNIV
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-04-01 至 2025-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10648527
• 关键词: Acceleration; Address; Adult; Affect; American; Anemia; Animal Model; Antioxidants; Award; Behavior; CX3CL1 gene; Cells; Characteristics; Chemotactic Factors; Chemotaxis; Child; Chronic; Chronic Kidney Failure; Clinical; Collecting Cell; Color; Data; Dialysis procedure; Disease Progression; End stage renal failure; Etiology; Evaluation; Excretory function; Exhibits; Fibrosis; Flow Cytometry; Functional disorder; Funding; Genes; Goals; Homeostasis; Human; Individual; Inflammatory; Injury to Kidney; Invaded; Iron; Kidney; Kidney Diseases; Kidney Failure; Kidney Transplantation; Knowledge; Life Cycle Stages; Macrophage; Molecular; Mus; Myelogenous; Myeloid Cells; Pathogenicity; Pathologic; Patients; Peripheral; Peripheral Blood Mononuclear Cell; Persons; Phenotype; Play; Population; Proteins; Renal function; Research Personnel; Role; Serum; Techniques; Testing; Therapeutic; Time; Tissues; Urine; Work; biobank; chemokine; chemokine receptor; experience; experimental study; human data; improved outcome; iron deficiency; iron metabolism; kidney fibrosis; migration; monocyte; novel; peripheral blood; preclinical study; receptor; repository; response; systemic inflammatory response; therapeutic target; transcriptomics

PROJECT SUMMARY Chronic kidney disease (CKD) affects 10% of the population worldwide. More than 37 million people are estimated to have CKD in the US, and 2 in every 1000 Americans need dialysis or a kidney transplant to survive. CKD has numerous systemic complications including anemia and dysfunctional systemic iron homeostasis. Kidney fibrosis is the final mechanism common for all progressive kidney disorders. Unfortunately, very few therapies are available to slow the progression of kidney fibrosis in patients with CKD. Kidney macrophages are one of the key cells implicated in the pathophysiology of kidney fibrosis. The majority of kidney macrophages originate from circulating monocytes. In his K08-funded project Dr. Akchurin elucidated the pathologic role of depletion of labile iron pool in kidney macrophages in propagation of kidney fibrosis. This R03 proposal will enhance his capabilities of deciphering the role of iron in pathologic behavior of myeloid cells in CKD by now, in addition to kidney macrophages focusing on their precursors, circulating monocytes. Based on the preliminary data, the central hypothesis of this proposal is that in patients with CKD, circulating monocytes, similar to kidney macrophages, exhibit pathologically depleted intracellular labile iron pool LIP, which alters their phenotype by inducing CX3CR1 and other receptors facilitating chemotaxis and tissue invasion. Repletion of this intracellular monocyte labile iron pool through correction of systemic iron imbalance will be associated with reduction of this pro-inflammatory differentiation. This hypothesis will be tested in two specific aims: (1) determine the intracellular iron status of classical, intermediate, and non-classical circulating monocytes in patients with CKD and (2) elucidate the role of systemic and intracellular iron status in the differentiation of monocytes in patients with CKD. To test this hypothesis, the applicant will use the existing repository of peripheral blood mononuclear cells collected from children with CKD with and without functional or absolute iron deficiency and from healthy control children. He will evaluate these cells using single cell transcriptomic and multicolor flow cytometry approaches with the focus on delineating the phenotypic features responsible for subsequent tissue invasion, such as expression relevant chemokine receptors, including CX3CR1. Furthermore, he will perform ex vivo evaluation of human monocytes collected from CKD patients to directly assess their relevant functional characteristics in the presence and absence of iron stimulation. Monocyte functional characteristics will be analyzed I the context of clinical parameters, levels of circulating and urine-excreted chemokines, and systemic parameters of iron homeostasis. Results techniques an kidney from t he outlined experiments will provide preliminary data, as well as experience in for the functional ex-vivo testing of uman monocytes, to support the K08 awardee's application for R01 proposal to evaluate the impact of iron metabolism and iron therapy on myeloid cells in the context of fibrosis and CKD progression. h

项目摘要 慢性肾脏疾病（CKD）影响全球10％的人口。超过3700万人 估计在美国有CKD，每1000名美国人中有2个需要透析或肾脏移植才能生存。 CKD具有许多全身并发症，包括贫血和功能失调的系统性铁稳态。 肾纤维化是所有进行性肾脏疾病常见的最终机制。不幸的是，很少 可用于减慢CKD患者肾脏纤维化的进展。肾脏巨噬细胞是 与肾纤维化的病理生理有关的关键细胞之一。大多数肾脏巨噬细胞 起源于循环单核细胞。 Akchurin博士在他的K08资助项目中阐明了 肾巨噬细胞中不稳定铁池的耗竭在肾纤维化传播中。这个R03提案将 现在，增强了他破译铁在CKD中髓样细胞病理行为中的作用的能力 除肾脏巨噬细胞外，还集中于其前体，循环单核细胞。基于初步 数据，该提议的中心假设是，在CKD，循环单核细胞的患者中，类似于肾脏 巨噬细胞，表现出病理耗尽的细胞内不稳定铁池嘴唇，从而改变其表型 诱导CX3CR1和其他受体促进趋化性和组织侵袭。这个细胞内的补充 单核细胞不稳定铁池通过校正全身铁不平衡将与此减少有关 促炎性分化。该假设将以两个特定的目的进行检验：（1）确定细胞内 CKD患者的经典，中间和非经典循环单核细胞的铁状态和（2） 阐明了全身和细胞内铁状态在CKD患者中单核细胞分化中的作用。 为了检验该假设，申请人将使用现有的外周血单核细胞的存储库 从有或没有功能性或绝对铁缺乏症的CKD儿童中收集 孩子们。他将使用单细胞转录组和多色流式细胞仪方法评估这些细胞 重点是描述负责随后的组织入侵的表型特征，例如 表达相关的趋化因子受体，包括CX3CR1。此外，他将进行离体评估 从CKD患者收集的人类单核细胞直接评估其相关功能特征 铁刺激的存在和不存在。将分析单核细胞功能特征 临床参数，循环和尿液的趋化因子的水平以及铁的全身参数 稳态。结果 技术 一个 肾 根据概述的实验将提供初步数据，以及 对于UMAN单核细胞的功能性活体测试，以支持K08获奖者的申请 R01提出的提议评估铁代谢和铁治疗对髓样细胞的影响 纤维化和CKD进展。 h