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 慢性肾脏病（CKD）影响着全球10%的人口。超过3700万人 据估计，在美国，每1000名美国人中就有2人需要透析或肾移植才能生存。 CKD具有许多全身性并发症，包括贫血和功能失调的全身性铁稳态。 肾纤维化是所有进行性肾脏疾病共同的最终机制。不幸的是， 目前已有减缓CKD患者肾纤维化进展的治疗方法。肾脏巨噬细胞 肾纤维化病理生理学中的关键细胞之一。大多数肾脏巨噬细胞 来源于循环单核细胞。在他的K08资助的项目中，Akchurin博士阐明了 在肾纤维化的传播中消耗肾巨噬细胞中的不稳定铁池。R03提案将 提高他破译铁在CKD骨髓细胞病理行为中的作用的能力， 此外，肾巨噬细胞集中在其前体，循环单核细胞。根据初步 数据，该建议的中心假设是，在CKD患者中，循环单核细胞，类似于肾脏 巨噬细胞，表现出病理性耗竭的细胞内不稳定铁池LIP，这改变了它们的表型， 诱导促进趋化性和组织侵袭的CX3CR1和其它受体。这种细胞内 通过纠正全身性铁失衡的单核细胞不稳定铁池将与这种减少有关。 促炎分化。这一假设将在两个特定的目标进行测试：（1）确定细胞内 CKD患者中经典型、中间型和非经典型循环单核细胞的铁状态和（2） 阐明全身和细胞内铁状态在CKD患者单核细胞分化中的作用。 为了验证这一假设，申请人将使用现有的外周血单核细胞库 收集自CKD伴或不伴功能性或绝对铁缺乏的儿童和健康对照组 孩子他将使用单细胞转录组学和流式细胞术方法来评估这些细胞 重点在于描绘负责随后的组织侵入的表型特征，例如 表达相关的趋化因子受体，包括CX3CR1。此外，他将进行体外评价， 从CKD患者收集的人单核细胞，以直接评估其在CKD患者中的相关功能特征。 有无铁刺激。单核细胞的功能特征将在以下背景下进行分析： 临床参数、循环和尿排泄趋化因子水平以及铁的全身参数 体内平衡结果 技术 一个 肾 从他概述的实验将提供初步数据，以及经验， 用于人单核细胞的功能性离体测试，以支持K08获奖者申请 R01建议在以下情况下评价铁代谢和铁治疗对骨髓细胞的影响： 纤维化和CKD进展。 H