Dissecting the heterogeneity and function of myeloid cells in lupus nephritis

剖析狼疮性肾炎骨髓细胞的异质性和功能

基本信息

批准号：
10588862
负责人：
Anne Davidson
金额：
$ 61.82万
依托单位：
FEINSTEIN INSTITUTE FOR MEDICAL RESEARCH
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-02-01 至 2027-12-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10588862
关键词：
Acceleration Address Adult Affect Aftercare Animal Model Antibodies Antigen-Antibody Complex Atlases Attenuated Autoantibodies Autoimmune Diseases Automobile Driving Biological Biological Response Modifier Therapy Biopsy Blood CRISPR library Cell Communication Cell Compartmentation Cell physiology Cells Child Clinical Trials Clustered Regularly Interspaced Short Palindromic Repeats Coculture Techniques Complication Data Dendritic Cells Deposition Development Diagnosis Disease Disease Progression Disease remission End stage renal failure FDA approved Fibrosis Flare Flow Cytometry Frequencies Functional disorder Gene Silencing Goals Heterogeneity Histologic Homeostasis Human Immune Immune system Immunosuppressive Agents Impairment In Vitro Infiltration Inflammation Inflammatory Injury to Kidney Intervention Studies Kidney Kidney Diseases Kidney Failure Lupus Lupus Nephritis Macrophage Maps Mediating Medicine Methods Modeling Molecular Profiling Mouse Strains Mus Myelogenous Myeloid Cells Nephritis Organ Outcome Pathogenicity Pathway interactions Patients Peripheral Pharmaceutical Preparations Pharmacotherapy Phase Physiological Prevention Prognosis Protocols documentation Remission Induction Remission Induction Therapy Renal function Stromal Cells Systemic Lupus Erythematosus Testing Therapeutic Intervention Therapeutic immunosuppression Time Tissues belimumab cytokine design human data human disease human model immune cell infiltrate improved in vivo individual patient interest kidney biopsy kidney cell member mouse model new technology new therapeutic target novel therapeutic intervention novel therapeutics outcome prediction personalized medicine prevent programs renal damage repair function responders and non-responders response single cell analysis single-cell RNA sequencing standard of care therapeutic target tissue repair transcription factor

项目摘要

PROJECT SUMMARY Lupus nephritis (LN) is a common manifestation of systemic lupus erythematosus that can lead to irreversible renal impairment. Current immunosuppressive therapies fail to reverse disease in more than half of treated patients and only 2 new drugs have been approved treatment, both of which confer only modest improvement. Renal myeloid cells are involved in renal injury in LN, both in humans and animal models. Macrophages are highly plastic and can mediate both pro-inflammatory and reparative functions. Because there are so many subsets of myeloid cells and some of them change their function over time, analysis of single cells is crucial to the study of these cells. Initial studies in Phase 1 and early Phase 2 of the Accelerating Medicines Partnerships-SLE (AMP-SLE), of which we are all members, have shown that there are multiple subsets of macrophages in LN kidneys that are analogous to myeloid subsets in mice but the function of these subsets and whether they are pathogenic, or protective is still not known. Study of kidney tissues in LN patients is hampered by the small size of the kidney biopsy and the infrequency of these biopsies in individual patients over the course of their disease. Therefore, animal models, when carefully and rigorously employed, are especially useful for addressing hypotheses generated by examining human data. Our overall hypothesis is that we will identify human-relevant myeloid cell subsets in LN models that reflect differences in disease pathophysiology, disease stage and responsiveness to treatments and that this information will help identify new pathways for therapeutic intervention and direct personalized treatment. Our approach is focused on identifying and testing the function of human-relevant cell subsets and pathways in vitro and under physiologic conditions in vivo. In Aim 1 we will complete the integration of single cell RNASeq analysis of myeloid cells from 5 different models of LN with that from AMP-SLE Phase 2 to map both shared and unique macrophage sub-populations. We will follow the fate of peripheral myeloid cells as they transition from the blood into the kidneys to define how the nephritis-specific profile of each subset and how they are related to each other. We will then determine whether there is a particular myeloid cell profile that is associated with response or non-response of LN patients to standard of care therapy. In Aim 2 we will determine how renal myeloid cells interact with renal stromal cells both in vitro and in vivo. We will analyze the fate of renal myeloid cells (in responder and non-responder mice) in human-relevant mouse models treated with remission induction therapies to identify reparative subsets and pathways. Our focus here will be on both standard of care therapy and on belimumab (anti-BAFF) since this is the only biologic drug so far approved for treatment of LN and for prevention of further renal flares. In Aim 3 we will use CRISPR libraries for in vivo targeting of 13 transcription factors we have identified in preliminary studies to screen for new therapeutic approaches to protect LN kidneys from disease progression, fibrosis, and end stage renal disease.

项目概要狼疮性肾炎（LN）是系统性红斑狼疮的常见表现，可导致不可逆的肾功能损害。目前的免疫抑制疗法未能逆转一半以上的疾病接受治疗的患者中，只有 2 种新药被批准用于治疗，这两种药物都只能提供有限的治疗效果改进。在人类和动物模型中，肾髓细胞参与 LN 的肾损伤。巨噬细胞具有高度可塑性，可以介导促炎和修复功能。因为骨髓细胞有很多亚群，其中一些亚群会随着时间的推移而改变其功能，分析单细胞对于这些细胞的研究至关重要。第一阶段和第二阶段早期的初步研究我们都是 SLE 加速药物合作伙伴关系 (AMP-SLE) 的成员，该合作伙伴关系表明， LN 肾脏中的多个巨噬细胞亚群类似于小鼠的骨髓亚群，但功能这些子集的种类以及它们是否具有致病性或保护性仍不清楚。肾组织的研究 LN 患者因肾活检体积小且活检频率低而受到阻碍个别患者在其病程中。因此，动物模型，当仔细和严格所采用的方法对于解决通过检查人类数据产生的假设特别有用。我们的总体假设是，我们将在 LN 模型中识别与人类相关的骨髓细胞亚群，这些亚群反映了疾病病理生理学、疾病阶段和对治疗的反应的差异，并且这信息将有助于确定治疗干预和直接个性化治疗的新途径。我们的该方法的重点是识别和测试与人类相关的细胞亚群和途径的功能体外和体内生理条件下。目标1我们将完成单细胞RNASeq的整合对 5 种不同 LN 模型与 AMP-SLE 第 2 期模型的骨髓细胞进行分析，以绘制两者共享的图谱和独特的巨噬细胞亚群。我们将追踪外周骨髓细胞转变时的命运从血液到肾脏，以确定每个子集的肾炎特异性特征以及它们的情况彼此相关。然后我们将确定是否存在相关的特定骨髓细胞谱 LN 患者对标准护理治疗有反应或无反应。在目标 2 中，我们将确定如何肾骨髓细胞在体外和体内均与肾基质细胞相互作用。我们来分析肾的命运经缓解治疗的人类相关小鼠模型中的骨髓细胞（有反应小鼠和无反应小鼠）诱导疗法以确定修复子集和途径。我们这里的重点将放在以下两个标准上：护理疗法和贝利木单抗（抗 BAFF），因为这是迄今为止唯一被批准用于治疗的生物药物 LN 并用于预防进一步的肾病发作。在目标 3 中，我们将使用 CRISPR 文库进行体内靶向 13 我们在初步研究中确定了转录因子，以筛选新的治疗方法保护 LN 肾脏免受疾病进展、纤维化和终末期肾病的影响。