Leveraging single index cell RNA sequencing to study macrophage populations in frozen biopsies of dermatomyositis

利用单标签细胞 RNA 测序研究皮肌炎冷冻活检中的巨噬细胞群

• 批准号: 10289088
• 负责人: Vladimir M Liarski
• 金额: $ 18.04万
• 依托单位: UNIVERSITY OF CHICAGO
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-07-21 至 2022-04-01
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10289088
• 关键词: Adrenal Cortex Hormones; Affect; Age; Animal Model; Anti-Inflammatory Agents; Antibodies; Applications Grants; Assessment tool; Autoimmune; Autoimmune Diseases; Biological; Biopsy; Biopsy Specimen; Breast; CD14 gene; CD80 gene; CD86 gene; CSF2RA gene; Cell Separation; Cells; Chromium; Clinical; Clinical Course of Disease; Coupled; Cryopreserved Tissue; Data; Dermatomyositis; Diagnosis; Diagnostic; Disease; FCGR3B gene; FDA approved; Flow Cytometry; Freezing; Fresh Tissue; Future; Goals; Grant; HLA-DR Antigens; Health; Human; IFNGR1 gene; ITGAM gene; ITGAX gene; Idiopathic Inflammatory Myopathies; Immune; Immune system; Immunology; In Situ; Inclusion Body Myositis; Infiltration; Inflammation; Inflammatory; Knowledge; Letters; Link; Location; Lupus Nephritis; Methods; Microscopy; Mus; Muscle; Myopathy; Myositis; Outcome; Pathogenesis; Pathway interactions; Patient-Focused Outcomes; Patients; Pharmaceutical Preparations; Phenotype; Plasma; Polymyositis; Population; Procedures; Protocols documentation; Questionnaires; Race; Rare Diseases; Recovery; Research; Risk; SELL gene; Sampling; Serum; Severities; Severity of illness; Skeletal Muscle; Sorting - Cell Movement; Stains; Stimulus; Symptoms; TNFRSF5 gene; Testing; Time; Tissues; Validation; Viral; Yolk Sac; adaptive immune response; base; cell type; clinical application; cohort; design; differential expression; experimental study; genetic technology; improved; indexing; innovation; macrophage; malignant breast neoplasm; monocyte; mortality; new therapeutic target; nonhuman primate; off-label use; power analysis; preservation; recruit; repository; sex; single cell analysis; single-cell RNA sequencing; tissue biomarkers; transcriptome sequencing

PROJECT SUMMARY/ABSTRACT Dermatomyositis (DM) is an acquired autoimmune muscle disorder with poorly understood causes. DM has no specific FDA-approved treatments and off-label use of immunosuppressive medications used in other autoimmune conditions has proven ineffective or infeasible. Up to 20% of patients die of their disease within 5 years, similar to stage III breast cancer. The goals of this grant proposal are to gain an understanding of which parts of the immune system are involved in causing DM and find which features predict the course of disease in each patient. Our long-term goals are to better understand how DM begins, identify which parts of the immune system are involved, predict the course of disease in each patient, and find new treatments to improve patient outcomes. The objective of this grant is to identify if macrophages – a specific immune cell type - participate in muscle inflammation in DM. Our central hypothesis is that macrophages recruit other immune cells into the muscles of patients with DM, causing disease. Our rationale is that by identifying the specific type of macrophages involved in inflammation, we will be able to better understand the main immune pathways involved in DM. This will identify new targets for future therapies that will improve patient outcomes. Our specific aims will test the following hypotheses: (Aim 1) Identify macrophage populations in normal skeletal muscle and dermatomyositis; (Aim 2) Elucidate macrophage signatures of DM severity. Upon conclusion of this project we will, for the first time, have the ability to isolate immune cells from frozen patient biopsies. This advance is necessary as current single cell analyses depend on fresh biopsy samples, which are not available due to the rarity of DM. Using this, we will understand the main macrophage types involved in DM and find out which specific muscle biopsy feature(s) influence clinical disease course. This contribution is significant because it will, for the first time, not only open up many frozen tissues for research purposes, but will also provide a broad overview of all macrophages participating in inflammation, identifying specific new targets for therapy. The proposed research is innovative as we will use cutting edge immunology and genetic technologies, previously limited to the use of fresh tissues, to achieve these goals. In addition, we will confirm a panel of commercial antibodies to use in immunofluorescent microscopy to identify specific macrophages associated with worse DM disease. This method will be clinically applicable and may have a large impact on current approaches to treatment.

项目摘要/摘要 皮肤炎（DM）是一种可获得的自身免疫性肌肉障碍，较差的原因是DM 没有特定的FDA批准治疗和标签的使用其他用于其他用于其他免疫抑制药物的方法 自身免疫性的疾病无效或不可行。 年，类似于第三阶段的乳腺癌。 免疫系统的一部分参与引起DM，并找到哪些特征可以预测 每个患者。 我们的长期目标是更好地了解DM如何开始，确定免疫的哪些部分 涉及系统，预测每个患者的疾病进程，并找到新的治疗方法以改善患者 结果。 DM中的肌肉炎症是巨噬细胞募集其他免疫细胞 DM患者的肌肉，导致疾病。 涉及炎症的巨噬细胞，我们将能够更好地了解主要的免疫途径 参与DM 具体目的将检验以下假设：（目标1）识别正常的巨噬细胞种群 骨骼肌和皮肌炎； 在该项目结束后，我们将首次具有分离免疫细胞的能力 冻结的患者活检是必要的。 样品，由于DM的稀有性，我们将了解巨噬细胞。 DM涉及的类型并找出哪些特定的肌肉活检特征影响临床疾病课程。 这种贡献很重要，因为它首次开放了许多冷冻组织 研究目的，但还将为所有参与炎症的巨噬细胞提供广泛的概述， 确定特定的治疗目标。 免疫学和遗传技术以前限于新鲜组织的使用 此外，我们还将确认一组用于免疫荧光显微镜的商业抗体，以鉴定 特定的巨噬细胞与Wourse DM疾病有关。 对当前的治疗方法有很大影响。