Resolution of ocular inflammation: the role of type 1 conventional dendritic cells

解决眼部炎症：1 型常规树突状细胞的作用

• 批准号: 10449898
• 负责人: Lynn M Hassman
• 金额: $ 21.82万
• 依托单位: WASHINGTON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-06-01 至 2027-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10449898
• 关键词: Acute; Address; Adopted; Adrenal Cortex Hormones; Affect; Anti-Inflammatory Agents; Automobile Driving; Bioinformatics; Biological Markers; Biological Response Modifiers; Blindness; CD8-Positive T-Lymphocytes; Cells; Chronic; Classification; Clinical; Data; Dendritic Cells; Development; Disease; Disease model; Experimental Models; Future; Gene Expression; Gene Expression Profile; Gene Expression Profiling; Genetic Transcription; Genomics; Goals; Heterogeneity; Human; Immune; Immune Targeting; Immunosuppression; Immunotherapy; In Vitro; Individual; Inflammation; Inflammatory; Inflammatory Response; Investigation; Knowledge; Liquid substance; Methodology; Methods; Modeling; Molecular; Mus; Myeloid Cells; Ophthalmology; Outcome; Pathogenicity; Patients; Persons; Pharmaceutical Preparations; Phenotype; Play; Public Health; Research; Resolution; Role; Sampling; Severities; Shapes; Signal Transduction; Surrogate Markers; Syndrome; Techniques; Testing; Therapeutic; Time; Tissue-Specific Gene Expression; Training; Translating; United States; Uveitis; Validation; adaptive immune response; aged; aqueous; arm; base; candidate marker; cohort; diagnostic tool; disease classification; disorder subtype; experience; exposed human population; human disease; immunoregulation; improved; in vivo; ineffective therapies; innovation; insight; large datasets; molecular diagnostics; molecular marker; mouse model; novel; peripheral blood; precision medicine; predictive signature; scaffold; side effect; single cell analysis; single-cell RNA sequencing; therapeutic target; tool; treatment strategy

Project Summary Non-infectious uveitis is comprised of a heterogeneous group of clinically-defined diseases for which empiric therapy fails many patients. Molecular characterization of uveitis, on the other hand, would result in more precise classification of disease subtypes, while simultaneously providing pathophysiologic insight with therapeutic potential. This could spare patients unnecessary toxic side effects or loss of vision due to therapeutic inefficacy. Consequently, there is a critical need to better classify uveitis subtypes in order to discover more effective strategies for targeted immune suppression. The long-term goal is to develop a platform for molecular characterization of uveitis that will eventually facilitate precision medicine treatment strategies. The approach is to utilize validated experimental techniques to explore novel observations obtained from single cell RNA-Sequencing (scRNA-Seq), a high-definition gene expression analysis of ocular immune cells. Specifically, the central hypothesis for this proposal, that type 1 conventional dendritic cells (DC1s) promote the resolution of ocular inflammation, is based on primary observations of human ocular DC1s obtained via scRNA-Seq. The overall objectives of this proposal are to a) ascertain the role of DC1s in ocular inflammation, and b) to determine how patient-specific alterations in DC1s contribute to the chronicity of uveitis. Toward these ends, the following specific aims will be pursued: 1) Test the hypothesis that the ocular microenvironment promotes pro-resolution DC1s. 2) Test the hypothesis that DC1s promote the resolution of ocular inflammation. 3) Test the hypothesis that patient-specific DC1 gene expression signatures predict uveitis chronicity. In aim 1, the effect of the ocular microenvironment on DC1s will be tested by a) analyzing aqueous-exposed human DC1s in vitro and b) analyzing murine DC1s in 2 distinct models of ocular inflammation in vivo. In aim 2, the specific effects of DC1s on disease chronicity during ocular inflammation will be explored using mice deficient in DC1s. Finally, in aim 3, the transcriptional signatures that best predict chronic and acute uveitis will be validated and used to a) identify peripheral blood biomarkers and b) identify potential therapeutic targets. This contribution will be significant because it will improve our ability to define uveitis entities based on pathophysiologic mechanisms that can then be appropriately targeted by precision application of immune therapies. This proposal is innovative in the use of scRNA-Seq to generate testable hypotheses from in-depth analysis of patient samples. Ultimately, improved molecular characterization of uveitis based on identification and validation of differential gene expression provides a scaffold for future analyses that will facilitate a precision medicine approach to immune therapy.

项目摘要 非感染葡萄膜炎由一组异质性临床定义的疾病组成，经验性疾病 治疗失败了许多患者。另一方面，葡萄膜炎的分子表征将导致更多 疾病亚型的精确分类，同时提供病理生理洞察力 治疗潜力。这可能使患者免于不必要的毒性副作用或由于 治疗效率低下。因此，迫切需要更好地对葡萄膜炎进行分类以便 发现针对靶向免疫抑制的更有效的策略。长期目标是开发 葡萄膜炎分子表征的平台，最终将促进精密医学治疗 策略。该方法是利用经过验证的实验技术来探索获得的新观测 从单细胞RNA - 序列（SCRNA-SEQ）中，眼部免疫的高定义基因表达分析 细胞。具体而言，该提案的中心假设，该型1型常规树突状细胞（DC1） 促进眼部炎症的分辨率是基于人眼DC1的主要观察结果 通过scrna-seq获得。该提案的总体目标是a）确定DC1在眼中的作用 炎症，b）确定DC1中患者特异性的变化如何促进 葡萄膜炎。在这些目的方面，将追求以下特定目标：1）检验以下假设。 微环境促进了促分辨率DC1。 2）检验DC1促进分辨率的假设 眼部炎症。 3）测试患者特异性DC1基因表达特征预测的假设 葡萄膜炎慢性。在AIM 1中，将通过A）分析来测试眼微环境对DC1的影响 在体外暴露于水性的人DC1和b）分析2种不同模型的鼠DC1 体内炎症。在AIM 2中，DC1对眼部炎症期间疾病慢性的具体影响将 使用DC1中缺乏的小鼠进行探索。最后，在AIM 3中，最能预测的转录签名 慢性和急性葡萄膜炎将得到验证并用于a）识别外围血液生物标志物，b）识别 潜在的治疗靶标。这项贡献将是重要的，因为它将提高我们定义的能力 基于病理生理机制的葡萄膜炎实体，然后可以通过精确对准这些机制 免疫疗法的应用。该建议在使用SCRNA-SEQ生成可测试的方面具有创新性 对患者样品的深入分析的假设。最终，改进的分子表征 基于鉴定和差异基因表达验证的葡萄膜炎为将来提供了脚手架 分析将促进免疫治疗的精确医学方法。