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 非感染性葡萄膜炎由一组异质性的临床定义的疾病组成， 治疗使许多患者失败。另一方面，葡萄膜炎的分子特征将导致更多的 疾病亚型的精确分类，同时提供病理生理学见解， 治疗潜力这可以避免患者不必要的毒副作用或视力丧失， 治疗无效。因此，迫切需要更好地对葡萄膜炎亚型进行分类，以便 发现更有效的靶向免疫抑制策略。长期目标是开发一个 用于葡萄膜炎分子表征的平台，最终将促进精确的药物治疗 战略布局该方法是利用经过验证的实验技术来探索所获得的新观测结果 从单细胞RNA测序（scRNA-Seq），一种高清晰度的眼部免疫基因表达分析， 细胞具体来说，这项提议的核心假设是，1型传统树突状细胞（DC 1） 促进眼部炎症的消退，是基于对人类眼部DC 1的初步观察 通过scRNA-Seq.本提案的总体目标是：a）确定DC 1在眼内 B）确定DC 1的患者特异性改变如何导致慢性炎症， 葡萄膜炎为了达到这些目的，将追求以下具体目标：1）测试假设， 微环境促进亲分辨率DC 1。2)测试DC 1促进以下问题解决的假设 眼部炎症3)检验患者特异性DC 1基因表达特征预测 慢性葡萄膜炎在目的1中，将通过以下方式测试眼睛微环境对DC 1的影响：a）分析 B）在两种不同的眼部模型中分析鼠DC 1 体内炎症。在目的2中，在眼部炎症期间DC 1对疾病慢性化的特异性作用将 使用DC 1缺陷的小鼠进行研究。最后，在目标3中，最能预测 将对慢性和急性葡萄膜炎进行验证，并用于a）鉴定外周血生物标志物和B）鉴定 潜在的治疗目标。这一贡献将是重大的，因为它将提高我们的能力， 基于病理生理学机制的葡萄膜炎实体，然后可以通过精确定位适当靶向 免疫疗法的应用。该提议在使用scRNA-Seq来产生可测试的 对患者样本进行深入分析的假设。最终，改进的分子表征 基于差异基因表达的鉴定和验证的葡萄膜炎为未来的研究提供了一个框架。 这些分析将有助于免疫治疗的精确医学方法。