Delineation of the immunobiology of sarcoidosis and characterization of the effects of Janus kinase inhibition

项目摘要 这份K08申请表中描述的学习和职业发展活动是为了让威廉博士 达姆斯基，首席研究员，在人类翻译和基础免疫学方面拥有经验和专业知识 以便成为这些领域的独立调查员。研究提案的重点是结节病， 一种特发性炎症性疾病，其特征是在受影响的组织中形成肉芽肿。结节病 导致显著的发病率和死亡率，并不成比例地影响美国的非裔美国人的治疗 对结节病的治疗目前并不令人满意。我们发现Janus Kinase(JAK)抑制剂可以用于 在其他药物无效且目前正在进行临床试验的情况下治疗结节病 以进一步评估这一点。根据我们的初步研究，我们假设JAK抑制剂通过阻断 致病的CD4T细胞产生的特异性JAK依赖的细胞因子的活性进而激活 巨噬细胞。在这项提案中，我们描述了如何使用皮肤结节病活检组织和单细胞rna。 测序以创建基于受体-配体的细胞-细胞相互作用组图以解卷卷信号驱动肉芽肿 结节病的形成。将特别关注依赖JAK的细胞因子。接下来，使用生物资源库 对10例系统性结节病患者在使用Janus治疗前和治疗期间的皮肤和血液样本进行检测 我们将使用多种方法来确定JAK抑制在这个过程中的作用机制 并评估基线和对JAK抑制剂的反应的免疫学特征的异质性。 我们还将使用蛋白质组学方法来分析10名患者在治疗前和治疗期间的血浆细胞因子水平。 JAK抑制剂治疗以确定潜在的生物标志物的反应和治疗对炎症的影响 一个系统性的层面。最后，我们将对来自多个器官(淋巴)的结节病组织进行空间转录 结节和肺)来评估我们的假设，即核心细胞因子信号推动肉芽肿的形成 结节病在不同的器官中很大程度上是保守的。总而言之，本提案的研究部分将 评估一种有效的疾病新治疗方法的分子机制 目前缺乏已获批准的治疗方法。该提案还将支持一段时间的职业发展。 我将在理查德博士的实验室接受基础免疫学和翻译免疫学方面的额外培训 Flavell在耶鲁大学医学院免疫生物学系工作。这是高度协作和支持的 研究环境与以计算为重点的定向额外职业发展活动相结合 方法、数量病理学、空间转录学和学术领导力将使我能够 作为一名内科科学家，成功地实现了研究独立，并开设了自己的实验室，这样我就可以 使用基础免疫学和翻译免疫学方法研究包括结节病在内的炎症性疾病。

Project Summary The studies and career development activities described in this K08 application are designed to equip Dr. William Damsky, the Principal Investigator, with experience and expertise in human translational and basic immunology in order to become in independent investigator in these areas. The focus of the research proposal is sarcoidosis, an idiopathic inflammatory disorder characterized by the formation of granulomas in affected tissues. Sarcoidosis causes significant morbidity and mortality and disproportionally affects African Americans in the U.S. Treatments for sarcoidosis are currently unsatisfactory. We have discovered that Janus kinase (JAK) inhibitors can be used to treat sarcoidosis in contexts where other medications have failed and are currently performing a clinical trial to evaluate this further. Based on our preliminary studies, we hypothesize that JAK inhibitors work by blocking the activity of specific JAK-dependent cytokines produced by pathogenic CD4+ T cells that in turn activate macrophages. In this proposal, we describe how we will use cutaneous sarcoidosis biopsies and single cell RNA sequencing to create a receptor-ligand based cell-cell interactome map to deconvolute signals driving granuloma formation in sarcoidosis. Special attention will be given to JAK-dependent cytokines. Next, using a biorepository of skin and blood samples from 10 patients with systemic sarcoidosis before and during treatment with a Janus kinase inhibitor, we will use multiple approaches to determine the mechanism of action of JAK inhibition in this disease and evaluate heterogeneity in immunologic characteristics at baseline and in response to a JAK inhibitor. We will also use proteomic approaches to profile plasma cytokine levels in the 10 patients before and during JAK inhibitor treatment to identify potential biomarkers of response and effects of treatment on inflammation at a systemic level. Last, we will perform spatial transcriptomics on sarcoidosis tissues from multiple organs (lymph nodes and lungs) to evaluate our hypothesis that core cytokine signals that drive granuloma formation in sarcoidosis are largely conserved among different organs. In summary, the research portion of this proposal will evaluate molecular mechanisms for a promising new treatment approach for a disease in which effective approved therapies are currently lacking. The proposal will also support a period of career development during which I will receive additional training in basic and translational immunology in the laboratory of Dr. Richard Flavell in the Department of Immunobiology at Yale School of Medicine. This highly collaborative and supportive research environment combined with directed additional career development activities focused on computational approaches, quantitative pathology, spatial transcriptomics, and academic leadership will allow me to successfully achieve research independence as a physician scientist and open my own laboratory so that I can use basic and translational immunologic approaches to study inflammatory disorders including sarcoidosis.