The NFkB System in Dendritic Cells

项目摘要/摘要 树突状细胞(DC)在针对病原体的靶向免疫反应中发挥关键作用，两者都通过以下方式发挥作用 通过分泌强大的先天免疫和炎性细胞因子。他们的区别在于 从在骨髓中发现的造血干细胞转变为多个亚型 炎症/适应性免疫或更多的先天免疫功能。对这些差异的错误调控 过程是包括急性髓系白血病(AML)在内的骨髓增生性疾病(MPD)的根本原因， 和炎症性疾病朗格汉斯细胞组织细胞增生症(LCH)。这些分化过程可能是 概述了关键的髓系分化和生长因子GM-CSF和Flt3L的体外实验，以及先前的工作 表明NFkappaB信号系统是一个重要的调节因子。然而，考虑到这种多方面的复杂性- 转录因子，多调节信号系统，其在亚型分化、增殖和蛋白合成中的作用 人们对这种疾病仍然知之甚少。 在拟议的项目中，我们将研究NFkappaB控制在协调 树突状细胞分为两条发育途径。基于我们的初步结果，我们建议 最重要的假设是在DC过程中NFkappaB信号系统的正确逐步组装 分化途径对细胞增殖和成熟是至关重要的，而经典的IkappaBalpha，-β，-epsilon起中介作用 暂时性NFkappaB炎症反应，最近描述的IkappaBome发挥了关键的缓冲和 在DC分化中的协调作用。对IkappaBome的错误调控导致了严重但令人惊讶的不同 DC两条发育途径的表型。 我们将结合一个经过实验验证的NFkappaB信号在树突状细胞期间的数学模型 分化和功能，用定量生化、流式细胞仪和活细胞荧光 显微镜、干涉测量和谱系追踪研究，涉及敲门报告小鼠，以及一些新的 遗传性小鼠品系揭示与骨髓增殖症相关的NFkappaB调控失调的分子基础 疾病(MPD)和朗格汉斯细胞组织细胞增生症(LCH)。我们将应用这些关于IkappaBome的见解 检查患者样本和潜在药物靶标的有效性和风险的法规。

Project Summary/Abstract Dendritic cells (DCs) play key roles in targeting immune responses to pathogens, both by functioning as antigen presenting cells and by secreting potent innate immune and inflammatory cytokines. They differentiate from hematopoietic stem cells found in the bone marrow into multiple subtypes that have more inflammatory/adaptive immune or more innate immune functions. Misregulation of these differentiation processes is the root cause of myeloproliferative disorders (MPD) including acute myeloid leukemia (AML), and the inflammatory disease Langerhans Cell Histiocytosis (LCH). These differentiation processes may be recapitulated ex vivo with key myeloid differentiation and growth factors GM-CSF and Flt3L, and prior work has shown that the NFkappaB signaling system is an important regulator. Yet given the complexity of this multi- transcription factor, multi-regulator signaling system, its roles in subtype-differentiation, proliferation, and in disease remain poorly understood. In the proposed project we will examine the role of NFkappaB control in coordinating differentiation programs of dendritic cells into two developmental pathways. Based on our preliminary results we propose the overarching hypothesis that proper stepwise assembly of the NFkappaB signaling system during DC differentiation pathways is critical for phasing proliferation and maturation; while classical IkappaBalpha, -Beta, -epsilon mediate transient NFkappaB inflammatory responses, the recently described IkappaBsome plays a critical buffering and coordinating role in DC differentiation. Misregulation of the IkappaBsome leads to severe but surprisingly different phenotypes in the two DC developmental pathways. We will combine an experimentally validated mathematical model of NFkappaB signaling during dendritic cell differentiation and function, with quantitative biochemical, flow cytometric, and live cell fluorescence microscopy, interferometry and lineage tracking studies involving knockin reporter mice, and a number of novel genetic mouse strains to uncover the molecular basis of NFkappaB misregulation associated with myeloprolferative disorders (MPD) and Langerhans Cell Histiocytosis (LCH). We will apply these insights about the IkappaBsome regulation to examine patient samples and the efficacy and risks of potential drug targets.