A novel regulator of dendritic cell differentiation

树突状细胞分化的新型调节剂

基本信息

批准号：
10189518
负责人：
Boris Reizis
金额：
$ 21.19万
依托单位：
NEW YORK UNIVERSITY SCHOOL OF MEDICINE
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-06-11 至 2022-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10189518
关键词：
Animals Antigens Autoimmunity CRISPR/Cas technology Cell Differentiation process Cell Line Cell physiology Cells Cholesterol Complex Coupling Dendritic Cells Development Disease Dissection FLT3 ligand FRAP1 gene Genes Genetic Genetic Screening Growth Factor Immune system In Vitro Interferon Type I Libraries Ligands Lipids Lysosomes Malignant Neoplasms Mediating Molecular Mus Mutagenesis NPC1 gene Nutrient Pathologic Pathway interactions Pattern recognition receptor Pharmacology Process Production Receptor Protein-Tyrosine Kinases Regulation Role Sentinel Signal Pathway Signal Transduction Signaling Molecule Supraoptic Vertical Ophthalmoplegia Surface T-Lymphocyte TSC1 gene Testing Virus adaptive immune response base cell growth cytokine forward genetics genetic approach genome-wide in vivo insight novel null mutation pathogen prevent progenitor protein complex response therapeutic target

项目摘要

ABSTRACT Dendritic cells (DCs) are key sentinel cells of the immune system that detect pathogens through pattern recognition receptors and orchestrate innate and adaptive immune responses. Conventional DCs (cDCs) efficiently present antigen to T lymphocytes, whereas plasmacytoid DCs (pDCs) specialize in virus-induced production of type I interferon. All DCs and their progenitors express the tyrosine kinase receptor Flt3, and its cytokine ligand Flt3L is necessary and sufficient for DC development. However, relatively little is known about the signaling pathways and molecules that regulate DC differentiation downstream of Flt3L. To dissect the molecular basis of DC differentiation, we implemented an unbiased forward genetics approach utilizing CRISPR/Cas9-based mutagenesis. In addition to known regulators, this approach yielded components of a protein complex that is enriched in DCs but has not been previously implicated in DC differentiation. The proposed project will explore the potential role of this complex in DC differentiation in vitro and in experimental animals (Aim 1) as well as the mechanism of its activity in DCs (Aim 2). If successful, these studies would provide novel insights into the molecular underpinnings of the DC differentiation, including lineage-specific signaling pathways that could be amenable to therapeutic targeting.

抽象的树突状细胞 (DC) 是免疫系统的关键前哨细胞，可通过模式检测病原体识别受体并协调先天性和适应性免疫反应。传统 DC (cDC) 有效地将抗原呈递给 T 淋巴细胞，而浆细胞样 DC (pDC) 专门负责病毒诱导的抗原生产I型干扰素。所有 DC 及其祖细胞均表达酪氨酸激酶受体 Flt3，并且其细胞因子配体 Flt3L 对于 DC 发育是必要且充分的。然而，人们知之甚少关于调节 Flt3L 下游 DC 分化的信号通路和分子。到为了剖析 DC 分化的分子基础，我们实施了一种无偏见的正向遗传学方法利用基于 CRISPR/Cas9 的诱变。除了已知的监管机构之外，这种方法还产生了蛋白质复合物的成分，富含 DC，但之前并未与 DC 相关差异化。拟议的项目将探索该复合物在 DC 分化中的潜在作用体外和实验动物（目标 1）及其在 DC 中的活性机制（目标 2）。如果如果成功，这些研究将为 DC 的分子基础提供新的见解分化，包括可能适合治疗的谱系特异性信号通路瞄准。