Regulation of Nfil3 in innate lymphocyte-mediated host immunity

Nfil3 在先天淋巴细胞介导的宿主免疫中的调节

• 批准号: 9895620
• 负责人: Joseph Chai-Yuen Sun
• 金额: $ 56.76万
• 依托单位: SLOAN-KETTERING INST CAN RESEARCH
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-04-01 至 2022-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9895620
• 关键词: Antigens; B-Lymphocytes; Bacterial Infections; Binding; Binding Sites; Biological Assay; Blood Circulation; CRISPR/Cas technology; Cell Compartmentation; Cell Line; Cell physiology; Cells; ChIP-seq; Characteristics; Citrobacter rodentium; Clinical; Clostridium difficile; Common Lymphoid Progenitor; Communicable Diseases; Data; Defect; Development; E4BP4; Education; Enhancers; Epithelial; Epithelium; Exhibits; Family; Gene Targeting; Generations; Genes; Genetic Transcription; Goals; Grant; Growth; Health; Herpesviridae Infections; Host Defense; Human; Immunity; Infection; Interleukin 2 Receptor Gamma; Interleukin-15; Interleukin-2; Interleukin-7; Intestines; Introns; Invaded; Luciferases; Lung; Lymphocyte; Lymphoid Cell; Mammals; Mass Spectrum Analysis; Mediating; Memory; Molecular; Mouse Strains; Mucous Membrane; Murid herpesvirus 1; Mus; Names; Natural Killer Cells; Nippostrongylus; Nucleic Acid Regulatory Sequences; Organ; Play; Predisposition; Process; Proteomics; Regulation; Research; Research Project Grants; Role; Sentinel; Signal Transduction; Skin; Stat5 protein; Surface; T-Lymphocyte; Testing; Therapeutic; Tissues; Transcriptional Regulation; Transgenic Mice; Untranslated RNA; Viral; Virus; Virus Diseases; Work; chromatin immunoprecipitation; cytokine; deep sequencing; differential expression; fighting; gastrointestinal; genome editing; helminth infection; human model; influenzavirus; member; mouse model; novel; pathogen; progenitor; promoter; protein complex; public health relevance; response; stem cells; transcription factor; transcriptome sequencing

ABSTRACT Innate lymphocytes represent early sentinels critical in host immunity against pathogens. The best studied innate lymphocyte is the natural killer (NK) cell, which specifically targets virally-infected host cells. Humans deficient in NK cells have severe health complications due to susceptibility to herpesvirus infections. In addition to NK cells, a family of innate lymphoid cells (ILC) has recently been demonstrated to play an important role in protection against pathogen infection at barrier surfaces such as the intestine, lung, and skin. Our long term goals are to understand the the molecular mechanisms underlying the development of these innate lymphocytes, which will increase our understanding of the function of these cells during infection. To this end, we have recently identified a novel role for the transcription factor Nfil3 (also known as E4BP4) in development of NK cells and all members of the ILC family. Using mouse pathogens that accurately model human infectious diseases, along with newly-generated transgenic mouse models that ablate either the Nfil3 gene or evolutionarily-conserved regulatory domains, this R01 grant proposes to investigate how Nfil3 is controling the development and anti-pathogen functions of NK cells and ILCs. In Aim 1, we will investigate how Nfil3 is transcriptionally regulated in innate lymphocyte progenitors by chromatin immunoprecipitation (ChIP) for the transcription factors STAT5, PU.1, TCF-1, and Ets-1 followed by qPCR or deep sequencing (seq). In Aim 2, we will use two novel mouse strains containing a CRISPR/Cas9-targeted deletion of conserved noncoding sequences (termed Nfil3 CNS1 and CNS2) to determine the influence of intronic regulatory regions on innate lymphocyte development and effector function against virus, helminth, and bacterial infection. In Aim 3, we will use ChIP-seq, RNA-seq, and mass spectrometry to determine the genes that Nfil3 is controlling, and binding partners that influence Nfil3-mediated regulation of these target genes. Together, the studies in this proposal will not only increase our understanding of the general molecular mechanisms whereby NK cells and ILCs develop and contribute to host defense during pathogen invasion, but also establish novel clinical paradigms for how the innate lymphocyte compartment may be harnessed for therapeutic strategies against infectious disease.

抽象的 先天淋巴细胞代表了宿主对病原体的免疫至关重要的早期哨兵。研究得最好的 先天淋巴细胞是自然杀伤（NK）细胞，专门针对病毒感染的宿主细胞。人类 NK 细胞缺乏的人由于易受疱疹病毒感染而导致严重的健康并发症。此外 对于 NK 细胞来说，先天性淋巴细胞 (ILC) 家族最近被证明在 防止肠、肺和皮肤等屏障表面的病原体感染。我们的长期 目标是了解这些先天发育的分子机制 淋巴细胞，这将增加我们对这些细胞在感染过程中功能的了解。为此， 我们最近发现了转录因子 Nfil3（也称为 E4BP4）在发育中的新作用 NK 细胞和 ILC 家族的所有成员。使用小鼠病原体准确模拟人类感染 疾病，以及新产生的转基因小鼠模型，该模型消除了 Nfil3 基因或 进化保守的调控域，这项 R01 资助计划研究 Nfil3 如何控制 NK 细胞和 ILC 的发育和抗病原体功能。在目标 1 中，我们将研究 Nfil3 的作用 通过染色质免疫沉淀 (ChIP) 在先天淋巴细胞祖细胞中进行转录调节 转录因子 STAT5、PU.1、TCF-1 和 Ets-1，然后进行 qPCR 或深度测序 (seq)。在目标 2 中，我们 将使用两种含有 CRISPR/Cas9 保守非编码区靶向删除的新型小鼠品系 序列（称为 Nfil3 CNS1 和 CNS2）以确定内含子调控区对先天的影响 淋巴细胞的发育和针对病毒、蠕虫和细菌感染的效应功能。在目标 3 中，我们将 使用 ChIP-seq、RNA-seq 和质谱法确定 Nfil3 控制和结合的基因 影响 Nfil3 介导的这些靶基因调节的伙伴。总的来说，本提案中的研究 不仅会增加我们对 NK 细胞和 ILC 的一般分子机制的理解 在病原体入侵期间发展并有助于宿主防御，同时也建立新的临床范例 了解如何利用先天淋巴细胞区室来制定针对感染性的治疗策略 疾病。