Neuroprotective protein ADNP-dependent histone remodeling complex promotes Th2 immune cell differentiation (ATAC-Seq). Neuroprotective protein ADNP-dependent histone remodeling complex promotes Th2 immune cell differentiation (ATAC-Seq)

Type-2 immune responses are critical in tissue homeostasis, anti-helminth immunity and allergy. Th2 cells produce interleukin (IL)-4, IL-5 and IL-13 from the type-2 gene cluster under regulation by transcription factors (TFs) including GATA3. To better understand this system we performed CRISPR-Cas9 screens targeting 1131 TFs during Th2 cell differentiation. We discovered that ADNP is indispensable for immune reactions to allergen. Mechanistically, ADNP performs a previously unappreciated role in gene activation forming a critical bridge in the transition from pioneer TFs to chromatin remodelling by recruiting CHD4 and BRG1. Although GATA3 and AP-1 bound the type-2 cytokine locus in the absence of ADNP, they were unable to initiate histone acetylation or DNA accessibility, resulting in highly impaired type-2 cytokine expression. Similar ADNP-dependent regulation was observed for the Maf and Il4ra genes which are known regulators of Th2 differentiation. Our results demonstrate an important role for ADNP in promoting immune cell specialization. Overall design: ATACseq data comparing Cd4Cre with Cd4CreAdnpflox Th2 cells

2型免疫应答（Type-2 immune responses）在组织稳态、抗蠕虫免疫及过敏反应中发挥关键调控作用。Th2细胞（Th2 cells）在包括GATA3在内的转录因子（transcription factors, TFs）的调控下，从2型细胞因子基因簇中表达白细胞介素（interleukin, IL）-4、IL-5及IL-13。为进一步解析该免疫调控系统，我们在Th2细胞分化过程中开展了针对1131个转录因子的CRISPR-Cas9筛选（CRISPR-Cas9 screens）。研究发现ADNP（活动依赖性神经保护蛋白，Activity-Dependent Neuroprotective Protein）对过敏原免疫反应不可或缺。从机制上看，ADNP在基因激活中具有此前未被阐明的重要功能：它通过招募CHD4与BRG1，在先驱转录因子（pioneer TFs）到染色质重塑的过程中搭建了关键桥梁。尽管在ADNP缺失的情况下，GATA3与AP-1仍可结合至2型细胞因子基因座，但二者无法启动组蛋白乙酰化（histone acetylation）或DNA可及性（DNA accessibility）的建立，最终导致2型细胞因子的表达严重受损。我们在已知的Th2细胞分化调控因子Maf基因与Il4ra基因中，也观察到了类似的ADNP依赖性调控模式。本研究结果证实，ADNP在免疫细胞特化过程中具有重要的调控功能。实验设计：采用ATAC-seq（转座酶可及性染色质测序，Assay for Transposase-Accessible Chromatin using sequencing）技术对比Cd4Cre与Cd4CreAdnpflox型Th2细胞的测序数据。