■肾素表达细胞是对维持体内平衡至关重要的肌内分泌细胞。肾素受cAMP调节,p300(组蛋白乙酰转移酶p300)/CBP(CREB结合蛋白),和Brd4(含溴结构域蛋白4)蛋白和相关途径。然而,抑制这些途径后发生的具体调控变化尚不清楚.
■我们用3种针对肾素转录所需的不同因子的抑制剂处理了As4.1细胞(来自组成型表达肾素的小鼠近球细胞的肿瘤细胞):H-89-二盐酸盐,PKA(蛋白激酶A)抑制剂;JQ1,Brd4溴结构域抑制剂;和A-485,p300/CBP抑制剂。我们执行了ATAC-seq,单细胞RNA测序,CUT&Tag,和染色质免疫沉淀测序H3K27ac和p300结合在处理和对照As4.1细胞的生物复制上。
■在对每种抑制剂的反应中,Ren1表达显著降低并且在洗出时是可逆的。Ren1基因座的染色质可及性没有显着变化,但在远端元件处整体降低。抑制PKA导致在Ren1超增强子区域内特异性结合H3K27ac和p300的显著降低。Further,我们确定了每个抑制性治疗共有的富集TF(转录因子)基序。最后,我们确定了一组9个基因,在3个肾素调节途径中的每一个中都有推定的作用,并观察到每个基因都表现出差异可接近的染色质,基因表达,H3K27ac,和p300在它们各自的基因座处结合。
■在组成型合成和释放肾素的细胞中肾素表达的抑制受到表观遗传开关从活性状态到平衡状态的调节,所述表观遗传开关与减少的细胞-细胞通讯和上皮-间质转化相关。这项工作突出并有助于定义肾素细胞在肌内分泌和收缩表型之间交替所必需的因素。
UNASSIGNED: Renin-expressing cells are myoendocrine cells crucial for the maintenance of homeostasis. Renin is regulated by cAMP, p300 (histone acetyltransferase p300)/CBP (CREB-binding protein), and Brd4 (bromodomain-containing protein 4) proteins and associated pathways. However, the specific regulatory changes that occur following inhibition of these pathways are not clear.
UNASSIGNED: We treated As4.1 cells (tumoral cells derived from mouse juxtaglomerular cells that constitutively express renin) with 3 inhibitors that target different factors required for renin transcription: H-89-dihydrochloride, PKA (protein kinase A) inhibitor; JQ1, Brd4 bromodomain inhibitor; and A-485, p300/CBP inhibitor. We performed assay for transposase-accessible chromatin with sequencing (ATAC-seq), single-cell RNA sequencing, cleavage under targets and tagmentation (CUT&Tag), and chromatin immunoprecipitation sequencing for H3K27ac (acetylation of lysine 27 of the histone H3 protein) and p300 binding on biological replicates of treated and control As4.1 cells.
UNASSIGNED: In response to each inhibitor, Ren1 expression was significantly reduced and reversible upon washout. Chromatin accessibility at the Ren1 locus did not markedly change but was globally reduced at distal elements. Inhibition of PKA led to significant reductions in H3K27ac and p300 binding specifically within the Ren1 super-enhancer region. Further, we identified enriched TF (transcription factor) motifs shared across each inhibitory treatment. Finally, we identified a set of 9 genes with putative roles across each of the 3 renin regulatory pathways and observed that each displayed differentially accessible chromatin, gene expression, H3K27ac, and p300 binding at their respective loci.
UNASSIGNED: Inhibition of renin expression in cells that constitutively synthesize and release renin is regulated by an epigenetic switch from an active to poised state associated with decreased cell-cell communication and an epithelial-mesenchymal transition. This work highlights and helps define the factors necessary for renin cells to alternate between myoendocrine and contractile phenotypes.