DataSheet_3_Impact of Interleukin 10 Deficiency on Intestinal Epithelium Responses to Inflammatory Signals.docx

Interleukin 10 (IL-10) is a pleiotropic, anti-inflammatory cytokine that has a major protective role in the intestine. Although its production by cells of the innate and adaptive immune system has been extensively studied, its intrinsic role in intestinal epithelial cells is poorly understood. In this study, we utilised both ATAC sequencing and RNA sequencing to define the transcriptional response of murine enteroids to tumour necrosis factor (TNF). We identified that the key early phase drivers of the transcriptional response to TNF within intestinal epithelium were NFκB transcription factor dependent. Using wild-type and Il10−/− enteroid cultures, we showed an intrinsic, intestinal epithelium specific effect of IL-10 deficiency on TNF-induced gene transcription, with significant downregulation of identified NFκB target genes Tnf, Ccl20, and Cxcl10, and delayed overexpression of NFκB inhibitor encoding genes, Nfkbia and Tnfaip3. IL-10 deficiency, or immunoblockade of IL-10 receptor, impacted on TNF-induced endogenous NFκB activity and downstream NFκB target gene transcription. Intestinal epithelium-derived IL-10 appears to play a crucial role as a positive regulator of the canonical NFκB pathway, contributing to maintenance of intestinal homeostasis. This is particularly important in the context of an inflammatory environment and highlights the potential for future tissue-targeted IL-10 therapeutic intervention.

白细胞介素10（Interleukin 10，IL-10）是一种多效性抗炎细胞因子，在肠道中发挥关键保护作用。尽管先天免疫与适应性免疫细胞分泌该细胞因子的机制已被广泛研究，但肠上皮细胞中IL-10的内在功能仍未得到充分阐明。本研究利用转座酶可及性测序（ATAC sequencing）与RNA测序（RNA sequencing），明确了小鼠肠类器官（murine enteroids）对肿瘤坏死因子（tumour necrosis factor，TNF）的转录应答特征。研究发现，肠上皮细胞内TNF诱导转录应答的早期核心调控因子依赖于核因子κB转录因子（NFκB transcription factor）。通过野生型与Il10基因敲除（Il10⁻/⁻）肠类器官培养体系，本研究证实IL-10缺失对TNF诱导的基因转录具有肠上皮细胞特异性的内在影响：可显著下调已鉴定的NFκB靶基因Tnf、Ccl20及Cxcl10的表达，并延迟NFκB抑制编码基因Nfkbia与Tnfaip3的过度表达。IL-10缺失或IL-10受体免疫阻断，均可影响TNF诱导的内源性NFκB活性及其下游靶基因的转录。肠上皮来源的IL-10似乎作为经典核因子κB通路的正向调控因子发挥关键作用，有助于维持肠道稳态。这一作用在炎性环境中尤为关键，也为未来开发组织靶向性IL-10治疗干预手段提供了潜在可能。