Long noncoding RNA TINCR is a novel regulator of human bronchial epithelial cell differentiation state.
Long noncoding RNA TINCR is a novel regulator of human bronchial epithelial cell differentiation state.
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DOI：
10.14814/phy2.14727
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发表时间：
2021-03
影响因子：
2.5
通讯作者：
Kaminski N
中科院分区：
文献类型：
--
作者：
Omote N;Sakamoto K;Li Q;Schupp JC;Adams T;Ahangari F;Chioccioli M;DeIuliis G;Hashimoto N;Hasegawa Y;Kaminski N
长非编码RNA(LncRNAs)具有多种控制细胞分化和组织发育的生物学功能。关于lncRNAs在人类肺中的作用的了解仍然有限。在这里，我们发现了末端分化诱导的lncRNA(TINCR)在支气管细胞分化中的调节作用。RNA原位杂交结果显示，TINCR主要在正常人肺组织的支气管上皮细胞中表达。我们对正常人支气管上皮细胞(NHBECs)进行了TINCR抑制前后的RNA测序分析，发现603个基因在细胞黏附和迁移、伤口愈合、细胞外基质组织、组织发育和分化方面具有丰富的差异表达。为了研究TINCR在NHBECs分化中的作用，我们采用了气液界面培养和3D类有机物形成实验。在分化过程中，TINCR表达上调，TINCR的缺失显著诱导了早期的基底样细胞表型(TP63)和晚期的纤毛细胞分化(FOXJ1)，而TINCR过表达抑制了基底细胞的表型和向纤毛细胞的分化。SOX2和NOTCH基因(NOTCH1、HES1和JAG1)等关键的分化调控因子在TINCR抑制后显著上调，而在TINCR过表达后显著下调。RNA免疫沉淀分析表明，Staufen1蛋白与SOX2、HES1和JAG1mRNA直接结合需要TINCR。Staufen1的缺失诱导了TP63、SOX2、NOTCH1、HES1和JAG1mRNA的表达，而TINCR的过表达部分抑制了这些基因的表达。总之，TINCR是一种新的支气管细胞分化规则，影响下游调控基因，如SOX2和Noch基因，可能与Staufen1协同作用。提出的模型代表TINCR和STAU1共同控制着支气管细胞分化的关键调控因子的表达，并维持正常的分化状态。
Long‐noncoding RNAs (lncRNAs) have numerous biological functions controlling cell differentiation and tissue development. The knowledge about the role of lncRNAs in human lungs remains limited. Here we found the regulatory role of the terminal differentiation‐induced lncRNA (TINCR) in bronchial cell differentiation. RNA in situ hybridization revealed that TINCR was mainly expressed in bronchial epithelial cells in normal human lung. We performed RNA sequencing analysis of normal human bronchial epithelial cells (NHBECs) with or without TINCR inhibition and found the differential expression of 603 genes, which were enriched for cell adhesion and migration, wound healing, extracellular matrix organization, tissue development and differentiation. To investigate the role of TINCR in the differentiation of NHBECs, we employed air–liquid interface culture and 3D organoid formation assay. TINCR was upregulated during differentiation, loss of TINCR significantly induced an early basal‐like cell phenotype (TP63) and a ciliated cell differentiation (FOXJ1) in late phase and TINCR overexpression suppressed basal cell phenotype and the differentiation toward to ciliated cells. Critical regulators of differentiation such as SOX2 and NOTCH genes (NOTCH1, HES1, and JAG1) were significantly upregulated by TINCR inhibition and downregulated by TINCR overexpression. RNA immunoprecipitation assay revealed that TINCR was required for the direct bindings of Staufen1 protein to SOX2, HES1, and JAG1 mRNA. Loss of Staufen1 induced TP63, SOX2, NOTCH1, HES1, and JAG1 mRNA expressions, which TINCR overexpression suppressed partially. In conclusion, TINCR is a novel regular of bronchial cell differentiation, affecting downstream regulators such as SOX2 and NOTCH genes, potentially in coordination with Staufen1. Proposed model representing that TINCR together with STAU1 controls the expression of critical regulators of bronchial cell differentiation and maintains the normal differentiation state.