Transcriptional and Proteomic Characterization of Telomere-Induced Senescence in a Human Alveolar Epithelial Cell Line.
Transcriptional and Proteomic Characterization of Telomere-Induced Senescence in a Human Alveolar Epithelial Cell Line.
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DOI：
10.3389/fmed.2021.600626
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发表时间：
2021
影响因子：
3.9
通讯作者：
Alder JK
中科院分区：
文献类型：
--
作者：
Sullivan DI;Jiang M;Hinchie AM;Roth MG;Bahudhanapati H;Nouraie M;Liu J;McDyer JF;Mallampalli RK;Zhang Y;Kass DJ;Finkel T;Alder JK
端粒功能障碍导致的细胞衰老已被假设在包括特发性肺纤维化（IPF）在内的年龄相关疾病中发挥作用。据推测，来自衰老肺泡上皮的旁分泌介质向周围的间充质细胞发出信号，并参与疾病的发病。然而，端粒诱导的肺泡上皮衰老的小鼠模型不能显示在衰老的人类细胞中观察到的典型衰老相关分泌表型（SASP）。为了了解人类对端粒功能障碍的特异性反应，我们在人类肺泡上皮细胞系中模拟了端粒功能障碍诱导的衰老。我们假设该系统将使我们能够探测体外转录和蛋白质组学衰老途径的差异，并鉴定可能导致IPF发病机制的新分泌蛋白（分泌组）变化。在诱导端粒功能障碍后，观察到强烈的衰老表型。衰老细胞的RNA-seq分析揭示了SASP，并与先前的小鼠数据进行比较，突出了对端粒功能障碍的反应差异。我们使用一种能够标记分泌蛋白的新型生物素连接酶对衰老细胞进行了蛋白质组学分析。然后在IPF和对照患者血浆中评估从转录和分泌组数据中选择的候选生物标志物。发现四种新蛋白在患者组之间存在差异表达：stanniocalcin-1、contactin-1、tenascin C和总抑制素。我们的数据表明，人类端粒诱导的肺泡上皮衰老导致转录SASP，这与在类似的小鼠细胞中看到的不同。我们的研究结果表明，考虑到对端粒功能障碍的物种特异性反应的可能性，动物模型的研究应该仔细验证。我们还描述了一种实用的方法来研究人类端粒诱导的肺泡上皮细胞衰老的后果。
Cellular senescence due to telomere dysfunction has been hypothesized to play a role in age-associated diseases including idiopathic pulmonary fibrosis (IPF). It has been postulated that paracrine mediators originating from senescent alveolar epithelia signal to surrounding mesenchymal cells and contribute to disease pathogenesis. However, murine models of telomere-induced alveolar epithelial senescence fail to display the canonical senescence-associated secretory phenotype (SASP) that is observed in senescent human cells. In an effort to understand human-specific responses to telomere dysfunction, we modeled telomere dysfunction-induced senescence in a human alveolar epithelial cell line. We hypothesized that this system would enable us to probe for differences in transcriptional and proteomic senescence pathways in vitro and to identify novel secreted protein (secretome) changes that potentially contribute to the pathogenesis of IPF. Following induction of telomere dysfunction, a robust senescence phenotype was observed. RNA-seq analysis of the senescent cells revealed the SASP and comparisons to previous murine data highlighted differences in response to telomere dysfunction. We conducted a proteomic analysis of the senescent cells using a novel biotin ligase capable of labeling secreted proteins. Candidate biomarkers selected from our transcriptional and secretome data were then evaluated in IPF and control patient plasma. Four novel proteins were found to be differentially expressed between the patient groups: stanniocalcin-1, contactin-1, tenascin C, and total inhibin. Our data show that human telomere-induced, alveolar epithelial senescence results in a transcriptional SASP that is distinct from that seen in analogous murine cells. Our findings suggest that studies in animal models should be carefully validated given the possibility of species-specific responses to telomere dysfunction. We also describe a pragmatic approach for the study of the consequences of telomere-induced alveolar epithelial cell senescence in humans.