RNA-seq profiling of thymic epithelial cells from Mcl1 knockout and wildtype mice. Mus musculus
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T cell differentiation is governed by interactions with thymic epithelial cells (TECs) and defects in this process undermine immune function and tolerance. To uncover new strategies to restore thymic function and adaptive immunity in immunodeficiency, we sought to determine the molecular mechanisms that control life and death decisions in TEC. We created a mouse model which specifically deleted the pro-survival gene Mcl1 in TEC. We found that while BCL-2 and BCL-XL were dispensable for TEC homeostasis, MCL-1 deficiency impacted on TEC as early as E15.5, resulting in early thymic atrophy and T cell lymphopenia, with near complete loss of thymic tissue by 2 months of age. MCL-1 was not necessary for TEC differentiation but was continually required for the survival of medullary TEC, including autoimmune regulator (AIRE) expressing TECs and the maintenance of overall thymic architecture. To understand the molecular mechanisms in more detail, RNA-seq profiling was undertaken of cortical and medullary thymic epithelial cells (cTECs and mTECs) from wildtype and knockout mice. Overall design: The number of biological replicates was n=4 for WT cTECs, n=2 for WT mTECs, n=1 for KO cTECs and n=1 for KO mTECs.
T细胞分化受与胸腺上皮细胞（thymic epithelial cells, TECs）的相互作用调控，该过程的缺陷会损害免疫功能与免疫耐受。为探索在免疫缺陷状态下恢复胸腺功能及适应性免疫的新策略，本研究旨在阐明调控胸腺上皮细胞生死抉择的分子机制。我们构建了在胸腺上皮细胞中特异性敲除促存活基因Mcl1的小鼠模型。研究发现，尽管BCL-2与BCL-XL对胸腺上皮细胞稳态非必需，但MCL-1缺失早在胚胎发育第15.5天（E15.5）便对胸腺上皮细胞造成影响，引发早期胸腺萎缩与T细胞淋巴细胞减少症，至小鼠2月龄时胸腺组织近乎完全缺失。MCL-1对胸腺上皮细胞分化并非必需，但持续参与维持髓质胸腺上皮细胞（medullary TECs, mTECs）的存活——包括表达自身免疫调节因子（autoimmune regulator, AIRE）的胸腺上皮细胞——以及整体胸腺结构的稳态维持。为更深入地解析其分子机制，本研究对野生型（Wildtype, WT）与基因敲除型（Knockout, KO）小鼠的皮质胸腺上皮细胞（cortical thymic epithelial cells, cTECs）及髓质胸腺上皮细胞（mTECs）开展了RNA测序（RNA-seq）转录组分析。实验整体设计：野生型皮质胸腺上皮细胞（WT cTECs）的生物学重复数为n=4，野生型髓质胸腺上皮细胞（WT mTECs）为n=2，基因敲除型皮质胸腺上皮细胞（KO cTECs）与基因敲除型髓质胸腺上皮细胞（KO mTECs）均为n=1。
