Histone Deacetylase 3 is required for efficient T cell development

Hdac3 is an important target of HDAC inhibitors used in the treatment of cutaneous T cell lymphoma. In order to gain an understanding of Hdac3 function in T cells,we deleted Hdac3 from early mouse thymocytes using LCK-Cre. Hdac3 deletion resulted in a loss of single positive thymocytes due to a defect in positive selection at the double positive (DP) stage of thymocyte development. To better characterize this defect, we sorted the DP1 and DP2 populations to for gene expression profiling. Total RNA was extracted from DP1 (GFP+CD4+CD8+CD5loTCRblo) or DP2 (GFP+CD4+CD8+CD5hiTCRbint) thymocytes isolated by FACS from Hdac3+/+ or Hdac3F/F LCK-Cre+ animals. Libraries were constructed from rRNA-depleted total RNA pools to identify altered gene expression in DP populations following Hdac3 deletion.Hdac3是用于治疗皮肤T细胞淋巴瘤的组蛋白去乙酰化酶（HDAC）抑制剂的重要作用靶点。为阐明Hdac3在T细胞中的功能，我们借助LCK-Cre重组酶系统，从小鼠早期胸腺细胞中敲除了Hdac3基因。Hdac3基因敲除会导致单阳性胸腺细胞丢失，该表型源于胸腺细胞发育双阳性（double positive, DP）阶段的阳性选择缺陷。为更好地表征这一发育缺陷，我们分选出DP1与DP2细胞群以开展基因表达谱分析。我们从经荧光激活细胞分选（FACS）分离自Hdac3+/+或Hdac3F/F LCK-Cre+小鼠的DP1（绿色荧光蛋白（GFP）+CD4+CD8+CD5loTCRblo）或DP2（GFP+CD4+CD8+CD5hiTCRbint）胸腺细胞中提取总RNA。随后，我们以经核糖体RNA去除（rRNA-depleted）处理的总RNA混合样本构建测序文库，以此鉴定Hdac3基因敲除后双阳性细胞群中出现表达异常的基因谱。