Epigenetic Rescue of Autism-like Social Deficits in Shank3-Deficient Mouse Models

To determine whether histone deacetylase (HDAC) inhibition has genome-wide effects on gene expression, we used RNA sequencing to analyze the mRNA profile in prefrontal cortex (PFC) of wild-type (WT) and Shank3-deficient (Shank3+/ΔC) mice treated with romidespin or saline control. We mapped the sequences to 24,420 mouse genes. Compared to WT, 365 genes in saline-treated Shank3+/ΔC mice showed a change in expression values of at least 1.2 fold and p ≤ 0.01 (213 downregulated, 152 upregulated). In romidepsin-treated Shank3+/ΔC mice, ~88% of the downregulated genes (n=187 genes) were normalized to control values. Further analysis of the romidepsin-restored genes revealed enrichment in actin cytoskeleton-mediated transport, signal transduction pathways, and developmental processes. Comparison of gene expression in prefrontal cortex of wild-type mice, saline-treated Shank3-deficient mice and romidepsin-treated mice Heterozygous Shank3+/ΔC mice (5-8 weeks old, male) expressing C-terminal (exon 21) deleted Shank3 (Jackson Labs), which exhibited the significant loss of full-length Shank3 expression, and age-matched wild-type littermates (male), were used in the experiments.

为探究组蛋白去乙酰化酶（histone deacetylase, HDAC）抑制是否会对基因表达产生全基因组范围的影响，我们采用RNA测序技术，分析了经罗米地辛（romidespin）或生理盐水对照处理的野生型（wild-type, WT）与Shank3基因缺陷型（Shank3-deficient, Shank3+/ΔC）小鼠前额叶皮层（prefrontal cortex, PFC）的mRNA表达谱。我们将测序序列比对至24420个小鼠基因。与野生型小鼠相比，生理盐水处理的Shank3+/ΔC小鼠中有365个基因的表达量变化至少达1.2倍，且p值≤0.01（其中213个基因表达下调，152个基因表达上调）。在经罗米地辛处理的Shank3+/ΔC小鼠中，约88%的下调基因（n=187）的表达水平恢复至对照水平。对罗米地辛恢复表达的基因进行进一步分析发现，其显著富集于肌动蛋白细胞骨架（actin cytoskeleton）介导的运输、信号转导通路（signal transduction pathways）以及发育过程中。本实验对野生型小鼠、生理盐水处理的Shank3基因缺陷型小鼠及罗米地辛处理的Shank3基因缺陷型小鼠的前额叶皮层基因表达进行了比较：所用实验动物为5~8周龄雄性杂合子Shank3+/ΔC小鼠——该小鼠表达C端（exon 21，外显子21）缺失的Shank3蛋白（购自杰克逊实验室（Jackson Labs）），其全长Shank3蛋白的表达量显著降低；同时使用同月龄雄性野生型同窝小鼠作为对照。