Developmental exposure to chlordecone induces transgenerational effects in somatic prostate tissue which are associated with epigenetic histone trimethylation changes
Developmental exposure to chlordecone induces transgenerational effects in somatic prostate tissue which are associated with epigenetic histone trimethylation changes
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DOI：
10.1016/j.envint.2021.106472
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发表时间：
2021-03-10
影响因子：
11.8
通讯作者：
Smagulova, Fatima
中科院分区：
文献类型：
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作者：
Legoff, Louis;D'Cruz, Shereen Cynthia;Smagulova, Fatima
背景：十氯酮（CD）又称甲酮（Kepone），是一种有机氯杀虫剂，在法属西印度群岛的香蕉作物中使用。由于土壤和水的长期污染，人口仍然暴露于CD。成年期暴露于CD与前列腺癌（PCa）风险增加相关。目的：我们研究了CD对小鼠前列腺组织的跨代效应。方法：我们将怀孕的瑞士小鼠暴露于CD。分析了直接暴露（F1）和未暴露（F3）雄性后代的前列腺。我们使用免疫荧光、RNA-seq和ChIP-seq技术对前列腺中的染色质状态进行综合分析。结果：在F1和F3代中，我们观察到前列腺上皮内瘤变表型（PIN）增加。使用RNA-seq在CD衍生的F1和F3前列腺中进行的转录组学分析显示，F1中的970个基因和F3中的218个基因差异表达。两个数据集中的差异表达基因可以根据共同的生物学过程进行聚类，？细胞分化？?发展过程？?信号的调节，这表明在两代中，类似的过程受到了干扰。我们检测到，在两个数据集中，几个Hox基因上调;在F1中，主要在Hoxb和Hoxd中检测到表达，在F3中，在Hoxa家族基因中检测到表达。使用大量的生物学重复和RT-qPCR，我们发现与睾酮合成有关的基因（Akr 1b 3，Cyp 11 a1，Cyp 17 a1，Srd 5a 1）在PIN样品中显著上调; Cyp 19 a1，将睾酮转化为雌二醇也升高。我们发现，在F1和F3前列腺含有PIN的Esr 2表达显着增加。含有PIN的样品具有自我更新相关基因（Nanog、Tbx 3、Sox 2、Sox 3、Rb 1）表达的强烈增加。我们观察到肝脏的变化，F1 CD暴露雄性在F1中与DNA修复、基质胶原和炎症相关途径相关的基因表达增加，但在F3成人CD衍生肝脏中则没有。RNA转录的变化与表观遗传变化有关。具体而言，我们发现F1小鼠前列腺中H3 K4三甲基化（H3 K4 me 3）的总体增加和H3 K27三甲基化（H3 K27 me 3）的减少。ChIP-seq分析显示，F1中的129个区域和F3中的240个区域在CD衍生的前列腺中获得了改变的H3 K4 me 3占有率，包括两个数据集中Hoxa家族基因的几个启动子的最高增加。H3 K4 me 3的突变在两个世代中重叠了73个基因，包括参与增殖调节的基因Tbx 2、Stat 3、Stat 5a、Pou 2f 3和同源异型盒基因Hoxa 13、Hoxa 9。
Background: Chlordecone (CD), also known as Kepone, is an organochlorine insecticide that has been used in banana crops in the French West Indies. Due to long-term contamination of soils and water, the population is still exposed to CD. Exposure to CD in adulthood is associated with an increased risk of prostate cancer (PCa). Objectives: We examined the transgenerational effects of CD on murine prostate tissue. Methods: We exposed pregnant Swiss mice to CD. The prostates from directly exposed (F1) and non-exposed (F3) male progeny were analyzed. We used immunofluorescence, RNA-seq and ChIP-seq techniques for the comprehensive analyses of chromatin states in prostate. Results: We observed an increased prostatic intraepithelial neoplasia phenotype (PIN) in both F1 and F3 generations. Transcriptomic analysis in CD-derived F1 and F3 prostate using RNA-seq revealed that 970 genes in F1 and 218 in F3 genes were differentially expressed. The differentially expressed genes in both datasets could be clustered accordingly to common biological processes, ?cell differentiation?, ?developmental process?, ?regulating of signaling?, suggesting that in both generations similar processes were perturbed. We detected that in both datasets several Hox genes were upregulated; in F1, the expression was detected mainly in Hoxb and Hoxd, and in F3, in Hoxa family genes. Using a larger number of biological replicates and RT-qPCR we showed that genes implicated in testosterone synthesis (Akr1b3, Cyp11a1, Cyp17a1, Srd5a1) were dramatically upregulated in PIN samples; Cyp19a1, converting testosterone to estradiol was elevated as well. We found a dramatic increase in Esr2 expression both in F1 and F3 prostates containing PIN. The PIN-containing samples have a strong increase in expression of self-renewal-related genes (Nanog, Tbx3, Sox2, Sox3, Rb1). We observed changes in liver, F1 CDexposed males have an increased expression of genes related to DNA repair, matrix collagen and inflammation related pathways in F1 but not in F3 adult CD-derived liver. The changes in RNA transcription were associated with epigenetic changes. Specifically, we found a global increase in H3K4 trimethylation (H3K4me3) and a decrease in H3K27 trimethylation (H3K27me3) in prostate of F1 mice. ChIP-seq analysis showed that 129 regions in F1 and 240 in F3 acquired altered H3K4me3 occupancy in CD-derived prostate, including highest increase at several promoters of Hoxa family genes in both datasets. The alteration in H3K4me3 in both generations overlap 73 genes including genes involved in proliferation regulation, Tbx2, Stat3, Stat5a, Pou2f3 and homeobox genes Hoxa13, Hoxa9.