Abstract：
OBJECTIVE: To determine if engineered HMGA2 overexpressing uterine primary myometrial cells recapitulate the transcriptional and epigenomic features of HMGA2-subtype leiomyomas.
METHODS: Isolated primary, \"normal\" myometrial cells from 3 patients were engineered to overexpress HMGA2 to determine how HMGA2 establishes transcriptomic and epigenomic features of HMGA2-overexpressing leiomyoma.
METHODS: Academic research laboratory PATIENTS: Primary myometrial cells were isolated from normal myometrium obtained from three patients undergoing hysterectomy.
METHODS: Not applicable MAIN OUTCOME MEASURES: Determined genome-wide transcriptomic and epigenomic features of engineered HMGA2-overexpressing uterine primary myometrial cells.
RESULTS: Engineered HMGA2-V5-overexpressing primary myometrial cells approximated the HMGA2 expression level observed in HMGA2-overexpression subtype leiomyoma. HMGA2-V5 expression resulted in differential expression of 1612 genes (FDR < 0.05) which were found to be enriched in pathways associated with leiomyoma formation, including extracellular matrix organization. Comparative gene expression analysis between HMGA2-V5 engineered primary cells and HMGA2-overexpression subtype leiomyoma revealed significant overlap of differentially expressed genes. Mechanistically, HMGA2-V5 overexpression resulted in 41,323 regions with differential H3K27ac deposition (FDR < 0.05) and 205,605 regions of altered chromatin accessibility (FDR < 0.05). Transcription factor binding site analysis implicated the AP-1 family of transcription factors.
UNASSIGNED: • What clinical problem is addressed by these studies? About 10-15% of the uterine leiomyoma cases exhibit increased expression of HMGA2; our study addresses the primary role of HMGA2 overexpression in the pathogenesis of leiomyoma. • What are the key findings? Ectopic expression of HMGA2 in primary myometrial cells transforms transcriptional and epigenomic machinery in these cells which to some extent resemble features of leiomyoma. • How do these findings apply to human fertility or the reproductive process? Our study provides a comprehensive analysis into the changes occurring in epigenome of myometrial cells when HMGA2 is overexpressed. Because epigenome can be targeted with drugs, therefore understanding alterations in epigenome provides new avenues for treating leiomyoma which affects reproductive age women.
METHODS: Isolated primary, \"normal\" myometrial cells from 3 patients were engineered to overexpress HMGA2 to determine how HMGA2 establishes transcriptomic and epigenomic features of HMGA2-overexpressing leiomyoma.
METHODS: Academic research laboratory PATIENTS: Primary myometrial cells were isolated from normal myometrium obtained from three patients undergoing hysterectomy.
METHODS: Not applicable MAIN OUTCOME MEASURES: Determined genome-wide transcriptomic and epigenomic features of engineered HMGA2-overexpressing uterine primary myometrial cells.
RESULTS: Engineered HMGA2-V5-overexpressing primary myometrial cells approximated the HMGA2 expression level observed in HMGA2-overexpression subtype leiomyoma. HMGA2-V5 expression resulted in differential expression of 1612 genes (FDR < 0.05) which were found to be enriched in pathways associated with leiomyoma formation, including extracellular matrix organization. Comparative gene expression analysis between HMGA2-V5 engineered primary cells and HMGA2-overexpression subtype leiomyoma revealed significant overlap of differentially expressed genes. Mechanistically, HMGA2-V5 overexpression resulted in 41,323 regions with differential H3K27ac deposition (FDR < 0.05) and 205,605 regions of altered chromatin accessibility (FDR < 0.05). Transcription factor binding site analysis implicated the AP-1 family of transcription factors.
UNASSIGNED: • What clinical problem is addressed by these studies? About 10-15% of the uterine leiomyoma cases exhibit increased expression of HMGA2; our study addresses the primary role of HMGA2 overexpression in the pathogenesis of leiomyoma. • What are the key findings? Ectopic expression of HMGA2 in primary myometrial cells transforms transcriptional and epigenomic machinery in these cells which to some extent resemble features of leiomyoma. • How do these findings apply to human fertility or the reproductive process? Our study provides a comprehensive analysis into the changes occurring in epigenome of myometrial cells when HMGA2 is overexpressed. Because epigenome can be targeted with drugs, therefore understanding alterations in epigenome provides new avenues for treating leiomyoma which affects reproductive age women.
摘要：
目的：确定工程改造的HMGA2过表达子宫原代子宫肌层细胞是否能重现HMGA2亚型平滑肌瘤的转录和表观基因组特征。
方法：隔离主要，来自3名患者的“正常”子宫肌层细胞被改造为过表达HMGA2，以确定HMGA2如何建立过表达HMGA2的平滑肌瘤的转录组和表观基因组特征。
方法：学术研究实验室患者：从3名接受子宫切除术的患者的正常子宫肌层中分离原代子宫肌层细胞。
方法：不适用主要结果指标：确定了工程化HMGA2过表达子宫原代子宫肌层细胞的全基因组转录组和表观基因组特征。
结果：工程化的HMGA2-V5过表达的原代子宫肌层细胞接近HMGA2过表达亚型平滑肌瘤中观察到的HMGA2表达水平。HMGA2-V5表达导致1612个基因（FDR<0.05）的差异表达，发现这些基因在与平滑肌瘤形成相关的途径中富集。包括细胞外基质组织。HMGA2-V5工程化原代细胞和HMGA2过表达亚型平滑肌瘤之间的比较基因表达分析揭示了差异表达基因的显着重叠。机械上,HMGA2-V5过表达导致具有差异的H3K27ac沉积的41,323个区域(FDR<0.05)和改变的染色质可及性的205,605个区域(FDR<0.05)。转录因子结合位点分析涉及转录因子的AP-1家族。
■•这些研究解决了什么临床问题？约10-15％的子宫平滑肌瘤病例表现出HMGA2表达增加；我们的研究探讨了HMGA2过度表达在平滑肌瘤发病机理中的主要作用。•关键发现是什么？HMGA2在原代子宫肌层细胞中的异位表达转化了这些细胞中的转录和表观基因组机制，在某种程度上类似于平滑肌瘤的特征。•这些发现如何适用于人类的生育能力或生殖过程？我们的研究提供了一个全面的分析，当HMGA2过表达时，子宫肌层细胞的表观基因组发生的变化。因为表观基因组可以被药物靶向，因此，了解表观基因组的改变为治疗影响育龄女性的平滑肌瘤提供了新的途径。
方法：隔离主要，来自3名患者的“正常”子宫肌层细胞被改造为过表达HMGA2，以确定HMGA2如何建立过表达HMGA2的平滑肌瘤的转录组和表观基因组特征。
方法：学术研究实验室患者：从3名接受子宫切除术的患者的正常子宫肌层中分离原代子宫肌层细胞。
方法：不适用主要结果指标：确定了工程化HMGA2过表达子宫原代子宫肌层细胞的全基因组转录组和表观基因组特征。
结果：工程化的HMGA2-V5过表达的原代子宫肌层细胞接近HMGA2过表达亚型平滑肌瘤中观察到的HMGA2表达水平。HMGA2-V5表达导致1612个基因（FDR<0.05）的差异表达，发现这些基因在与平滑肌瘤形成相关的途径中富集。包括细胞外基质组织。HMGA2-V5工程化原代细胞和HMGA2过表达亚型平滑肌瘤之间的比较基因表达分析揭示了差异表达基因的显着重叠。机械上,HMGA2-V5过表达导致具有差异的H3K27ac沉积的41,323个区域(FDR<0.05)和改变的染色质可及性的205,605个区域(FDR<0.05)。转录因子结合位点分析涉及转录因子的AP-1家族。
■•这些研究解决了什么临床问题？约10-15％的子宫平滑肌瘤病例表现出HMGA2表达增加；我们的研究探讨了HMGA2过度表达在平滑肌瘤发病机理中的主要作用。•关键发现是什么？HMGA2在原代子宫肌层细胞中的异位表达转化了这些细胞中的转录和表观基因组机制，在某种程度上类似于平滑肌瘤的特征。•这些发现如何适用于人类的生育能力或生殖过程？我们的研究提供了一个全面的分析，当HMGA2过表达时，子宫肌层细胞的表观基因组发生的变化。因为表观基因组可以被药物靶向，因此，了解表观基因组的改变为治疗影响育龄女性的平滑肌瘤提供了新的途径。
