目的:评估子宫腺肌病的细胞和分子景观。
方法:子宫内膜匹配组织的全基因组mRNA表达(scRNA-seq)的单细胞分析,子宫腺肌病,和使用相对大量的活细胞的子宫肌层。
方法:接受弥漫性子宫腺肌病子宫切除术的患者(n=3,年龄范围40-44岁)使用scRNA-seq并比较这些组织中不同细胞类型以鉴定疾病特异性细胞群,来自同一子宫的子宫内膜和子宫肌层组织。异常基因表达和通路激活,和间充质-上皮相互作用。
结果:子宫内膜中最大的细胞群由紧密聚集的成纤维细胞组成,占所有细胞的36%,似乎起源于分化为表达雌激素/孕激素受体的子宫内膜基质细胞的周细胞祖细胞。相比之下,子宫腺肌病中的整个成纤维细胞群体占所有细胞的较大部分(50%),并且与任何周细胞祖细胞无关。腺病毒成纤维细胞最终分化为表达细胞外基质蛋白的成纤维细胞和平滑肌细胞。mRNA表达的分层聚类揭示了一个独特的腺体成纤维细胞群体,其转录上与子宫内膜成纤维细胞聚类,提示子宫内膜基质细胞群作为子宫腺肌病的祖细胞。具有疾病特异性转录组的其他四个腺样体成纤维细胞簇与子宫内膜或子宫肌层成纤维细胞不同。天然WNT抑制剂的mRNA水平,named,与子宫内膜成纤维细胞簇相比,这4个腺体成纤维细胞簇中分泌的卷曲相关蛋白(SFRP)1、2和4较高。此外,我们发现多个WNTs,起源于成纤维细胞和目标纤毛和无纤毛的上皮细胞和内皮细胞,在腺病毒组织中构成关键的旁分泌信号网络。与子宫内膜组织相比,子宫腺肌病中的无纤毛和纤毛上皮细胞占该组织的比例明显较小,并表现出孕酮抵抗和雌激素信号调节减弱的分子证据。
结论:我们发现在子宫腺肌成纤维细胞样细胞中存在高度异质性。涉及SFRP差异表达的WNT信号,作为WNTs的诱饵受体,成纤维细胞可能在该疾病的病理生理中起关键作用。
To assess the cellular and molecular landscape of adenomyosis.
Single-cell analysis of genome-wide messenger RNA (mRNA) expression (single-cell RNA sequencing) of matched tissues of endometrium, adenomyosis, and myometrium using relatively large numbers of viable cells.
Not applicable.
Patients (n = 3, age range 40-44 years) undergoing hysterectomy for diffuse adenomyosis.
Definition of the molecular landscape of matched adenomyotic, endometrial and myometrial tissues from the same uterus using single-cell RNA sequencing and comparison of distinct cell types in these tissues to identify disease-specific cell populations, abnormal gene expression and pathway activation, and mesenchymal-epithelial interactions.
The largest cell population in the endometrium was composed of closely clustered fibroblast groups, which comprise 36% of all cells and seem to originate from pericyte progenitors differentiating to estrogen/progesterone receptor-expressing endometrial stromal- cells. In contrast, the entire fibroblast population in adenomyosis comprised a larger (50%) portion of all cells and was not linked to any pericyte progenitors. Adenomyotic fibroblasts eventually differentiate into extracellular matrix protein-expressing fibroblasts and smooth muscle cells. Hierarchical clustering of mRNA expression revealed a unique adenomyotic fibroblast population that clustered transcriptomically with endometrial fibroblasts, suggestive of an endometrial stromal cell population serving as progenitors of adenomyosis. Four other adenomyotic fibroblast clusters with disease-specific transcriptomes were distinct from those of endometrial or myometrial fibroblasts. The mRNA levels of the natural WNT inhibitors, named, secreted frizzled-related proteins 1, 2, and 4, were higher in these 4 adenomyotic fibroblast clusters than in endometrial fibroblast clusters. Moreover, we found that multiple WNTs, which originate from fibroblasts and target ciliated and unciliated epithelial cells and endothelial cells, constitute a critical paracrine signaling network in adenomyotic tissue. Compared with endometrial tissue, unciliated and ciliated epithelial cells in adenomyosis comprised a significantly smaller portion of this tissue and exhibited molecular evidence of progesterone resistance and diminished regulation of estrogen signaling.
We found a high degree of heterogeneity in fibroblast-like cells in the adenomyotic uterus. The WNT signaling involving differential expression of secreted frizzled-related proteins, which act as decoy receptors for WNTs, in adenomyotic fibroblasts may have a key role in the pathophysiology of this disease.