MicroRNA-193a Regulates the Transdifferentiation of Human Parietal Epithelial Cells toward a Podocyte Phenotype
MicroRNA-193a Regulates the Transdifferentiation of Human Parietal Epithelial Cells toward a Podocyte Phenotype
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DOI：
10.1681/asn.2014020190
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发表时间：
2015-06-01
影响因子：
13.6
通讯作者：
Meyer-Schwesinger, Catherine
中科院分区：
文献类型：
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作者：
Kietzmann, Leonie;Guhr, Sebastian S. O.;Meyer-Schwesinger, Catherine
壁上皮细胞已被确定为肾小球再生的潜在祖细胞，但这一过程的分子机制尚未完全确定。在这里，我们建立了一个永生化的多克隆人壁上皮细胞（hPEC）线从幼稚的人鲍曼氏囊细胞分离的机械显微解剖。这些hPEC表达高水平的PEC特异性蛋白和microRNA-193 a（miR-193 a），其是通过下调小鼠中的Wilms肿瘤1来抑制足细胞分化的抑制因子。然后，我们研究了nniR-193 a在足细胞和PEC身份建立中的功能，并确定nniR-193 a的抑制是否影响肾小球疾病中佩奇的行为。在稳定敲除miR-193 a后，hPEC采用足细胞样形态和标记物表达，PEC标记物的表达水平降低。在小鼠中，通过互补锁核酸抑制miR-193 a导致足细胞蛋白synaptopodin和Wilms肿瘤1的上调。相反，体内miR-193 a的过表达导致PEC标记物的上调和分离肾小球中足细胞标记物的丢失。在肾毒性肾炎小鼠模型中抑制miR-193 a导致新月体形成减少和蛋白尿减少。总之，这些结果显示了人PEC系的建立，并表明nniR-193 a起主开关的作用，使得具有高水平的miR-193 a的肾小球上皮细胞采用PEC表型，而具有低水平的nniR-193 a的细胞采用足细胞表型。miR-193 a介导的佩奇维持在未分化的反应状态可能是PEC在新月体形成中增殖和迁移的先决条件。
Parietal epithelial cells have been identified as potential progenitor cells in glomerular regeneration, but the molecular mechanisms underlying this process are not fully defined. Here, we established an immortalized polyclonal human parietal epithelial cell (hPEC) line from naive human Bowman's capsule cells isolated by mechanical nnicrodissection. These hPECs expressed high levels of PEC-specific proteins and microRNA-193a (miR-193a), a suppressor of podocyte differentiation through downregulation of Wilms' tumor 1 in mice. We then investigated the function of nniR-193a in the establishment of podocyte and PEC identity and determined whether inhibition of nniR-193a influences the behavior of PECs in glomerular disease. After stable knockdown of miR-193a, hPECs adopted a podocyte-like morphology and marker expression, with decreased expression levels of PEC markers. In mice, inhibition of miR-193a by complementary locked nucleic acids resulted in an upregulation of the podocyte proteins synaptopodin and Wilms' tumor 1. Conversely, overexpression of miR-193a in vivo resulted in the upregulation of PEC markers and the loss of podocyte markers in isolated glomeruli. Inhibition of miR-193a in a mouse model of nephrotoxic nephritis resulted in reduced crescent formation and decreased proteinuria. Together, these results show the establishment of a human PEC line and suggest that nniR-193a functions as a master switch, such that glomerular epithelial cells with high levels of miR-193a adopt a PEC phenotype and cells with low levels of nniR-193a adopt a podocyte phenotype. miR-193a mediated maintenance of PECs in an undifferentiated reactive state might be a prerequisite for PEC proliferation and migration in crescent formation.