Targeting perlecan in human keratinocytes reveals novel roles for perlecan in epidermal formation
Targeting perlecan in human keratinocytes reveals novel roles for perlecan in epidermal formation
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DOI：
10.1074/jbc.m509500200
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发表时间：
2006-02-24
影响因子：
4.8
通讯作者：
Ron, D
中科院分区：
文献类型：
--
作者：
Sher, I;Zisman-Rozen, S;Ron, D
肝素结合生长因子对人表皮的形成至关重要，但硫酸乙酰肝素蛋白多糖在此过程中的作用知之甚少。在这里，我们研究了硫酸乙酰肝素蛋白聚糖，串珠素，在人表皮形成中的作用，通过利用体外工程化的人皮肤。通过破坏串珠素在真皮或表皮中的表达，我们发现表皮衍生的串珠素对于表皮形成是必需的。缺乏串珠素的角质形成细胞由于过早凋亡和未能完成其分层程序而形成了非常薄且组织不良的表皮。外源性串珠素完全恢复表皮形成。基底膜区的串珠素沉积与多层表皮的形成有关。然而，串珠素缺乏对主要基底膜成分的衬里和沉积没有影响，层粘连蛋白和IV型胶原蛋白的连续线性染色证明了这一点。类似地，串珠素缺乏不影响β 1整合素的分布。添加串珠素配体，成纤维细胞生长因子7，保护串珠素缺乏的角质形成细胞免于细胞死亡，并改善表皮的厚度。综上所述，我们的研究结果揭示了串珠素在表皮形成中的新作用。串珠素调节角质形成细胞的存活和终末分化步骤。我们的研究结果提出了一个模型，其中串珠素调节这些过程通过控制串珠素结合的可溶性因子参与表皮形态发生的生物利用度。
Heparin-binding growth factors are crucial for the formation of human epidermis, but little is known about the role of heparan sulfate proteoglycans in this process. Here we investigated the role of the heparan sulfate proteoglycan, perlecan, in the formation of human epidermis, by utilizing in vitro engineered human skin. By disrupting perlecan expression either in the dermis or the epidermis, we found that epidermally derived perlecan is essential for epidermal formation. Perlecan-deficient keratinocytes formed a strikingly thin and poorly organized epidermis because of premature apoptosis and failure to complete their stratification program. Exogenous perlecan fully restored epidermal formation. Perlecan deposition in the basement membrane zone correlated with formation of multilayered epidermis. Perlecan deficiency, however, had no effect on the lining and deposition of major basement membrane components as was evident by a continuous linear staining of laminin and collagen IV. Similarly, perlecan deficiency did not affect the distribution of beta 1 integrin. Addition of the perlecan ligand, fibroblast growth factor 7, protected perlecan-deficient keratinocytes from cell death and improved the thickness of the epidermis. Taken together, our results revealed novel roles for perlecan in epidermal formation. Perlecan regulates both the survival and terminal differentiation steps of keratinocytes. Our results suggested a model whereby perlecan regulates these processes via controlling the bioavailability of perlecan-binding soluble factors involved in epidermal morphogenesis.