Niche formed by bone morphogenetic protein antagonists gremlin 1 and gremlin 2 in human hair follicles.
Niche formed by bone morphogenetic protein antagonists gremlin 1 and gremlin 2 in human hair follicles.
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DOI：
10.1002/hsr2.486
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发表时间：
2022-03
影响因子：
2
通讯作者：
Kishimoto J
中科院分区：
文献类型：
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作者：
Niiyama S;Ishimatsu-Tsuji Y;Kishimoto J
有人提出，毛球中的细胞因子生态位在毛发周期的精确调节中发挥着重要作用。最近报道了对 WNT 相关因子形成的休止期-毛发生长初期阶段的生态位的值得注意的分析。 1, 2 在本研究中，我们分析了可能参与生态位形成的骨形态发生蛋白 (BMP) 相关因子的定位。 BMP 家族为毛囊 (HF) 循环提供一些最关键的信号，据报道可以抑制细胞增殖并促进上皮细胞的分化和迁移。 3, 4 此前，我们通过转录组分析发现，BMP 拮抗剂 gremlin 2 (GREM2) 在人 HF 真皮鞘杯 (DSC) 中的表达高于真皮乳头 (DP) 和真皮鞘上部区域。 5 DSC 是 DS 的球周组成部分，从临床角度来看，它具有独特的特征，即源自 DSC 的细胞在啮齿动物模型和人类临床研究中具有毛发诱导潜力。 6, 7 本研究获得了参与本研究的捐赠者的书面知情同意书，该研究是在东邦大学大桥医学中心和资生堂全球创新中心伦理委员会批准下进行的。含有枕部头皮 HF 的皮肤片取自一名 37 岁的女性捐赠者，患有女性图案大胆症。对石蜡包埋的组织切片进行原位杂交。 GREM1-、GREM2-、BMP4-、淋巴增强子结合因子 1 (LEF1)-、SRY 盒转录因子 2 (SOX2) 特异性探针根据 Entrez 基因 ID26585、64 388、51 776、6657、652 的序列信息，基于 View RNATM ISH Tissue 1-Plex 检测试剂盒中的高度特异性分支 DNA 信号放大技术由 Thermo Fisher Scientific Inc（马萨诸塞州沃尔瑟姆）提供。用固红底物检测信号，然后进行荧光观察。使用 Hoechst 33342 (Thermo Fischer Scientific Inc) 进行核复染。使用 LSM880 共焦显微镜（Carl Zeiss Microscopy GmbH，Oberkochen，德国）进行图像采集。所有实验均使用不同的头皮组织重复进行，以确认再现性。为了验证结果，在来自不同男性供体的 10 多个 HF 中确认了 GREM2 的表达，并且在每两个或更多个体中至少有 3 个 HF 中发现了 GREM1、BMP4、LEF1 和 SOX2 的表达（数据未显示）。我们通过组织切片上的原位杂交检查了 GREM2 和其他 BMP 相关基因的定位。在毛发生长初期、毛发生长早期和小型毛发生长初期 HF 的毛发周期中，GREM2 的表达在 DSC 中表现强劲（图 1A、D、E），其仅限于 DSC，并且在整个 HF 中的 DS 上部区域和漏斗部中不存在。接下来，我们分析了毛球中其他 BMP 相关分子的定位。与 GREM2 信号不同，在正常和小型生长期 HF 的 DSC 和上皮基质中观察到另一种 gremlin 亚型 gremlin 1 (GREM1) 的信号。在退行期的 DP 以及 DSC 和上皮基质中观察到了一些 GREM1 信号（图 1B、F）。 BMP4 在上皮基质、内根鞘 (IRS) 和生长期毛囊的 DP 中表达（图 1C）。在与 DP 相邻的 DSC 限制区域中也观察到 BMP4 的表达相对较低。 BMP4 的这种表达模式在退行期早期得以维持（图 1G）。
It has been proposed that cytokine niches in the hair bulb play an important role in the precise regulation of the hair cycle. Noteworthy analyses of the niche in the telogen-anagen phase formed by WNT-related factors have been reported recently. 1, 2 In the present study, we analyzed the localization of bone-morphogenic-protein (BMP)-related factors that might be involved in niche formation. The BMP family provides some of the most critical signals for the cycling of hair follicles (HFs), which has been reported to suppress cell proliferation and promote differentiation and migration of epithelial cells. 3, 4 Previously, we found that expression of a BMP antagonist, gremlin 2 (GREM2), in the dermal sheath cup (DSC) was higher than that in the dermal papilla (DP) and upper area of the dermal sheath in human HFs by transcriptome analysis. 5 The DSC is the peribulbar component of the DS, which has unique characteristics from the clinical viewpoint that cells derived from the DSC possessed a hairinductive potential in a rodent model and clinical study in humans. 6, 7 Written informed consent was obtained from the donors participating in this study that was conducted with the approval of the Ethics Committee of Toho University Ohashi Medical Center and Shiseido Global Innovation Center. Skin pieces containing occipital scalp HFs were obtained from a 37-year-old female donor, suffering from female pattern boldness. In situ hybridization was performed on paraffin-embedded tissue sections. GREM1-, GREM2-, BMP4-, lymphoid enhancer-binding factor 1 (LEF1)-, SRY box transcription factor 2 (SOX2)-specific probes according to the sequence information of Entrez gene ID26585, 64 388, 51 776, 6657, 652 based on highly specific branched DNA signal amplification technology in a View RNATM ISH Tissue 1-Plex Assay kit were supplied from Thermo Fisher Scientific Inc (Waltham, Massachusetts). Signals were detected with Fast Red substrate, followed by fluorescence observation. Nuclear counterstaining was performed with Hoechst 33342 (Thermo Fischer Scientific Inc). Image acquisition was performed using an LSM880 confocal microscope (Carl Zeiss Microscopy GmbH, Oberkochen, Germany). All experiments were performed repeatedly using different scalp tissues to confirm reproducibility. For the validation of the results, the expression of GREM2 was confirmed in more than 10 HFs derived from different male donors, and expression of GREM1, BMP4, LEF1, and SOX2 was seen in at least three HFs per two individuals or more (data not shown). We examined the localization of GREM2 and other BMP-related genes by in situ hybridization on tissue sections. GREM2 expression was robust in the DSC during the hair cycle in anagen, early catagen, and the miniaturized anagen HF (Figure 1A, D, E), which was restricted to the DSC and absent in the upper area of the DS and the infundibulum in the entire HF.Next, we analyzed the localization of other BMP-related molecules in the hair bulb. Signals of another gremlin subtype, gremlin 1 (GREM1), were observed in the DSC and the epithelial matrix of normal and miniaturized anagen HFs, unlike GREM2 signals. A few signals of GREM1 were observed in the DP as well as the DSC and epithelial matrix in the catagen phase (Figure 1B, F). BMP4 was expressed in the epithelial matrix, inner root sheath (IRS), and DP of anagen follicles (Figure 1C). Relatively low expression of BMP4 was also observed in a restricted area of the DSC adjacent to the DP. This expression pattern of BMP4 was maintained in the early stage of catagen (Figure 1G).