Nikhila Leemon, Surya Suresh, Najeeb Shahul Hameed, Binumon Thankachan Mangalathettu, Gupta Aayush, Gopalakrishnan Sandeep, Dabagh Mahsa, Sreejith Parameswara Panicker
Department of Zoology and Advanced Centre for Regenerative Medicine and Stem cell in Cutaneous Research (AcREM-Stem), University of Kerala, Thiruvananthapuram, Kerala, India.
Dr. D.Y. Patil Medical College Hospital and Research Center, Dr. D Y Patil Vidyapeeth, Pimpri, Pune, India.
PLoS One. 2025 Sep 15;20(9):e0308455. doi: 10.1371/journal.pone.0308455. eCollection 2025.
Hair follicle cycle and the functioning of stem cells in alopecia are influenced by the suppression of cholesterol synthesis and the accumulation of sterol intermediates.
This investigation aims to elucidate the regulatory function of disrupted cholesterol homeostasis in the functioning of hair follicle stem cells (HFSCs) and the cycling of hair follicles. Additionally, it seeks to provide an understanding of the fate of stem cells in primary cicatricial alopecia (PCA).
To evaluate the influence of cholesterol on the functionality of hair follicles, a study was conducted to analyse gene expression and pathways associated with hair follicle stem cell markers in scalp samples affected by PCA (LPP, FFA, CCCA, DC, DF, TF). To assess the influence of disrupted cholesterol homeostasis on HFSCs, we conducted experiments involving the administration of 7-dehydrocholesterol (7DHC) and BM15766 (Pharmacological inhibitor of cholesterol biosynthesis), to Human Hair Follicle Outer Root Sheath Cells (HORSCs), as well as C57BL/6 mice, and hair follicle organoid cultures. The study utilised reverse transcription polymerase chain reaction (RT-PCR) to assess gene expression, while immunofluorescence was employed to analyse protein expression. The tracking of stem cell fate was accomplished through the utilisation of a BrdU pulse-chase experiment, while the verification of apoptotic consequences was established by utilising the TUNEL assay. A statistical analysis was conducted to assess the statistical significance of the data.
There was a notable decrease in the expression of HFSC marker genes among patients afflicted with PCA. In vitro data further confirmed the cholesterol inhibition and sterol intermediate accumulation in stem cells, leading to stem cell characteristics' disruption. The experimental group of mice exposed to 7DHC and BM15766 also exhibited a notable inability to initiate hair regrowth. Consequently, this deficiency in hair regrowth resulted in the activation of apoptosis, specifically in the stem cells. Additionally, our confirmatory analysis, which was performed utilising organoid culture, consistently yielded comparable results. The results as mentioned above emphasise the significant importance of cholesterol production in preserving the integrity and functionality of HFSCs, hence providing novel insights into the progression of alopecia.
Individuals with aberrant cholesterol production, especially those affected by PCA, endure a lasting hindrance in hair regrowth stemming from the irreversible damage to their hair follicles. The observed phenomenon is hypothesized to result from the depletion of hair follicle stem cells. Our study reveals further insights that unveil the previously unrecognized role of sterol intermediates in governing hair follicle cycling and stem cell function in PCA. The modulation of cholesterol production and the accumulation of sterol intermediates significantly influence the hair follicle cycle and the role of stem cells in alopecia.
胆固醇合成的抑制以及固醇中间体的积累会影响毛囊周期和脱发中干细胞的功能。
本研究旨在阐明胆固醇稳态破坏对毛囊干细胞(HFSCs)功能和毛囊周期的调节作用。此外,旨在了解原发性瘢痕性脱发(PCA)中干细胞的命运。
为评估胆固醇对毛囊功能的影响,开展了一项研究,分析受PCA影响的头皮样本(LPP、FFA、CCCA、DC、DF、TF)中与毛囊干细胞标志物相关的基因表达和信号通路。为评估胆固醇稳态破坏对HFSCs的影响,我们对人毛囊外根鞘细胞(HORSCs)、C57BL/6小鼠以及毛囊类器官培养物进行了实验,分别给予7-脱氢胆固醇(7DHC)和BM15766(胆固醇生物合成的药理学抑制剂)。该研究利用逆转录聚合酶链反应(RT-PCR)评估基因表达,同时采用免疫荧光分析蛋白质表达。通过BrdU脉冲追踪实验追踪干细胞命运,利用TUNEL检测确定凋亡结果。进行统计分析以评估数据的统计学意义。
PCA患者中HFSC标志物基因的表达显著降低。体外数据进一步证实了干细胞中胆固醇抑制和固醇中间体积累,导致干细胞特性破坏。暴露于7DHC和BM15766的小鼠实验组也表现出明显无法启动毛发生长。因此,这种毛发生长缺陷导致了凋亡的激活,特别是在干细胞中。此外,我们利用类器官培养进行的验证性分析始终得出类似结果。上述结果强调了胆固醇生成在维持HFSCs完整性和功能方面的重要意义,从而为脱发的进展提供了新的见解。
胆固醇生成异常的个体,尤其是受PCA影响的个体,由于毛囊受到不可逆损伤,毛发生长持续受阻。据推测,观察到的现象是由于毛囊干细胞耗竭所致。我们的研究揭示了进一步的见解,揭示了固醇中间体在PCA中调控毛囊周期和干细胞功能方面以前未被认识的作用。胆固醇生成的调节和固醇中间体的积累显著影响毛囊周期以及干细胞在脱发中的作用。