Ruiz M, González S, Bonnet C, Deng S X
Cornea Division, Stein Eye Institute, University of California, 100 Stein Plaza, Los Angeles, CA, 90095, USA.
Cornea Department, Paris University, Cochin Hospital, AP-HP, F-75014, Paris, France.
Biomark Res. 2022 May 31;10(1):36. doi: 10.1186/s40364-022-00384-2.
Dysfunction or loss of limbal stem cells can result in limbal stem cell deficiency (LSCD), a disease that cause corneal opacity, pain, and loss of vision. Cultivated limbal epithelial transplantation (CLET) can be used to restore stem cell niche homeostasis and replenish the progenitor pool. Transplantation has been reported with high success rate, but there is an unmet need of prognostic markers that correlate with clinical outcomes. To date, the progenitor content in the graft is the only parameter that has been retrospectively linked to success.
In this study, we investigate extracellular micro RNAs (miRNAs) associated with stem/progenitor cells in cultivated limbal epithelial cells (cLECs). Using micro RNA sequencing and linear regression modelling, we identify a miRNA signature in cultures containing high proportion of stem/progenitor cells. We then develop a robust RNA extraction workflow from culture media to confirm a positive miRNA correlation with stem/progenitor cell proportion.
miR-6723-5p is associated with cultures containing high proportion of stem/progenitor cells, and is detected in the basal layer of corneal epithelium.
These results indicate that miR-6723-5p could potentially serve as a stem/progenitor cell marker in cLECs.
角膜缘干细胞功能障碍或缺失可导致角膜缘干细胞缺乏症（LSCD），这是一种可引起角膜混浊、疼痛和视力丧失的疾病。培养的角膜缘上皮移植（CLET）可用于恢复干细胞微环境的稳态并补充祖细胞池。据报道，该移植成功率很高，但仍需要与临床结果相关的预后标志物。迄今为止，移植物中的祖细胞含量是唯一与成功存在回顾性关联的参数。
在本研究中，我们研究了培养的角膜缘上皮细胞（cLECs）中与干细胞/祖细胞相关的细胞外微小RNA（miRNA）。通过微小RNA测序和线性回归建模，我们在含有高比例干细胞/祖细胞的培养物中鉴定出一种miRNA特征。然后，我们开发了一种从培养基中提取RNA的可靠工作流程，以确认miRNA与干细胞/祖细胞比例呈正相关。
miR-6723-5p与含有高比例干细胞/祖细胞的培养物相关，并在角膜上皮基底层中被检测到。
这些结果表明，miR-6723-5p可能作为cLECs中的干细胞/祖细胞标志物。
