Analysis Of The Effect And Molecular Mechanism Of Estrogen On Endometriosis Based On The Results Of Single-cell Transcriptome Sequencing

Endometriosis(EMs)is reguarded as a gynacology disease that the endometrial tissues appear and survive in the outside uterine,causing abdominal pain,dysmenorrhea,sexual intercourse uncomfortable,even infertility symptoms.The incidence of EMs is 6%-10%,and the recurrence rate even reaches 50%,which were harmful to patients and their families.The pathogenesis of EMs is not clear,thus comprehensive analysis of endometriotic lesions formation and development process are greatly significant for investing the mechanism.In this paper,we analyzed the changed of cell type and function of endometriotic lesions in the single-cell level,explored the modulation of eutopic tissues microenvironment,and investigated the effect and mechanism of estrogen on the formation of endometriotic lesions.In this study,single-cell RNA sequencing(scRNA Seq)was employed to analyze the changes of composition and function between the eutopic endometria and matched endometriotic lesion from three EMs patients.As the results of scRNA Seq indicated that a total of 15,300 cells were captured and 7 different cell types were clustered.And the physicological function of ectopic cells were obvious different to eutopic cells.Ectopic endometrial cells were characteried of epithelial-mesenchymal transition,and increased secretion of cytokines,changed expression of extracellular matrix,and enhancedd expression of estrogen synthase.Therefore,ectopic endometrial cells resulted changes of the extracellular microenvironment of lesions and chemotaxis of immune cells.Ectopic vascular endothelial cells were analysed that the angiopoietic ability were increased,and the expression chemokines and adhesion molecules were further upregulated which induced the infiltration of immune cells to the ectopic lesions.The expression of inflammatory factors in ectopic macrophages were enhanced,which further promotd the formation of an inflammatory microenvironment.However,the number of T cell from endometriotic was reduced,and its cytokine expression and the cell killing function were significantly decreased,which might result in the inability to eliminate ectopic lesions.Therefore,scRNA Seq revealed the difference of cell number and physiology obetween eutopic endometria and endometriotic lesions,and further analysis elucidated the effects of different cells on the microenvironment of endometriotic lesions.This research further focused on the effect and mechanism of estrogen on the formation of ectopic lesion.Estrogen and its receptor ERα obviously promoted the proliferation,migration,invasion and EMT of ectopic epithelial cells.The expression change of ERa can affect the function of estrogen,indicating that ERa mediates the effect of estrogen.Both of estrogen and ERa could promoted the expression of RhoA,ROCK1 and ROCK2,resulting the activation of RhoA/ROCK pathway.And changing the expression of RhoA could affect the function of estrogen and ERa on cell proliferation and EMT,indicating that the RhoA/ROCK pathway acts as a downstream signal transferring the signaling of estrogen and ERα.By the establishment of EMs mouse model by using BALB/c mice.Thus,we found estrogen could promoted the expression of proteins in RhoA/ROCK pathway and promote the formation of ectopic lesions in mice.To sum up,this study analyzed the cellular heterogeneity of eutopic endometria and ectopic lesions in patients with EMs via scRNA seq,and found changes in the number and function of cells in ectopic lesions.According to the abnormal cell function,the changes of the extracellular microenvironment such as the inflammatory microenvironment,extracellular matrix changes and high estrogen of the lesion tissue were analyzed.At the same time,this paper also discovered the molecular mechanism of estrogen regulating RhoA/ROCK signaling pathway through ERa to affect the formation of ectopic lesions,which provides new ideas for the treatment of EMs.