Target Identification And Drug Discovery By Data-driven Hypothesis And Experimental Validation In Ovarian Endometriosis

Objective: To identify targets and discover drugs for ovarian endometriosis(OE).Design: A basic study based on data-driven hypothesis and experimental validation.Setting: Center for Reproductive Medicine.Patients/Animals: A total of 14 patients with OE and 7 healthy donors were recruited from the Department of Gynecology and Obstetrics,Union Hospital,Huazhong University of Science and Technology.Fifteen female C57/BL6 mice were purchased from Charles River Laboratory Animal Technology Co.Ltd.(Beijing,China).Interventions: OE lesions and normal endometrium were obtained from patients and healthy donors,respectively.The ITPR1-knockdowned ectopic human endometrial stromal cells(HESCs)were subjected to RNA-sequencing,CCK-8 assay,Edu staining,and flow cytometry analysis.Camptothecin was administrated in HESCs and OE mouse model.Main outcome measures: ITPR1 expression in OE lesions and normal endometrium,cell proliferation and apoptosis of the ITPR1-knockdowned HESCs and the camptothecin-treat HESCs,autograft volume in OE mouse model.Results: Based on the gene expression profiles of OE patients,two significant OE-relevant gene modules were identified to involve the PI3K/Akt and the aging-relevant pathways.Fifteen hub genes were identified and confirmed in the validation dataset as well,among which the most significant gene ITPR1 was robustly elevated in OE lesions(n=14)compared to normal endometrium(n=7).RNA-sequencing revealed that the ITPR1 is highly relevant to cell proliferation and apoptosis,which was further confirmed by CCK-8 assay,Edu staining,and flow cytometry analysis.The knock-down of ITPR1 inhibited cell proliferation and induced apoptosis of ectopic human endometrial stromal cells(HESCs).Moreover,the candidate drugs targeting these modules were screened by a cumulatively-scoring algorithm,among which camptothecin and irinotecan were identified as the promising drugs.Camptothecin and irinotecan both suppressed the proliferation and induce apoptosis of HESCs,meanwhile the expression of ITPR1 was suppressed by camptothecin.The therapeutic effect of camptothecin was also confirmed in OE mouse model.Conclusion(s): This study identified the therapeutic targets and the promising drugs for OE and shed light of the application of camptothecin in OE treatment.