Chemosensitivity and chemoresistance in endometriosis – differences for ectopic versus eutopic cells

Abstract

RESEARCH QUESTION Endometriosis is a common gynecological disease defined by the presence of endometrium-like tissue outside uterus. This complex disease, often accompanied by severe pain and infertility, causes significant medical and socioeconomic burden; hence, novel strategies are sought for treatment of endometriosis. Here, we set out to explore cytotoxic effects of a panel of compounds to find toxins with different efficiency in eutopic versus ectopic cells, thus highlighting alterations in the corresponding molecular pathways. DESIGN Effect of 14 compounds on cellular viability was established in a cohort of paired eutopic and ectopic endometrial stromal cell samples from 11 patients. The biological targets covered by the panel included pro-survival enzymes, cytoskeleton proteins, proteasome, and cell repair machinery. RESULTS We showed that protein kinase inhibitors GSK690693, ARC-775, and sorafenib, proteasome inhibitor bortezomib, and microtubule-depolymerizing toxin MMAE, were more effective in eutopic cells. In contrast, 10 M anthracycline toxin doxorubicin caused cellular death in ectopic cells more effectively than in eutopic cells. The large-scale sequencing of mRNA isolated from doxorubicin-treated and control cells indicated different survival strategies in eutopic versus ectopic endometrium. CONCLUSIONS Overall, our results confirm the evidence of large-scale metabolic reprogramming in endometriotic cells, which underlies the observed differences in sensitivity towards toxins. The enhanced efficiency of doxorubicin interfering with redox equilibria and/or DNA repair mechanisms pinpoints key players that can be potentially used for selective targeting of ectopic lesions in endometriosis.

Viability fingerprint of glioblastoma cell lines: roles of mitotic, proliferative, and epigenetic targets

“…The cells were incubated with individual compounds or mixtures for 48 h, and viability assay was then carried out according to the previously published protocol 77 (see details under the Supplementary Methods ).…”

“…The final volume per well was 1 mL, and the concentration of DMSO in the treated wells was 0.1% by volume. Following the post-treatment, the cells were grown for another 48 h, and viability assay was then carried out according to the previously published protocol 77 (see details under the Supplementary Methods ).…”

Revisiting the Resazurin-Based Sensing of Cellular Viability: Widening the Application Horizon

“…The general protocol of the resazurin assay was published previously [10]; the experimental details are listed below.…”

“…The spectra were recorded following the viability studies (as described in Section 2.2.1) at 30 • C or 37 • C using the following parameters: (A) fluorescence intensity-excitation at 540 (10) nm, emission scanned within the range of 570 to 700 nm or 580 to 700 nm (step of 2 nm) using a monochromator, top optics, gain 50, and a read height of 8.5 mm; (B) absorbance-within the range of 300 to 700 nm (step of 5 nm) using a monochromator.…”

Homing Peptide-Based Targeting of Tenascin-C and Fibronectin in Endometriosis

“…We have also recently demonstrated cellpenetrating peptide and siRNA-mediated therapeutic effects on endometriosis models and proposed a novel therapeutic target, ribonucleotide reductase for the treatment of endometriosis [57]. In addition, we have previously identified miRNAs that are differentially expressed in ectopic endometriotic stromal cells [58] and found that some anticancer drugs, such as doxorubicin, are selectively cytotoxic to ectopic endometrial cells [59]. Affinity-targeting of such drugs using nanoformulations could be an efficacious strategy to potentiate their activity of specifically depleting endometriotic lesions.…”