Study question: in the present work, we investigate if luteinizing hormone (LH), through its known effects on ovogenesis, is able to exert a protective action on oocytes against cisplatin (CS) and epirubicin (Epi), an other drug commonly used in the therapy of breast cancer. Summary answer: the obtained results point to LH as a potential protective factor against the deleterious effect of some chemotherapics on the oocyte reserve. What is known already: Chemotherapy treatments can frequently cause in women cancer patients a reduction of the ovarian reserve. as a consequence of DNA damage inducing oocytes cell death. Our recent studies on prepuberal mice have shown that CS treatment causes a rapid depletion of the ovarian reserve and loss of fertility. By inhibiting a c-Abl/p63-dependent cell death pathway, we demonstrate that it was possible to significantly reduce oocyte depletion and preserve fertility. Study design, size, duration: LH was added to the culture at different dosage (10, 50, 100mUI), 2h before CS (7.5μM) or Epi (0.3, 0.5, 1μM). At the end of the culture time (about 24h), oocytes were scored. Materials, setting, methods: Ovaries from transgenic c-Kit/GFP mice were sliced in 8 fragments and cultured in appropriate medium. After 3 day of culture, we added LH, CS and Epi. Only GFP-positive oocytes with a normal morphology and a diameter >18-20μm were scored. Main results and the role of chance: The results showed that while LH alone didn’t affect the survival of oocytes within primordial follicles, 7.5μM CS and 1μM Epi caused degeneration of about 50-60% of the oocytes. Interestingly, we observed a significant reduction of oocyte depletion induced by CS in the presence of all LH concentration whereas the hormone was unable to counteract the effect of Epi (oocytes survival= CS, 32+2.9; CS+LH, 54+5.2; EPI, 47+4.9; EPI+LH, 49+6.6; mean+ SE, three independent experiments). Limitations, reasons for caution: A better understanding of the mechanism of LH regulation in this assay is needed. Wider inplications of the findings: These results suggest that the in vitro culture system employed in the present work could be suitable for a rapid and easy screening of chemotherapics potentially harmful for the ovarian reserve, the identification of the triggered cell death pathways and of compounds able to interfere with such pathways. Moreover, they point to LH as a potential protective factor against the deleterious effect of some chemotherapics on the oocyte reserve. Study funding: This research was supported by Merck Seono S.p.a. Keywords: ovarian reserve, LH, chemotherapy, fertility preservation
Development of a predictive in vitro assay for prevention of chemiotherapic effect on the oocytes of the ovarian reserve
Klinger F.G.;
2013-01-01
Abstract
Study question: in the present work, we investigate if luteinizing hormone (LH), through its known effects on ovogenesis, is able to exert a protective action on oocytes against cisplatin (CS) and epirubicin (Epi), an other drug commonly used in the therapy of breast cancer. Summary answer: the obtained results point to LH as a potential protective factor against the deleterious effect of some chemotherapics on the oocyte reserve. What is known already: Chemotherapy treatments can frequently cause in women cancer patients a reduction of the ovarian reserve. as a consequence of DNA damage inducing oocytes cell death. Our recent studies on prepuberal mice have shown that CS treatment causes a rapid depletion of the ovarian reserve and loss of fertility. By inhibiting a c-Abl/p63-dependent cell death pathway, we demonstrate that it was possible to significantly reduce oocyte depletion and preserve fertility. Study design, size, duration: LH was added to the culture at different dosage (10, 50, 100mUI), 2h before CS (7.5μM) or Epi (0.3, 0.5, 1μM). At the end of the culture time (about 24h), oocytes were scored. Materials, setting, methods: Ovaries from transgenic c-Kit/GFP mice were sliced in 8 fragments and cultured in appropriate medium. After 3 day of culture, we added LH, CS and Epi. Only GFP-positive oocytes with a normal morphology and a diameter >18-20μm were scored. Main results and the role of chance: The results showed that while LH alone didn’t affect the survival of oocytes within primordial follicles, 7.5μM CS and 1μM Epi caused degeneration of about 50-60% of the oocytes. Interestingly, we observed a significant reduction of oocyte depletion induced by CS in the presence of all LH concentration whereas the hormone was unable to counteract the effect of Epi (oocytes survival= CS, 32+2.9; CS+LH, 54+5.2; EPI, 47+4.9; EPI+LH, 49+6.6; mean+ SE, three independent experiments). Limitations, reasons for caution: A better understanding of the mechanism of LH regulation in this assay is needed. Wider inplications of the findings: These results suggest that the in vitro culture system employed in the present work could be suitable for a rapid and easy screening of chemotherapics potentially harmful for the ovarian reserve, the identification of the triggered cell death pathways and of compounds able to interfere with such pathways. Moreover, they point to LH as a potential protective factor against the deleterious effect of some chemotherapics on the oocyte reserve. Study funding: This research was supported by Merck Seono S.p.a. Keywords: ovarian reserve, LH, chemotherapy, fertility preservationI documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.
