Sphingosine-1-phosphate prevents chemotherapy-induced human primordial follicle death.
OBJECTIVE
Can Sphingosine-1-phosphate (S1P), a ceramide-induced death pathway inhibitor, prevent cyclophosphamide (Cy) or doxorubicin (Doxo) induced apoptotic follicle death in human ovarian xenografts?


CONCLUSIONS
S1P can block human apoptotic follicle death induced by both drugs, which have differing mechanisms of cytotoxicity.


BACKGROUND
S1P has been shown to decrease the impact of chemotherapy and radiation on germinal vesicle oocytes in animal studies but no human translational data exist.


METHODS
Experimental human ovarian xenografting to test the in vivo protective effect of S1P on primordial follicle survival in the chemotherapy setting. The data were validated by assessing the same protective effect in the ovaries of xenografted mice in parallel.


METHODS
Xenografted mice were treated with Cy (75 mg/kg), Cy+S1P (200 μM), Doxo (10 mg/kg), Doxo+S1P or vehicle only (Control). S1P was administered via continuous infusion using a mini-osmotic pump beginning 24 h prior to and ending 72 h post-chemotherapy. Grafts were then recovered and stained with anti-caspase 3 antibody for the detection of apoptosis in primordial follicles. The percentage of apoptotic to total primordial follicles was calculated in each group.


RESULTS
Both Cy and Doxo resulted in a significant increase in apoptotic follicle death in human ovarian xenografts compared with controls (62.0 ± 3.9% versus 25.7 ± 7.4%, P < 0.01 and 76.7 ± 7.4% versus 25.7 ± 7.4%, P < 0.01, respectively). This chemotherapy-induced apoptotic death was reduced both in the Cy+S1P (32.7 ± 4.4%, P < 0.01) and the Doxo+S1P group (27.1 ± 7.6%, P < 0.01) compared with Cy and Doxo groups, respectively. In the Doxo+S1P and Cy+S1P groups, the percentages of apoptotic follicles were similar to those of vehicle-treated controls (P > 0.05). The findings from the ovaries of the severe combined immunodeficient mice mirrored the findings with human tissue.


CONCLUSIONS
The functionality of the rescued human ovarian follicles needs to be evaluated in future studies though the studies in rodents showed that rescued oocytes can result in healthy offspring. In addition, the impact of S1P on cancer cells should be further studied.


CONCLUSIONS
S1P and its future analogs hold promise for preserving fertility by pharmacological means for patients undergoing chemotherapy.


BACKGROUND
This research is supported by NIH's NICHD and NCI (5R01HD053112-06 and 5R21HD061259-02) and the Flemish Foundation for Scientific Research (FWO-Vlaanderen, grant number FWO G0.065.11N10). The authors have no conflicts of interest to disclose.
Laboratory of Molecular Reproduction and Fertility Preservation, Obstetrics & Gynecology, New York Medical College, 40 Sunshine Cottage Road, Valhalla, NY 10595, USA.
Fang
F
Li
Volkan
V
Turan
Sylvie
S
Lierman
Claude
C
Cuvelier
Petra
P
De Sutter
Kutluk
K
Oktay
D000293:Adolescent; D000328:Adult; D000818:Animals; D017209:Apoptosis / Q000187:drug effects*; D053148:Caspase 3 / Q000378:metabolism; D003520:Cyclophosphamide / Q000037:antagonists & inhibitors*; D004317:Doxorubicin / Q000037:antagonists & inhibitors*; D005260:Female; D064593:Heterografts; D006801:Humans; D008246:Lysophospholipids / Q000494:pharmacology*; D051379:Mice; D016513:Mice, SCID; D006080:Ovarian Follicle / Q000187:drug effects* / Q000473:pathology; D010053:Ovary / Q000502:physiology / Q000637:transplantation*; D013110:Sphingosine / Q000031:analogs & derivatives* / Q000494:pharmacology
D008246:Lysophospholipids; C060506:sphingosine 1-phosphate; D004317:Doxorubicin; D003520:Cyclophosphamide; C505407:CASP3 protein, human; C505409:Casp3 protein, mouse; D053148:Caspase 3; D013110:Sphingosine
Sphingosine-1-phosphate; apoptosis; cyclophosphamide; doxorubicin; human ovarian xenograft