title: GnRH agonist leuprolide acetate does not confer any protection against ovarian damage induced by chemotherapy and radiation in vitro.
abstract: OBJECTIVE
  Is there any in vitro evidence for or against ovarian protection by co-administration of a GnRH agonist with chemotherapy in human?
  
  
  CONCLUSIONS
  The co-administration of GnRH agonist leuprolide acetate with cytotoxic chemotherapy agents does not preserve ovarian reserve in vitro.
  
  
  BACKGROUND
  Randomized controlled trials of the co-administration of gonadotrophin-releasing hormone (GnRH) agonists with adjuvant chemotherapy to preserve ovarian function have shown contradictory results. This fact, together with the lack of a proven molecular mechanism of action for ovarian protection with GnRH agonist (GnRHa) places this approach as a fertility preservation strategy under scrutiny. We therefore aimed in this study to provide in vitro evidence for or against the role of GnRHa in the prevention of chemotherapy-induced damage in human ovary.
  This translational research study of ex vivo and in vitro models of human ovary and granulosa cells was conducted in a university hospital between 2013 and 2015.
  
  
  METHODS
  Ovarian cortical pieces (n = 15, age 14-37) and mitotic non-luteinized (COV434 and HGrC1) and non-mitotic luteinized human granulosa cells (HLGC) expressing GnRH receptor were used for the experiments. The samples were treated with cyclophosphamide, cisplatin, paclitaxel, 5-FU, or TAC combination regimen (docetaxel, adriamycin and cyclophosphamide) with and without GnRHa leuprolide acetate for 24 h. DNA damage, apoptosis, follicle reserve, hormone markers of ovarian function and reserve (estradiol (E2), progesterone (P) and anti-mullerian hormone (AMH)) and the expression of anti-apoptotic genes (bcl-2, bcl-xL, bcl-2L2, Mcl-1, BIRC-2 and XIAP) were compared among control, chemotherapy and chemotherapy + GnRHa groups.
  
  
  RESULTS
  The greatest magnitude of cytotoxicity was observed in the samples treated with cyclophosphamide, cisplatin and TAC regimen. Exposure to these drugs resulted in DNA damage, apoptosis and massive follicle loss along with a concurrent decline in the steroidogenic activity of the samples. GnRHa co-administered with chemotherapy agents stimulated its receptors and raised intracellular cAMP levels. But it neither activated anti-apoptotic pathways nor prevented follicle loss, DNA damage and apoptosis induced by these drugs.
  
  
  CONCLUSIONS
  Our findings do not conclusively rule out the possibility that GnRHa may offer protection, if any, through some other mechanisms in vivo.
  
  
  CONCLUSIONS
  GnRH agonist treatment with chemotherapy does not prevent or ameliorate ovarian damage and follicle loss in vitro. These data can be useful when consulting a young patient who may wish to receive GnRH treatment with chemotherapy to protect her ovaries from chemotherapy-induced damage.
authors.affiliation: The Graduate School of Health Sciences, Koc University, Istanbul, Turkey.
authors.forename: Gamze
authors.initials: G
authors.lastname: Bildik
authors.affiliation: The Graduate School of Health Sciences, Koc University, Istanbul, Turkey.
authors.forename: Nazlı
authors.initials: N
authors.lastname: Akin
authors.affiliation: The Graduate School of Health Sciences, Koc University, Istanbul, Turkey.
authors.forename: Filiz
authors.initials: F
authors.lastname: Senbabaoglu
authors.affiliation: The Graduate School of Health Sciences, Koc University, Istanbul, Turkey.
authors.forename: Gizem Nur
authors.initials: GN
authors.lastname: Sahin
authors.affiliation: Department of Histology and Embryology, Koc University School of Medicine, Istanbul, Turkey.
authors.forename: Sercin
authors.initials: S
authors.lastname: Karahuseyinoglu
authors.affiliation: Department of Pathology, Acibadem University School of Medicine, Istanbul, Turkey.
authors.forename: Umit
authors.initials: U
authors.lastname: Ince
authors.affiliation: Department of Obstetrics and Gynecology, Koc University School of Medicine, Istanbul, Turkey Comprehensive Cancer Care and Fertility Preservation Programs American Hospital, Istanbul, Turkey.
authors.forename: Cagatay
authors.initials: C
authors.lastname: Taskiran
authors.affiliation: Comprehensive Cancer Care and Fertility Preservation Programs American Hospital, Istanbul, Turkey Department of Radiation Oncology, Koc University School of Medicine, Istanbul, Turkey Department of Radiation Oncology, University of Texas MD Anderson Cancer Center, Houston, TX, USA.
authors.forename: Ugur
authors.initials: U
authors.lastname: Selek
authors.affiliation: Department of Obstetrics and Gynecology, Koc University School of Medicine, Istanbul, Turkey.
authors.forename: Kayhan
authors.initials: K
authors.lastname: Yakin
authors.affiliation: Assisted Reproduction Unit, Women' Health Center American Hospital, Istanbul, Turkey.
authors.forename: Yilmaz
authors.initials: Y
authors.lastname: Guzel
authors.affiliation: Assisted Reproduction Unit, Women' Health Center American Hospital, Istanbul, Turkey.
authors.forename: Cem
authors.initials: C
authors.lastname: Ayhan
authors.affiliation: Assisted Reproduction Unit, Women' Health Center American Hospital, Istanbul, Turkey.
authors.forename: Ebru
authors.initials: E
authors.lastname: Alper
authors.affiliation: Comprehensive Cancer Care and Fertility Preservation Programs American Hospital, Istanbul, Turkey Department of Hematology and Oncology, Koc University School of Medicine, Istanbul, Turkey.
authors.forename: Mustafa
authors.initials: M
authors.lastname: Cetiner
authors.affiliation: Assisted Reproduction Unit, Women' Health Center American Hospital, Istanbul, Turkey.
authors.forename: Basak
authors.initials: B
authors.lastname: Balaban
authors.affiliation: Comprehensive Cancer Care and Fertility Preservation Programs American Hospital, Istanbul, Turkey Department of Medical Oncology, Koc University School of Medicine, Istanbul, Turkey.
authors.forename: Nil Molinas
authors.initials: NM
authors.lastname: Mandel
authors.affiliation: Comprehensive Cancer Care and Fertility Preservation Programs American Hospital, Istanbul, Turkey Department of Urology and Urologic Oncology, Koc University School of Medicine, Istanbul, Turkey.
authors.forename: Tarık
authors.initials: T
authors.lastname: Esen
authors.affiliation: Department of Obstetrics and Gynecology, Nagoya University, Nagoya, Japan.
authors.forename: Akira
authors.initials: A
authors.lastname: Iwase
authors.affiliation: Department of Obstetrics and Gynecology, Koc University School of Medicine, Istanbul, Turkey Comprehensive Cancer Care and Fertility Preservation Programs American Hospital, Istanbul, Turkey Assisted Reproduction Unit, Women' Health Center American Hospital, Istanbul, Turkey.
authors.forename: Bulent
authors.initials: B
authors.lastname: Urman
authors.affiliation: Department of Obstetrics and Gynecology, Koc University School of Medicine, Istanbul, Turkey Comprehensive Cancer Care and Fertility Preservation Programs American Hospital, Istanbul, Turkey Assisted Reproduction Unit, Women' Health Center American Hospital, Istanbul, Turkey ooktem@ku.edu.tr.
authors.forename: Ozgur
authors.initials: O
authors.lastname: Oktem
mesh_terms: D000293:Adolescent; D000328:Adult; D000970:Antineoplastic Agents / Q000494:pharmacology*; D017209:Apoptosis / Q000187:drug effects / Q000528:radiation effects; D004249:DNA Damage / Q000187:drug effects / Q000528:radiation effects; D005260:Female; D005300:Fertility Agents, Female / Q000008:administration & dosage*; D059247:Fertility Preservation / Q000379:methods; D006107:Granulosa Cells / Q000187:drug effects* / Q000528:radiation effects; D006801:Humans; D016729:Leuprolide / Q000008:administration & dosage*; D065851:Ovarian Reserve / Q000187:drug effects* / Q000528:radiation effects; D010053:Ovary / Q000187:drug effects* / Q000528:radiation effects; D020011:Protective Agents / Q000008:administration & dosage*; D055815:Young Adult
chemical_list: D000970:Antineoplastic Agents; D005300:Fertility Agents, Female; D020011:Protective Agents; D016729:Leuprolide
keywords: GnRH agonist; apoptosis; chemotherapy; follicle reserve; granulosa cells; ovarian failure; steroidogenic activity