title: New methods to improve the safety assessment of cryopreserved ovarian tissue for fertility preservation in breast cancer patients.
abstract: OBJECTIVE
  To develop a novel molecular panel of markers to detect breast cancer (BC) disseminated malignant cells in ovarian tissue, and to improve the safety of ovarian tissue transplantation.
  
  
  METHODS
  Experimental study.
  
  
  METHODS
  University hospital.
  
  
  METHODS
  Ten ovarian biopsies from healthy patients, 13 biopsies with diagnosed BC metastasis, and 4 biopsies from primary BC tumor for designing a diagnostic panel of BC cell contamination; 60 ovarian biopsies from BC patients undergoing fertility preservation for validating the panel.
  
  
  METHODS
  Female nude mice.
  
  
  METHODS
  A novel panel for BC malignant cell detection by reverse-transcription polymerase chain reaction (RT-PCR), inmmunohistochemical analysis, in vitro invasion assay and xenotransplantation assayed in ovarian tissue from BC patients.
  
  
  METHODS
  Expression of GCDFP15, MGB1, SBEM, MUC1, WT-1, and NY-BR-01, selected as markers, assessed by quantitative RT-PCR in samples with confirmed BC metastasis. The most sensitive markers were confirmed by immunohistochemistry, and tested in vitro and in vivo.
  
  
  RESULTS
  GCDFP15, MGB1, and SBEM were the most sensitive and specific markers to detect BC metastatic cells when at least one was expressed by quantitative RT-PCR. The panel was validated in 60 patients and confirmed in an in vitro invasion assay, where no invasive cells were observed. Samples negative for BC cells cannot develop disease when xenografted.
  
  
  CONCLUSIONS
  GCDFP15, MGB1, and SBEM were the most sensitive molecules to create a diagnostic panel for BC malignant cell contamination, which may make ovarian tissue cryopreservation and transplantation a safe technique for fertility preservation in BC patients.
authors.affiliation: Departamento de Pediatría, Obstetricia y Ginecología Facultad de Medicina, Universidad de Valencia, Valencia, Spain; Grupo de investigación de Medicina Reproductiva, Instituto de Investigación Sanitario La Fe, Valencia, Spain; Present address: IGENOMIX, Universidad de Valencia, Valencia, Spain.
authors.forename: Beatriz
authors.initials: B
authors.lastname: Rodríguez-Iglesias
authors.affiliation: Grupo de investigación de Medicina Reproductiva, Instituto de Investigación Sanitario La Fe, Valencia, Spain; Unidad de Genética, Hospital Universitario y Politécnico La Fe, Valencia, Spain.
authors.forename: Edurne
authors.initials: E
authors.lastname: Novella-Maestre
authors.affiliation: Departamento de Pediatría, Obstetricia y Ginecología Facultad de Medicina, Universidad de Valencia, Valencia, Spain; Grupo de investigación de Medicina Reproductiva, Instituto de Investigación Sanitario La Fe, Valencia, Spain; Fundación IVI-Universidad de Valencia, INCLIVA, Valencia, Spain; Unidad de Preservación de la Fertilidad, Área de Salud de la Mujer, Hospital Universitario y Politécnico La Fe, Valencia, Spain. Electronic address: sonia.herraiz@ivi.es.
authors.forename: Sonia
authors.initials: S
authors.lastname: Herraiz
authors.affiliation: Departamento de Pediatría, Obstetricia y Ginecología Facultad de Medicina, Universidad de Valencia, Valencia, Spain; Grupo de investigación de Medicina Reproductiva, Instituto de Investigación Sanitario La Fe, Valencia, Spain; Unidad de Preservación de la Fertilidad, Área de Salud de la Mujer, Hospital Universitario y Politécnico La Fe, Valencia, Spain.
authors.forename: César
authors.initials: C
authors.lastname: Díaz-García
authors.affiliation: Fundación IVI-Universidad de Valencia, INCLIVA, Valencia, Spain.
authors.forename: Nuria
authors.initials: N
authors.lastname: Pellicer
authors.affiliation: Departamento de Pediatría, Obstetricia y Ginecología Facultad de Medicina, Universidad de Valencia, Valencia, Spain; Grupo de investigación de Medicina Reproductiva, Instituto de Investigación Sanitario La Fe, Valencia, Spain; Fundación IVI-Universidad de Valencia, INCLIVA, Valencia, Spain; Unidad de Preservación de la Fertilidad, Área de Salud de la Mujer, Hospital Universitario y Politécnico La Fe, Valencia, Spain.
authors.forename: Antonio
authors.initials: A
authors.lastname: Pellicer
mesh_terms: D000328:Adult; D000368:Aged; D000818:Animals; D014408:Biomarkers, Tumor / Q000235:genetics / Q000378:metabolism; D001706:Biopsy; D001943:Breast Neoplasms / Q000235:genetics / Q000378:metabolism / Q000473:pathology* / Q000628:therapy; D002352:Carrier Proteins / Q000235:genetics / Q000378:metabolism; D016022:Case-Control Studies; D015925:Cryopreservation; D005260:Female; D005298:Fertility; D059247:Fertility Preservation / Q000379:methods*; D006023:Glycoproteins / Q000235:genetics / Q000378:metabolism; D064593:Heterografts; D006801:Humans; D007150:Immunohistochemistry; D007247:Infertility, Female / Q000209:etiology / Q000503:physiopathology / Q000628:therapy*; D060785:Mammaglobin A / Q000235:genetics / Q000378:metabolism; D026901:Membrane Transport Proteins; D008819:Mice, Nude; D008875:Middle Aged; D009077:Mucins / Q000235:genetics / Q000378:metabolism; D009361:Neoplasm Invasiveness; D061206:Neoplasm Micrometastasis; D010051:Ovarian Neoplasms / Q000235:genetics / Q000378:metabolism / Q000556:secondary*; D010053:Ovary / Q000378:metabolism / Q000473:pathology* / Q000637:transplantation; D011237:Predictive Value of Tests; D011247:Pregnancy; D015203:Reproducibility of Results; D027724:Reproductive Techniques, Assisted / Q000009:adverse effects; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012307:Risk Factors
chemical_list: D014408:Biomarkers, Tumor; D002352:Carrier Proteins; D006023:Glycoproteins; C511061:MUCL1 protein, human; D060785:Mammaglobin A; D026901:Membrane Transport Proteins; D009077:Mucins; C488892:PIP protein, human; C556448:SCGB2A2 protein, human
keywords: Breast cancer; GCDFP15; MGB1; SBEM; ovarian cortex cryopreservation