title: Treatment with human, recombinant FSH improves sperm DNA fragmentation in idiopathic infertile men depending on the FSH receptor polymorphism p.N680S: a pharmacogenetic study.
abstract: Does the sperm DNA fragmentation index (DFI) improve depending on the FSH receptor (FSHR) genotype as assessed by the nonsynonymous polymorphisms rs6166 (p.N680S) after 3 months of recombinant FSH treatment in men with idiopathic infertility?
  
  
  
  FSH treatment significantly improves sperm DFI only in idiopathic infertile men with the p.N680S homozygous N FSHR.
  
  
  
  FSH, fundamental for spermatogenesis, is empirically used to treat male idiopathic infertility and several studies suggest that DFI could be a candidate predictor of response to FSH treatment, in terms of probability to conceive. Furthermore, it is known that the FSHR single nucleotide polymorphism (SNP) rs6166 (p.N680S) influences ovarian response in women and testicular volume in men.
  
  
  
  A multicenter, longitudinal, prospective, open-label, two-arm clinical trial was performed. Subjects enrolled were idiopathic infertile men who received 150 IU recombinant human FSH s.c. every other day for 12 weeks and were followed-up for a further 12 weeks after FSH withdrawal. Patients were evaluated at baseline, at the end of treatment and at the end of follow-up.
  
  
  
  Eighty-nine men with idiopathic infertility carrier of the FSHR p.N680S homozygous N or S genotype, FSH ≤ 8 IU/l and DFI >15%, were enrolled. A total of 66 patients had DFI analysis completed on at least two visits. DFI was evaluated in one laboratory by TUNEL/PI (propidium iodide) assay coupled to flow cytometry, resolving two different fractions of sperm, namely the 'brighter' and 'dimmer' sperm DFI fractions.
  
  
  
  Thirty-eight men (57.6%) were carriers of the p.N680S homozygous N and 28 (42.4%) of the homozygous S FSHR. Sperm concentration/number was highly heterogeneous and both groups included men ranging from severe oligozoospermia to normozoospermia. Total DFI was significantly lower at the end of the study in homozygous carriers of the p.N680S N versus p.N680S S allele (P = 0.008). Total DFI decreased significantly from baseline to the end of the study (P = 0.021) only in carriers of the p.N680S homozygous N polymorphism, and this decrease involved the sperm population containing vital sperm (i.e. brighter sperm) (P = 0.008). The dimmer sperm DFI fraction, including only nonvital sperm, was significantly larger in p.N680S S homozygous patients than in homozygous N men (P = 0.018). Total DFI was inversely related to total sperm number (P = 0.020) and progressive sperm motility (P = 0.014). When patients were further stratified according to sperm concentration (normoozospermic versus oligozoospermic) or -211G>T polymorphism in the FSHB gene (rs10835638) (homozygous G versus others), the significant improvement of sperm DFI in FSHR p.N680S homozygous N men was independent of sperm concentration and associated with the homozygous FSHB -211G>T homozygous G genotype.
  
  
  
  The statistical power of the study is 86.9% with alpha error 0.05. This is the first pharmacogenetic study suggesting that FSH treatment induces a significant improvement of total DFI in men carriers of the p.N680S homozygous N FSHR; however, the results need to be confirmed in larger studies using a personalized FSH dosage and treatment duration.
  
  
  
  The evaluation of sperm DFI as a surrogate marker of sperm quality, and of the FSHR SNP rs6166 (p.N680S), might be useful to predict the response to FSH treatment in men with idiopathic infertility.
  
  
  
  The study was supported by an unrestricted grant to M.S. and H.M.B. from Merck Serono that provided the drug used in the study. MS received additional grants from Merck Serono and IBSA as well as honoraria from Merck Serono. The remaining authors declare that no conflicts of interest are present.
  
  
  
  EudraCT number 2010-020240-35.
authors.affiliation: Department of Biomedical, Metabolic and Neural Sciences, University of Modena & Reggio Emilia, Modena, Italy Azienda USL of Modena, Modena, Italy Center for Genomic Research, University of Modena & Reggio Emilia, Modena, Italy.
authors.forename: Manuela
authors.initials: M
authors.lastname: Simoni
authors.affiliation: Department of Biomedical, Metabolic and Neural Sciences, University of Modena & Reggio Emilia, Modena, Italy Azienda USL of Modena, Modena, Italy santi.daniele@gmail.com.
authors.forename: Daniele
authors.initials: D
authors.lastname: Santi
authors.affiliation: Department of Gynecology, Division of Gynecology and Reproductive Medicine, Humanitas Fertility Center, Humanitas Research Hospital, Rozzano (Milan), Italy.
authors.forename: Luciano
authors.initials: L
authors.lastname: Negri
authors.affiliation: Center for Reproductive Medicine and Andrology, Martin Luther University Halle-Wittenberg, Halle, Germany.
authors.forename: Ivan
authors.initials: I
authors.lastname: Hoffmann
authors.affiliation: Department of Experimental and Clinical Biomedical Sciences, Sexual Medicine and Andrology Unit, University of Florence, Firenze, Italy.
authors.forename: Monica
authors.initials: M
authors.lastname: Muratori
authors.affiliation: Department of Experimental and Clinical Biomedical Sciences, Sexual Medicine and Andrology Unit, University of Florence, Firenze, Italy.
authors.forename: Elisabetta
authors.initials: E
authors.lastname: Baldi
authors.affiliation: Department of Experimental and Clinical Biomedical Sciences, Sexual Medicine and Andrology Unit, University of Florence, Firenze, Italy.
authors.forename: Marta
authors.initials: M
authors.lastname: Cambi
authors.affiliation: Center for Reproductive Medicine and Andrology, Martin Luther University Halle-Wittenberg, Halle, Germany.
authors.forename: Marios
authors.initials: M
authors.lastname: Marcou
authors.affiliation: Center for Reproductive Medicine and Andrology, Martin Luther University Halle-Wittenberg, Halle, Germany.
authors.forename: Thomas
authors.initials: T
authors.lastname: Greither
authors.affiliation: Department of Laboratory Medicine and Pathological Anatomy, Azienda USL of Modena, Modena, Italy.
authors.forename: Enrica
authors.initials: E
authors.lastname: Baraldi
authors.affiliation: Department of Laboratory Medicine and Pathological Anatomy, Azienda USL of Modena, Modena, Italy.
authors.forename: Simonetta
authors.initials: S
authors.lastname: Tagliavini
authors.affiliation: Department of Laboratory Medicine and Pathological Anatomy, Azienda USL of Modena, Modena, Italy.
authors.forename: Daniela
authors.initials: D
authors.lastname: Carra
authors.affiliation: Department of Experimental Medicine, University of Rome "La Sapienza", Roma, Italy.
authors.forename: Francesco
authors.initials: F
authors.lastname: Lombardo
authors.affiliation: Department of Experimental Medicine, University of Rome "La Sapienza", Roma, Italy.
authors.forename: Loredana
authors.initials: L
authors.lastname: Gandini
authors.affiliation: Department of Experimental Medicine, University of Rome "La Sapienza", Roma, Italy.
authors.forename: Francesco
authors.initials: F
authors.lastname: Pallotti
authors.affiliation: Department of Experimental and Clinical Biomedical Sciences, Sexual Medicine and Andrology Unit, University of Florence, Firenze, Italy.
authors.forename: Csilla
authors.initials: C
authors.lastname: Krausz
authors.affiliation: Department of Experimental and Clinical Biomedical Sciences, Sexual Medicine and Andrology Unit, University of Florence, Firenze, Italy.
authors.forename: Giulia
authors.initials: G
authors.lastname: Rastrelli
authors.affiliation: Department of Medicine, Unit of Andrology and Reproductive Medicine, University of Padova, Padova, Italy.
authors.forename: Alberto
authors.initials: A
authors.lastname: Ferlin
authors.affiliation: Department of Medicine, Unit of Andrology and Reproductive Medicine, University of Padova, Padova, Italy.
authors.forename: Massimo
authors.initials: M
authors.lastname: Menegazzo
authors.affiliation: Department of Biomedical, Metabolic and Neural Sciences, University of Modena & Reggio Emilia, Modena, Italy Center for Genomic Research, University of Modena & Reggio Emilia, Modena, Italy.
authors.forename: Elisa
authors.initials: E
authors.lastname: Pignatti
authors.affiliation: Department of Biomedical, Metabolic and Neural Sciences, University of Modena & Reggio Emilia, Modena, Italy Azienda USL of Modena, Modena, Italy.
authors.forename: Francesca
authors.initials: F
authors.lastname: Linari
authors.affiliation: Department of Biomedical, Metabolic and Neural Sciences, University of Modena & Reggio Emilia, Modena, Italy Center for Genomic Research, University of Modena & Reggio Emilia, Modena, Italy.
authors.forename: Marco
authors.initials: M
authors.lastname: Marino
authors.affiliation: Department of Gynecology, Division of Gynecology and Reproductive Medicine, Humanitas Fertility Center, Humanitas Research Hospital, Rozzano (Milan), Italy.
authors.forename: Renzo
authors.initials: R
authors.lastname: Benaglia
authors.affiliation: Department of Gynecology, Division of Gynecology and Reproductive Medicine, Humanitas Fertility Center, Humanitas Research Hospital, Rozzano (Milan), Italy.
authors.forename: Paolo E
authors.initials: PE
authors.lastname: Levi-Setti
authors.affiliation: Center for Reproductive Medicine and Andrology, Martin Luther University Halle-Wittenberg, Halle, Germany.
authors.forename: Hermann M
authors.initials: HM
authors.lastname: Behre
mesh_terms: D000328:Adult; D000483:Alleles; D053938:DNA Fragmentation / Q000187:drug effects*; D043373:Follicle Stimulating Hormone, Human / Q000494:pharmacology* / Q000627:therapeutic use; D005838:Genotype; D006801:Humans; D007248:Infertility, Male / Q000188:drug therapy* / Q000235:genetics; D008297:Male; D000071185:Pharmacogenomic Testing; D020641:Polymorphism, Single Nucleotide; D011962:Receptors, FSH / Q000235:genetics*; D013081:Sperm Motility / Q000187:drug effects; D013091:Spermatogenesis / Q000235:genetics; D013094:Spermatozoa / Q000187:drug effects; D016896:Treatment Outcome
chemical_list: D043373:Follicle Stimulating Hormone, Human; D011962:Receptors, FSH
keywords: FSH; FSH receptor; FSHB -211G/T; FSHR pN680S; male infertility; sperm DNA fragmentation index