Mol. Hum. Reprod. Advance Access originally published online on December 14, 2007
Molecular Human Reproduction 2008 14(1):9-15; doi:10.1093/molehr/gam077
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Toward gene therapy of primary ovarian failure: adenovirus expressing human FSH receptor corrects the Finnish C566T mutation
1Department of Obstetrics and Gynecology, University of Texas Medical Branch, Galveston, TX 77555, US 2Department of Genetics, School of Medicine, Tehran University of Medical Sciences, Tehran, Islamic Republic of Iran 3Department of Obstetrics and Gynecology and Reproductive Sciences, University of Saskatchewan, Saskatoon, Saskatchewan, Canada 4Present address: Department of Obstetrics and Gynecology, Center for Women Health Research, Meharry Medical College, Nashville, TN 37208, USA
5 Correspondence address. Tel: +615-963-3148; Fax: +615-327-6982; E-mail: ahendy{at}mmc.edu
Resistance ovarian syndrome is a heterogeneous disorder inherited as a Mendelian recessive trait and characterized by infertility, primary amenorrhea, normal karyotype and elevated serum FSH and LH levels. An inactivating mutation, C566T, in FSH receptor gene (FSHR) has been identified initially in Finland. We investigated if an adenovirus expressing a normal copy of human FSHR (Ad-hFSHR) has the ability to: (i) transfect granulosa cell lines, (ii) render the transfected cell lines responsive to FSH stimulation and (iii) transcomplement the malfunctioning form of human FSHR gene with C566T mutation. COS-7, JC-410, JC-410-P450-scc-luc and JC-410-StAR-luc cell lines were infected by Ad-hFSHR followed by treatment with FSH. Functional activity of the Ad-hFSHR was tested by measuring cyclic adenosine monophosphate (cAMP) or luciferase activity in response to FSH stimulation, and showed 2–4.6-fold increases in Ad-hFSHR transfected cells compared with untransfected or Ad-LacZ transfected cells, indicating that Ad-hFSHR is functionally active and expressing hFSHR. Generation of cAMP in cells expressing only mutated hFSHR-T566 showed minimal increase after FSH stimulation. Co-transfection of Ad-hFSHR in these cells carrying the malfunction form of human FSHR caused significant increases of 2.2–7.4-fold in FSH dependent cAMP generation (P = 0.0007). We concluded that adenovirus expressing a normal human FSHR can compensate the inactivating human FSHR-C566T mutation and restore FSH responsiveness.
Key words: Finnish C566T mutation/follicle-stimulating hormone receptor/gene therapy of ovarian failure/infertility/primary amenorrhea
Submitted on August 25, 2007; resubmitted on October 10, 2007; accepted on October 19, 2007.