Toward gene therapy of premature ovarian failure: intraovarian injection of adenovirus expressing human FSH receptor restores folliculogenesis in FSHR(-/-) FORKO mice

Abstract

A homozygous missense mutation, C566T, in the follicle stimulation hormone receptor (FSHR) gene has been linked to premature ovarian failure. The disease leads to infertility in a normal karyotype female with an elevated follicle stimulating hormone (FSH) and decreased serum estrogen level. Female mice carrying mutated FSHR gene, called follitropin receptor knockout (FORKO), display similar phenotype and are sterile because of a folliculogenesis block at a primary stage. We investigated the effects of bilateral intra-ovarian injection of an adenovirus expressing a normal copy of human FSHR on the reproductive system of 6-10 weeks female FORKO mice. Ad-LacZ was injected directly into each ovary of the control group. Animals were sacrificed at 2, 4, 8 and 12 weeks post-injection and tissues collected for evaluation. Treated mice showed estrogenic changes in daily vaginal smear whereas control animals remained fixated in the diestrus stage. Histological evaluation showed on average 26 +/- 4 follicles/ovary in treated group with 8 +/- 2 follicles at the antral stage compared with only 5 +/- 2 with zero follicles at antral stage in Ad-LacZ control mice. There was no significant change in serum level of progesterone, however, estrogen level increased 2-3-fold (P < 0.02) and FSH decreased by up to 50% (P < 0.04) in treated animals. FSHR mRNA was detected in the ovaries of the treated group. In conclusion, intra-ovarian injection of an adenovirus expressing human FSHR gene is able to restore FSH responsiveness and reinitiate ovarian folliculogenesis as well as resume estrogen production in female FORKO mice. Ad-LacZ injections indicate the absence of systemic viral dissemination or germ line transmission of adenovirus DNA to offspring.

Development of a technique for efficient gene transfer to antral follicular cells in the mouse ovary

“…In this context, an ovary appears to be a good target for gene delivery to test the function of a GOI, as previously shown by others [Matsumoto et al 1999;Shimizu et al 2003;Shimizu et al 2004;Shimizu et al 2008;Ghadami et al 2010].…”

Genetics of primary ovarian insufficiency: a review

“…Women who are homozygous for an antisense mutation in this gene are diagnosed with POI given the gonadotropin-resistant ovary syndrome, in which the follicular reserve remains, but is not functional. The possibility of gene therapy using adenovirus expressing the correct copy to restore fertility is under study, which has proven successful in mice [41]. In Finland, the mutation is a common cause of POI in 46,XX females, but this does not seem to apply to other countries like USA, Germany, Brazil or Mexico [10].…”

“…Locus Other diseases associated FSHR [41,10] 2p21-p16 Ovarian hyperstimulations yndrome, mucinous cystadenocarcinoma LHR [43] 2p21 Precocious puberty, Leydig cells hypoplasia FSHβ [4] 11p13 Follicle-stimulating hormone deficiency, isolated, pandas LHβ [4] 19q13.32 Frozen shoulder, hypogonadism GNAS [21] 20q13.3 Pseudohypoparathyroidism, Mccune-albrightsyndrome FOXL2 [44] 3q22-q23 Blepharophimosis, blepharophimosis, ptosis and epicanthus inversus syndrome GALT [46] 9p13 Galactosemia, galactokinasedeficiency AIRE [47] 21q22.3 Candidiasis, autoinmunepolyendocrinesyndrome StAR [48] 8p11.2 Lipoid adrenal hyperplasia CYP17A1 [49] 10q24.3 17-alpha-hydroxilase-deficient congenitaladrenalhyperplasia CYP19A1 [71], [50] 15q21.1 Aromatasedeficiency, Leydig cell tumor…”

Genes involved in human premature ovarian failure

“…The p.A189V mutation appears to be particularly frequent only in the Finnish population, and it was not found in most other populations, suggesting a founder effect (da Fonte Kohek et al 1998, Jiang et al 1998, Layman et al 1998, Loutradis et al 2006, Prakash et al 2009. Ghadami et al (2008Ghadami et al ( , 2010 succeeded in restoring FSH responsiveness in various cell lines expressing the mutated FSHR through the transfection of the normal human FSHR gene carried by an adenovirus vector and, very recently, also in the FSHR K/K mouse. These are the first attempts to develop a gene therapy approach for this type of ovarian failure.…”