Toward gene therapy of premature ovarian failure: intraovarian injection of adenovirus expressing human FSH receptor restores folliculogenesis in FSHR(-/-) FORKO mice
Toward gene therapy of premature ovarian failure: intraovarian injection of adenovirus expressing human FSH receptor restores folliculogenesis in FSHR(-/-) FORKO mice
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DOI：
10.1093/molehr/gaq003
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发表时间：
2010-04-01
影响因子：
4
通讯作者：
Al-Hendy, A.
中科院分区：
文献类型：
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作者：
Ghadami, M.;El-Demerdash, E.;Al-Hendy, A.
促卵泡激素受体（FSHR）基因中的纯合错义突变C566 T与卵巢早衰有关。这种疾病导致正常核型女性不孕，卵泡刺激素（FSH）升高，血清雌激素水平降低。携带突变的FSHR基因的雌性小鼠，称为促卵泡素受体敲除（FORKO），表现出相似的表型，并且由于在初级阶段卵泡发生阻滞而不育。我们研究了双侧卵巢内注射表达正常拷贝人FSHR的腺病毒对6-10周雌性FORKO小鼠生殖系统的影响。对照组将Ad-LacZ直接注射于卵巢内。在注射后2、4、8和12周处死动物，并收集组织用于评价。治疗组小鼠在每日阴道涂片中显示雌激素变化，而对照组动物在间情期保持固定。组织学评价显示，治疗组中平均26 +/- 4个卵泡/卵巢，窦期8 +/- 2个卵泡，而Ad-LacZ对照小鼠中仅5 +/- 2个卵泡，窦期0个卵泡。血清孕酮水平无明显变化，而雌激素水平升高2-3倍（P < 0.02），FSH下降50%（P < 0.04）。FSHR mRNA在治疗组卵巢中表达。总之，卵巢内注射表达人FSHR基因的腺病毒能够恢复FSH反应性，重新启动卵巢卵泡发生，并恢复雌性FORKO小鼠的雌激素产生。Ad-LacZ注射表明不存在腺病毒DNA向后代的系统性病毒传播或生殖系传播。
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.