Batchvarov Iordan Stefanov, Taylor Rachel Williamson, Bustamante-Marín Ximena, Czerwinski Michael, Johnson Erika Segear, Kornbluth Sally, Capel Blanche
Department of Cell Biology, Duke University Medical Center, Durham, NC 27710, USA.
School of Medicine, Duke University Medical Center, Durham, NC 27710, USA.
Mol Hum Reprod. 2016 Dec;22(12):842-851. doi: 10.1093/molehr/gaw064. Epub 2016 Oct 3.
Can host fertility be rescued by grafting of a fragment of a healthy ovary soon after chemotherapy?
We found that grafting a green fluorescent protein (GFP)-positive fragment from a healthy isogenic ovary to the left ovary of a chemo-treated host rescued function and fertility of the grafted host ovary, and resulted in the production of host-derived offspring as late as the sixth litter after chemotherapy (CTx) treatment, whereas none of the ungrafted controls produced a second litter.
In women and girls undergoing chemotherapy, infertility and premature ovarian failure are frequent outcomes. There are accumulating reports of improved endocrine function after autotransplantation of an ovarian fragment, raising the possibility that the transplant is beneficial to the endogenous ovary.
STUDY DESIGN, SIZE, DURATION: We first established a CTx treatment regimen that resulted in the permanent loss of fertility in 100% of female mice of the FVB inbred strain. We grafted an isogenic ovary fragment from a healthy female homozygous for a GFP transgene to the left ovary of 100 CTx-treated hosts, and compared fertility to 39 ungrafted controls in 6 months of continuous matings, using GFP to distinguish offspring derived from the graft, and those derived from the host.
PARTICIPANTS/MATERIALS, SETTING, METHODS: Immunofluoresece and western blot analysis of 39 treated ovaries during and 15 days after CTx treatment revealed elevated apoptosis, rapid loss of granulosa cells and an increased recruitment of growing follicles. Using immunofluorescence and confocal imaging, we tracked the outcome of the grafted tissue over 4 months and its effect on the adjacent and contralateral ovary of the host.
Fifty-three percent of grafted females produced a second litter whereas none of the ungrafted females produced a second litter. The likelihood that this could occur by chance is very low (P < 0.0001).
LIMITATIONS, REASONS FOR CAUTION: These results are shown only in mice, and whether or how they might apply to chemotherapy patients subjected to different CTx regimens is not yet clear.
Our experiments prove that rescue of a chemo-treated ovary is possible, and establish a system to investigate the mechanism of rescue and to identify the factors responsible with the long-term goal of developing therapies for preservation of ovarian endocrine function and fertility in women undergoing chemotherapy.
No large datasets were produced.
STUDY FUNDING/COMPETING INTERESTS: Duke University Medical Center Chancellor's Discovery Grant to BC; ESJ was supported by an NRSA 5F31CA165545; SK was supported by NIH RO1 GM08033; RWT was supported by the Duke University School of Medicine Ovarian Cancer Research Fellowship; XBM was supported by CONICYT. The authors have no conflicts of interest to declare.
化疗后不久移植健康卵巢片段能否挽救宿主的生育能力？
我们发现，将来自健康同基因卵巢的绿色荧光蛋白（GFP）阳性片段移植到化疗后宿主的左卵巢，可挽救移植宿主卵巢的功能和生育能力，并导致化疗（CTx）治疗后第六窝仍能产生宿主来源的后代，而未移植的对照组没有产生第二窝后代。
在接受化疗的女性和女孩中，不孕和卵巢早衰是常见的后果。越来越多的报道称卵巢片段自体移植后内分泌功能有所改善，这增加了移植对卵巢有益的可能性。
研究设计、规模、持续时间：我们首先建立了一种CTx治疗方案，该方案导致100%的FVB近交系雌性小鼠永久性丧失生育能力。我们将来自GFP转基因纯合健康雌性小鼠的同基因卵巢片段移植到100只接受CTx治疗的宿主的左卵巢，并在6个月的连续交配中，将其生育能力与39只未移植的对照组进行比较，利用GFP区分来自移植卵巢和宿主的后代。
参与者/材料、环境、方法：对39只治疗后的卵巢在CTx治疗期间及治疗后15天进行免疫荧光和蛋白质印迹分析，结果显示细胞凋亡增加、颗粒细胞迅速丢失以及生长卵泡募集增加。利用免疫荧光和共聚焦成像，我们追踪了移植组织4个月内的情况及其对宿主相邻和对侧卵巢的影响。
53%的移植雌性小鼠产下一窝后代，而未移植的雌性小鼠均未产下一窝后代。这种情况偶然发生的可能性非常低（P<0.0001）。
局限性、谨慎原因：这些结果仅在小鼠中得到证实，它们是否适用于接受不同CTx方案的化疗患者，或者如何适用，目前尚不清楚。
我们的实验证明化疗后卵巢的挽救是可能的，并建立了一个系统来研究挽救机制，识别相关因素，其长期目标是开发针对接受化疗女性的卵巢内分泌功能和生育能力保存的治疗方法。
未产生大规模数据集。
研究资金/利益冲突：杜克大学医学中心校长发现奖授予BC；ESJ由NRSA 5F31CA165545资助；SK由NIH RO1 GM08033资助；RWT由杜克大学医学院卵巢癌研究奖学金资助；XBM由智利国家科学技术研究委员会资助。作者声明无利益冲突。
