Inhibitors of apoptosis protect the ovarian reserve from cyclophosphamide
Inhibitors of apoptosis protect the ovarian reserve from cyclophosphamide
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DOI：
10.1530/joe-18-0370
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发表时间：
2019-02-01
影响因子：
4
通讯作者：
Kim, So-Youn
中科院分区：
文献类型：
--
作者：
Luan, Yi;Edmonds, Maxwell E.;Kim, So-Youn
癌症治疗可能会导致脱靶效应，包括卵巢损伤，这可能导致女孩和绝经前妇女出现原发性卵巢功能不全。卵巢储备中的卵泡丧失会导致卵巢内分泌功能障碍和生育能力受损。环磷酰胺 (CPA) 是一种常用的化疗和免疫抑制剂，是一种性腺毒性药物，通过交联 DNA 破坏卵巢细胞。为了保护卵巢免受 CPA 损伤，我们试图精确绘制 CPA 耗尽卵巢储备的机制。我们发现，CPA 特异性地消耗原始卵泡，而不影响体内三种独立小鼠品系（CD-1、C57BL/6J 和 BALB/cJ）的初级和次级卵泡。我们直接在体外测试了CPA活性代谢物1μM 4-羟基过氧环磷酰胺(4-HC)的作用，并证实了原始卵母细胞的损失，但初级和次级卵泡的数量没有变化。我们证明，注射 CPA 3 天后，磷酸化 AKT (p-AKT) 和裂解的 PARP (cPARP) 存在于原始卵母细胞中，这与这些标记物作为细胞凋亡级联的一部分的作用一致。有趣的是，p-AKT 阳性原始卵母细胞共表达 cPARP。用细胞凋亡途径成分的特异性抑制剂 ETP46464 和 CHK2 治疗动物，可在体外阻断 4-HC 诱导的 DNA 损伤。这些数据表明，CPA 通过刺激细胞凋亡途径来靶向卵巢储备中的原始生殖细胞。保护原始生殖细胞免受 CPA 脱靶效应影响的辅助疗法可能会降低 POI 的风险。
Cancer therapy can cause off-target effects including ovarian damage, which may result in primary ovarian insufficiency in girls and premenopausal women. Loss of ovarian follicles within the ovarian reserve leads to ovarian endocrine dysfunction and impaired fertility. Cyclophosphamide (CPA), a commonly used chemotherapeutic and immunosuppressant agent, is a gonadotoxic agent that destroys ovarian cells by crosslinking DNA. To protect the ovary against CPA damage, we sought to precisely map the mechanism by which the ovarian reserve is depleted by CPA. We found that CPA specifically depletes primordial follicles without affecting primary and secondary follicles in three independent murine strains (CD-1, C57BL/6J and BALB/cJ) in vivo. We directly tested the effect of the active metabolite of CPA, 1 mu M 4-hydroxyperoxycyclophophamide (4-HC), in vitro and confirmed the loss of primordial oocytes but no change in the number of primary and secondary follicles. We demonstrated that phospho-AKT (p-AKT) and cleaved PARP (cPARP) are present in primordial oocytes 3 days after CPA injection, consistent with the role of these markers as part of the apoptotic cascade. Interestingly, p-AKT positive primordial oocytes co-expressed cPARP. Treatment of animals with specific inhibitors of apoptotic pathway components, ETP46464 and CHK2, blocked 4-HC-induced DNA damage in vitro. These data suggest that CPA targets primordial germ cells in the ovarian reserve by stimulating apoptosis pathways. Adjuvant therapies to protect primordial germ cells from the off-target effects of CPA may reduce the risk of POI.