Spermatogonial quantity in human prepubertal testicular tissue collected for fertility preservation prior to potentially sterilizing therapy
Spermatogonial quantity in human prepubertal testicular tissue collected for fertility preservation prior to potentially sterilizing therapy
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DOI：
10.1093/humrep/dey240
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发表时间：
2018-09-01
影响因子：
6.1
通讯作者：
Jahnukainen, K.
中科院分区：
文献类型：
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作者：
Stukenborg, J. -B.;Alves-Lopes, J. P.;Jahnukainen, K.
研究问题：化疗暴露(含或不含烷化剂)或初步诊断是否影响人类青春期前睾丸组织中的精原细胞数量？总结回答：在接受烷化剂治疗或使用羟基脲治疗镰状细胞病的青春期前男孩的睾丸中，精原细胞数量显著减少。已知的是：精原干细胞冷冻保存，治疗后移植到睾丸，是恢复儿童生育能力的一种建议的临床选择。这一方法背后的关键临床考虑是足够数量的健康冷冻精原细胞。然而，由于大多数患有恶性肿瘤的男孩开始治疗时使用的不是潜在的绝育药物，他们在考虑睾丸组织冷冻保存之前已经接受了一些化疗。研究设计、大小、持续时间：我们检查了青春期前睾丸组织的组织切片，以阐明化疗暴露或初步诊断是否会影响精原细胞的数量。作为NORDFERTIL研究的一部分，从2014年到2017年连续32名男孩(平均年龄6.3+/-3.8岁)和来自内部生物库的14名对照样本(来自内部生物库的14名对照样本)收集的石蜡包埋睾丸组织样本的治疗特征和组织学标准参考值，评估了每一横切小管的精原细胞数量(S/T)。芬兰和冰岛面临着与极高不孕症风险相关的治疗，他们被提供了睾丸冷冻保存的实验程序。排除标准为睾丸体积10毫升和高出血或感染风险。恶性肿瘤诊断18例，非恶性诊断14例。20例患者在化疗后1~45天进行了睾丸活检，12例患者未接受任何化疗。此外，从卡罗林斯卡大学医院病理科内部生物库获得的14例无睾丸病理报告的患者的睾丸组织样本，除了从最近发表的荟萃分析中获得的参考值之外，还被作为对照样本。通过形态和免疫组织化学分析来评估精原细胞的数量。MAIN结果和机会的作用：主要发现，在接受烷化剂或羟基尿素治疗镰状细胞病的男孩中，精原细胞数量显著减少。接触烷化剂男童(0.2+/-0.3，n=6)和镰状细胞病合并羟基脲接触男童(0.3+/-0.6，n=6)的S/T比值(P=0.003和P=0.008)显著低于非烷化剂接触组和生物库对照组(分别为1.7+/-1.0，n=8和4.1+/-4.6，n=14)。生物库对照组和暴露于非烷基化剂的患者组的睾丸组织样本的平均S/T值均在最近公布的标准参考值范围内。限制，谨慎理由：本研究纳入的正常睾丸组织样本来自卡罗林斯卡大学医院内部生物库。样本被认为是正常的，如果在分析的样本中没有报告睾丸病理，则包括在研究中。然而，关于以前的药物治疗和包括在这个生物库中的患者的睾丸体积的详细信息是不可获得的。研究结果的广泛意义：这项研究首次总结了青春期前患者队列中潜在绝育治疗前后的精原细胞数量。面临癌症和细胞毒治疗的男孩被认为是将从新的生育保护技术中受益的主要群体。以前没有报告将精原细胞数量与烷化剂和蒽环类药物(非烷化剂)的累积暴露联系起来，也没有关于这一特定患者队列中细胞毒性暴露的时间的信息。对于有保留生育能力的青春期前男孩，应该在开始用烷化剂进行化疗之前获取睾丸组织，而对于患有镰状细胞病并接受羟基尿素治疗的男孩，这种方法可能不可行。研究资金/竞争兴趣(S)：这项研究得到了瑞典儿童癌症基金会的资助(PR2016-0124；TJ2016-0093；PR2015-0073，TJ2015-0046)(J.-B.S.和K.J.)，简和丹·奥尔森基金会(2016-33)(J.-B.S.)，芬兰癌症学会(K.J.)，儿科研究基金会(J.-B.S.)，Kronprinsessan Lovisas Forvising for Barnasjukvard/Stiftelsen Axel Tielmann Minnerfond，Samariten Foundation(J.-B.S.)，Vare基金会儿科癌症研究(K.J.)和瑞典研究理事会(2012-6352)(O.S.)。R.T.M.得到了惠康信托奖学金的支持(09822)。J.P.A.-L.和M.K.得到了ITN玛丽·居里项目“成长精子”(EU-fp7-People-2013-ITN 603568)的支持。作者声明没有利益冲突。
STUDY QUESTION: Does chemotherapy exposure (with or without alkylating agents) or primary diagnosis affect spermatogonial quantity in human prepubertal testicular tissue?SUMMARY ANSWER: Spermatogonial quantity is significantly reduced in testes of prepubertal boys treated with alkylating agent therapies or with hydroxyurea for sickle cell disease.WHAT IS KNOWN ALREADY: Cryopreservation of spermatogonial stem cells, followed by transplantation into the testis after treatment, is a proposed clinical option for fertility restoration in children. The key clinical consideration behind this approach is a sufficient quantity of healthy cryopreserved spermatogonia. However, since most boys with malignancies start therapy with agents that are not potentially sterilizing, they will have already received some chemotherapy before testicular tissue cryopreservation is considered.STUDY DESIGN, SIZE, DURATION: We examined histological sections of prepubertal testicular tissue to elucidate whether chemotherapy exposure or primary diagnosis affects spermatogonial quantity. Quantity of spermatogonia per transverse tubular cross-section (S/T) was assessed in relation to treatment characteristics and normative reference values in histological sections of paraffin embedded testicular tissue samples collected from 32 consecutive boy patients (aged 6.3 +/- 3.8 [mean +/- SD] years) between 2014 and 2017, as part of the NORDFERTIL study, and in 14 control samples (from boys aged 5.6 +/- 5.0 [mean +/- SD] years) from an internal biobank.PARTICIPANTS/MATERIALS, SETTING, METHODS: Prepubertal boys in Sweden, Finland and Iceland who were facing treatments associated with a very high risk of infertility, were offered the experimental procedure of testicular cryopreservation. Exclusion criteria were testicular volumes > 10 ml and high bleeding or infection risk. There were 18 patients with a diagnosis of malignancy and 14 patients a nonmalignant diagnosis. While 20 patients had the testicular biopsy performed 1-45 days after chemotherapy, 12 patients had not received any chemotherapy. In addition, 14 testicular tissue samples of patients with no reported testicular pathology, obtained from the internal biobank of the Department of Pathology at Karolinska University Hospital, were included as control samples in addition to reference values obtained from a recently published meta-analysis. The quantity of spermatogonia was assessed by both morphological and immunohistochemical analysis.MAIN RESULTS AND THE ROLE OF CHANCE: The main finding was a significant reduction in spermatogonial cell counts in boys treated with alkylating agents or with hydroxyurea for sickle cell disease. The mean S/T values in boys exposed to alkylating agents (0.2 +/- 0.3, n = 6) or in boys with sickle cell disease and exposed to hydroxyurea (0.3 +/- 0.6, n = 6) were significantly lower (P = 0.003 and P = 0.008, respectively) than in a group exposed to non-alkylating agents or in biobank control samples (1.7 +/- 1.0, n = 8 and 4.1 +/- 4.6, n = 14, respectively). The mean S/T values of the testicular tissue samples included in the biobank control group and the patient group exposed to nonalkylating agents were within recently published normative reference values.LIMITATIONS, REASONS FOR CAUTION: Normal testicular tissue samples included in this study were obtained from the internal biobank of Karolinska University Hospital. Samples were considered normal and included in the study if no testicular pathology was reported in the analysed samples. However, detailed information regarding previous medical treatments and testicular volumes of patients included in this biobank were not available.WIDER IMPLICATIONS OF THE FINDINGS: This study summarizes, for the first time, spermatogonial quantity in a prepubertal patient cohort just before and after potentially sterilizing treatments. Boys facing cancer and cytotoxic therapies are regarded as the major group who will benefit from novel fertility preservation techniques. There are no previous reports correlating spermatogonial quantity to cumulative exposure to alkylating agents and anthracyclines (non-alkylating agents) and no information about the timing of cytotoxic exposures among this particular patient cohort. For prepubertal boys in whom fertility preservation is indicated, testicular tissue should be obtained before initiation of chemotherapy with alkylating agents, whilst for those with sickle cell disease and treated with hydroxyurea, this approach to fertility preservation may not be feasible.STUDY FUNDING/COMPETING INTEREST(S): This study was supported by grants from The Swedish Childhood Cancer Foundation (PR2016-0124; TJ2016-0093; PR2015-0073, TJ2015-0046) (J.-B.S. and K.J.), the Jane and Dan Olssons Foundation (2016-33) (J.-B.S.), the Finnish Cancer Society (K.J.), the Foundation for Paediatric Research (J.-B.S.), Kronprinsessan Lovisas Forening For Barnasjukvard/Stiftelsen Axel Tielmans Minnesfond, Samariten Foundation (J.-B.S.), the Vare Foundation for Paediatric Cancer Research (K.J.) and the Swedish Research Council (2012-6352) (O.S.). R.T.M. was supported by a Wellcome Trust Fellowship (09822). J.P.A.-L. and M.K. were supported by the ITN Marie Curie program 'Growsperm' (EU-FP7-PEOPLE-2013-ITN 603568). The authors declare no conflicts of interest.