MRC Centre for Reproductive Health, The Queen's Medical Research Institute, The University of Edinburgh, 47 Little France Crescent, Edinburgh, EH16 4TJ, Scotland, UK.
KK Women's and Children's Hospital, Bukit Timah Rd, 100, Singapore, 229899, Singapore.
BMC Med. 2020 Dec 4;18(1):374. doi: 10.1186/s12916-020-01844-y.
Clinical studies indicate chemotherapy agents used in childhood cancer treatment regimens may impact future fertility. However, effects of individual agents on prepubertal human testis, necessary to identify later risk, have not been determined. The study aimed to investigate the impact of cisplatin, commonly used in childhood cancer, on immature (foetal and prepubertal) human testicular tissues. Comparison was made with carboplatin, which is used as an alternative to cisplatin in order to reduce toxicity in healthy tissues.
We developed an organotypic culture system combined with xenografting to determine the effect of clinically-relevant exposure to platinum-based chemotherapeutics on human testis. Human foetal and prepubertal testicular tissues were cultured and exposed to cisplatin, carboplatin or vehicle for 24 h, followed by 24-240 h in culture or long-term xenografting. Survival, proliferation and apoptosis of prepubertal germ stem cell populations (gonocytes and spermatogonia), critical for sperm production in adulthood, were quantified.
Cisplatin exposure resulted in a significant reduction in the total number of germ cells (- 44%, p < 0.0001) in human foetal testis, which involved an initial loss of gonocytes followed by a significant reduction in spermatogonia. This coincided with a reduction (- 70%, p < 0.05) in germ cell proliferation. Cisplatin exposure resulted in similar effects on total germ cell number (including spermatogonial stem cells) in prepubertal human testicular tissues, demonstrating direct relevance to childhood cancer patients. Xenografting of cisplatin-exposed human foetal testicular tissue demonstrated that germ cell loss (- 42%, p < 0.01) persisted at 12 weeks. Comparison between exposures to human-relevant concentrations of cisplatin and carboplatin revealed a very similar degree of germ cell loss at 240 h post-exposure.
This is the first demonstration of direct effects of chemotherapy exposure on germ cell populations in human foetal and prepubertal testis, demonstrating platinum-induced loss of all germ cell populations, and similar effects of cisplatin or carboplatin. Furthermore, these experimental approaches can be used to determine the effects of established and novel cancer therapies on the developing testis that will inform fertility counselling and development of strategies to preserve fertility in children with cancer.
临床研究表明，儿童癌症治疗方案中使用的化疗药物可能会影响未来的生育能力。然而，尚未确定个别药物对青春期前人类睾丸的影响，而这对于确定日后的风险是必要的。本研究旨在调查顺铂（一种常用于儿童癌症的药物）对未成熟（胎儿和青春期前）人类睾丸组织的影响。并将其与卡铂（一种用于降低健康组织毒性的顺铂替代品）进行比较。
我们开发了一种器官型培养系统，结合异种移植，以确定临床相关浓度的铂类化疗药物对人类睾丸的影响。将胎儿和青春期前的睾丸组织进行培养，并分别用顺铂、卡铂或载体处理 24 小时，然后在培养物中培养 24-240 小时或进行长期异种移植。定量分析了青春期前生殖干细胞群（精原细胞和精母细胞）的存活、增殖和凋亡情况，这些细胞对于成年后的精子生成至关重要。
顺铂暴露导致人类胎儿睾丸中的生殖细胞总数减少（-44%，p<0.0001），这涉及到精原细胞的初始丢失，随后精母细胞数量显著减少。这与生殖细胞增殖减少（-70%，p<0.05）相一致。顺铂暴露对青春期前人类睾丸组织中的总生殖细胞数量（包括精原干细胞）也有类似的影响，这与儿童癌症患者直接相关。顺铂暴露的人类胎儿睾丸组织的异种移植显示，12 周时生殖细胞丢失仍持续（-42%，p<0.01）。将人类暴露于顺铂和卡铂的相关浓度进行比较，结果表明，在暴露后 240 小时，生殖细胞丢失程度非常相似。
这是首次证明化疗药物暴露对人类胎儿和青春期前睾丸生殖细胞群的直接影响，表明铂类药物诱导所有生殖细胞群丢失，顺铂和卡铂的作用相似。此外，这些实验方法可用于确定现有的和新型癌症疗法对发育中睾丸的影响，为癌症患儿的生育咨询和保留生育能力策略的制定提供信息。
