Evaluation of minimal disseminated disease in cryopreserved ovarian tissue from bone and soft tissue sarcoma patients
Evaluation of minimal disseminated disease in cryopreserved ovarian tissue from bone and soft tissue sarcoma patients
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DOI：
10.1093/humrep/dew193
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发表时间：
2016-10-01
影响因子：
6.1
通讯作者：
Poirel, H.
中科院分区：
文献类型：
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作者：
Dolmans, M. M.;Iwahara, Y.;Poirel, H.
在肉瘤患者冷冻保存的卵巢组织中发现恶性细胞的风险是什么？在26例患者的冷冻-解冻卵巢组织中，采用任何敏感的方法均未检测到微小播散性疾病（MDD）。在白血病的情况下，恶性细胞通过移植物传播的风险是众所周知的，并被广泛记录。然而，对于骨癌，如尤文肉瘤或骨肉瘤，只有少数病例报告已发表。这些癌症通常影响青春期前的女孩，其中卵巢组织冷冻保存和移植是保存生育能力的唯一选择。骨/软组织肉瘤患者冷冻保存的卵巢组织中恶性细胞的存在与疾病特异性标记物一起研究，使用免疫组织化学（IHC），FISH和实时定量RT-PCR（qPCR），48名肉瘤患者参加了这项研究，其中12人随后死亡。在每种情况下，研究原发性肿瘤的组织，以确定标记物（免疫组织化学和/或分子），以逐个分析卵巢组织。无法评价骨肉瘤（n = 15）、脂肪肉瘤（n = 1）和未分化肉瘤（n = 5）患者的卵巢组织，因为在其任何原发性肿瘤组织中均未通过FISH或敏感性IHC检测到特异性标志物。1例Li-Fraumeni综合征患者也从研究中排除。因此，对26名患者的卵巢组织进行了IHC分析，对19名患者进行了qPCR分析。涉及的原发性肿瘤是尤文肉瘤家族肿瘤（n = 14）、横纹肌肉瘤（n = 7）、滑膜肉瘤（n = 2）、透明细胞肉瘤（n = 2）和恶性外周神经鞘瘤（n = 1）ADD在该最大报告系列中使用敏感技术在26个分析样品中的任何一个中均未检测到。甚至来自随后死亡的患者和/或出现转移的患者（11/26），因此是最具侵袭性的骨癌形式。事实上，对尤文肉瘤家族肿瘤患者（n = 14）进行的抗CD 99 IHC和PCR在所有病例中均为阴性。在软组织肉瘤患者（n = 12）中，原始肿瘤标记物通过IHC检测到，并且在卵巢组织中为阴性。PCR只能在6/12的患者中进行，再次证明为阴性。用于移植的冷冻保存的卵巢碎片无法进行检测，因此这种恶性细胞的分析不能保证所有冷冻保存的碎片都不含有任何播散性疾病。此外，并非所有类型的肿瘤都能轻易获得分子标记物。这些结果对于卵巢恶性细胞移植的风险是令人放心的，因为该研究涉及包括不同类型肉瘤的大系列。我们相信，这将有助于临床医生在他们的病人咨询生育能力的保存和恢复。这项工作是由国家科学研究基金会的支持下，赠款编号7.4578.14（T，l，vie到MS）和5/4/150/5到MMD。作者声明没有竞争性的经济利益。
What is the risk of finding malignant cells in cryopreserved ovarian tissue from sarcoma patients?Minimal disseminated disease (MDD) was not detected in frozen-thawed ovarian tissue from 26 patients by any of the sensitive methods applied.In case of leukemia, the risk of malignant cell transmission through the graft is well known and widely documented. However, for bone cancer, like Ewing sarcoma or osteosarcoma, only a small number of case reports, have been published. These cancers often affect prepubertal girls, in whom ovarian tissue cryopreservation and transplantation is the only option to preserve fertility.The presence of malignant cells in cryopreserved ovarian tissue from patients with bone/soft tissue sarcoma was investigated with disease-specific markers for each patient, using immunohistochemistry (IHC), FISH and real-time quantitative RT-PCR (qPCR), with the original tumor serving as a positive control.Forty-eight sarcoma patients were enrolled in the study, 12 of whom subsequently died. In each case, tissue from the primary tumor was investigated in order to identify markers (immunohistochemical and/or molecular) to analyze the ovarian tissue case by case. Ovarian tissue from osteosarcoma (n = 15), liposarcoma (n = 1) and undifferentiated sarcoma (n = 5) patients could not be evaluated, as no specific markers were detected by FISH or sensitive IHC in any of their primary tumoral tissue. One patient with Li-Fraumeni syndrome was also excluded from the study. IHC analyses were therefore performed on ovarian tissue from 26 patients and qPCR on 19. The primary tumors involved were Ewing sarcoma family of tumors (n = 14), rhabdomyosarcoma (n = 7), synovial sarcoma (n = 2), clear cell sarcoma (n = 2) and a malignant peripheral nerve sheath tumor (n = 1).MDD was not detected in any of the 26 analyzed samples using sensitive techniques in this largest reported series, even from patients who subsequently died and/or those who presented with metastasis (11/26), hence the most aggressive forms of bone cancer. Indeed, anti-CD99 IHC and PCR performed on patients presenting with Ewing sarcoma family of tumors (n = 14) was negative in all cases. In patients with soft tissue sarcoma (n = 12) primitive tumor markers were detected by IHC and were negative in ovarian tissue. PCR could only be performed in 6/12 of these patients, again proving negative.Cryopreserved ovarian fragments to be transplanted cannot be tested, so this analysis of malignant cells cannot guarantee that all cryopreserved fragments will not contain any disseminated disease. Moreover, molecular markers are not readily available for all types of tumors.These results are reassuring regarding the risk of malignant cells in the ovary for transplantation, as the study involves a large series including different types of sarcomas. We believe this will help clinicians in their patient counseling for fertility preservation and restoration.This work was supported by the Fonds National de la Recherche Scientifique de Belgique-FNRS under Grants Nos 7.4578.14 (T,l,vie to MS) and 5/4/150/5 to MMD. The authors declare no competing financial interests.