Dias Nunes Joana, Ntemou Elissavet, Van den Steen Géraldine, Findikli Necati, Fastrez Maxime, Delbaere Anne, Lambertini Matteo, Devos Melody, Demeestere Isabelle
Research Laboratory on Human Reproduction, Université Libre de Bruxelles (ULB), Brussels, Belgium.
Fertility Clinic, HUB Erasme Hospital, Brussels, Belgium.
Hum Reprod. 2025 Jul 11. doi: 10.1093/humrep/deaf133.
Are ovarian tissue fragments from patients with BReast CAncer gene 1 (BRCA1)-mutated breast cancer (BC) more sensitive to carboplatin and/or paclitaxel exposure compared to those from non-mutated patients with BC?
Carboplatin and paclitaxel treatment showed similar gonadotoxicity, irrespective of the genetic background.
Studies have shown that mutations of BRCA1 gene negatively impact the ovarian reserve due to defects in DNA repair mechanisms. As a result, patients with BRCA germline mutations might be more vulnerable to chemotherapy-induced gonadotoxicity. Carboplatin and paclitaxel are known moderately gonadotoxic drugs, but the impact of their combination on fertility remains unclear, particularly in BRCA-mutated patients.
STUDY DESIGN, SIZE, DURATION: Cryopreserved ovarian tissue fragments from patients with BC, either carrying a BRCA1 germline mutation (n = 4) or not (n = 4), were exposed to chemotherapy using two models: (i) in vitro culture or (ii) in vivo xenotransplantation model. First, thawed ovarian tissue fragments were cultured for 3 days with carboplatin (10 µg/ml), paclitaxel (1 µM), carboplatin, and paclitaxel or vehicle (dimethyl sulfoxide). Next, ovarian tissue fragments from the same patients were xenografted into the peritoneum of immunodeficient mice, followed by 3-week injections with either carboplatin (50 mg/kg/week) and paclitaxel (10 mg/kg/every 3 days) or saline solution as a control.
PARTICIPANTS/MATERIALS, SETTING, METHODS: Ovarian cortex was processed for histological analyses to assess follicle activation and survival in both experimental models. Follicle counting and morphological assessment were performed to evaluate the rates of follicles at different developmental stages, as well as the rate of atretic follicles. Immunostainings were performed for follicle activation (KL and p-RPS6), apoptosis (Terminal deoxynucleotidyl transferase dUTP Nick-End Labeling (TUNEL) assay), and DNA repair mechanisms (γH2AX, RAD51, and DNA PKcs).
While chemotherapy exposure did not significantly affect the proportion of primordial follicles in vitro, an increase in the proportion of quiescent follicles was observed after xenografting in the treated conditions compared to their respective controls, regardless of the presence of a BRCA mutation (BRCA+: 79.6 ± 5.07% versus 35.4 ± 8.26%, P = 0.0003; BRCA-: 81.8 ± 10.50% versus 17.9 ± 21.93%, P = 0.0014), reflecting the massive destruction of the pool of growing follicles. No difference was observed in the rate of atretic follicles, but the TUNEL assay revealed that chemotherapy, alone or in combination, increased DNA fragmentation rates (BRCA+: 37-49%; BRCA-: 43-55%) compared to the control conditions (BRCA+: 13-19%; BRCA-: 17-23%) both in vitro and in vivo. DNA repair mechanisms were affected following chemotherapy exposure, as evidenced by a significant increase in γH2AX-stained follicles in vitro (both populations) and in vivo (BRCA-mutated tissue) compared to the respective controls. Finally, chemotherapy had a similar impact on follicular atresia and apoptosis in both populations. However, BRCA-mutated tissue had lower rates of apoptotic (41% in BRCA+ versus 56% in BRCA-; P = 0.0184) and γH2AX-positive follicles (20% in BRCA+ versus 42% in BRCA-; P = 0.0142) than non-mutated fragments when exposed to carboplatin alone in vitro.
N/A.
LIMITATIONS, REASONS FOR CAUTION: Experiments on human tissue have limitations, particularly with rare material such as ovarian tissue from young BRCA-mutated patients donated for research. The study is limited by the small sample size and high intra- and inter-patient variability.
Studies on BRCA-mutated ovarian tissue are essential to better understand how BRCA mutations impact the ovarian reserve and respond to gonadotoxic treatments. Unexpectedly, this study suggested that follicles from young BRCA-mutated patients below 35 are not more sensitive to chemotherapy than the one from non-mutated patients, although DNA repair mechanisms seems to be differentially affected. Altogether, the results can help to provide appropriate fertility preservation counseling in BRCA1-mutated patients.
STUDY FUNDING/COMPETING INTEREST(S): This study is funded by Télévie, a grant of the Fonds National de la Recherche Scientifique-FNRS (Grant Nos 7.4531.22 and 7.6509.24), the Fondation contre le Cancer (Grant No. FAF-C/2018/1274), and supported by Fonds Erasme.
N/A.
与非乳腺癌（BC）基因突变患者的卵巢组织片段相比，携带乳腺癌1号基因（BRCA1）突变的乳腺癌患者的卵巢组织片段对卡铂和/或紫杉醇暴露是否更敏感？
无论基因背景如何，卡铂和紫杉醇治疗显示出相似的性腺毒性。
研究表明，BRCA1基因突变由于DNA修复机制缺陷而对卵巢储备产生负面影响。因此，携带BRCA种系突变的患者可能更容易受到化疗引起的性腺毒性影响。卡铂和紫杉醇是已知的中度性腺毒性药物，但它们联合使用对生育能力的影响仍不清楚，尤其是在携带BRCA突变的患者中。
研究设计、规模、持续时间：采用两种模型对携带BRCA1种系突变（n = 4）或不携带（n = 4）的乳腺癌患者的冷冻保存卵巢组织片段进行化疗暴露：（i）体外培养或（ii）体内异种移植模型。首先，将解冻的卵巢组织片段与卡铂（10 µg/ml）、紫杉醇（1 µM）、卡铂和紫杉醇或溶剂（二甲基亚砜）一起培养3天。接下来，将来自同一患者的卵巢组织片段异种移植到免疫缺陷小鼠的腹膜中，然后每周注射卡铂（50 mg/kg）和紫杉醇（10 mg/kg，每3天一次）或生理盐水作为对照，持续3周。
参与者/材料、设置、方法：对卵巢皮质进行组织学分析，以评估两种实验模型中的卵泡激活和存活情况。进行卵泡计数和形态学评估，以评估不同发育阶段的卵泡率以及闭锁卵泡率。对卵泡激活（KL和p-RPS6）、细胞凋亡（末端脱氧核苷酸转移酶dUTP缺口末端标记法（TUNEL）检测）和DNA修复机制（γH2AX、RAD51和DNA-PKcs）进行免疫染色。
虽然化疗暴露在体外对原始卵泡的比例没有显著影响，但在治疗条件下异种移植后，与各自的对照组相比，静止卵泡的比例有所增加，无论是否存在BRCA突变（BRCA+：79.6±5.07%对35.4±8.26%，P = 0.0003；BRCA-：81.8±10.50%对17.9±21.93%，P = 0.0014），这反映了生长卵泡池的大量破坏。闭锁卵泡率没有差异，但TUNEL检测显示，与对照条件相比，化疗单独或联合使用均增加了DNA片段化率（BRCA+：37 - 49%；BRCA-：43 - 55%），在体外和体内均如此（BRCA+：13 - 19%；BRCA-：十七 - 23%）。化疗暴露后DNA修复机制受到影响，与各自的对照组相比，体外（两个群体）和体内（BRCA突变组织）中γH2AX染色的卵泡显著增加证明了这一点。最后，化疗对两个群体的卵泡闭锁和细胞凋亡有相似的影响。然而，在体外单独暴露于卡铂时，BRCA突变组织的凋亡率（BRCA+中为41%，BRCA-中为56%；P = 0.0184）和γH2AX阳性卵泡率（BRCA+中为20%，BRCA-中为42%；P = 0.0142）低于非突变片段。
无。
局限性、注意事项：人体组织实验存在局限性，特别是对于为研究而捐赠的年轻BRCA突变患者的卵巢组织等稀有材料。该研究受样本量小以及患者内和患者间高变异性的限制。
对BRCA突变的卵巢组织进行研究对于更好地理解BRCA突变如何影响卵巢储备以及对性腺毒性治疗的反应至关重要。出乎意料的是，本研究表明，35岁以下携带BRCA突变的年轻患者的卵泡对化疗的敏感性并不比非突变患者的卵泡更高，尽管DNA修复机制似乎受到不同程度的影响。总之，这些结果有助于为携带BRCA1突变的患者提供适当的生育力保存咨询。
研究资金/利益冲突：本研究由Télévie资助，这是国家科学研究基金 - FNRS的一项拨款（拨款号7.4531.22和7.6509.24），由抗癌基金会（拨款号FAF-C/2018/1274）资助，并得到伊拉斯谟基金的支持。
无。
Zotero 插件