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GDF15单抗筛选及其联合卡铂逆转卵巢癌铂耐药的研究
Screening of anti-GDF15 monoclonal antibodies and their combination with carboplatin to reverse platinum resistance in ovarian cancer
目的 探讨生长分化因子15（growth differentiation factor 15，GDF15）单克隆抗体在逆转卵巢癌铂耐药中的作用，并筛选疗效最佳的候选抗体，同时阐明其潜在分子机制。方法 于2025年4月至2026年1月在中国医学科学院化学研究所和中国医学科学院北京协和医院肿瘤医院构建SK-OV-3-R铂耐药细胞系及患者来源异种移植（patient-derived xenograft，PDX）模型。体内外平行筛选5种GDF15单克隆抗体以确定最优候选。采用二苯基四氮唑溴盐（MTT）法检测细胞活力，异硫氰酸荧光素标记的膜联蛋白V/碘化丙啶（Annexin V-FITC/PI）双染结合流式细胞术评估细胞凋亡；通过γ-H2AX流式检测评估DNA损伤水平；采用蛋白激酶B（AKT）激动剂和抑制剂干预验证相关信号通路。PDX模型中监测肿瘤体积并计算肿瘤生长抑制率（tumor growth inhibition，TGI）。结果 筛选确定CTL-002为最优GDF15单克隆抗体。在SK-OV-3-R细胞中，CTL-002联合卡铂可显著增强抗肿瘤效应，半数抑制浓度（IC50）由29.47 μmol/L降低至13.60 μmol/L，凋亡率升高至（26.85±1.6）%。同时，联合治疗能显著提高γ-H2AX平均荧光强度，提示DNA损伤水平增加。机制研究表明，AKT抑制剂可模拟CTL-002的增敏作用，而AKT激动剂可部分逆转其效应。在PDX模型中，联合治疗组终点肿瘤体积为（170.37±17.72）mm³，肿瘤体积抑制率（TGI）达66.3%，显著优于卡铂单药组（47.9%）。结论 GDF15单克隆抗体CTL-002可显著增强卡铂对铂耐药卵巢癌的体外抑制效应并提高体内抑瘤效果，提示GDF15单克隆抗体具有逆转卵巢癌铂耐药的潜在应用价值。
Objective To investigate the role of Growth Differentiation Factor 15（GDF15）monoclonal antibodies in reversing platinum resistance in ovarian cancer，to identify the most effective candidate antibody，and to explore the underlying molecular mechanisms. Methods This study was carried out from April 2025 to January 2026 at the Institute of Chemistry, Chinese Academy of Sciences, and the Cancer Hospital, Chinese Academy of Medical Sciences & Peking Union Medical College. A platinum-resistant ovarian cancer cell line（SK-OV-3-R）and a patient-derived xenograft（PDX）model were established. Five GDF15 monoclonal antibodies were screened in parallel in vitro and in vivo to determine the optimal candidate. Cell viability was assessed by MTT assay，and apoptosis was evaluated using Annexin V-FITC/PI staining followed by flow cytometry. DNA damage was measured by γ-H2AX flow cytometry. AKT activator and inhibitor were applied to investigate the involvement of the PI3K/AKT signaling pathway. Tumor volume and tumor growth inhibition（TGI）were assessed in the PDX model. Results CTL-002 was identified as the most effective GDF15 monoclonal antibody. In SK-OV-3-R cells，CTL-002 combined with carboplatin significantly enhanced antitumor activity，reducing the IC50 from 29.47 μmol/L to 13.60 μmol/L and increasing the apoptosis rate to（26.85±1.6）%. Meanwhile，the combination treatment markedly increased γ-H2AX mean fluorescence intensity，indicating elevated DNA damage. Mechanistically，AKT inhibition mimicked the sensitizing effect of CTL-002，whereas AKT activatior partially reversed it. In the PDX model，the combination therapy significantly suppressed tumor growth，with a final tumor volume of（170.37±17.72）mm³ and a TGI of 66.3%，which was superior to carboplatin monotherapy（47.9%）. Conclusion The anti-GDF15 monoclonal antibody CTL-002 can significantly enhance the in vitro inhibitory effect of carboplatin against platinum-resistant ovarian cancer and improve its antitumor efficacy in vivo，suggesting that anti-GDF15 monoclonal antibodies have potential clinical value for reversing platinum resistance in ovarian cancer.
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Background: The standard treatment of ovarian cancer is surgery followed by a chemotherapeutic combination consisting of a platinum agent, such as cisplatin and a taxane-like paclitaxel. We previously observed that patients with ovarian cancer wild-type for p53 had a poorer survival rate than did those with p53 mutations. Thus, a better understanding of the molecular changes of epithelial ovarian cancer cells with wild-type p53 in response to treatment with cisplatin could reveal novel mechanisms of chemoresistance. Methods: Gene expression profiling was performed on an ovarian cancer cell line A2780 with wild-type p53 treated with cisplatin. A gene encoding a secretory protein growth differentiation factor 15 (GDF15) was identified to be highly induced by cisplatin treatment in vitro. This was further validated in a panel of wild-type and mutant p53 ovarian cancer cell lines, as well as in mouse orthotopic models. The mouse tumor tissues were further analyzed by histology and RNA-seq. Results: GDF15 was identified as one of the highly induced genes by cisplatin or carboplatin in ovarian cancer cell lines with wild-type p53. The wild-type p53-induced expression of GDF15 and GDF15-confered chemotherapy resistance was further demonstrated in vitro and in vivo. This study also discovered that GDF15-knockdown (GDF15-KD) tumors had less stromal component and had different repertoires of activated and inhibited canonical pathways in the stromal cell and cancer cell components from that of the control tumors after cisplatin treatment. Conclusions: GDF15 expression from the wild-type p53 cancer cells can modulate the canonical pathways in the tumor microenvironment in response to cisplatin, which is a possible mechanism of chemoresistance.
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高维怡, 杨欣, 王闻楚. 生长分化因子15在肿瘤化疗耐药中的研究进展[J]. 中国肿瘤生物治疗杂志, 2025, 32(11):1202-1207.
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Platinum-based drugs cisplatin, carboplatin, and oxaliplatin are widely used for chemotherapeutic eradication of cancer. However, the side effects of platinum drugs, such as lack of selectivity, high systemic toxicity, and drug resistance, seriously limit their clinical application. With advancements in nanotechnology and chemical synthesis, Pt-based anti-cancer drugs have made great progress in cancer therapy in recent years. Many strategies relied on the anti-cancer mechanism similar to cisplatin and achieved some success by modifying existing platinum drugs. Pt-based nanodrugs, such as platinum nanoclusters, have novel anti-cancer mechanisms and great potential in tumor-targeted therapy and have shown promising results in clinical application. In this review, we systematically explored the development of first-line platinum chemotherapy drugs in the clinic and their anti-cancer mechanisms. We also summarize the progress of Pt-based anti-cancer drug application in cancer therapy, emphasizing their modification to enhance the anti-tumor effect. Finally, we address challenges faced by platinum chemotherapy drugs, especially Pt nanocluster-based nanodrugs, in cancer treatment. The new platinum drugs and their targeted modifications undoubtedly provide a promising prospect for improving the current anti-cancer treatments.© The author(s).
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Platinum-based antineoplastic drugs, such as cisplatin, are commonly used to induce tumor cell death. Cisplatin is believed to induce apoptosis as a result of cisplatin-DNA adducts that inhibit DNA and RNA synthesis. Although idea that DNA damage underlines anti-proliferative effects of cisplatin is dominant in cancer research, there is a poor correlation between the degree of the cell sensitivity to cisplatin and the extent of DNA platination. Here, we examined possible effects of cisplatin on post-transcriptional gene regulation that may contribute to cisplatin-mediated cytotoxicity. We show that cisplatin suppresses formation of stress granules (SGs), pro-survival RNA granules with multiple roles in cellular metabolism. Mechanistically, cisplatin inhibits cellular translation to promote disassembly of polysomes and aggregation of ribosomal subunits. As SGs are in equilibrium with polysomes, cisplatin-induced shift towards ribosomal aggregation suppresses SG formation. Our data uncover previously unknown effects of cisplatin on RNA metabolism.Copyright © 2021 The Authors. Published by Elsevier Masson SAS.. All rights reserved.
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Cancer cachexia is a highly prevalent condition associated with poor quality of life and reduced survival. Tumor-induced perturbations in the endocrine, immune and nervous systems drive anorexia and catabolic changes in adipose tissue and skeletal muscle, hallmarks of cancer cachexia. However, the molecular mechanisms driving cachexia remain poorly defined, and there are currently no approved drugs for the condition. Elevation in circulating growth differentiation factor 15 (GDF15) correlates with cachexia and reduced survival in patients with cancer, and a GDNF family receptor alpha like (GFRAL)-Ret proto-oncogene (RET) signaling complex in brainstem neurons that mediates GDF15-induced weight loss in mice has recently been described. Here we report a therapeutic antagonistic monoclonal antibody, 3P10, that targets GFRAL and inhibits RET signaling by preventing the GDF15-driven interaction of RET with GFRAL on the cell surface. Treatment with 3P10 reverses excessive lipid oxidation in tumor-bearing mice and prevents cancer cachexia, even under calorie-restricted conditions. Mechanistically, activation of the GFRAL-RET pathway induces expression of genes involved in lipid metabolism in adipose tissues, and both peripheral chemical sympathectomy and loss of adipose triglyceride lipase protect mice from GDF15-induced weight loss. These data uncover a peripheral sympathetic axis by which GDF15 elicits a lipolytic response in adipose tissue independently of anorexia, leading to reduced adipose and muscle mass and function in tumor-bearing mice.
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利益冲突 所有作者均声明不存在利益冲突
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