Autophagy-mediated ID1 turnover dictates chemo-resistant fate in ovarian cancer stem cells.

IF 11.4 1区医学 Q1 ONCOLOGY Journal of Experimental & Clinical Cancer Research Pub Date : 2024-08-10 DOI:10.1186/s13046-024-03147-z

Pratham Phadte, Aniketh Bishnu, Pranay Dey, Manikandan M, Megha Mehrotra, Prerna Singh, Shritama Chakrabarty, Rounak Majumdar, Bharat Rekhi, Malay Patra, Abhijit De, Pritha Ray

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Abstract

Background: The mechanisms enabling dynamic shifts between drug-resistant and drug-sensitive states in cancer cells are still underexplored. This study investigated the role of targeted autophagic protein degradation in regulating ovarian cancer stem cell (CSC) fate decisions and chemo-resistance.

Methods: Autophagy levels were compared between CSC-enriched side population (SP) and non-SP cells (NSP) in multiple ovarian cancer cell lines using immunoblotting, immunofluorescence, and transmission electron microscopy. The impact of autophagy modulation on CSC markers and differentiation was assessed by flow cytometry, immunoblotting and qRT-PCR. In silico modeling and co-immunoprecipitation identified ID1 interacting proteins. Pharmacological and genetic approaches along with Annexin-PI assay, ChIP assay, western blotting, qRT-PCR and ICP-MS were used to evaluate effects on cisplatin sensitivity, apoptosis, SLC31A1 expression, promoter binding, and intracellular platinum accumulation in ID1 depleted backdrop. Patient-derived tumor spheroids were analyzed for autophagy and SLC31A1 levels.

Results: Ovarian CSCs exhibited increased basal autophagy compared to non-CSCs. Further autophagy stimulation by serum-starvation and chemical modes triggered proteolysis of the stemness regulator ID1, driving the differentiation of chemo-resistant CSCs into chemo-sensitive non-CSCs. In silico modeling predicted TCF12 as a potent ID1 interactor, which was validated by co-immunoprecipitation. ID1 depletion freed TCF12 to transactivate the cisplatin influx transporter SLC31A1, increasing intracellular cisplatin levels and cytotoxicity. Patient-derived tumor spheroids exhibited a functional association between autophagy, ID1, SLC31A1, and platinum sensitivity.

Conclusions: This study reveals a novel autophagy-ID1-TCF12-SLC31A1 axis where targeted autophagic degradation of ID1 enables rapid remodeling of CSCs to reverse chemo-resistance. Modulating this pathway could counter drug resistance in ovarian cancer.

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自噬介导的ID1周转决定了卵巢癌干细胞耐化疗的命运。

背景：癌细胞在耐药和药敏状态之间动态转变的机制仍未得到充分探索。本研究探讨了靶向自噬蛋白降解在调控卵巢癌干细胞（CSC）命运决定和化疗耐药性中的作用：方法：使用免疫印迹、免疫荧光和透射电子显微镜比较了多种卵巢癌细胞系中富含CSC的侧群（SP）和非SP细胞（NSP）的自噬水平。流式细胞术、免疫印迹和 qRT-PCR 评估了自噬调节对 CSC 标记和分化的影响。硅学建模和共免疫沉淀确定了与ID1相互作用的蛋白。药理学和遗传学方法以及Annexin-PI检测、ChIP检测、Western印迹、qRT-PCR和ICP-MS被用来评估ID1缺失背景对顺铂敏感性、细胞凋亡、SLC31A1表达、启动子结合和细胞内铂积累的影响。对源自患者的肿瘤球体进行了自噬和 SLC31A1 水平分析：结果：与非卵巢干细胞相比，卵巢干细胞的基础自噬能力增强。血清饥饿和化学模式对自噬的进一步刺激引发了干性调节因子ID1的蛋白水解，促使耐化疗的CSCs分化为对化疗敏感的非CSCs。硅学建模预测TCF12是ID1的有效互作因子，这一点通过共免疫沉淀得到了验证。去除了 ID1，TCF12 就能反式激活顺铂流入转运体 SLC31A1，从而提高细胞内顺铂水平和细胞毒性。源自患者的肿瘤球体显示了自噬、ID1、SLC31A1和铂敏感性之间的功能关联：这项研究揭示了一种新的自噬-ID1-TCF12-SLC31A1轴，在这一轴上，ID1的靶向自噬降解能够快速重塑造血干细胞，从而逆转化疗耐药性。调节这一通路可对抗卵巢癌的耐药性。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Journal of Experimental & Clinical Cancer Research 医学-肿瘤学

CiteScore

18.20

自引率

1.80%

发文量

333

审稿时长

1 months

期刊介绍： The Journal of Experimental & Clinical Cancer Research is an esteemed peer-reviewed publication that focuses on cancer research, encompassing everything from fundamental discoveries to practical applications. We welcome submissions that showcase groundbreaking advancements in the field of cancer research, especially those that bridge the gap between laboratory findings and clinical implementation. Our goal is to foster a deeper understanding of cancer, improve prevention and detection strategies, facilitate accurate diagnosis, and enhance treatment options. We are particularly interested in manuscripts that shed light on the mechanisms behind the development and progression of cancer, including metastasis. Additionally, we encourage submissions that explore molecular alterations or biomarkers that can help predict the efficacy of different treatments or identify drug resistance. Translational research related to targeted therapies, personalized medicine, tumor immunotherapy, and innovative approaches applicable to clinical investigations are also of great interest to us. We provide a platform for the dissemination of large-scale molecular characterizations of human tumors and encourage researchers to share their insights, discoveries, and methodologies with the wider scientific community. By publishing high-quality research articles, reviews, and commentaries, the Journal of Experimental & Clinical Cancer Research strives to contribute to the continuous improvement of cancer care and make a meaningful impact on patients' lives.