PO.ET03.04 · 实验与分子治疗
慢性 1 型干扰素暴露在高级别浆液性卵巢癌中表型模拟慢性顺铂暴露
Chronic type 1 interferon exposure phenocopies chronic cisplatin exposure in high grade serous ovarian cancer
作者与单位 Authors & Affiliations
摘要 Abstract
中文摘要
尽管最初对铂类治疗有反应，高级别浆液性卵巢癌（HGSOC）仍是最致命的妇科恶性肿瘤之一，其原因在于高度的分子和遗传异质性，这导致了获得性耐药机制和快速复发。该疾病的一线治疗在 30 年间基本保持不变，CA125 在过去 40 年中一直是唯一确立的生物标志物，而 PARP 抑制剂是为数不多的可用靶向治疗之一。迫切需要更好地理解可能预测治疗反应和肿瘤侵袭性、从而促成复发和耐药的因素。我们此前发表过关于在一对同源的治疗敏感和耐药的患者来源 HGSOC 细胞中，干性与治疗耐药相分离的研究。RNA 测序揭示了化疗耐药（CR）细胞中 JAK/STAT 信号的富集以及与 1 型干扰素产生和信号相关基因特征的上调。我们假设，慢性、低水平的 IFN-1 信号通过启动 DNA 损伤相关的 IFN-1 反应，促进 HGSOC 治疗耐药的发展。使用 RT-qPCR 和流式细胞术验证了关注的通路和靶点。发现 IFN-1 和促肿瘤的干扰素相关 DNA 损伤特征（IRDS）基因在 CR 细胞中显著上调。急性顺铂处理在所有细胞中富集了 IFN-1 相关蛋白表达。CR 和 SE 细胞对急性顺铂暴露表现出不同的 IFN-1 和 IRDS RNA 时间表达模式，SE 细胞对顺铂表现出急性 IFN-1 反应，而 CR 细胞则以延迟的 IFN-1 诱导作出反应。为确定 IFN-β 是否为耐药发展的驱动因素，用慢性、低水平 IFN-β 处理细胞数周。与高剂量细胞因子处理 24 小时后相比，慢性、低水平 IFN-β 未诱导抗病毒 ISG 表达。然而，经过 6 次传代后，该处理钝化了 CR 和 SE 细胞中的 ISG 表达。IFN-β 处理表型模拟了慢性顺铂暴露的若干方面：对顺铂耐药性增加、上皮样形态变化以及增殖降低。尽管发生了这些变化，IFN 反应却被抑制。未来的研究旨在探索通过影响 IFN-I 信号来克服化疗耐药的策略。
查看英文原文 English abstract
Despite initial response to platinum-based therapeutics, high grade serous ovarian carcinoma (HGSOC) remains one of the deadliest gynecologic malignancies, due to high molecular and genetic heterogeneity that contributes to acquired drug resistance mechanisms and rapid recurrence. Frontline treatment for the disease has remained largely unchanged for 30 years, with CA125 serving as the only established biomarker for the past 4 decades and PARP inhibitors representing one of the few available targeted therapies. There is an urgent need for improved understanding of factors that may predict therapy response and tumor aggression, thus contributing to recurrence and resistance. We previously published on the dissociation of stemness and therapy resistance in an isogenic pair of therapy sensitive and resistant patient-derived HGSOC cells. RNA sequencing revealed enrichment of JAK/STAT signaling and an upregulation of gene signatures associated with type 1 interferon production and signaling in the chemo-resistant (CR) cells. We hypothesize that chronic, low-level IFN-1 signaling promotes the development of therapy resistance in HGSOC, via the priming of a DNA damage associated IFN-1 response. RT-qPCR and flow cytometry were used to validate the pathways and targets of interest. IFN-1 and pro-tumor interferon-related DNA damage signature (IRDS) genes were found to be significantly upregulated in the CR cells. Acute cisplatin treatment enriched IFN-1-related protein expression in all cells. CR and SE cells exhibited different temporal expression patterns of IFN-1 and IRDS RNA in response to acute cisplatin exposure, with SE cells displaying an acute IFN-1 response to cisplatin and CR cells responding with delayed IFN-1 induction. To determine if IFN-beta is a driving factor in the development of resistance, cells were treated with chronic, low-level IFN-beta for several weeks. Chronic, low-level IFN-beta did not induce anti-viral ISG expression when compared to high dose cytokine after 24 hours. However, after 6 passages the treatment blunted ISG expression in both CR and SE cells. IFN-beta treatment phenocopied several aspects of chronic cisplatin exposure: increased resistance to cisplatin, epithelial-like morphology changes, and reduced proliferation. Despite these changes, IFN response was dampened. Future studies aim to explore strategies for surmounting chemotherapy resistance by impacting IFN-I signaling.
利益披露 Disclosure
A. Conant, None..
K. McGivney, None..
T. Suzuki, None..
S. Asariah, None..
B. Vazquez, None..
J. Deng, None..
H. Cheon, None.