Inhibition of PERK-mediated unfolded protein response acts as a switch for reversal of residual senescence and as senolytic therapy in glioblastoma.

IF 16.4 1区 医学 Q1 CLINICAL NEUROLOGY Neuro-oncology Pub Date : 2024-11-04 DOI:10.1093/neuonc/noae134
Madhura Ketkar, Sanket Desai, Pranav Rana, Rahul Thorat, Sridhar Epari, Amit Dutt, Shilpee Dutt
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Abstract

Background: Glioblastoma due to recurrence is clinically challenging with 10-15 months overall survival. Previously we showed that therapy-induced senescence (TIS) in glioblastoma reverses causing recurrence. Here, we aim to delineate the TIS reversal mechanism for potential therapeutic intervention to prevent glioblastoma (GBM) recurrence.

Methods: Residual senescent (RS) and end of residual senescence (ERS) cells were captured from GBM patient-derived primary-cultures and cell lines mimicking clinical scenarios. RNA-sequencing, transcript/protein validations, knock-down/inhibitor studies, ChIP RT-PCR, biochemical assays, and IHCs were performed for the mechanistics of TIS reversal. In vivo validations were conducted in GBM orthotopic mouse model.

Results: Transcriptome analysis showed co-expression of endoplasmic reticulum (ER) stress-unfolded protein response (UPR) and senescence-associated secretory phenotype (SASP) with TIS induction and reversal. Robust SASP production and secretion by RS cells could induce senescence, Reactive oxygen specis (ROS), DNA damage, and ER stress in paracrine fashion independent of radiation. Neutralization of most significantly enriched cytokine from RS-secretome IL1β, suppressed SASP, and delayed senescence reversal. Mechanistically, with SASP and massive protein accumulation in ER, RS cells displayed stressed ER morphology, upregulated ER stress markers, and PERK pathway activation via peIF2α-ATF4-CHOP which was spontaneously resolved in ERS. ChIP RT-PCR showed CHOP occupancy at CXCL8/IL8, CDKN1A/p21, and BCL2L1/BCLXL aiding survival. PERK knockdown/inhibition with GSK2606414 in combination with radiation led to sustained ER stress and senescence without SASP. PERKi in RS functioned as senolytic via apoptosis and prevented recurrence in vitro and in vivo ameliorating overall survival.

Conclusion: We demonstrate that PERK-mediated UPR regulates senescence reversal and its inhibition can be exploited as a potential seno-therapeutic option in glioblastoma.

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抑制 PERK 介导的 UPR 可作为逆转残余衰老的开关和胶质母细胞瘤的衰老疗法。
背景:胶质母细胞瘤因复发导致的总生存期仅为 10-15 个月,在临床上具有挑战性。此前,我们发现胶质母细胞瘤的治疗诱导衰老(TIS)可逆转导致复发。在此,我们旨在阐明TIS逆转机制,为预防GBM复发提供潜在的治疗干预:残余衰老(RS)细胞和残余衰老末期(ERS)细胞来自 GBM 患者来源的原代培养物和模拟临床情况的细胞系。进行了 RNA 测序、转录本/蛋白质验证、基因敲除/抑制剂研究、ChIP RT-PCR、生化检测和 IHC,以了解 TIS 逆转的机理。在 GBM 正位小鼠模型中进行了体内验证:转录组分析显示,ER应激-UPR和衰老相关分泌表型(SASP)与TIS诱导和逆转共同表达。RS细胞产生和分泌的大量SASP能以旁分泌方式诱导衰老、ROS、DNA损伤和ER应激,与辐射无关。中和RS分泌组中最重要的细胞因子IL1β可抑制SASP并延缓衰老逆转。从机理上讲,随着 SASP 和内质网中大量蛋白质的积累,RS 细胞显示出受压的 ER 形态、上调的 ER 应激标志物和通过 peIF2α-ATF4-CHOP 激活的 PERK 通路,而这在 ERS 中会自发地得到解决。ChIP RT-PCR 显示,CHOP 在 CXCL8/IL8、CDKN1A/p21 和 BCL2L1/BCLXL 上的占位有助于存活。用GSK2606414敲除/抑制PERK,并与辐射结合使用,可导致持续的ER应激和衰老,但无SASP。PERKi在RS中通过细胞凋亡起到了溶解衰老的作用,并在体外和体内防止了复发,改善了总生存率:我们证明了 PERK 介导的 UPR 调节衰老逆转,其抑制作用可被用作胶质母细胞瘤的潜在衰老治疗方案。
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来源期刊
Neuro-oncology
Neuro-oncology 医学-临床神经学
CiteScore
27.20
自引率
6.30%
发文量
1434
审稿时长
3-8 weeks
期刊介绍: Neuro-Oncology, the official journal of the Society for Neuro-Oncology, has been published monthly since January 2010. Affiliated with the Japan Society for Neuro-Oncology and the European Association of Neuro-Oncology, it is a global leader in the field. The journal is committed to swiftly disseminating high-quality information across all areas of neuro-oncology. It features peer-reviewed articles, reviews, symposia on various topics, abstracts from annual meetings, and updates from neuro-oncology societies worldwide.
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