The oxido-metabolic driver ATF4 enhances temozolamide chemo-resistance in human gliomas

Abstract

Malignant gliomas are devastating neoplasia with limited curative treatment options. Temozolomide (TMZ, Temcat^®, Temodal^® or Temodar^®) is a first-line treatment for malignant gliomas but the development of drug resistance remains a major concern. Activating transcription factor 4 (ATF4) is a critical oxido-metabolic regulator in gliomas, and its role in the pathogenesis of TMZ-resistance remains elusive. We investigated the effect of TMZ on human glioma cells under conditions of enhanced ATF4 expression (ATF4^OE) and ATF4 knock down (ATF4^KD). We monitored cell survival, ATF4 mRNA expression of ATF4 and xCT (SLC7a11) regulation within human gliomas. TMZ treatment induces a transcriptional response with elevated expression of ATF4, xCT and Nrf2, as a sign of ER stress and toxic cell damage response. ATF4 overexpression (ATF4^OE) fosters TMZ resistance in human gliomas and inhibits TMZ-induced autophagy. Conversely, ATF4 suppression by small interfering RNAs (ATF4^KD) leads to increased TMZ susceptibility and autophagy in comparison to wild type gliomas. ATF4^OE gliomas show reduced cell cycle shift and apoptotic cell death, whereas ATF4^KD gliomas reveal higher susceptibility towards cell cycle rearrangements. Hence, the migration capacity of ATF4^OE glioma cells is almost not affected by TMZ treatment. In contrast, ATF4^KD gliomas show a migratory stop following TMZ application. Mechanistically, xCT elevation is a consequence of ATF4 activation and increased levels of xCT amplifies ATF4-induced TMZ resistance. Our data show that ATF4 operates as a chemo-resistance gene in gliomas, and the tumor promoting function of ATF4 is mainly determined by its transcriptional target xCT. Therefore, therapeutic inactivation of ATF4 can be a promising strategy to overcome chemo-resistance and promote drug efficacy in human gliomas.