2. Academic Validation
  3. Functionally-instructed modifiers of response to ATR inhibition in experimental glioma

Functionally-instructed modifiers of response to ATR inhibition in experimental glioma

  • J Exp Clin Cancer Res. 2024 Mar 12;43(1):77. doi: 10.1186/s13046-024-02995-z.
Bianca Walter 1 2,  Sophie Hirsch 1 2,  Laurence Kuhlburger 1 2 3 4,  Aaron Stahl 5,  Leonard Schnabel 1,  Silas Wisser 1,  Lara A Haeusser 1 2 6,  Foteini Tsiami 1 2,  Sarah Plöger 5,  Narges Aghaallaei 7,  Advaita M Dick 7,  Julia Skokowa 7,  Christian Schmees 2 5,  Markus Templin 5,  Katja Schenke-Layland 2 5 8,  Marcos Tatagiba 9 10,  Sven Nahnsen 2 3 4,  Daniel J Merk 1 2,  Ghazaleh Tabatabai 11 12 13 14
Affiliations

Affiliations

  • 1 Department of Neurology & Interdisciplinary Neuro-Oncology, University Hospital Tübingen, Hertie Institute for Clinical Brain Research, Eberhard Karls University Tübingen, 72076, Tübingen, Germany.
  • 2 Cluster of Excellence (EXC 2180) "Image Guided and Functionally Instructed Tumor Therapies", Eberhard Karls University Tübingen, 72076, Tübingen, Germany.
  • 3 Quantitative Biology Center, Eberhard Karls University Tübingen, 72076, Tübingen, Germany.
  • 4 Biomedical Data Science, Department of Computer Science, Eberhard Karls University Tübingen, 72076, Tübingen, Germany.
  • 5 NMI Natural and Medical Sciences Institute, University of Tübingen, 72770, Reutlingen, Germany.
  • 6 German Consortium for Translational Cancer Research (DKTK), Partner Site Tübingen, 72076, Tübingen, Germany.
  • 7 Division of Translational Oncology, Department of Internal Medicine II, University Hospital Tübingen, Eberhard Karls University Tübingen, 72076, Tübingen, Germany.
  • 8 Institute of Biomedical Engineering, Department for Medical Technologies and Regenerative Medicine, Eberhard Karls University Tübingen, 72076, Tübingen, Germany.
  • 9 Center for Neuro-Oncology, Comprehensive Cancer Center Tübingen-Stuttgart, Eberhard Karls University Tübingen, 72076, Tübingen, Germany.
  • 10 Department of Neurosurgery, University Hospital Tübingen, Eberhard Karls University Tübingen, Tübingen, Germany.
  • 11 Department of Neurology & Interdisciplinary Neuro-Oncology, University Hospital Tübingen, Hertie Institute for Clinical Brain Research, Eberhard Karls University Tübingen, 72076, Tübingen, Germany. [email protected].
  • 12 Cluster of Excellence (EXC 2180) "Image Guided and Functionally Instructed Tumor Therapies", Eberhard Karls University Tübingen, 72076, Tübingen, Germany. [email protected].
  • 13 German Consortium for Translational Cancer Research (DKTK), Partner Site Tübingen, 72076, Tübingen, Germany. [email protected].
  • 14 Center for Neuro-Oncology, Comprehensive Cancer Center Tübingen-Stuttgart, Eberhard Karls University Tübingen, 72076, Tübingen, Germany. [email protected].
PMID: 38475864 DOI: 10.1186/s13046-024-02995-z
Abstract

Background: The DNA damage response (DDR) is a physiological network preventing malignant transformation, e.g. by halting cell cycle progression upon DNA damage detection and promoting DNA repair. Glioblastoma are incurable primary tumors of the nervous system and DDR dysregulation contributes to acquired treatment resistance. Therefore, DDR targeting is a promising therapeutic anti-glioma strategy. Here, we investigated Ataxia telangiectasia and Rad3 related (ATR) inhibition (ATRi) and functionally-instructed combination therapies involving ATRi in experimental glioma.

Methods: We used acute cytotoxicity to identify treatment efficacy as well as RNAseq and DigiWest protein profiling to characterize ATRi-induced modulations within the molecular network in glioma cells. Genome-wide CRISPR/Cas9 functional genomic screens and subsequent validation with functionally-instructed compounds and selected shRNA-based silencing were employed to discover and investigate molecular targets modifying response to ATRi in glioma cell lines in vitro, in primary cultures ex vivo and in zebrafish and murine models in vivo.

Results: ATRi monotherapy displays anti-glioma efficacy in vitro and ex vivo and modulates the molecular network. We discovered molecular targets by genome-wide CRISPR/Cas9 loss-of-function and activation screens that enhance therapeutic ATRi effects. We validated selected druggable targets by a customized drug library and functional assays in vitro, ex vivo and in vivo.

Conclusion: In conclusion, our study leads to the identification of novel combination therapies involving ATRi that could inform future preclinical studies and early phase clinical trials.

Keywords

Combination therapies; DNA damage response pathway; DigiWest; Functional genomics.

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