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CRISPRi screening identifies CASP8AP2 as an essential viability factor in lung cancer controlling tumor cell death

Ksenia Myacheva1, Andrew Walsh2, Marisa Riester3

  • 1Division of Cancer Research, Department of Thoracic Surgery, Medical Center - University of Freiburg, Faculty of Medicine, University of Freiburg, German Cancer Consortium (DKTK) - Partner Site Freiburg, Germany; Division of RNA Biology & Cancer, German Cancer Research Center (DKFZ), Heidelberg, Germany.

Cancer Letters
|October 17, 2022
PubMed

Insights

A CRISPR screen identified CASP8AP2 as a key gene for lung adenocarcinoma (LUAD) survival. Its knockdown triggers cell death via autophagy and apoptosis, mediated by the AP-1 transcription factor.

Area of Science:

  • Oncology
  • Molecular Biology
  • Genetics

Background:

  • Lung cancer, particularly lung adenocarcinoma (LUAD), is a leading global cause of cancer mortality.
  • Novel therapeutic targets are urgently needed to improve LUAD patient outcomes.

Purpose of the Study:

  • To identify novel genes regulating LUAD cell viability using a large-scale CRISPR interference (CRISPRi) loss-of-function screen.
  • To elucidate the molecular mechanisms underlying LUAD cell death induced by targeting key regulatory genes.

Main Methods:

  • Conducted a CRISPRi loss-of-function screen targeting 2098 deregulated genes in eight LUAD cell lines.
  • Utilized a customized algorithm to analyze gene functionality at the transcriptional start site (TSS) level.
  • Performed knockdown (KD) of candidate genes and analyzed downstream effects on cell viability, autophagy, and apoptosis.
  • Conducted systematic expression profiling to identify transcription factors involved in gene KD-induced cell death.

Main Results:

  • CASP8AP2 was identified as the sole gene significantly impacting the viability of all screened LUAD cell lines, with minimal effect on non-transformed lung cells.
  • CASP8AP2 KD induced both autophagy and apoptotic cell death pathways in LUAD cells.
  • The AP-1 transcription factor was linked to CASP8AP2 KD-induced cancer cell death, and its inhibition reversed the observed phenotype.

Conclusions:

  • The CASP8AP2 - AP-1 axis is a critical mediator of lung adenocarcinoma cell viability.
  • CASP8AP2 represents a potential therapeutic target for LUAD, with its modulation influencing cancer cell death pathways.
Keywords:
AutophagyCRISPRCaspaseFLASHLung adenocarcinomaNSCLC

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