Protein disulfide-isomerase A4 confers glioblastoma angiogenesis promotion capacity and resistance to anti-angiogenic therapy

J Exp Clin Cancer Res. 2023 Mar 30;42(1):77. doi: 10.1186/s13046-023-02640-1.

Abstract

Introduction: Increasing evidence has revealed the key activity of protein disulfide isomerase A4 (PDIA4) in the endoplasmic reticulum stress (ERS) response. However, the role of PDIA4 in regulating glioblastoma (GBM)-specific pro-angiogenesis is still unknown.

Methods: The expression and prognostic role of PDIA4 were analyzed using a bioinformatics approach and were validated in 32 clinical samples and follow-up data. RNA-sequencing was used to search for PDIA4-associated biological processes in GBM cells, and proteomic mass spectrum (MS) analysis was used to screen for potential PDIA4 substrates. Western blotting, real-time quantitative polymerase chain reaction (RT-qPCR), and enzyme-linked immunosorbent assays (ELISA) were used to measure the levels of the involved factors. Cell migration and tube formation assays determined the pro-angiogenesis activity of PDIA4 in vitro. An intracranial U87 xenograft GBM animal model was constructed to evaluate the pro-angiogenesis role of PDIA4 in vivo.

Results: Aberrant overexpression of PDIA4 was associated with a poor prognosis in patients with GBM, although PDIA4 could also functionally regulate intrinsic GBM secretion of vascular endothelial growth factor-A (VEGF-A) through its active domains of Cys-X-X-Cys (CXXC) oxidoreductase. Functionally, PDIA4 exhibits pro-angiogenesis activity both in vitro and in vivo, and can be upregulated by ERS through transcriptional regulation of X-box binding protein 1 (XBP1). The XBP1/PDIA4/VEGFA axis partially supports the mechanism underlying GBM cell survival under ER stress. Further, GBM cells with higher expression of PDIA4 showed resistance to antiangiogenic therapy in vivo.

Conclusions: Our findings revealed the pro-angiogenesis role of PDIA4 in GBM progression and its potential impact on GBM survival under a harsh microenvironment. Targeting PDIA4 might help to improve the efficacy of antiangiogenic therapy in patients with GBM.

Keywords: Angiogenesis; Endoplasmic reticulum stress (ERS); Glioblastoma (GBM); Protein disulfide-isomerase A4 (PDIA4); X-box binding protein 1 (XBP1).

MeSH terms

  • Animals
  • Cell Line, Tumor
  • Enzyme-Linked Immunosorbent Assay
  • Glioblastoma* / drug therapy
  • Glioblastoma* / genetics
  • Humans
  • Protein Disulfide-Isomerases* / genetics
  • Protein Disulfide-Isomerases* / metabolism
  • Proteomics
  • Tumor Microenvironment
  • Vascular Endothelial Growth Factor A / metabolism

Substances

  • Protein Disulfide-Isomerases
  • Vascular Endothelial Growth Factor A
  • PDIA4 protein, human