P4HA1 mediates YAP hydroxylation and accelerates collagen synthesis in temozolomide-resistant glioblastoma

Chin Med J (Engl). 2025 Aug 20;138(16):1991-2005. doi: 10.1097/CM9.0000000000003679. Epub 2025 Jun 9.

Abstract

Background: Temozolomide (TMZ) resistance is a significant challenge in treating glioblastoma (GBM). Collagen remodeling has been shown to be a critical factor for therapy resistance in other cancers. This study aimed to investigate the mechanism of TMZ chemoresistance by GBM cells reprogramming collagens.

Methods: Key extracellular matrix components, including collagens, were examined in paired primary and recurrent GBM samples as well as in TMZ-treated spontaneous and grafted GBM murine models. Human GBM cell lines (U251, TS667) and mouse primary GBM cells were used for in vitro studies. RNA-sequencing analysis, chromatin immunoprecipitation, immunoprecipitation-mass spectrometry, and co-immunoprecipitation assays were conducted to explore the mechanisms involved in collagen accumulation. A series of in vitro and in vivo experiments were designed to assess the role of the collagen regulators prolyl 4-hydroxylase subunit alpha 1 (P4HA1) and yes-associated protein (YAP) in sensitizing GBM cells to TMZ.

Results: This study revealed that TMZ exposure significantly elevated collagen type I (COL I) expression in both GBM patients and murine models. Collagen accumulation sustained GBM cell survival under TMZ-induced stress, contributing to enhanced TMZ resistance. Mechanistically, P4HA1 directly binded to and hydroxylated YAP, preventing ubiquitination-mediated YAP degradation. Stabilized YAP robustly drove collagen type I alpha 1 ( COL1A1) transcription, leading to increased collagen deposition. Disruption of the P4HA1-YAP axis effectively reduced COL I deposition, sensitized GBM cells to TMZ, and significantly improved mouse survival.

Conclusion: P4HA1 maintained YAP-mediated COL1A1 transcription, leading to collagen accumulation and promoting chemoresistance in GBM.

Keywords: Chemoresistance; Collagen; Glioblastoma; Prolyl 4-hydroxylase subunit alpha 1; Temozolomide; Yes-associated protein.

MeSH terms

  • Adaptor Proteins, Signal Transducing* / metabolism
  • Animals
  • Antineoplastic Agents, Alkylating / therapeutic use
  • Cell Line, Tumor
  • Collagen Type I / metabolism
  • Collagen* / biosynthesis
  • Dacarbazine* / analogs & derivatives
  • Dacarbazine* / pharmacology
  • Dacarbazine* / therapeutic use
  • Drug Resistance, Neoplasm / genetics
  • Glioblastoma* / drug therapy
  • Glioblastoma* / metabolism
  • Humans
  • Hydroxylation
  • Mice
  • Prolyl Hydroxylases* / metabolism
  • Temozolomide*
  • Transcription Factors* / metabolism
  • YAP-Signaling Proteins

Substances

  • Temozolomide
  • YAP-Signaling Proteins
  • Dacarbazine
  • Adaptor Proteins, Signal Transducing
  • Transcription Factors
  • Collagen
  • YAP1 protein, human
  • Collagen Type I
  • Prolyl Hydroxylases
  • Antineoplastic Agents, Alkylating