Glioblastoma Cells Express and Secrete Alternatively Spliced Transcripts of Coagulation Factor X

Area of Science:

• Oncology
• Hematology
• Molecular Biology

Background:

• Cancer patients often develop a prothrombotic state, increasing risks of venous and arterial thrombosis.
• Glioblastoma multiforme (GBM) shows a high association with thrombosis, yet the causes remain unclear.
• Understanding the prothrombotic mechanisms in GBM is crucial for improving patient outcomes.

Purpose of the Study:

• To investigate the expression of coagulation factor X (FX) and its splice variants in glioblastoma.
• To analyze FX production and secretion in glioblastoma cell lines under various conditions.
• To determine the functional activity of FX derived from glioblastoma.

Main Methods:

• Quantification of F10 mRNA and FX protein in glioblastoma tissues and cell lines.
• Analysis of F10 transcript isoforms using Sanger and amplicon sequencing.
• Assessment of FX activity via chromogenic assays and examination of secretion in conditioned media.
• Stimulation of glioblastoma cells with lipopolysaccharide (LPS) or oxygen/glucose starvation.

Main Results:

• Glioblastoma tissues and cell lines express high levels of full-length and alternatively spliced F10 mRNA, producing truncated FX.
• The ratio of full-length to truncated F10 transcripts is lower in glioblastoma than in normal brain tissue.
• Secreted FX from glioblastoma cells is enzymatically active and its expression/secretion is upregulated by LPS or nutrient deprivation.

Conclusions:

• Glioblastoma cells synthesize and secrete active FX, contributing to thrombin generation.
• This FX production represents a novel mechanism underlying the prothrombotic state in glioblastoma patients.
• Targeting FX may offer therapeutic strategies to mitigate thrombosis risk in GBM.