Riboflavin and ultraviolet illumination affects selected platelet mRNA transcript amounts differently

Area of Science:

• Biotechnology
• Hematology
• Transfusion Medicine

Background:

• Pathogen inactivation (PI) techniques, using UV light, aim to eliminate pathogens in blood products.
• Platelets (PLTs) contain RNA and can synthesize proteins, but the effect of PI on PLT RNA and protein synthesis is not well understood.
• Altered protein synthesis in PLTs could affect their overall quality and function.

Purpose of the Study:

• To investigate the impact of riboflavin and UV illumination PI on platelet (PLT) RNA.
• To quantify the reduction in specific messenger RNA (mRNA) transcripts after PI treatment.
• To determine the relationship between mRNA transcript length and susceptibility to PI.

Main Methods:

• Paired apheresis PLT concentrates were treated with riboflavin and UV illumination or left untreated.
• Quantitative polymerase chain reaction (qPCR) and absorbance were used to measure total RNA and specific mRNA levels.
• Quantified mRNAs included those for glycoproteins (GP)IIIa, GPIIb, GPIb, platelet factor 4 (PF4), osteonectin, thrombospondin (TSP), and glyceraldehyde-3-phosphate dehydrogenase (GAPDH).

Main Results:

• All analyzed mRNAs were significantly reduced (p < 0.05) after PI treatment, with varying degrees of reduction.
• Transcripts for GAPDH and PF4 were less affected, with approximately 70% remaining 1 hour post-treatment.
• Less than 15% of GPIIIa and TSP mRNA remained post-treatment, and a correlation (R² = 0.85) was observed between transcript length and remaining mRNA.
• Total RNA levels were stable, reflecting the normal PLT lifespan of 10-11 days.

Conclusions:

• Riboflavin and UV illumination PI significantly affects all platelet mRNA transcripts.
• The degree of mRNA reduction is dependent on transcript length, with shorter transcripts being more susceptible.
• These findings highlight a potential impact of PI on platelet protein synthesis and overall quality.