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Updated: Aug 9, 2026
LC-MS Analysis of Human Platelets as a Platform for Studying Mitochondrial Metabolism
Published on: April 4, 2016
Effect of the psoralen-based photochemical pathogen inactivation on mitochondrial DNA in platelets
I Bruchmüller1, R Lösel, P Bugert
1Institute of Transfusion Medicine and Immunology, Red Cross Blood Service Baden-Württemberg-Hessen, Germany. i.bruchmuller@blutspende.de
Abstract:
Photochemical treatment (PCT) of platelet concentrates, using amotosalen HCl and UVA-light, inactivates pathogens by forming adducts between amotosalen and nucleic acids. The impact of the photochemical treatment on pathogens and leukocytes has been studied extensively. Yet little is known about the effect of PCT on nucleic acids in platelets. Platelets contain viable mitochondria and mitochondrial DNA (mtDNA) and this study aimed at evaluating the amotosalen modifications on platelet mtDNA. We applied two independent but complementary molecular assays to investigate qualitative as well as quantitative aspects of the psoralen-mediated DNA modifications in platelet mtDNA. The amotosalen-DNA modification density was measured using (14)C-labeled amotosalen. Amotosalen (150 microM) yielded 4.0 +/- 1.2 psoralen adducts per 1,000 bp in mtDNA after irradiation with 3 J/cm(2) UVA. Furthermore, we tested if the PCT-induced DNA modifications could be detected by a PCR assay. On the basis of PCR inhibition due to amotosalen-DNA adducts, mtDNA-specific PCR assays were developed and tested for their specificity and sensitivity. Our data revealed that mtDNA in platelets is substantially modified by PCT and that these modifications can be documented by a PCR inhibition system.
Insights
Photochemical treatment (PCT) modifies platelet mitochondrial DNA (mtDNA) by forming amotosalen adducts. These DNA modifications, caused by amotosalen and UVA light, can be detected using PCR inhibition assays.
Area of Science:
- Biochemistry
- Molecular Biology
- Hematology
Background:
- Platelet concentrates undergo photochemical treatment (PCT) with amotosalen HCl and UVA light to inactivate pathogens.
- While PCT's effects on pathogens and leukocytes are known, its impact on platelet nucleic acids, particularly mitochondrial DNA (mtDNA), is less understood.
Purpose of the Study:
- To evaluate amotosalen modifications on platelet mtDNA following PCT.
- To investigate both qualitative and quantitative aspects of psoralen-mediated DNA modifications in platelet mtDNA.
Main Methods:
- Utilized (14)C-labeled amotosalen to measure amotosalen-DNA modification density.
- Developed and tested mtDNA-specific PCR assays based on PCR inhibition due to amotosalen-DNA adducts.
Main Results:
- Amotosalen (150 microM) resulted in approximately 4.0 adducts per 1,000 bp in platelet mtDNA after 3 J/cm(2) UVA irradiation.
- PCT-induced amotosalen-DNA modifications in platelet mtDNA were successfully detected using a PCR inhibition system.
Conclusions:
- Photochemical treatment significantly modifies platelet mtDNA.
- The developed PCR inhibition system effectively documents amotosalen-induced modifications in platelet mtDNA.
