Peter Schubert1,2,3, Brankica Culibrk1,2, Simrath Karwal1,2, Raymond P Goodrich4, Dana V Devine1,2,3. 1. Centre for Innovation, Canadian Blood Services, University of British Columbia, Vancouver, BC, Canada. 2. Centre for Blood Research, University of British Columbia, Vancouver, BC, Canada. 3. Department of Pathology and Laboratory Medicine, University of British Columbia, Vancouver, BC, Canada. 4. TerumoBCT Biotechnologies, Lakewood, CO, USA.
Abstract
PURPOSE: Pathogen inactivation technologies (PITs) were introduced into blood banking to further improve the safety of blood products. However, the UV light used in PITs to terminate pathogen growth might alter the functionality of the cells in the blood product as well as the protein profile of the blood components. This study employed proteomic approaches to assess changes in the platelet proteome and translatome. EXPERIMENTAL DESIGN: Apheresis-derived platelet concentrates treated with riboflavin/UV light or untreated controls were analyzed throughout blood bank storage by quantitative proteomics using iTRAQ and puromycin-associated nascent chain (PUNCH) proteomics. RESULTS: Quantitative proteomic analysis identified 408 individual proteins including 26 unique proteins that changed in the treated arm during storage. Proteomic results were confirmed using immunoblot analyses and results suggested a translational control of the protein expression profile. PUNCH proteomic analysis of day 7 samples from illuminated units identified 52 unique platelet proteins that incorporated puromycin, including proteins involved in the cytoskeleton, metabolism, and signaling. CONCLUSION AND CLINICAL RELEVANCE: This study demonstrates for the first time that platelets can synthesize proteins despite the riboflavin and UV treatment and suggests that platelets may possess a mechanism to protect their mRNA from damage by the PI treatment.
PURPOSE: Pathogen inactivation technologies (PITs) were introduced into blood banking to further improve the safety of blood products. However, the UV light used in PITs to terminate pathogen growth might alter the functionality of the cells in the blood product as well as the protein profile of the blood components. This study employed proteomic approaches to assess changes in the platelet proteome and translatome. EXPERIMENTAL DESIGN: Apheresis-derived platelet concentrates treated with riboflavin/UV light or untreated controls were analyzed throughout blood bank storage by quantitative proteomics using iTRAQ and puromycin-associated nascent chain (PUNCH) proteomics. RESULTS: Quantitative proteomic analysis identified 408 individual proteins including 26 unique proteins that changed in the treated arm during storage. Proteomic results were confirmed using immunoblot analyses and results suggested a translational control of the protein expression profile. PUNCH proteomic analysis of day 7 samples from illuminated units identified 52 unique platelet proteins that incorporated puromycin, including proteins involved in the cytoskeleton, metabolism, and signaling. CONCLUSION AND CLINICAL RELEVANCE: This study demonstrates for the first time that platelets can synthesize proteins despite the riboflavin and UV treatment and suggests that platelets may possess a mechanism to protect their mRNA from damage by the PI treatment.