Ex Vivo Stored Blood Component Safety and Quality: Evaluation of Novel Methods for Pathogen Reduction and Functional Regulation in Blood Components
C.D. Atreya, Ph.D.
Office of Blood Research and Review
Division of Blood Components and Devices
Laboratory of Cellular Hematology
Dr. C.D. Atreya, PhD is the Associate Director for research, Office of Blood Research and Review, Center for Biologics Research and Review at the U.S. Food and Drug Administration. He has published more than 80 scientific publications in peer-reviewed journals and serves on the editorial board for a number of peer-review scientific journals.
Dr. Atreya received a PhD from University of Hyderabad, India in the field of molecular genetics, followed by postdoctoral training at Wayne State University, Detroit, MI. He then became an assistant professor of research at the University of Kentucky, Lexington, KY. Subsequently, Dr. Atreya joined CBER/FDA in 1993.
Ex vivo stored blood components save millions of lives in the US and globally. Research in my laboratory addresses some of the unmet critical public health needs to help facilitate new approaches to pathogen reduction in the blood components, while preserving the components' functionality and understanding the quality changes during storage of products (platelets and RBCs) by using miRNAs as markers and regulators of changes.
1) Blood Component Safety: Development and evaluation of novel approaches to microbial reduction in stored human blood components (plasma, platelets, and red blood cells [RBCs]).
Due to steps that are inherent to drawing blood from a healthy donor and proceeding toward final storage, plasma, platelets, and RBCs may be exposed to microbial contamination. Platelets are stored at room temperature, which promotes bacterial growth and poses the additional risk of a life-threatening bacterial infection called sepsis ("blood infection," causing chills, fever, and other symptoms); rarely, recipients of platelet transfusions experience other complications or even die. Therefore, platelets are only allowed to be stored for five – seven days, and platelets are tested for contamination before being transfused into patients.
Current FDA-approved first-generation pathogen reduction technologies (PRTs) are limited to treating stored platelets and plasma only, and no PRTs are available yet in the U.S. for stored whole blood (WB) and RBCs. Hence, this is an unmet need. While approved PRTs for stored platelets and plasma are a good start, pathogen inactivation has unintended consequences on the treated blood components. Therefore, the objective of this research is to identify and evaluate methods or technologies that may be more selective, inactivating pathogens while preserving the full functional spectrum of the treated blood product.
2) Cellular blood Component Quality: Evaluation of platelets and red blood cells (RBC) in storage towards enhancing their quality.
During storage, the blood cells undergo physiological and biochemical changes which negatively impact product quality and subsequent performance in transfused patients. These changes are collectively known as "storage lesions” (SL). FDA has approved a few additive solutions for blood cells to preserve their quality during storage. However, developing ideal storage media for these cellular products has been challenging, because mechanisms underlying the product deterioration during storage are not fully understood.
We also need in vitro product quality markers to predict blood component performance in transfused patients. Further, efforts to enhance blood component shelf-life to longer periods especially during disasters, when blood collection from donors will be drastically disrupted, will be critical to patient care. Increasing the product shelf-life will also safeguard donors' health, as donation frequency could be reduced. We began to study blood cells in storage to understand molecular mechanisms underlying the product deterioration and to identify markers of product quality. The success of these studies will strengthen FDA's science base and facilitate review of new regulatory applications relevant to stored cellular blood components in the future.
1) Blood component Safety: Development and evaluation of novel approaches to microbial reduction in stored platelets.
In collaboration with Michelle Maclean at the University of Strathclyde, Glasgow, UK, we have demonstrated that 405 nm visible blue light is effective on selected bacteria spiked in stored plasma and platelets. The treated platelets show survival and recovery in SCID mice similar to that of untreated platelets. Subsequently, we demonstrated that a blood-borne parasite, Trypanosoma cruzi (which causes Chagas disease) can be completely inactivated by 405 nm light, while preserving the functions of the platelets and plasma protein integrity. Based on some of this work, a prototype apparatus to treat blood components was developed (patent pending, DHHS Ref. No. E-153-2014/0-PCT-02).
2) Blood Component Quality: Study of platelets in storage towards enhancing their quality.
MicroRNAs (miRNAs) have been found to regulate expression of many genes via messenger RNAs (mRNAs). We and others have identified altered expression of miRNAs in RBCs and platelets during storage. The regulatory functions of miRNA are more relevant to stored platelets and RBCs, as these cells are enucleated and have lost their nucleus-driven regulation. Therefore, these cells depend on available cytoplasmic regulatory mechanisms for their survival (i.e., mRNA regulation by miRNAs). We are investigating the regulation of messenger RNAs (mRNAs) by miRNAs relevant to platelet and RBC functions during storage. To date, we have experimentally demonstrated the role of miRNAs in platelet activation, aggregation, and apoptosis.
Taken together, the information we obtain from these studies will help identify storage markers of the product quality (platelet and RBC) and decipher key cellular regulatory pathways to enhance product quality of ex vivo stored platelets and RBCs.
- ORCID ID: 0000-0001-7393-2157
- Photochem Photobiol 2021 Dec 21 [Epub ahead of print]
Violet-blue 405-nm light-based photoinactivation for pathogen reduction of human plasma provides broad antibacterial efficacy without visible degradation of plasma proteins.
Stewart CF, Tomb RM, Ralston HJ, Armstrong J, Anderson JG, MacGregor SJ, Atreya CD, Maclean M
- Microrna 2021 Jun;10(2):123-9
MiRNA-103b downregulates ITGB3 and mediates apoptosis in ex vivo stored human platelets.
Dahiya N, Atreya C
- Front Med 2020 Nov 24;7:617373
Complete inactivation of blood borne pathogen Trypanosoma cruzi in stored human platelet concentrates and plasma treated with 405 nm violet-blue light.
Jankowska KI, Nagarkatti R, Acharyya N, Dahiya N, Stewart CF, Macpherson RW, Wilson MP, Anderson JG, MacGregor SJ, Maclean M, Dey N, Debrabant A, Atreya CD
- Blood Transfus 2021 Sep;19(4):403-12
Analysis of the mechanism of damage produced by thiazole orange photoinactivation in apheresis platelets.
Gough P, Getz T, De Paoli S, Wagner S, Atreya C
- Int J Mol Sci 2020 Aug 6;21(16):E5621
A foundational study for normal F8-containing mouse models for the miRNA regulation of hemophilia A: identification and analysis of mouse miRNAS that downregulate the murine F8 gene.
Jankowska KI, Chattopadhyay M, Sauna ZE, Atreya CD
- Front Cell Dev Biol 2020 Jul 30;8:669
Further evidence that microRNAs can play a role in Hemophilia A disease manifestation: F8 gene downregulation by miR-19b-3p and miR-186-5p.
Jankowska KI, McGill J, Pezeshkpoor B, Oldenburg J, Sauna ZE, Atreya CD
- Microrna 2020;9(3):240-6
MiR-181a reduces platelet activation via the inhibition of endogenous RAP1B.
Dahiya N, Atreya CD
- Transfusion 2020 Feb;60(2):401-13
Clinical manifestation of hemophilia A in the absence of mutations in the F8 gene that encodes FVIII: role of microRNAs.
Jankowska KI, McGill J, Pezeshkpoor B, Oldenburg J, Atreya CD, Sauna ZE
- Front Med 2020 Jan 15;6:331
Non-ionizing 405 nm light as a potential bactericidal technology for platelet safety: evaluation of in vitro bacterial inactivation and in vivo platelet recovery in severe combined immunodeficient mice.
Maclean M, Gelderman MP, Kulkarni S, Tomb RM, Stewart CF, Anderson JG, MacGregor SJ, Atreya CD
- Transfusion 2019 Sep;59(9):3002-25
Proceedings of the Food and Drug Administration public workshop on pathogen reduction technologies for blood safety 2018.
Atreya C, Glynn S, Busch M, Kleinman S, Snyder E, Rutter S, AuBuchon J, Flegel W, Reeve D, Devine D, Cohn C, Custer B, Goodrich R, Benjamin RJ, Razatos A, Cancelas J, Wagner S, Maclean M, Gelderman M, Cap A, Ness P
- Microrna 2019;8(1):36-42
RAP1 downregulation by miR-320c reduces platelet activation in ex vivo storage.
Atreya C, Dahiya N
- Microrna 2018;7(3):223-8
MicroRNA-223 regulates Septin-2 and Septin-6 in stored platelets.
Chattopadhyay M, Dahiya N, Atreya C
- Transfusion 2018 Aug;58(8):2013-21
Antimicrobial peptides: an effective approach to prevent bacterial biofilm formation in platelet concentrates.
Alabdullatif M, Atreya CD, Ramirez-Arcos S
- Transfusion 2017 Dec;57(12):2995-3000
Analysis of Argonaute 2-microRNA complexes in ex vivo stored red blood cells.
Vu L, Ragupathy V, Kulkarni S, Atreya C
- Food Environ Virol 2017 Jun;9(2):159-67
New proof-of-concept in viral inactivation: virucidal efficacy of 405 nm light against feline calicivirus as a model for norovirus decontamination.
Tomb RM, Maclean M, Coia JE, Graham E, McDonald M, Atreya CD, MacGregor SJ, Anderson JG
- Platelets 2017 Jan;28(1):74-81
miR-570 interacts with mitochondrial ATPase subunit g (ATP5L) encoding mRNA in stored platelets.
Dahiya N, Sarachana T, Kulkarni S, Wood III WH, Zhang Y, Becker KG, Wang BD, Atreya CD
- J Blood Transfusion 2016;2016:2920514
A new proof of concept in bacterial reduction: antimicrobial action of violet-blue light (405 nm) in ex vivo stored plasma.
Maclean M, Anderson JG, MacGregor SJ, White T, Atreya CD
- Transfusion 2015 Nov;55(11):2672-83
Evaluation of small noncoding RNAs in ex vivo stored human mature red blood cells: changes in noncoding RNA levels correlate with storage lesion events.
Sarachana T, Kulkarni S, Atreya CD
- Transfus Med Rev 2015 Oct;29(4):215-9
Platelet microRNAs: an overview.
Dahiya N, Sarachana T, Vu L, Becker KG, Wood WH 3rd, Zhang Y, Atreya CD
- PLoS One 2015 Jul 15;10(7):e0132433
Small ncRNA expression-profiling of blood from hemophilia A patients identifies miR-1246 as a potential regulator of Ffctor 8 gene.
Sarachana T, Dahiya N, Simhadri VL, Pandey GS, Saini S, Guelcher C, Guerrera MF, Kimchi-Sarfaty C, Sauna ZE, Atreya CD