Principal Investigator: C. D. Atreya, PhD
Office / Division / Lab: OBRR / DBCD / LCH

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General Overview

Platelets are cells in the blood that trigger blood clotting. Doctors use platelets to prevent or treat bleeding in individuals with dangerously low platelet counts, such as those undergoing chemotherapy for cancer, as well those who suffer major blood loss due to trauma or surgery, and individuals who do not produce adequate numbers of platelets.

However, donated, stored platelets contaminated with bacteria pose the risk of a life-threatening infection in patients who receive them. Therefore, platelets are stored for no more than five days and tested for contamination before being transfused into patients.

In response to this problem, first we are developing novel ways to reduce the levels of pathogenic bacteria and viruses in stored platelets. This work is important because growth of bacteria in room temperature-stored platelet concentrates (PCs) can cause sepsis ("blood infection," causing chills, fever, and other symptoms) and other complications in recipients of transfusions. Thus, reducing bacterial growth in platelets and other blood products, including bacteria that are resistant to conventional antibiotics, would significantly enhance blood and blood product safety. Our laboratory is studying the ability of certain molecules called anti-microbial peptides (AMPs) to reduce pathogens in PCs and other blood products. The initial goal of this project is to evaluate selected AMPs against a panel of bacteria that are potential threats to platelet safety. Subsequently, we will extend our studies to the protection of red blood cells.

We are also studying the potential usefulness of tiny pieces of genetic material called cellular micro-RNAs (miRs) in their value in identifying whole blood or blood products that are contaminated with infectious microorganisms. This will also contribute to biodefense efforts aimed at protecting the blood supply. In addition, screening for the cellular miRs might also help to predict whether these stored therapeutic products will function properly after being transfused into patients.

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Scientific Overview

Pathogen reduction: A single, easy-to-use, and cost-effective pathogen inactivation approach would help to improve the safety of our nation's blood supply, including red blood cells, plasma, and platelets (PLTs). Several methods and technologies are currently being studied to help reduce bacterial contamination of blood components.

Our laboratory is introducing a novel proof of concept that uses known therapeutic antimicrobial peptides (AMPs) as bactericidal agents for platelet concentrates (PCs) stored at room temperature. We tested nine synthetic AMPs (four from PLT microbicidal protein-derived peptides (PD1-4) and five Arg-Trp (RW) repeat peptides containing one to five repeats) for bactericidal activity in plasma and PC samples spiked with Staphylococcus aureus, S. epidermidis, Escherichia coli, Pseudomonas aeruginosa, Klebsiella pneumoniae, and Bacillus cereus. A 3-log reduction of viable bacteria was considered effective bactericidal activity for a given peptide.

In both plasma alone and PCs, RW3 peptide demonstrated bactericidal activity against S. aureus, S. epidermidis, E. coli, P. aeruginosa, and K. pneumoniae; PD4 and RW2 against P. aeruginosa; and RW4 against K. pneumoniae. The activity of each of these four peptides against the remaining bacterial species in the test panel resulted in less than a 3-log reduction in the number of viable bacteria and hence considered ineffective.

These findings suggest a new approach to improving the safety of blood components, demonstrating the potential usefulness of screening therapeutic AMPs against selected bacteria to identify suitable bactericidal agents for stored plasma, and blood cells.

Micro-RNAs as biomarkers of infection and stored blood and blood cells: In addition to the above described studies, we are also studying cellular micro-RNAs (miRs) to determine a) if they can be used as early indicators of infection when whole blood or cellular blood components are contaminated with pathogens--work that will also contribute to biodefense efforts aimed at protecting the blood supply and b) whether cellular miR profiles of stored blood cells (platelets and red blood cells) can be useful as markers of storage that are predictive of their function following transfusion into patients.

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Publications

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