PB0762 - Storage and Characterization of mRNA-Lipid Nanoparticle Engineered Platelets

Monday, June 24, 2024

13:45 – 14:45 ICT

Room: Exhibition Hall

Presenting Author(s)

EK

Emma Kang

Student
University of British Columbia
Vancouver, British Columbia, Canada

Co-Author(s)

CS

Colton Strong

PhD Candidate
The University of British Columbia
Vancouver, British Columbia, Canada
Jerry Leung, PhD

Postdoctoral Research Fellow
The University of British Columbia
Vancouver, British Columbia, Canada
KB

Katherine Badior

Postdoctoral Research Fellow
Versiti Blood Research Institute
Milwaukee, Wisconsin, United States
MR

Madelaine Robertson

Graduate Student
The University of British Columbia
Vancouver, British Columbia, Canada
NP

Nicolas Pereyra

Graduate Student
The University of British Columbia
Vancouver, British Columbia, Canada
ER

Elyn Rowe

Graduate Student
The University of British Columbia
Vancouver, British Columbia, Canada
AW

Amanda Wietrzny

Graduate Student
Versiti Blood Research Institute
Milwaukee, Wisconsin, United States
BM

Brenda Ma

Undergraduate Student
The University of British Columbia
Vancouver, British Columbia, Canada
ZN

Zechariah Noronha

Student
University of British Columbia
Vancouver, British Columbia, Canada
DA

Deaglan Arnold

Employee
Nanovation Therapeutics
Vancouver, British Columbia, Canada
MC

Marco A. Ciufolini

Professor
The University of British Columbia
Vancouver, British Columbia, Canada
DD

Dana Devine

Professor
The University of British Columbia
Vancouver, British Columbia, Canada
EJ

Eric Jan

Professor
The University of British Columbia
Vancouver, British Columbia, Canada
PC

Pieter R. Cullis

Professor
The University of British Columbia
Vancouver, British Columbia, Canada
Christian J. Kastrup

Senior Investigator
Versiti Blood Research Institute
Milwaukee, Wisconsin, United States

Background: In cases of major hemorrhage, platelet transfusions are essential, but not always effective. One approach to overcome limitations of current platelet products is to enhance their innate function using lipid nanoparticles (LNP) to deliver exogenous RNA cargo. We have identified a mRNA-LNP formulation that is able to transfect platelets in plasma and plasma-based systems, however, it is unclear whether the mRNA-LNP dosing is scalable to physiological and supraphysiological concentrations and what their effects are on platelet function and storage.

Aims: To determine (1) if mRNA-LNP transfection can be scaled to physiological and supraphysiological platelet concentrations and (2) the effect of mRNA-LNPs on overall platelet physiology during storage.

Methods: In this study, human platelet units were processed at Canadian Blood Services. Platelets stored at physiological and supraphysiological concentrations were treated with different doses of mRNA-LNP encapsulating NanoLuciferase (nLuc) and characterized to assess function and quality. Transfection was evaluated by measuring total luminescence. Platelet function was measured as a responsiveness to various agonists using flow cytometry, rotational thromboelastometry (ROTEM), and imaging through transmission electron microscopy (TEM).

Results: An optimized mRNA-LNP achieved platelet transfection at physiological and supraphysiological concentrations in plasma and plasma-based systems. Increasing doses of mRNA-LNP did not significantly increase platelet activation and did not decrease their responsiveness to agonists (Figure 1). Moreover, the platelets transfected with mRNA-LNPs generated a similar clotting profile to untreated platelets, and found no significant differences in ROTEM parameters. The morphology of the treated and untreated platelets was visualized using TEM, and revealed no notable differences.

Conclusion(s): mRNA-LNP transfection in platelets is scalable to physiological and supraphysiological concentrations without impairing coagulability and agonist responsiveness. Further advancing genetically engineered platelets with clinical suitability holds the potential to broaden the therapeutic scope of platelets as a versatile cell therapy.