Resveratrol Enhances mRNA and siRNA Lipid Nanoparticles Primary CLL Cell Transfection.

Edo Kon,Arnon Nagler,Dan Peer,Daniel Rosenblum,Niv Cohen,Sushmita Chatterjee,Pia Raanani,Inbal Hazan-Halevy,Nuphar Veiga,Osnat Bairey,Ohad Benjamini

doi:10.3390/pharmaceutics12060520

Edo Kon, Arnon Nagler + Show 9 more

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https://doi.org/10.3390/pharmaceutics12060520

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Abstract

Chronic lymphocytic leukemia (CLL) is the most common adult leukemia in Western populations. Therapies such as mRNA and siRNA encapsulated in lipid nanoparticles (LNPs) represent a clinically advanced platform and are utilized for a wide variety of applications. Unfortunately, transfection of RNA into CLL cells remains a formidable challenge and a bottleneck for developing targeted therapies for this disease. Therefore, we aimed to elucidate the barriers to efficient transfection of RNA-encapsulated LNPs into primary CLL cells to advance therapies in the future. To this end, we transfected primary CLL patient samples with mRNA and siRNA payloads encapsulated in an FDA-approved LNP formulation and characterized the transfection. Additionally, we tested the potential of repurposing caffeic acid, curcumin and resveratrol to enhance the transfection of nucleic acids into CLL cells. The results demonstrate that the rapid uptake of LNPs is required for successful transfection. Furthermore, we demonstrate that resveratrol enhances the delivery of both mRNA and siRNA encapsulated in LNPs into primary CLL patient samples, overcoming inter-patient heterogeneity. This study points out the important challenges to consider for efficient RNA therapeutics for CLL patients and advocates the use of resveratrol in combination with RNA lipid nanoparticles to enhance delivery into CLL cells.

Highlights

Chronic lymphocytic leukemia (CLL) is the most common leukemia, with 15,000 new diagnoses and claiming 5000 lives per year in the United States alone [1,2,3,4]
We transfected primary CLL cells with si-CD44 and si-signal transducer and activator of transcription 3 (STAT3) and determined CD44 and STAT3 gene expression levels (Figure 4C,D). These results demonstrate that resveratrol enhances mRNA, and siRNA-lipid nanoparticles (LNPs) transfection
We demonstrate the capability of repurposing resveratrol as an RNA-LNP transfection enhancer for both mRNA and siRNA payloads, hinting at a possible shared mechanism

Summary

Introduction

Chronic lymphocytic leukemia (CLL) is the most common leukemia, with 15,000 new diagnoses and claiming 5000 lives per year in the United States alone [1,2,3,4]. The median age of CLL patients at diagnosis is above 70 years [1,3]. CLL cells can be found primarily in the peripheral blood, bone marrow, and lymph nodes. They express both B cell markers, such as cluster of differentiation (CD), together with non-B cell markers, such as CD5 and CD23 [3,4,5,6]. The standard treatment for CLL is a harsh chemo-immunotherapy regimen of fludarabine (purine analog), cyclophosphamide or chlorambucil (alkylating agents) and rituximab (α-CD20 antibody), known by its acronym FCR, which is not applicable for all patients [3,7]

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