Highly efficient CD4+ T cell targeting and genetic recombination using engineered CD4+ cell-homing mRNA-LNPs

Abstract

Nucleoside-modified messenger RNA (mRNA)-lipid nanoparticles (LNPs) are the basis for the first two EUA (Emergency Use Authorization) COVID-19 vaccines. The use of nucleoside-modified mRNA as a pharmacological agent opens immense opportunities for therapeutic, prophylactic and diagnostic molecular interventions. In particular, mRNA-based drugs may specifically modulate immune cells, such as T lymphocytes, for immunotherapy of oncologic, infectious and other conditions. The key challenge, however, is that T cells are notoriously resistant to transfection by exogenous mRNA. Here, we report that conjugating CD4 antibody to LNPs enables specific targeting and mRNA interventions to CD4+ cells, including T cells. After systemic injection in mice, CD4-targeted radiolabeled mRNA-LNPs accumulated in spleen, providing $30-fold higher signal of reporter mRNA in T cells isolated from spleen as compared with nontargeted mRNA-LNPs. Intravenous injection of CD4-targeted LNPs loaded with Cre recombinase-encoding mRNA provided specific dose-dependent loxP-mediated genetic recombination, resulting in reporter gene expression in about 60% and 40% of CD4+ T cells in spleen and lymph nodes, respectively. T cell phenotyping showed uniform transfection of T cell subpopulations, with no variability in uptake of CD4-targeted mRNA-LNPs in naive, central memory, and effector cells. The specific and efficient targeting and transfection of mRNA to T cells established in this study provides a platform technology for immunotherapy of devastating conditions and HIV cure.

Antisense Oligonucleotide-Based Therapy of Viral Infections

“…Inhalable or intranasal delivery of antiviral ASOs may be used for pulmonary delivery of ASOs targeting viral respiratory infections [141]. Finally, LNPs are one of the most promising carriers for ASO delivery and can be modified with cell-specific ligands or antibodies for selective ASO delivery to infected tissues-for example, GalNAc for liver cells and anti-CD4 for CD4 + T cells [142]. Growing knowledge and emerging technologies would benefit clinical translation of ASOs and other RNA-based therapies in the future.…”

The role of lipid components in lipid nanoparticles for vaccines and gene therapy

“…In the fast-growing field of cancer immunotherapy, the focus is on modifying and activating immune cells. A recent study [117] showed that conjugating CD4 antibody to LNPs enables specific targeting to and mRNA interventions in CD4+ cells, including T cells. After systemic injection in mice, CD4-targeted radiolabeled mRNA-LNPs accumulated in spleen, providing ∼30-fold higher signal from reporter mRNA in splenic T cells, as compared with non-targeted mRNA-LNPs.…”

Lipid Nanoparticle Delivery Systems to Enable mRNA-Based Therapeutics

“…Studies led by the Muzykantov laboratory have demonstrated the successful conjugation of antibodies targeting vascular adhesion molecule PECAM-1 or ICAM-1 and re-directing the LNP accumulation away from the liver to the lung [164,165] or VCAM-1 and targeting to the cerebral endothelium during brain edema [165]. A similar approach, using CD-4 antibody conjugated to the surface of the LNP, directed uptake into all T cells (naïve, central, memory, and effector) in both spleen and lymph enabling the possibility to conduct chimeric antigen receptor (CAR) T cell therapy in vivo [166]. Targeting ligands can re-distribute the LNP to extrahepatic sites; however, these examples are only demonstrated in small research scale settings.…”