Receptor-Targeted Nipah Virus Glycoproteins Improve Cell-Type Selective Gene Delivery and Reveal a Preference for Membrane-Proximal Cell Attachment

Abstract

Receptor-targeted lentiviral vectors (LVs) can be an effective tool for selective transfer of genes into distinct cell types of choice. Moreover, they can be used to determine the molecular properties that cell surface proteins must fulfill to act as receptors for viral glycoproteins. Here we show that LVs pseudotyped with receptor-targeted Nipah virus (NiV) glycoproteins effectively enter into cells when they use cell surface proteins as receptors that bring them closely enough to the cell membrane (less than 100 Å distance). Then, they were flexible in receptor usage as demonstrated by successful targeting of EpCAM, CD20, and CD8, and as selective as LVs pseudotyped with receptor-targeted measles virus (MV) glycoproteins, the current standard for cell-type specific gene delivery. Remarkably, NiV-LVs could be produced at up to two orders of magnitude higher titers compared to their MV-based counterparts and were at least 10,000-fold less effectively neutralized than MV glycoprotein pseudotyped LVs by pooled human intravenous immunoglobulin. An important finding for NiV-LVs targeted to Her2/neu was an about 100-fold higher gene transfer activity when particles were targeted to membrane-proximal regions as compared to particles binding to a more membrane-distal epitope. Likewise, the low gene transfer activity mediated by NiV-LV particles bound to the membrane distal domains of CD117 or the glutamate receptor subunit 4 (GluA4) was substantially enhanced by reducing receptor size to below 100 Å. Overall, the data suggest that the NiV glycoproteins are optimally suited for cell-type specific gene delivery with LVs and, in addition, for the first time define which parts of a cell surface protein should be targeted to achieve optimal gene transfer rates with receptor-targeted LVs.

Pseudotyped Lentiviral Vectors: One Vector, Many Guises

“…NiV-LVs retargeted to EpCAM, CD8, CD117, or GluA4 were used successfully. 93 These reports demonstrate that in the recent years, largely due to the tremendous flexibility of paramyxoviral envelopes, targeting of LVs to specific cell types has become more feasible and efficient.…”

Immune Modulatory Cell Therapy for Hemophilia B Based on CD20-Targeted Lentiviral Gene Transfer to Primary B Cells

“…Although this could be very efficient for ex vivo B cell transduction, this vector lacks selectivity for B cells, making direct in vivo application difficult. A recent paper demonstrated that the Nipah virus (NiV) glycoproteins can be engineered for receptor targeting of LV, resulting in substantially improved vector production yields, while retaining the high selectivity of the MV-based system 39 . In particular, when the same CD20-specific scFv that was used here on the MV glycoproteins was used on the NiV glycoproteins, vector production yields increased by more than two orders of magnitude, making a scale-up for the production of clinical vector lots conceivable.…”

Design and construction of a recombinant lentiviral vector with specific tropism to human epidermal growth factor‐overexpressed cancer cells: Developing a new retargeting system for lentivirus vectors

“…Therefore, NiV glycoproteins appear to be a more appropriate option for generating the pseudotyped LVs than MV or TPMV. However, the pseudotyped NiV‐LVs are highly sensitive to the position of the binding site to the target receptor and proximal positions of the membrane are preferred to the membrane distal, which may slightly limit their use …”

“…However, the pseudotyped NiV-LVs are highly sensitive to the position of the binding site to the target receptor and proximal positions of the membrane are preferred to the membrane distal, which may slightly limit their use. 27 Given the above explanations, we aimed to design a novel Figure 1A and B), using the lipofectamine reagent (Thermo Fisher Scientific, Waltham, MA, USA) as described previously. 29 The Figure 1B).…”