Nortriptyline-targeted siRNA-loaded liposomes: Design, affinity, biodistribution, and bioactivity in a murine Herpes Simplex Virus 1 corneal infection model

Doaa Jbara-Agbaria; Neethi C Thathapudi; Marc Groleau; Majd Agbaria; Marie-Claude Robert; Natalia Callai Da Silva; Sebastien Talbot; Janet Laganiere; May Griffith; Gershon Golomb

doi:10.1016/j.ejps.2025.107388

Nortriptyline-targeted siRNA-loaded liposomes: Design, affinity, biodistribution, and bioactivity in a murine Herpes Simplex Virus 1 corneal infection model

Eur J Pharm Sci. 2026 Feb 1:217:107388. doi: 10.1016/j.ejps.2025.107388. Epub 2025 Nov 27.

Authors

Affiliations

¹ Institute for Drug Research, School of Pharmacy, Faculty of Medicine, The Hebrew University of Jerusalem, Jerusalem, 9112001, Israel. Electronic address: doaa.jbara@mail.huji.ac.il.
² Department of Ophthalmology, Université de Montréal, and Maisonneuve-Rosemont Hospital Research Centre, Montreal, QC H1T 2M4, Canada; Institute of Biomedical Engineering, Université de Montréal, Montreal, QC H3C 3J7, Canada. Electronic address: neethi.thathapudi@umontreal.ca.
³ Department of Ophthalmology, Université de Montréal, and Maisonneuve-Rosemont Hospital Research Centre, Montreal, QC H1T 2M4, Canada; Department of Microbiology, Infectiology and Immunology, Université de Montréal, Montreal, QC H3C 3J7, Canada. Electronic address: marc.groleau@umontreal.ca.
⁴ Institute for Drug Research, School of Pharmacy, Faculty of Medicine, The Hebrew University of Jerusalem, Jerusalem, 9112001, Israel. Electronic address: majd.agbaria@mail.huji.ac.il.
⁵ Department of Ophthalmology, Université de Montréal, and Maisonneuve-Rosemont Hospital Research Centre, Montreal, QC H1T 2M4, Canada; CHUM Research Centre, Montreal, QC H2 × 0A9 Canada. Electronic address: marie-claude.robert.2@umontreal.ca.
⁶ Department of Ophthalmology, Université de Montréal, and Maisonneuve-Rosemont Hospital Research Centre, Montreal, QC H1T 2M4, Canada; Institute of Biomedical Engineering, Université de Montréal, Montreal, QC H3C 3J7, Canada. Electronic address: natalia.callai.da.silva@umontreal.ca.
⁷ Department of Biomedical and Molecular Sciences, Queen's University, Kingston, Ontario K7L 2V7, Canada; Department of Physiology and Pharmacology, Karolinska Institute, Stockholm 171 77, Sweden. Electronic address: sebastien.talbot@queensu.ca.
⁸ Department of Ophthalmology, Université de Montréal, and Maisonneuve-Rosemont Hospital Research Centre, Montreal, QC H1T 2M4, Canada. Electronic address: casteleau@gmail.com.
⁹ Department of Ophthalmology, Université de Montréal, and Maisonneuve-Rosemont Hospital Research Centre, Montreal, QC H1T 2M4, Canada; Institute of Biomedical Engineering, Université de Montréal, Montreal, QC H3C 3J7, Canada; CHUM Research Centre, Montreal, QC H2 × 0A9 Canada. Electronic address: may.griffith.montreal@gmail.com.
¹⁰ Institute for Drug Research, School of Pharmacy, Faculty of Medicine, The Hebrew University of Jerusalem, Jerusalem, 9112001, Israel; The Center for Nanoscience and Nanotechnology, The Hebrew University of Jerusalem, Jerusalem, 9190401, Israel. Electronic address: gershon.golomb@mail.huji.ac.il.

PMID: 41318007
DOI: 10.1016/j.ejps.2025.107388

Abstract

Herpes simplex virus type 1 (HSV1) establishes latency in the trigeminal ganglia and reactivates to cause recurrent infections and severe complications, including herpes simplex keratitis (HSK), the leading cause of infectious corneal blindness in developed countries. We developed and characterized a targeted siRNA delivery system designed to suppress HSV1 by silencing the immediate-early gene ICP0, which encodes a protein essential for lytic infection and viral reactivation. To enhance neuronal accumulation, siRNA-loaded liposomes were decorated with nortriptyline (NTP), a ligand with high affinity for neuronal cells, including those in the trigeminal ganglia. The liposomes were optimized to exhibit nanoscale size, low polydispersity, neutral surface charge, high siRNA loading, spherical morphology, and long-term shelf stability. Studies in cell lines confirmed efficient internalization with good tolerability, and targeted liposomes showed enhanced binding to differentiated PC‑12 neuronal cells, competitively inhibited by free NTP, supporting their targeting specificity. Following retro-orbital administration in mice, NTP-targeted siHSV1 liposomes preferentially accumulated in the trigeminal ganglia. Antiviral efficacy was further assessed by topical application of siHSV1‑loaded liposomes within a biocompatible hydrogel in a murine HSV1 corneal perforation model of HSK. In the corneal limbus, targeted liposomes produced a significant reduction in viral load, whereas in the contralateral trigeminal ganglia, they significantly reduced viral load with a pronounced trend toward decreased ICP0 expression. These results demonstrate the feasibility of using NTP-mediated targeting ligand and support the potential of this delivery platform for treating latent HSV1, warranting further optimization for improved therapeutic efficacy.

Keywords: Herpes simplex keratitis; Herpes simplex virus 1; ICP0; Nortriptyline targeted delivery; Trigeminal ganglia; siRNA/siHSV1 liposomes.

MeSH terms

Animals
Antiviral Agents* / administration & dosage
Antiviral Agents* / pharmacokinetics
Cornea / drug effects
Cornea / metabolism
Cornea / virology
Disease Models, Animal
Female
Herpesvirus 1, Human* / drug effects
Herpesvirus 1, Human* / genetics
Humans
Immediate-Early Proteins / genetics
Keratitis, Herpetic* / drug therapy
Keratitis, Herpetic* / metabolism
Keratitis, Herpetic* / virology
Liposomes
Mice
Mice, Inbred BALB C
Nortriptyline* / administration & dosage
Nortriptyline* / chemistry
Nortriptyline* / pharmacokinetics
PC12 Cells
RNA, Small Interfering* / administration & dosage
RNA, Small Interfering* / chemistry
RNA, Small Interfering* / pharmacokinetics
Rats
Tissue Distribution
Trigeminal Ganglion / metabolism

Substances

Liposomes
Nortriptyline
RNA, Small Interfering
Antiviral Agents
Immediate-Early Proteins