DRG-targeted helper-dependent adenoviruses mediate selective gene delivery for therapeutic rescue of sensory neuronopathies in mice
Tomoya Terashima, Kazuhiro Oka, Angelika B. Kritz, Hideto Kojima, Andrew H. Baker, Lawrence Chan
Tomoya Terashima, Kazuhiro Oka, Angelika B. Kritz, Hideto Kojima, Andrew H. Baker, Lawrence Chan
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Technical Advance Genetics

DRG-targeted helper-dependent adenoviruses mediate selective gene delivery for therapeutic rescue of sensory neuronopathies in mice

Abstract

Dorsal root ganglion (DRG) neuron dysfunction occurs in a variety of sensory neuronopathies for which there are currently no satisfactory treatments. Here we describe the development of a strategy to target therapeutic genes to DRG neurons for the treatment of these disorders. We genetically modified an adenovirus (Ad) to generate a helper virus (HV) that was detargeted for native adenoviral tropism and contained DRG homing peptides in the adenoviral capsid fiber protein; we used this HV to generate DRG-targeted helper-dependent Ad (HDAd). In mice, intrathecal injection of this HDAd produced a 100-fold higher transduction of DRG neurons and a markedly attenuated inflammatory response compared with unmodified HDAd. We also injected HDAd encoding the β subunit of β-hexosaminidase (Hexb) into Hexb-deficient mice, a model of the neuronopathy Sandhoff disease. Delivery of the DRG-targeted HDAd reinstated neuron-specific Hexb production, reversed gangliosidosis, and ameliorated peripheral sensory dysfunction. The development of DRG neuron–targeted HDAd with proven efficacy in a preclinical model may have implications for the treatment of sensory neuronopathies of diverse etiologies.

Authors

Tomoya Terashima, Kazuhiro Oka, Angelika B. Kritz, Hideto Kojima, Andrew H. Baker, Lawrence Chan

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Figure 5

Gene therapy for a mouse model of Sandhoff disease.

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Gene therapy for a mouse model of Sandhoff disease. (A) Hexb gene expres...
(A) Hexb gene expression in neuronal tissues. Various fiber-modified HDAd vectors (1 × 108 vp) were injected into Hexb–/– mice through subarachnoid space at the lumbar level, and neuronal tissues were isolated for extraction of cellular RNA 8 weeks after injection. Hexb mRNA was quantified by real-time RT-PCR, and expression was calculated relative to that in the brains of wild-type mice. Undetectable expression levels are denoted by “0.” (B–D) Hexosaminidase activities in nervous tissues from wild-type and Hexb–/– mice with fiber-modified HDAd vector injection. (B) Total hexosaminidase (HEX). (C) Hexa. (D) Hexb (calculated by subtracting Hexa activity from total hexosaminidase activity). (E) Immunofluorescence (IF) for Hexb protein in DRG. (F) In situ staining for hexosaminidase activity. Nuclei were stained with methyl green. Arrows indicate weak positive staining in neurons. Scale bars: 20 μm. *P < 0.001; **P < 0.01, ***P < 0.05. #P < 0.01, ##P < 0.05 versus empty vector–treated Hexb–/–.