Doyal, Mark F.
(2015)
Promoter-targeted anti-nociceptive HSV-1 vectors have differential effects on pain based on the neuronal population targeted.
Master's Thesis, University of Pittsburgh.
(Unpublished)
Abstract
Herpes simplex virus-1 (HSV-1) is a neurotropic virus capable of infecting sensory neurons through their termini at any site on the body. Upon infection, the HSV-1 genome persists in nuclei within sensory ganglia for the life of the host. Replication-defective HSV-1 vectors allow for the efficient transduction of diverse populations of sensory neurons without risk of herpetic disease and therefore represent an ideal gene therapy vehicle for the treatment of peripheral neurological diseases, including chronic pain. To evaluate the potential of HSV-1 vectors to treat pain, a set of replication-defective HSV-1 vectors was generated, driving the expression of an anti-nociceptive product (GlyRIS) from neuronal promoters to target expression to distinct neuronal populations. Specifically, the TRPV1 promoter was used to target heat-sensitive nociceptors, and the NF200 promoter was used to target large diameter Aβ-fibers which may be recruited for pain signaling after injury or inflammation. The ubiquitously expressed CMV promoter was used as a control. As expected, when these vectors were used to transduce cultured DRG cells, the neuronal promoters largely expressed in neuronal cells only, while the CMV promoter expressed in neuronal and support cells. For pain studies, vectors were injected under the skin of the right hind footpad in rats. After nine days, baseline thermal and mechanical withdrawal thresholds were taken before and after the application of the ligand ivermectin, activating vector-delivered GlyRIS. TRPV1-GlyRIS transduced rats and to a lesser extent CMV-GlyRIS transduced rats showed increased thermal withdrawal thresholds on the transduced side after ligand administration. Resiniferatoxin (RTX) was then injected to ablate TRPV1+ neurons and induce mechanical allodynia. After 20 days, all RTX-injected rats showed increased thermal withdrawal thresholds with a loss of dependence on vector injection and ligand administration, consistent with TRPV1+ neuron ablation. All RTX-injected rats developed bilateral mechanical allodynia, except for the NF200-GlyRIS transduced rats (Aβ-targeted) which demonstrated decreased mechanical allodynia on the transduced side relative to the contralateral side after ivermectin administration. These differential effects on nociception represent the functional outcome of differentially targeted anti-nociceptive HSV-1 vectors and support the use of promoter-targeting to express transgenes in specific neuronal subpopulations.
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Details
| Item Type: |
University of Pittsburgh ETD
|
| Status: |
Unpublished |
| Creators/Authors: |
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| ETD Committee: |
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| Date: |
24 April 2015 |
| Date Type: |
Publication |
| Defense Date: |
24 April 2015 |
| Approval Date: |
24 April 2015 |
| Submission Date: |
24 April 2015 |
| Access Restriction: |
3 year -- Restrict access to University of Pittsburgh for a period of 3 years. |
| Number of Pages: |
86 |
| Institution: |
University of Pittsburgh |
| Schools and Programs: |
School of Medicine > Molecular Virology and Microbiology |
| Degree: |
MS - Master of Science |
| Thesis Type: |
Master's Thesis |
| Refereed: |
Yes |
| Uncontrolled Keywords: |
Herpes simplex virus HSV HSV-1 gene therapy vector promoter transcriptional targeting pain nociception afferent nociceptor rodent rat resiniferatoxin RTX TRPV1 NF200 |
| Date Deposited: |
24 Apr 2015 20:31 |
| Last Modified: |
24 Apr 2018 05:15 |
| URI: |
http://d-scholarship.pitt.edu/id/eprint/25066 |
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