McClain, Lora
(2016)
HSV-1 infection in human induced pluripotent stem cell-derived neurons: cellular models for quiescence and drug discovery.
Doctoral Dissertation, University of Pittsburgh.
(Unpublished)
Abstract
Background: Herpes simplex virus, type 1 (HSV-1) establishes latency in human sensory ganglia following primary infection through mucosal tissues. Once latent, the virus persists for the host’s lifetime, with periodic reactivations that cause lytic lesions. First-line medications like acyclovir (ACV) abort lytic reactivations, but drug resistance has been reported and second line drugs may cause serious side effects. These facts, together with inefficacy of antivirals against latency, compel new drug screens; human neural tissues that model aspects of latency are arguably well-suited for such screens.
Studies:
Study 1. Human induced pluripotent stem cell-derived neurons (iPSC-neurons) were used to model HSV-1 infection employing an HSV-1 construct that incorporates dual fluorescent reporter genes from different kinetic expression groups. Lytic infection was demonstrated initially. ‘Quiescent’ infection was next established using protocols from animal models of latent infection. The quiescent infection fulfilled most criteria for latency, including viral gene expression and heterochromatization patterns.
Studies 2 and 3: The antiviral activity of two series of compounds was investigated using monkey epithelial cells and iPSC-neurons. Anti-lytic activity that reduced viral copy number and protein levels was induced by four compounds, of which two compounds also inhibited reactivation of HSV-1 from quiescence/latency. Four compounds were also efficacious against varicella zoster virus and/or human cytomegalovirus infections.
Study 4: Moderate throughput platforms for antiviral drug screens based on iPSC-neurons were designed, with readouts based on high content analysis and flow cytometry.
Conclusions: Lytic and latent HSV-1 infection was modeled in human iPSC-neurons, with features similar to animal models. Compounds with novel effects against HSV-1 infections were identified using the iPSC-neuron model. Platforms for moderate throughput drug screens are feasible using human iPSC-neuron models.
Public Health Significance: Herpesviruses are highly prevalent, with rates exceeding 95% in some populations. Recurrent HSV-1 eye infections are the leading cause of infectious corneal blindness in the USA. HSV-1 encephalitis that is particularly devastating to neonates can be fatal. Associations between HSV-1 seropositivity and cognitive impairment have been reported even without encephalitis. There are no effective HSV-1 vaccines and there are growing concerns about antiviral drug efficacy. In view of the enormous burden, there is a compelling need for novel drug screens.
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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: |
29 June 2016 |
| Date Type: |
Publication |
| Defense Date: |
2 March 2016 |
| Approval Date: |
29 June 2016 |
| Submission Date: |
30 March 2016 |
| Access Restriction: |
5 year -- Restrict access to University of Pittsburgh for a period of 5 years. |
| Number of Pages: |
290 |
| Institution: |
University of Pittsburgh |
| Schools and Programs: |
School of Public Health > Human Genetics |
| Degree: |
PhD - Doctor of Philosophy |
| Thesis Type: |
Doctoral Dissertation |
| Refereed: |
Yes |
| Uncontrolled Keywords: |
HSV-1, VZV, HCMV, antiviral, iPSC, neuron, drug discovery, gene expression |
| Date Deposited: |
29 Jun 2016 17:36 |
| Last Modified: |
01 May 2021 05:15 |
| URI: |
http://d-scholarship.pitt.edu/id/eprint/27447 |
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