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Principal Investigator: Ronald Goldstein
Organization: UNIVERSITY OF PITTSBURGH AT PITTSBURGH
Fiscal Year: 2021
Award: $184,530
Funding agency: National Institute of Allergy and Infectious Diseases
Abstract
As highlighted in PA-19-237, there is a gap in knowledge about non-coding RNAs and their functions
for many pathogens, and the human herpesvirus Varicella Zoster Virus (VZV) is one such pathogen. VZV is
one of only two of nine human herpesviruses in which microRNAs have not been reported. Our searches
identified a family of novel VZV-encoded small non-coding RNAs (VZVsncRNA), suggesting an exciting
possibility that VZV regulates its infectious process using sncRNA. Our overarching premise is that better
knowledge of sncRNA could form the basis for novel therapies for VZV disease, which is still a major public
health concern. While childhood “chickenpox” is no longer common in the US due to varicella vaccination,
most adults still harbor wild-type VZV in their ganglia in a latent state, with potential to reactivate. When it does,
Herpes Zoster develops, a painful morbid and debilitating disease encountered by a third in their lifetime that is
frequently complicated, most often by chronic pain. There is still some half of a million HZ cases annually.
We detected these VZVsncRNA in both infected epithelial cells and lytic infected neurons. Importantly,
cells transfected with locked nucleic acid (LNA) inhibitors to them alter VZV spread and growth. Here we focus
on four VZVsncRNA which are antisense to the VZV Latency Transcript (VLT) found in latently infected human
ganglia and productive infections, as LNA inhibitors to them reduce VZV virus production (indicating the
sncRNA promote infection). A combination of three LNA inhibitors effectively reduced VZV growth by more
than 90%, not only supporting a functional role in infection but also establishing a promising novel therapeutic
strategy. We hypothesize that these sncRNA regulate epithelial productive infections and in neurons, the
lytic/latency decisions that must be made. Aim 1 addresses how these four VZVsncRNA influence VZV growth
in epithelial cells using LNA modulators, in single and in combination, to determine how they change viral
expression patterns. To determine if they target virus or host, we will develop VZV altered in sequence
encoding the VZVsncRNA and using them in conjunction with epithelial cell lines expressing the VZVsncRNA.
Our hypothesis predicts that specific virus mutation will prevent expression of sncRNA from influencing VZV
infection; if not, it will indicate sncRNA may target host pathways important for VZV. Our primary hypothesis is
that sncRNA target VLT RNA to influence expression of the critical pro-lytic ORF61 protein. Our second aim
will test the hypotheses that the four VZVsncRNA promote lytic infection or enhance reactivation from latency
in neurons. We will exploit our human neuron culture system that hosts a model VZV latent state and
experimental reactivation from it. We will develop AAV to express VZVsncRNA in latently infected neurons
and determine if reactivation efficiency is favored. Globally, our studies will set the stage for understanding a
novel gene regulatory mechanism for VZV, their possible functions in interacting with VLT and the potential for
targeting sncRNA as a future therapeutic approach for treating Herpes zoster.
Terms: <21+ years old><AAV vector><Acids><Address><Adult><Adult Human><Affect><Agonist><Anti-Sense RNA><Antisense RNA><Antiviral Agents><Antiviral Drugs><Antivirals><Cadaver><Cell Body><Cell Line><Cell Mediated Immunology><Cell-Mediated Immunity><CellLine><Cells><Cellular Immunity><Chicken Pox><Chickenpox><Chickenpox Virus><Childhood><Chronic><Climacteric><Development><Disease><Disorder><ES cell><Elderly><Elements><Epithelial><Epithelial Cells><Family><Frequencies><Functional RNA><Future><Ganglia><Gene Expression><Gene Transcription><Generalized Growth><Genes><Genetic Alteration><Genetic Change><Genetic Transcription><Genetic defect><Goals><Growth><HHV-3><HHV3><Herpes Zoster><Herpes zoster Virus><Herpes zoster disease><Herpesviridae><Herpesvirus Type 3><Herpesvirus varicellae><Herpesviruses><Human><Impairment><Infection><Knowledge><Lytic><Lytic Cycle><Lytic Infection><Lytic Phase><Messenger RNA><Micro RNA><MicroRNAs><Modeling><Modern Man><Mutate><Mutation><Nerve Cells><Nerve Unit><Nervous System><Nervous system structure><Neural Cell><Neural Ganglion><Neurocyte><Neurologic Body System><Neurologic Organ System><Neurons><Non-Coding><Non-Coding RNA><Non-Polyadenylated RNA><Non-translated RNA><Noncoding RNA><Nontranslated RNA><Nucleotides><Ocular Herpes zoster Virus><Oligo><Oligonucleotides><Pain><Painful><Parents><Pathway interactions><Patients><Pattern><Process><Production><Proteins><Public Health><RNA><RNA Expression><RNA Gene Products><Regulator Genes><Regulatory Protein><Reporting><Ribonucleic Acid><Risk><Role><Shingles><Small RNA><Strains Cell Lines><System><Testing><Therapeutic><Tissue Growth><Transcript><Transcription><Transcriptional Regulatory Elements><Transfection><Untranslated RNA><VZ Virus><Vaccination><Varicella><Varicella-Zoster Virus><Viral><Viral Diseases><Viral Genome><Viral Latency><Virus><Virus Diseases><Virus Latency><Virus Replication><Work><Zona><Zoster><Zoster Vaccine><adeno-associated viral vector><adeno-associated virus vector><adulthood><advanced age><anti-viral agents><anti-viral drugs><anti-virals><chronic pain><cultured cell line><developmental><elders><embryonic stem cell><genetic regulatory protein><genome mutation><geriatric><global gene expression><global transcription profile><herpes virus><herpes zona><inhibitor><inhibitor/antagonist><interest><late life><latent infection><later life><life change><locked nucleic acid><mRNA><miRNA><miRNAs><mutant><neuronal><neuronal growth><new drug treatments><new drugs><new therapeutic approach><new therapeutic intervention><new therapeutic strategies><new therapeutics><new therapy><new therapy approaches><next generation therapeutics><noncoding><novel><novel drug treatments><novel drugs><novel therapeutic approach><novel therapeutic intervention><novel therapeutic strategies><novel therapeutics><novel therapy><novel therapy approach><nucleic acid inhibitor><older adult><older person><oligos><ontogeny><pathogen><pathway><pediatric><prevent><preventing><reactivation from latency><regulatory gene><regulatory gene product><senior citizen><social role><stem cell of embryonic origin><trans acting element><transcriptome><viral infection><viral multiplication><viral replication><virus genome><virus infection><virus multiplication><virus-induced disease>