Human Sensory Neurons Derived from Induced Pluripotent Stem Cells Support Varicella-Zoster Virus Infection

Damron, F. Heath; Zhou, Wenbo; Scott-McKean, Jonah J.; Emmerling, Kaitlin L.; Cai, Guang-Yun; Krah, David L.; Costa, Alberto C. S.; Freed, Curt R.; Levin, Myron J.

doi:10.1371/journal.pone.0053010

Cited by 73 publications

(61 citation statements)

References 60 publications

(65 reference statements)

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“…In addition, human neurons derived from induced pluripotent stem cells (iPSCs) (29), neuronal stem cells (30), and embryonic stem cells (ESCs) (31,32) have been explored as infection models for human alphaherpesvirus. While these human cells support HSV productive infection (24,(27)(28)(29)31), reliable models to study the establishment of latency and/or reactivation have not been achieved using these human cells.…”

mentioning

confidence: 99%

An Immortalized Human Dorsal Root Ganglion Cell Line Provides a Novel Context To Study Herpes Simplex Virus 1 Latency and Reactivation

Thellman

Botting

Madaj

et al. 2017

J Virol

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A defining characteristic of alphaherpesviruses is the establishment of lifelong latency in host sensory ganglia with occasional reactivation causing recurrent lytic infections. As an alternative to rodent models, we explored the use of an immortalized cell line derived from human dorsal root ganglia. HD10.6 cells proliferate by virtue of a transduced tetracycline-regulated v-myc oncogene. In the presence of doxycycline, HD10.6 cells mature to exhibit neuronal morphology and express sensory neuron-associated markers such as neurotrophin receptors TrkA, TrkB, TrkC, and RET and the sensory neurofilament peripherin. Infection of mature HD10.6 neurons by herpes simplex virus 1 (HSV-1) results in a delayed but productive infection. However, infection at a low multiplicity of infection (MOI) in the presence of acyclovir results in a quiescent infection resembling latency in which viral genomes are retained in a low number of neurons, viral gene expression is minimal, and infectious virus is not released. At least some of the quiescent viral genomes retain the capacity to reactivate, resulting in viral DNA replication and release of infectious virus. Reactivation can be induced by depletion of nerve growth factor; other commonly used reactivation stimuli have no significant effect.IMPORTANCE Infections by herpes simplex viruses (HSV) cause painful cold sores or genital lesions in many people; less often, they affect the eye or even the brain. After the initial infection, the virus remains inactive or latent in nerve cells that sense the region where that infection occurred. To learn how virus maintains and reactivates from latency, studies are done in neurons taken from rodents or in whole animals to preserve the full context of infection. However, some cellular mechanisms involved in HSV infection in rodents are different from those in humans. We describe the use of a human cell line that has the properties of a sensory neuron. HSV infection in these cultured cells shows the properties expected for a latent infection, including reactivation to produce newly infectious virus. Thus, we now have a cell culture model for latency that is derived from the normal host for this virus.KEYWORDS HSV-1, quiescent infection, sensory neurons H erpesviruses are unusual among viruses in that they employ two infection strategies, lytic (replicative or productive) infection and latency (quiescent) infection. Alphaherpesviruses typically replicate in epithelial cells and establish latency in the peripheral nervous system within the sensory neurons of the ganglia that innervate mucosa of the primary infection site (1), such as the trigeminal ganglia (TG) and dorsal root ganglia (DRG). Operationally, latency is defined as "the persistence of a viral genome within tissue where, at any given time, there is a population of cells that lack detectable infectious virus, viral proteins, or viral lytic transcripts that are dormant but have the capability of being reactivated" (2). During latency of herpes simplex viruses

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mentioning

confidence: 99%

An Immortalized Human Dorsal Root Ganglion Cell Line Provides a Novel Context To Study Herpes Simplex Virus 1 Latency and Reactivation

Thellman

Botting

Madaj

et al. 2017

J Virol

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“…The infection of the HFF by a suspension of neurons that had been infected with cell free-VZV strongly suggests that these neurons produced VZV that readily infect susceptible cells. It has been shown by ourselves and others that cultured neurons infected with cell-associated VZV release virions into the medium (Gowrishankar et al, 2007), (Markus et al, 2011), (Lee et al, 2012). In a second approach, we tested the ability of hESC-derived neurons infected by cell-free VZV to not only produce but release infectious virions.…”

Section: Resultsmentioning

confidence: 99%

“…It is important to note that cell-free VZV produced in a different laboratory (in China) has also recently been shown to generate productive infection (as shown by spread of infection, for example) of hESC-derived neurons and neuroblastoma-derived neurons (Selariu et al, 2012). Even more recently, incubation of human iPS-derived neurons with cell-free VZV was shown to result in release of infectious virus into the medium in (Lee et al, 2012). …”

Section: Discussionmentioning

confidence: 99%

Productive vs non-productive infection by cell-free varicella zoster virus of human neurons derived from embryonic stem cells is dependent upon infectious viral dose

2013

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Varicella Zoster virus (VZV) productively infects humans causing varicella upon primary infection and herpes zoster upon reactivation from latency in neurons. In-vitro studies using cell-associated VZV infection have demonstrated productive VZV-infection, while a few recent studies of human neurons derived from stem cells incubated with cell-free, vaccine-derived VZV did not result in generation of infectious virus. In the present study, 90%-pure human embryonic stem cell-derived neurons were incubated with recombinant cell-free pOka-derived made with an improved method or with VZV vaccine. We found that cell-free pOka and vOka at higher multiplicities of infection elicited productive infection in neurons followed by spread of infection, cytopathic effect and release of infectious virus into the medium. These results further validate the use of this unlimited source of human neurons for studying unexplored aspects of VZV interaction with neurons such as entry, latency and reactivation.

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“…The recent development of induced pluripotent stem cell (iPSC) and embryonic stem cell (ESC)-derived cells could theoretically enable this improved approach as multiple groups have shown that CNS derivatives can be derived from these cell populations [20,21]. One can envision using iPSCderived cells to make these disease models both cell-type and patient-specific lines so that the models express the cellular responses while also incorporating the patient's unique genetic profile.…”

Section: Modeling Disease Using Human Cells In General and Psc-derivementioning

confidence: 99%

Induced Pluripotent Stem Cell Models to Enable In Vitro Models for Screening in the Central Nervous System

Hunsberger

Efthymiou

Malik

et al. 2015

Stem Cells and Development

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There is great need to develop more predictive drug discovery tools to identify new therapies to treat diseases of the central nervous system (CNS). Current nonpluripotent stem cell-based models often utilize non-CNS immortalized cell lines and do not enable the development of personalized models of disease. In this review, we discuss why in vitro models are necessary for translational research and outline the unique advantages of induced pluripotent stem cell (iPSC)-based models over those of current systems. We suggest that iPSC-based models can be patient specific and isogenic lines can be differentiated into many neural cell types for detailed comparisons. iPSC-derived cells can be combined to form small organoids, or large panels of lines can be developed that enable new forms of analysis. iPSC and embryonic stem cell-derived cells can be readily engineered to develop reporters for lineage studies or mechanism of action experiments further extending the utility of iPSC-based systems. We conclude by describing novel technologies that include strategies for the development of diversity panels, novel genomic engineering tools, new three-dimensional organoid systems, and modified high-content screens that may bring toxicology into the 21st century. The strategic integration of these technologies with the advantages of iPSC-derived cell technology, we believe, will be a paradigm shift for toxicology and drug discovery efforts.Disease Modeling D iseases of the central nervous system (CNS) affect a large number of people, but therapeutic intervention is hampered by the lack of useful models for many of these diseases. Current research on human subjects, particularly for drug discovery for CNS diseases, is severely (and appropriately) limited by ethical guidelines. Therefore, surrogate models are needed that share important anatomical, physiological, and genetic features to advance new treatments and therapies for CNS diseases [1].Developing rapid and effective therapies for CNS diseases requires the availability of in vitro models that accurately recapitulate disease phenotypes and predict patient treatment response. A proper model must be both sensitive and predictive while reflecting both normal and disease processes. Equally important, these models should enable the investigation of genetic and environmental risk factors contributing to diseases in a rapid and economical way. Currently used models often do not reflect a typical human response [2-4], despite efforts underway to better characterize these models and increase their preclinical value in predicting safety and efficacy in the clinic [5,6]. Therefore, there is a great need to develop disease-and patient-specific models from cells directly affected in CNS disorders. These cellbased models, we envision, could either replace or supplement current animal models and enable the efficient translation of basic research into the clinical setting. Limitations with current CNS modelsCurrently, drug discovery relies on the use of animalbased or cell-based models, ...

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Human Sensory Neurons Derived from Induced Pluripotent Stem Cells Support Varicella-Zoster Virus Infection

Cited by 73 publications

References 60 publications

An Immortalized Human Dorsal Root Ganglion Cell Line Provides a Novel Context To Study Herpes Simplex Virus 1 Latency and Reactivation

An Immortalized Human Dorsal Root Ganglion Cell Line Provides a Novel Context To Study Herpes Simplex Virus 1 Latency and Reactivation

Productive vs non-productive infection by cell-free varicella zoster virus of human neurons derived from embryonic stem cells is dependent upon infectious viral dose

Induced Pluripotent Stem Cell Models to Enable In Vitro Models for Screening in the Central Nervous System

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