The mucosal surface is the site of entry of >90% of common infectious pathogens in humans. However, studies of virus entry and infection are largely limited to non-polarized cells and little is known regarding the mechanisms used by viral pathogens to infect polarized cell monolayers. Enteroviruses are ideal pathogens for the study of polarized cell-virus interactions as they have evolved highly efficient strategies to bypass both epithelial and endothelial barriers for infection. Our previous studies have established that coxsackievirus B (CVB) and poliovirus (PV) enter polarized cells by endocytic mechanisms that require activation of specific intracellular signaling molecules that drive actin cytoskeleton reorganization, junctional complex modulation, and eventual virus endocytosis. Our preliminary studies suggest that CVB endocytosis into polarized intestinal epithelia is distinct from that in polarized BBB endothelia and that the signaling molecules required to facilitate virus entry are disparate between these cell types. Based on these observations, we hypothesize that enterovirus entry into polarized epithelial and endothelial cells is mediated by cell-type-specific intracellular signals and occur through different endocytic mechanisms. In this proposal, we have designed experiments to more clearly define the cellular pathways hijacked by CVB to gain entry into polarized human brain microvascular endothelial cells (HBMEC), a model of the blood-brain barrier. Specifically, we will (1) define the endocytic pathway used by CVB to gain entry into HBMEC (Specific Aim 1), (2) characterize the role of intracellular kinases in facilitating CVB entry into HBMEC (Specific Aim 2), and (3) compare the intracellular signaling molecules required for enterovirus infection in a variety of cell types (Specific Aim 3). Given the diverse pathological complications associated with enterovirus infections, these studies will provide insights into how enteroviruses have evolved to infect polarized cells, and will serve as a model for how other viral pathogens circumvent the barriers presented by polarized cell monolayers. Furthermore, as there are currently no effective therapeutics to treat enterovirus infections, these studies may lead to the design of cell-type-specific anti-viral targets.

Public Health Relevance

Enteroviruses (which include coxsackievirus, poliovirus, and echovirus) account for as many as 15 million symptomatic infections in the United States each year. There are currently no effective therapeutics to combat enteroviral infections. The studies in this proposal will identify cellular genes required for mediating enterovirus infection and may lead to the design of cell-type-specific antiviral targets.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
5R01AI081759-02
Application #
7871369
Study Section
Virology - A Study Section (VIRA)
Program Officer
Park, Eun-Chung
Project Start
2009-07-01
Project End
2013-06-30
Budget Start
2010-07-01
Budget End
2011-06-30
Support Year
2
Fiscal Year
2010
Total Cost
$359,667
Indirect Cost
Name
University of Pittsburgh
Department
Physiology
Type
Schools of Medicine
DUNS #
004514360
City
Pittsburgh
State
PA
Country
United States
Zip Code
15213
Barrila, Jennifer; Crabbé, Aurélie; Yang, Jiseon et al. (2018) Modeling Host-Pathogen Interactions in the Context of the Microenvironment: Three-Dimensional Cell Culture Comes of Age. Infect Immun 86:
Platt, Derek J; Smith, Amber M; Arora, Nitin et al. (2018) Zika virus-related neurotropic flaviviruses infect human placental explants and cause fetal demise in mice. Sci Transl Med 10:
Wells, Alexandra I; Coyne, Carolyn B (2018) Type III Interferons in Antiviral Defenses at Barrier Surfaces. Trends Immunol 39:848-858
Lanik, Wyatt E; Mara, Madison A; Mihi, Belgacem et al. (2018) Stem Cell-Derived Models of Viral Infections in the Gastrointestinal Tract. Viruses 10:
Bramley, John C; Drummond, Coyne G; Lennemann, Nicholas J et al. (2017) A Three-Dimensional Cell Culture System To Model RNA Virus Infections at the Blood-Brain Barrier. mSphere 2:
Drummond, Coyne G; Bolock, Alexa M; Ma, Congrong et al. (2017) Enteroviruses infect human enteroids and induce antiviral signaling in a cell lineage-specific manner. Proc Natl Acad Sci U S A 114:1672-1677
Arora, Nitin; Sadovsky, Yoel; Dermody, Terence S et al. (2017) Microbial Vertical Transmission during Human Pregnancy. Cell Host Microbe 21:561-567
Corry, Jacqueline; Arora, Nitin; Good, Charles A et al. (2017) Organotypic models of type III interferon-mediated protection from Zika virus infections at the maternal-fetal interface. Proc Natl Acad Sci U S A 114:9433-9438
Drummond, Coyne G; Nickerson, Cheryl A; Coyne, Carolyn B (2016) A Three-Dimensional Cell Culture Model To Study Enterovirus Infection of Polarized Intestinal Epithelial Cells. mSphere 1:
Ouyang, Yingshi; Bayer, Avraham; Chu, Tianjiao et al. (2016) Isolation of human trophoblastic extracellular vesicles and characterization of their cargo and antiviral activity. Placenta 47:86-95

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