Zika Virus Infection Modulates Extracellular Vesicle Biogenesis and Morphology in Human Umbilical Cord Endothelial Cells: A Proteomic and Microscopic Analysis

Infection with Zika virus (ZIKV) is a perinatal health problem and a vertical infection that promotes neurological fetal damage. ZIKV infects different cellular components at the maternal–fetal interface, including umbilical cord endothelial cells (HUVECs). Extracellular vesicles (EVs) are cellular...

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Main Authors: Manuel Adrián Velázquez-Cervantes, Arturo Flores-Pliego, Yazmín Rocío Benitez-Zeferino, Victor Javier Cruz-Holguín, Luis Herrera Moro-Huitron, Addy Cecilia Helguera-Repetto, David Eduardo Meza-Sánchez, José Luis Maravillas-Montero, Nicolás Cayetano-Castro, Javier Mancilla-Ramírez, Aurora Casarrubias-Betancourt, Guadalupe León-Reyes, Macario Martínez-Castillo, Isabel Wong-Baeza, Luis Adrián De Jesús-González, María Isabel Baeza-Ramírez, Moisés León-Juárez
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Language:English
Published: MDPI AG 2025-06-01
Series:Microorganisms
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Online Access:https://www.mdpi.com/2076-2607/13/6/1402
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author Manuel Adrián Velázquez-Cervantes
Arturo Flores-Pliego
Yazmín Rocío Benitez-Zeferino
Victor Javier Cruz-Holguín
Luis Herrera Moro-Huitron
Addy Cecilia Helguera-Repetto
David Eduardo Meza-Sánchez
José Luis Maravillas-Montero
Nicolás Cayetano-Castro
Javier Mancilla-Ramírez
Aurora Casarrubias-Betancourt
Guadalupe León-Reyes
Macario Martínez-Castillo
Isabel Wong-Baeza
Luis Adrián De Jesús-González
María Isabel Baeza-Ramírez
Moisés León-Juárez
author_facet Manuel Adrián Velázquez-Cervantes
Arturo Flores-Pliego
Yazmín Rocío Benitez-Zeferino
Victor Javier Cruz-Holguín
Luis Herrera Moro-Huitron
Addy Cecilia Helguera-Repetto
David Eduardo Meza-Sánchez
José Luis Maravillas-Montero
Nicolás Cayetano-Castro
Javier Mancilla-Ramírez
Aurora Casarrubias-Betancourt
Guadalupe León-Reyes
Macario Martínez-Castillo
Isabel Wong-Baeza
Luis Adrián De Jesús-González
María Isabel Baeza-Ramírez
Moisés León-Juárez
author_sort Manuel Adrián Velázquez-Cervantes
collection DOAJ
description Infection with Zika virus (ZIKV) is a perinatal health problem and a vertical infection that promotes neurological fetal damage. ZIKV infects different cellular components at the maternal–fetal interface, including umbilical cord endothelial cells (HUVECs). Extracellular vesicles (EVs) are cellular components that mediate extracellular communication. Viruses have the capacity to hijack and modify the biogenesis machinery of EVs for their own benefit. The present work provides proteomic results (2D electrophoresis) that show the regulation of the expression of proteins involved in autophagy, oxidative stress, and exosome biogenesis in HUVECs infected with ZIKV. We confirmed that Alix and CD9 proteins were downregulated following the infection. Additionally, EVs isolated from infected cells showed the expression of Alix, and CD9 was increased in contrast to the mock condition. Interestingly, nanoparticle tracking and cryo-microscopy assays revealed that these EVs showed an increase in the quantity and size of ZIKV infection to differences in mock conditions. Furthermore, EVs isolated from infected cells showed infectivity, and both RNA and viral proteins were detected. Finally, our cryo-microscopy analysis revealed that the viral infection promoted morphological changes in these extracellular vesicles to identify vesicles with double and triple vesicles and electrodense and double membranes. In conclusion, our data suggest that ZIKV infection can modulate cellular factors involved in the formation and morphology of EVs in HUVECs. Furthermore, these EVs carry viral elements that may contribute to the dissemination of infection. Future studies aimed at the proteomic and lipidomic composition analyses of these EVs are needed to understand the biological implications in vertical infection.
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spelling doaj-art-48448462951a48cdacd3b1a7972f7e402025-08-20T02:21:11ZengMDPI AGMicroorganisms2076-26072025-06-01136140210.3390/microorganisms13061402Zika Virus Infection Modulates Extracellular Vesicle Biogenesis and Morphology in Human Umbilical Cord Endothelial Cells: A Proteomic and Microscopic AnalysisManuel Adrián Velázquez-Cervantes0Arturo Flores-Pliego1Yazmín Rocío Benitez-Zeferino2Victor Javier Cruz-Holguín3Luis Herrera Moro-Huitron4Addy Cecilia Helguera-Repetto5David Eduardo Meza-Sánchez6José Luis Maravillas-Montero7Nicolás Cayetano-Castro8Javier Mancilla-Ramírez9Aurora Casarrubias-Betancourt10Guadalupe León-Reyes11Macario Martínez-Castillo12Isabel Wong-Baeza13Luis Adrián De Jesús-González14María Isabel Baeza-Ramírez15Moisés León-Juárez16Laboratorio de Virología Perinatal y Diseño Molecular de Antígenos y Biomarcadores, Departamento de Inmunobioquimica, Instituto Nacional de Perinatología “Isidro Espinosa de los Reyes”, Mexico City 11000, MexicoDepartamento de Inmunobioquimica, Instituto Nacional de Perinatología “Isidro Espinosa de los Reyes”, Mexico City 11000, MexicoLaboratorio de Virología Perinatal y Diseño Molecular de Antígenos y Biomarcadores, Departamento de Inmunobioquimica, Instituto Nacional de Perinatología “Isidro Espinosa de los Reyes”, Mexico City 11000, MexicoLaboratorio de Virología Perinatal y Diseño Molecular de Antígenos y Biomarcadores, Departamento de Inmunobioquimica, Instituto Nacional de Perinatología “Isidro Espinosa de los Reyes”, Mexico City 11000, MexicoLaboratorio de Virología Perinatal y Diseño Molecular de Antígenos y Biomarcadores, Departamento de Inmunobioquimica, Instituto Nacional de Perinatología “Isidro Espinosa de los Reyes”, Mexico City 11000, MexicoDepartamento de Inmunobioquimica, Instituto Nacional de Perinatología “Isidro Espinosa de los Reyes”, Mexico City 11000, MexicoRed de Apoyo a la Investigación, Coordinación de la Investigación Científica, Instituto Nacional de Ciencias Médicas y Nutrición “Salvador Zubirán”, Universidad Nacional Autónoma de México, Mexico City 04510, MexicoDepartamento de Medicina Molecular y Bioprocesos, Instituto de Biotecnología, Universidad Autónoma de México, Cuernavaca City 62210, MéxicoCentro de Nanociencias y Micro y Nanotecnologías, Instituto Politécnico Nacional, Mexico City 07738, MexicoEscuela Superior de Medicina, Instiuto Politécnico Nacional, Mexico City 11340, MexicoHospital de la Mujer, Secretaria de Salud, Mexico City 11340, MexicoLaboratorio de Nutrigenética y Nutrigenómica, Instituto Nacional de Medicina Genómica (INMEGEN), Mexico City 14610, MexicoSección de Estudios de Posgrado e Investigación, Escuela Superior de Medicina, Instituto Politécnico Nacional, Mexico City 1340, MexicoLaboratorio de Biomembranas, Escuela Nacional de Ciencias Biológicas, Instituto Politécnico Nacional, Mexico City 11340, MexicoUnidad de Investigación Biomédica de Zacatecas, Instituto Mexicano del Seguro Social, Zacatecas 98000, MexicoLaboratorio de Biomembranas, Escuela Nacional de Ciencias Biológicas, Instituto Politécnico Nacional, Mexico City 11340, MexicoLaboratorio de Virología Perinatal y Diseño Molecular de Antígenos y Biomarcadores, Departamento de Inmunobioquimica, Instituto Nacional de Perinatología “Isidro Espinosa de los Reyes”, Mexico City 11000, MexicoInfection with Zika virus (ZIKV) is a perinatal health problem and a vertical infection that promotes neurological fetal damage. ZIKV infects different cellular components at the maternal–fetal interface, including umbilical cord endothelial cells (HUVECs). Extracellular vesicles (EVs) are cellular components that mediate extracellular communication. Viruses have the capacity to hijack and modify the biogenesis machinery of EVs for their own benefit. The present work provides proteomic results (2D electrophoresis) that show the regulation of the expression of proteins involved in autophagy, oxidative stress, and exosome biogenesis in HUVECs infected with ZIKV. We confirmed that Alix and CD9 proteins were downregulated following the infection. Additionally, EVs isolated from infected cells showed the expression of Alix, and CD9 was increased in contrast to the mock condition. Interestingly, nanoparticle tracking and cryo-microscopy assays revealed that these EVs showed an increase in the quantity and size of ZIKV infection to differences in mock conditions. Furthermore, EVs isolated from infected cells showed infectivity, and both RNA and viral proteins were detected. Finally, our cryo-microscopy analysis revealed that the viral infection promoted morphological changes in these extracellular vesicles to identify vesicles with double and triple vesicles and electrodense and double membranes. In conclusion, our data suggest that ZIKV infection can modulate cellular factors involved in the formation and morphology of EVs in HUVECs. Furthermore, these EVs carry viral elements that may contribute to the dissemination of infection. Future studies aimed at the proteomic and lipidomic composition analyses of these EVs are needed to understand the biological implications in vertical infection.https://www.mdpi.com/2076-2607/13/6/1402ZIKVEVsHUVEC2D electrophoresiscryo-TEM
spellingShingle Manuel Adrián Velázquez-Cervantes
Arturo Flores-Pliego
Yazmín Rocío Benitez-Zeferino
Victor Javier Cruz-Holguín
Luis Herrera Moro-Huitron
Addy Cecilia Helguera-Repetto
David Eduardo Meza-Sánchez
José Luis Maravillas-Montero
Nicolás Cayetano-Castro
Javier Mancilla-Ramírez
Aurora Casarrubias-Betancourt
Guadalupe León-Reyes
Macario Martínez-Castillo
Isabel Wong-Baeza
Luis Adrián De Jesús-González
María Isabel Baeza-Ramírez
Moisés León-Juárez
Zika Virus Infection Modulates Extracellular Vesicle Biogenesis and Morphology in Human Umbilical Cord Endothelial Cells: A Proteomic and Microscopic Analysis
Microorganisms
ZIKV
EVs
HUVEC
2D electrophoresis
cryo-TEM
title Zika Virus Infection Modulates Extracellular Vesicle Biogenesis and Morphology in Human Umbilical Cord Endothelial Cells: A Proteomic and Microscopic Analysis
title_full Zika Virus Infection Modulates Extracellular Vesicle Biogenesis and Morphology in Human Umbilical Cord Endothelial Cells: A Proteomic and Microscopic Analysis
title_fullStr Zika Virus Infection Modulates Extracellular Vesicle Biogenesis and Morphology in Human Umbilical Cord Endothelial Cells: A Proteomic and Microscopic Analysis
title_full_unstemmed Zika Virus Infection Modulates Extracellular Vesicle Biogenesis and Morphology in Human Umbilical Cord Endothelial Cells: A Proteomic and Microscopic Analysis
title_short Zika Virus Infection Modulates Extracellular Vesicle Biogenesis and Morphology in Human Umbilical Cord Endothelial Cells: A Proteomic and Microscopic Analysis
title_sort zika virus infection modulates extracellular vesicle biogenesis and morphology in human umbilical cord endothelial cells a proteomic and microscopic analysis
topic ZIKV
EVs
HUVEC
2D electrophoresis
cryo-TEM
url https://www.mdpi.com/2076-2607/13/6/1402
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