Antiretroviral drug therapy does not reduce neuroinflammation in an HIV-1 infection brain organoid model

Abstract Background HIV-1-associated neurocognitive impairment (HIV-1-NCI) is marked by ongoing and chronic neuroinflammation with loss and decline in neuronal function even when antiretroviral drug therapy (ART) successfully suppresses viral replication. Microglia, the primary reservoirs of HIV-1 i...

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Main Authors: Samuel Martinez-Meza, Thomas A. Premeaux, Stefano M. Cirigliano, Courtney M. Friday, Stephanie Michael, Sonia Mediouni, Susana T. Valente, Lishomwa C. Ndhlovu, Howard A. Fine, Robert L. Furler O’Brien, Douglas F. Nixon
Format: Article
Language:English
Published: BMC 2025-03-01
Series:Journal of Neuroinflammation
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Online Access:https://doi.org/10.1186/s12974-025-03375-w
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author Samuel Martinez-Meza
Thomas A. Premeaux
Stefano M. Cirigliano
Courtney M. Friday
Stephanie Michael
Sonia Mediouni
Susana T. Valente
Lishomwa C. Ndhlovu
Howard A. Fine
Robert L. Furler O’Brien
Douglas F. Nixon
author_facet Samuel Martinez-Meza
Thomas A. Premeaux
Stefano M. Cirigliano
Courtney M. Friday
Stephanie Michael
Sonia Mediouni
Susana T. Valente
Lishomwa C. Ndhlovu
Howard A. Fine
Robert L. Furler O’Brien
Douglas F. Nixon
author_sort Samuel Martinez-Meza
collection DOAJ
description Abstract Background HIV-1-associated neurocognitive impairment (HIV-1-NCI) is marked by ongoing and chronic neuroinflammation with loss and decline in neuronal function even when antiretroviral drug therapy (ART) successfully suppresses viral replication. Microglia, the primary reservoirs of HIV-1 in the central nervous system (CNS), play a significant role in maintaining this neuroinflammatory state. However, understanding how chronic neuroinflammation is generated and sustained by HIV-1, or impacted by ART, is difficult due to limited access to human CNS tissue. Methods We generated an in vitro model of admixed hematopoietic progenitor cell (HPC) derived microglia embedded into embryonic stem cell (ESC) derived Brain Organoids (BO). Microglia were infected with HIV-1 prior to co-culture. Infected microglia were co-cultured with brain organoids BOs to infiltrate the BOs and establish a model for HIV-1 infection, “HIV-1 M-BO”. HIV-1 M-BOs were treated with ART for variable directions. HIV-1 infection was monitored with p24 ELISA and by digital droplet PCR (ddPCR). Inflammation was measured by cytokine or p-NF-kB levels using multiplex ELISA, flow cytometry and confocal microscopy. Results HIV-1 infected microglia could be co-cultured with BOs to create a model for “brain” HIV-1 infection. Although HIV-1 infected microglia were the initial source of pro-inflammatory cytokines, astrocytes, neurons and neural stem cells also had increased p-NF-kB levels, along with elevated CCL2 levels in the supernatant of HIV-1 M-BOs compared to Uninfected M-BOs. ART suppressed the virus to levels below the limit of detection but did not decrease neuroinflammation. Conclusions These findings indicate that HIV-1 infected microglia are pro-inflammatory. Although ART significantly suppressed HIV-1 levels, neuronal inflammation persisted in ART-treated HIV-1 M-BOs. Together, these findings indicate that HIV-1 infection of microglia infiltrated into BOs provides a robust in vitro model to understand the impact of HIV-1 and ART on neuroinflammation.
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spelling doaj-art-736a839a8b974bd3b3e8b75fc5cd0d0b2025-08-20T01:57:44ZengBMCJournal of Neuroinflammation1742-20942025-03-0122111610.1186/s12974-025-03375-wAntiretroviral drug therapy does not reduce neuroinflammation in an HIV-1 infection brain organoid modelSamuel Martinez-Meza0Thomas A. Premeaux1Stefano M. Cirigliano2Courtney M. Friday 3Stephanie Michael4Sonia Mediouni5Susana T. Valente6Lishomwa C. Ndhlovu7Howard A. Fine8Robert L. Furler O’Brien9Douglas F. Nixon10Institute of Translational Research, Feinstein Institutes for Medical Research, Northwell HealthDivision of Infectious Diseases, Department of Medicine, Weill Cornell MedicineMeyer Cancer Center, Division of Neuro-Oncology, Department of Neurology, New York-Presbyterian Hospital/Weill Cornell MedicineDivision of Infectious Diseases, Department of Medicine, Weill Cornell MedicineInstitute of Translational Research, Feinstein Institutes for Medical Research, Northwell HealthDepartment of Immunology and Microbiology, The Herbert Wertheim UF Scripps Institute for Biomedical Innovation and TechnologyDepartment of Immunology and Microbiology, The Herbert Wertheim UF Scripps Institute for Biomedical Innovation and TechnologyDivision of Infectious Diseases, Department of Medicine, Weill Cornell MedicineMeyer Cancer Center, Division of Neuro-Oncology, Department of Neurology, New York-Presbyterian Hospital/Weill Cornell MedicineInstitute of Translational Research, Feinstein Institutes for Medical Research, Northwell HealthInstitute of Translational Research, Feinstein Institutes for Medical Research, Northwell HealthAbstract Background HIV-1-associated neurocognitive impairment (HIV-1-NCI) is marked by ongoing and chronic neuroinflammation with loss and decline in neuronal function even when antiretroviral drug therapy (ART) successfully suppresses viral replication. Microglia, the primary reservoirs of HIV-1 in the central nervous system (CNS), play a significant role in maintaining this neuroinflammatory state. However, understanding how chronic neuroinflammation is generated and sustained by HIV-1, or impacted by ART, is difficult due to limited access to human CNS tissue. Methods We generated an in vitro model of admixed hematopoietic progenitor cell (HPC) derived microglia embedded into embryonic stem cell (ESC) derived Brain Organoids (BO). Microglia were infected with HIV-1 prior to co-culture. Infected microglia were co-cultured with brain organoids BOs to infiltrate the BOs and establish a model for HIV-1 infection, “HIV-1 M-BO”. HIV-1 M-BOs were treated with ART for variable directions. HIV-1 infection was monitored with p24 ELISA and by digital droplet PCR (ddPCR). Inflammation was measured by cytokine or p-NF-kB levels using multiplex ELISA, flow cytometry and confocal microscopy. Results HIV-1 infected microglia could be co-cultured with BOs to create a model for “brain” HIV-1 infection. Although HIV-1 infected microglia were the initial source of pro-inflammatory cytokines, astrocytes, neurons and neural stem cells also had increased p-NF-kB levels, along with elevated CCL2 levels in the supernatant of HIV-1 M-BOs compared to Uninfected M-BOs. ART suppressed the virus to levels below the limit of detection but did not decrease neuroinflammation. Conclusions These findings indicate that HIV-1 infected microglia are pro-inflammatory. Although ART significantly suppressed HIV-1 levels, neuronal inflammation persisted in ART-treated HIV-1 M-BOs. Together, these findings indicate that HIV-1 infection of microglia infiltrated into BOs provides a robust in vitro model to understand the impact of HIV-1 and ART on neuroinflammation.https://doi.org/10.1186/s12974-025-03375-wHIV-1NeurocognitiveBrainOrganoidMicrogliaNeurons
spellingShingle Samuel Martinez-Meza
Thomas A. Premeaux
Stefano M. Cirigliano
Courtney M. Friday
Stephanie Michael
Sonia Mediouni
Susana T. Valente
Lishomwa C. Ndhlovu
Howard A. Fine
Robert L. Furler O’Brien
Douglas F. Nixon
Antiretroviral drug therapy does not reduce neuroinflammation in an HIV-1 infection brain organoid model
Journal of Neuroinflammation
HIV-1
Neurocognitive
Brain
Organoid
Microglia
Neurons
title Antiretroviral drug therapy does not reduce neuroinflammation in an HIV-1 infection brain organoid model
title_full Antiretroviral drug therapy does not reduce neuroinflammation in an HIV-1 infection brain organoid model
title_fullStr Antiretroviral drug therapy does not reduce neuroinflammation in an HIV-1 infection brain organoid model
title_full_unstemmed Antiretroviral drug therapy does not reduce neuroinflammation in an HIV-1 infection brain organoid model
title_short Antiretroviral drug therapy does not reduce neuroinflammation in an HIV-1 infection brain organoid model
title_sort antiretroviral drug therapy does not reduce neuroinflammation in an hiv 1 infection brain organoid model
topic HIV-1
Neurocognitive
Brain
Organoid
Microglia
Neurons
url https://doi.org/10.1186/s12974-025-03375-w
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