# Exploring HIV-associated Neurocognitive Disorder (HAND) and HIV Latency at the Single Cell Level in Cerebral Organoids

> **NIH NIH R01** · J. DAVID GLADSTONE INSTITUTES · 2022 · $708,000

## Abstract

Project Summary
Fully half of all HIV-infected individuals continue to display some (often milder) form of HIV-associated
neurocognitive disorder (HAND) despite the introduction of antiretroviral therapy (ART). More than one in 10 of
these individuals will exhibit progressive neurologic deterioration on ART. More severe forms of HAND, including
HIV-associated dementia, remain common in the developing world, especially in individuals not receiving ART.
HAND is likely caused by chronic inflammation in the brain leading to neuronal dysfunction. The conundrum is
how this inflammatory response is sustained despite effective suppression of viral replication with ART. We
believe latent HIV infection of microglia likely plays a central role. Microglia comprise 10-15% of all cells in the
CNS and serve as the brain's "constant gardeners" shaping neuronal plasticity through synaptic pruning and
stripping; microglia also participate in bidirectional signaling with closely intertwined neurons. How best to study
these microglia, their interplay with neurons, and the effects of HIV infection? We propose to coculture two iPSC-
derived sub-lines engineered to express doxycycline-inducible transcription factors that are sufficient to drive
differentiation into either microglia or excitatory neurons. When induced and cocultured in 3D conditions, these
cells form cerebral microorganoids (CMs) that recapitulate many of the cytoarchitectural features and functions
of the fetal brain. We will study these CMs in an unbiased manner using scRNA-seq to define gene expression
profiles and scATAC-sec to interrogate chromatin accessibility. Use of a combinatorial indexing system of
barcodes will allow measurement of these parameters in the same cell. We hypothesize that microglia are
latently infected and that sustained neuronal neurotransmitter signaling is likely sufficient to reactivate virus
expression plus exposure to opioids will further enhance reactivation (virus production is not impaired by ART).
Release of reactivated virions may directly trigger a chronic inflammatory response. Additionally, when these
viruses are transmitted cell-to-cell, an abortive form of HIV infection may ensue due to the action of the RT
inhibitors present in ART. The IFI16 DNA sensor may detect these RT products leading to inflammasome
assembly, caspase-1 activation, production of IL-1β and IL-18 and death by pyroptosis, a highly inflammatory
form of programmed cell death. Because pyroptosis breeds more pyroptosis, this feed-forward form of
inflammation could a create chronic inflammatory response resistant to ART. Finally, we are eager to explore
two CNS-tailored approaches for attacking the latent HIV reservoir in microglia. In the first, virus will be purged
with a CNS-penetrant LRA and cells producing viral RNA will be selectively killed by induction of RIG-I-dependent
apoptosis. In the second, durable, sequence-specific transcriptional silencing of HIV proviruses will be tested
using C...

## Key facts

- **NIH application ID:** 10466829
- **Project number:** 5R01DA049525-04
- **Recipient organization:** J. DAVID GLADSTONE INSTITUTES
- **Principal Investigator:** Melanie Maria Ott
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2022
- **Award amount:** $708,000
- **Award type:** 5
- **Project period:** 2019-09-15 → 2024-08-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10466829

## Citation

> US National Institutes of Health, RePORTER application 10466829, Exploring HIV-associated Neurocognitive Disorder (HAND) and HIV Latency at the Single Cell Level in Cerebral Organoids (5R01DA049525-04). Retrieved via AI Analytics 2026-05-22 from https://api.ai-analytics.org/grant/nih/10466829. Licensed CC0.

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