Summary
This project aims to develop and apply innovative in-situ imaging techniques, including optical tissue clearing and volumetric microscopy, to visualize and characterize HIV viral reservoirs in the central nervous system (CNS) of non-human primates, particularly in the context of substance abuse.
What they want
The project is divided into R61 (development) and R33 (application) phases. In the R61 phase, the team will optimize optical tissue clearing methods (e.g., Clarity, hydrogel formulations, lipid removal) for non-human primate whole tissue, ensuring excellent structural retention and compatibility with antibody immunostaining and in-situ hybridization of HIV RNA/DNA probes. This optimization will focus on novel CNS-associated reservoirs like the calvarial bone marrow, superior cervical lymph node, and key brain areas. In the R33 phase, the developed protocols will be utilized to investigate the establishment kinetics, infected cell types, and burden of HIV reservoirs in these CNS sites, specifically examining the impact of chronic opioid exposure and antiretroviral therapy (ART).
Deliverables
- Optimized protocols for optical tissue clearing in non-human primate whole tissue
- Analytical determinations for optical clearing parameters
- Evidence for novel HIV reservoirs in calvarial bone marrow, superior cervical lymph node, and key brain areas
- Data on HIV reservoir establishment, infected cell types, and reservoir burden as a function of chronic opioid exposure or ART
- Critical information on CNS associated HIV reservoirs using novel in-situ imaging methodologies
Technical requirements
- Optical tissue clearing (Clarity)
- Volumetric deep tissue microscopy
- Machine learning image analysis
- Antibody immunostaining
- In-situ hybridization of HIV RNA/DNA probes
- Use of non-human primate whole tissue
