Table 3.

Summary of preliminary lessons and corresponding next steps for Last Gift and collaborators

Last Gift lessons about CNS reservoirs to date	Next steps
Heterogeneous distribution of HIV proviral DNA even within different cortical regions	1) Cell sorting, single-cell, and spatial omics analyses to determine cellular characteristics of the different reservoirs and immunologic microenvironments (microglial macrophage phenotypes, CD4 + subtypes) 2) Compare more CNS sites (multiple cortical areas, brain stem, spinal cord, and CSF)
CNS compartmentalization and differential bNAb susceptibility in brain tissue	1) Expand analysis with larger sample size and more CNS sites 2) Measure ART concentrations, resistance mutations 3) Tropism assays
Migration within brain and bidirectionally across BBB by HIV DNA sequence analysis and statistical modeling	1) Using single-cell techniques and spatial omics analyses to identify which cell types are migrating 2) Tropism
CNS has provirus with intact full length envelope gene, supportive of potential for viral rebound from CNS reservoirs	1) Adapted quantitative viral outgrowth assays, full-length HIV genome sequencing, and insertion site analysis to better characterize replication competence and clonality 2) Tropism of replication-competent virus