Multifunctional Nanotherapeutics for the Treatment of NeuroHIV

Abstract

Even though the emergence of highly active antiretroviral therapy (HAART) significantly has helped in reducing the prevalence and bettering the quality of life of HIV‐infected individuals. The combined antiretroviral therapy (cART) mainly target active HIV‐1 infection but fails to target/ eradicate the latent virus from viral reservoirs [(e.g., brain due to limited penetration across the blood‐brain barrier (BBB)]. Also, due to neurocognitive issues (e.g. HIV associated neurocognitive disorder‐HAND) during the later stage of the disease leads to the issue of medication adherence. To overcome the BBB drug delivery, recently magnetic nanoparticles (MNPs) based sustained drug delivery systems have been successfully explored to deliver the drugs across the brain for targeting active and latent HIV. The main advantage of sustained MNP‐nanoformulation is that they can deliver drugs across BBB, target HIV infected cells and release the drugs at a constant rate for a longer period to increase therapeutic adherence and can help eradicate the HIV reservoir without hampering the integrity of the BBB. As most of cART drug component can’t target the latent HIV reservoirs, recently, broadly neutralizing antibodies (bNAbs) have shown good efficacy in targeting the latent reservoir via binding the site of HIV‐1 virus and neutralizes HIV‐1 potently both in vitro and in vivo without causing apparent off‐targeting effects peripherally but no report for CNS HIV‐1 targeting is yet available. So, the objective of our work is to develop a combination of two different sustained release nanoformulation (NF) consisting of NF: 1 bNAb (3BNC117 or N6) and NF‐2: anti‐HIV drugs (Emt, TAF) alone or in combination (NF1+NF‐2) for the treatment of NeuroHIV (Fig. 1, Rationale for dual NF development). The anti‐HIV drugs and bNAbs were assembled on MNPs using Layer‐by‐Layer (LbL) nanoassembly (Fig. 2). The NF will be tested for size, shape, charge, drug/bNABs loading, in‐vitro release profile, BBB transmigration assay along with in‐vitro efficacy and cytotoxicity using HIV‐1 infected human primary microglial cells (p24 & LTR assay). Results showed overall NF size of ~150 nm (loaded with 3 layers of bNAbs/cART), drug/bNAbs was released in a sustained manner up to 7 days and able to reduce the HIV‐1 infectivity up to ~50% (>20 ng/mL to <50 ng/mL) continuously using single NF treatment. The magnetic treatment (0.8 T) was able to transport (25.8% ± 3.0%) NF effectively without inducing any cytotoxic effects (MTT assay) due to NF presence in the brain. In conclusion, the developed NF can provide a better approach for the HIV‐1 cure and a foundation for future HIV‐1 purging strategies from the CNS using nanotechnology platform.Support or Funding InformationThe Campbell Foundation and TTUHSC start up fundsRationale for dual NF development NF: 1 anti‐HIV drugs (Emt, TAF) and NF‐2: bNAb (3BNC117 or N6) alone or in combination (NF1+NF‐2)Figure 1Step‐by‐step development nanoformulation (NF)[Tenofovir alafenamide (TAF), Emtricitabine (FTC)] and bNAb was assembled on MNPs. To load a higher amount of individual drugs or bNAbs or ART, we have used LbL self‐assembly technique and for CNS cell‐specific targeting (Microglial specific antibody: anti‐TMEM119 Ab; ab209064, Abcam) was usedFigure 2