Abstract
Using nanoparticle-based RNA interference (RNAi), we have previously shown that silencing the host autophagic protein, Beclin1, in HIV-infected human microglia and astrocytes restricts HIV replication and its viral-associated inflammatory responses. Here, we confirmed the efficacy of Beclin1 small interfering RNA (siBeclin1) as an adjunctive antiviral and anti-inflammatory therapy in myeloid human microglia and primary human astrocytes infected with HIV, both with and without exposure to combined antiretroviral (cART) drugs. To specifically target human microglia and human astrocytes, we used a nanoparticle (NP) comprised of linear cationic polyethylenimine (PEI) conjugated with mannose (Man) and encapsulated with siBeclin1. The target specificity of the PEI-Man NP was confirmed in vitro using human neuronal and glial cells transfected with the NP encapsulated with fluorescein isothiocyanate (FITC). PEI-Man-siBeclin1 NPs were intranasally delivered to healthy C57BL/6 mice in order to report the biodistribution of siBeclin1 in different areas of the brain, measured using stem-loop RT-PCR. Postmortem brains recovered at 1–48 h post-treatment with the PEI-Man-siRNA NP showed no significant changes in the secretion of the chemokines regulated on activation, normal T cell expressed and secreted (RANTES) and monocyte chemotactic protein-1 (MCP-1) and showed significant decreases in the secretion of the cytokines interleukin 6 (IL-6) and tumor necrosis factor alpha (TNF-α) when compared to phosphate-buffered saline (PBS)-treated brains. Nissl staining showed minimal differences between the neuronal structures when compared to PBS-treated brains, which correlated with no adverse behavioral affects. To confirm the brain and peripheral organ distribution of PEI-siBeclin1 in living mice, we used the In vivo Imaging System (IVIS) and demonstrated a significant brain accumulation of siBeclin1 through intranasal administration.
Highlights
With the blood–brain barrier (BBB) being a major obstacle for the entry of a drug into the brain, intranasal drug delivery has emerged as a reliable method to bypass the BBB and treat neurological diseases
Modulation of the autophagy pathway has been been considered as a therapeutic target for several diseases, including Alzheimer’s disconsidered as a therapeutic target for several diseases, including Alzheimer’s disease, ease, cardiac diseases, and cancer [41,42,43,44]
In terms of HIV, modulating autophagy has been proposed as a potential alternative approach to of HIV, modulating autophagy has been proposed as a potential alternative approach to reduced HIV infection [45,46,47,48,49]
Summary
With the blood–brain barrier (BBB) being a major obstacle for the entry of a drug into the brain, intranasal drug delivery has emerged as a reliable method to bypass the BBB and treat neurological diseases. CART is not completely effective in controlling HIV replication in brain reservoirs (including microglia and, to a lesser degree, astrocytes) and does not directly target the inflammatory cascades that are believed to be the primary cause of neuronal injury or dysfunction related to HIV-associated neurological pathology [3,4,5]. Considering these limitations, many alternative methods to developing anti-HIV drugs are under investigation [1,6,7,8,9]. We synthesized a siBeclin1-PEI NP conjugated with mannose (PEI-Man) particles to target mannose-expressing brain cells in vitro and in vivo via intranasal delivery in C57BL/6 mice
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