Magnetoelectric Extracellular Vesicle Latency-Targeting (MELT) Nanotherapeutic for the Block-Lock-and-Kill HIV Eradication Strategy
Background: Human immunodeficiency virus (HIV) establishes latent infections in cellular reservoirs, including microglia. HC69 cells, a microglial model of HIV latency, contain an HIV promoter long terminal repeat (LTR)-GFP reporter and were used for testing the efficacy of a two-step magnetoelectri...
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2025-01-01
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| author | Mickensone Andre Nagesh Kolishetti Adriana Yndart Arti Vashist Madhavan Nair Andrea D. Raymond |
| author_facet | Mickensone Andre Nagesh Kolishetti Adriana Yndart Arti Vashist Madhavan Nair Andrea D. Raymond |
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| description | Background: Human immunodeficiency virus (HIV) establishes latent infections in cellular reservoirs, including microglia. HC69 cells, a microglial model of HIV latency, contain an HIV promoter long terminal repeat (LTR)-GFP reporter and were used for testing the efficacy of a two-step magnetoelectric nanoparticle (MENP) and extracellular vesicle (xEV) latency-targeting (MELT) nanotherapeutic. GFP expression in HC69 at rest is low (GFP<sup>Lo</sup>), and upon exposure to LTR, transcription-activating agents (i.e., TNF-α) are induced to be high expressing (GFP<sup>Hi</sup>). Methods: The first step of MELT utilized ZL0580, an HIV Tat inhibitor loaded into EVs (80%) via incubation. ZL0580-EVs were taken up by GFP<sup>Lo</sup> and blocked LTR transcriptional reactivation by 50% and were 90% less toxic than ZL0580 alone. The second step in MELT involved conjugation of monomethyl auristatin E (MMAE) to MENPs. HPLC measurements showed 80% MMAE attachment to MENPs. Flow cytometry-based measurements of the membrane potential indicated that the membranes of GFP<sup>Hi</sup> HC69 were 60% more polarized than GFP<sup>Lo</sup> HC69 cells. More MMAE–MENPs were internalized by GFP<sup>Lo</sup> HC69. Results: Using a mixed-cell blood–brain barrier (BBB) Transwell model, we demonstrated that 20% of MELT crossed the BBB, was taken up by HC69 cells, and reduced LTR reactivation by 10%. Conclusions: Overall, this study demonstrated that MELT can potentially be utilized as a nanotherapeutic to target HIV latency in microglia. |
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| spelling | doaj-art-581cda7172134b39af0b4dcdca3be8422025-01-24T13:24:11ZengMDPI AGBiomedicines2227-90592025-01-0113114710.3390/biomedicines13010147Magnetoelectric Extracellular Vesicle Latency-Targeting (MELT) Nanotherapeutic for the Block-Lock-and-Kill HIV Eradication StrategyMickensone Andre0Nagesh Kolishetti1Adriana Yndart2Arti Vashist3Madhavan Nair4Andrea D. Raymond5Herbert Wertheim College of Medicine, Cellular and Molecular Medicine, Florida International University, Miami, FL 33199, USAHerbert Wertheim College of Medicine, Cellular and Molecular Medicine, Florida International University, Miami, FL 33199, USAHerbert Wertheim College of Medicine, Cellular and Molecular Medicine, Florida International University, Miami, FL 33199, USAHerbert Wertheim College of Medicine, Cellular and Molecular Medicine, Florida International University, Miami, FL 33199, USAHerbert Wertheim College of Medicine, Cellular and Molecular Medicine, Florida International University, Miami, FL 33199, USAHerbert Wertheim College of Medicine, Cellular and Molecular Medicine, Florida International University, Miami, FL 33199, USABackground: Human immunodeficiency virus (HIV) establishes latent infections in cellular reservoirs, including microglia. HC69 cells, a microglial model of HIV latency, contain an HIV promoter long terminal repeat (LTR)-GFP reporter and were used for testing the efficacy of a two-step magnetoelectric nanoparticle (MENP) and extracellular vesicle (xEV) latency-targeting (MELT) nanotherapeutic. GFP expression in HC69 at rest is low (GFP<sup>Lo</sup>), and upon exposure to LTR, transcription-activating agents (i.e., TNF-α) are induced to be high expressing (GFP<sup>Hi</sup>). Methods: The first step of MELT utilized ZL0580, an HIV Tat inhibitor loaded into EVs (80%) via incubation. ZL0580-EVs were taken up by GFP<sup>Lo</sup> and blocked LTR transcriptional reactivation by 50% and were 90% less toxic than ZL0580 alone. The second step in MELT involved conjugation of monomethyl auristatin E (MMAE) to MENPs. HPLC measurements showed 80% MMAE attachment to MENPs. Flow cytometry-based measurements of the membrane potential indicated that the membranes of GFP<sup>Hi</sup> HC69 were 60% more polarized than GFP<sup>Lo</sup> HC69 cells. More MMAE–MENPs were internalized by GFP<sup>Lo</sup> HC69. Results: Using a mixed-cell blood–brain barrier (BBB) Transwell model, we demonstrated that 20% of MELT crossed the BBB, was taken up by HC69 cells, and reduced LTR reactivation by 10%. Conclusions: Overall, this study demonstrated that MELT can potentially be utilized as a nanotherapeutic to target HIV latency in microglia.https://www.mdpi.com/2227-9059/13/1/147extracellular vesiclesHIV latencyMENPsmicrogliaMELT |
| spellingShingle | Mickensone Andre Nagesh Kolishetti Adriana Yndart Arti Vashist Madhavan Nair Andrea D. Raymond Magnetoelectric Extracellular Vesicle Latency-Targeting (MELT) Nanotherapeutic for the Block-Lock-and-Kill HIV Eradication Strategy Biomedicines extracellular vesicles HIV latency MENPs microglia MELT |
| title | Magnetoelectric Extracellular Vesicle Latency-Targeting (MELT) Nanotherapeutic for the Block-Lock-and-Kill HIV Eradication Strategy |
| title_full | Magnetoelectric Extracellular Vesicle Latency-Targeting (MELT) Nanotherapeutic for the Block-Lock-and-Kill HIV Eradication Strategy |
| title_fullStr | Magnetoelectric Extracellular Vesicle Latency-Targeting (MELT) Nanotherapeutic for the Block-Lock-and-Kill HIV Eradication Strategy |
| title_full_unstemmed | Magnetoelectric Extracellular Vesicle Latency-Targeting (MELT) Nanotherapeutic for the Block-Lock-and-Kill HIV Eradication Strategy |
| title_short | Magnetoelectric Extracellular Vesicle Latency-Targeting (MELT) Nanotherapeutic for the Block-Lock-and-Kill HIV Eradication Strategy |
| title_sort | magnetoelectric extracellular vesicle latency targeting melt nanotherapeutic for the block lock and kill hiv eradication strategy |
| topic | extracellular vesicles HIV latency MENPs microglia MELT |
| url | https://www.mdpi.com/2227-9059/13/1/147 |
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