Limits...
Opposing regulation of endolysosomal pathways by long-acting nanoformulated antiretroviral therapy and HIV-1 in human macrophages.

Araínga M, Guo D, Wiederin J, Ciborowski P, McMillan J, Gendelman HE - Retrovirology (2015)

Bottom Line: NanoART and native ATV treated uninfected cells showed limited effects.The data was confirmed by Western blot.DAVID and KEGG bioinformatics analyses of proteomic data showed relationships between secretory, mobility and phagocytic cell functions and virus and particle trafficking.

View Article: PubMed Central - PubMed

Affiliation: Departments of Pharmacology and Experimental Neuroscience, University of Nebraska Medical Center, 985880 Nebraska Medical Center, Omaha 68198-5880, NE, USA. hegendel@unmc.edu.

ABSTRACT

Background: Long-acting nanoformulated antiretroviral therapy (nanoART) is designed to improve patient regimen adherence, reduce systemic drug toxicities, and facilitate clearance of human immunodeficiency virus type one (HIV-1) infection. While nanoART establishes drug depots within recycling and late monocyte-macrophage endosomes, whether or not this provides a strategic advantage towards viral elimination has not been elucidated.

Results: We applied quantitative SWATH-MS proteomics and cell profiling to nanoparticle atazanavir (nanoATV)-treated and HIV-1 infected human monocyte-derived macrophages (MDM). Native ATV and uninfected cells served as controls. Both HIV-1 and nanoATV engaged endolysosomal trafficking for assembly and depot formation, respectively. Notably, the pathways were deregulated in opposing manners by the virus and the nanoATV, likely by viral clearance. Paired-sample z-scores, of the proteomic data sets, showed up- and down- regulation of Rab-linked endolysosomal proteins. NanoART and native ATV treated uninfected cells showed limited effects. The data was confirmed by Western blot. DAVID and KEGG bioinformatics analyses of proteomic data showed relationships between secretory, mobility and phagocytic cell functions and virus and particle trafficking.

Conclusions: We posit that modulation of endolysosomal pathways by antiretroviral nanoparticles provides a strategic path to combat HIV infection.

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Related in: MedlinePlus

Endolysosomal proteins in HIV-1, HIV + nanoATV, nanoATV and native ATV treated MDM.
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Fig3: Endolysosomal proteins in HIV-1, HIV + nanoATV, nanoATV and native ATV treated MDM.

Mentions: The proteins relative to functional pathways were further investigated using the KEGG database, which indicated phagosomes as one of the main pathways related to HIV-1 infection and nanoATV treatment. First we identified the role of HIV-1 infection as compared to HIV-1 infected nanoART-treated MDM on the phagosome network (Figure 1A and B, respectively). Few proteins were deregulated with nanoATV treatment. However, more proteins were deregulated during HIV-1 infection and nanoATV treatment. Notably, there was an opposite regulation for proteins within the phagosome and endosomal compartment between HIV-1-infected and HIV-1-infected and nanoATV-treated MDM. Up-regulation of Rab5 and −7 proteins was observed in HIV-1 infected cells; in contrast these same proteins were down-regulated in nanoATV-treated HIV-1 infected cells (pink = increased expression, blue = decreased expression). A similar pattern for LAMP1 was also observed. Moreover, DAVID functional enrichment clustering gave similar enrichment results for lysosomes in HIV-1-infected and HIV-1-infected and nanoATV-treated cells by filtering the data sets at a P value <0.01 (Additional files 4, 5, 6). There was a down-regulation of endosomal and lysosomal proteins in the uninfected cells treated with native ATV (Figure 2A) or nanoATV. The latter showed few down-regulated proteins in parallel endosomal compartments (Figure 2B). HIV-1 and native ATV treatment was similar to HIV-1 alone (i.e. high number of altered proteins). There were less numbers of oppositely regulated proteins compared to HIV and nanoATV (data not shown). A composite of these protein network changes are summarized in Figure 3.Figure 3


Opposing regulation of endolysosomal pathways by long-acting nanoformulated antiretroviral therapy and HIV-1 in human macrophages.

Araínga M, Guo D, Wiederin J, Ciborowski P, McMillan J, Gendelman HE - Retrovirology (2015)

Endolysosomal proteins in HIV-1, HIV + nanoATV, nanoATV and native ATV treated MDM.
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4307176&req=5

Fig3: Endolysosomal proteins in HIV-1, HIV + nanoATV, nanoATV and native ATV treated MDM.
Mentions: The proteins relative to functional pathways were further investigated using the KEGG database, which indicated phagosomes as one of the main pathways related to HIV-1 infection and nanoATV treatment. First we identified the role of HIV-1 infection as compared to HIV-1 infected nanoART-treated MDM on the phagosome network (Figure 1A and B, respectively). Few proteins were deregulated with nanoATV treatment. However, more proteins were deregulated during HIV-1 infection and nanoATV treatment. Notably, there was an opposite regulation for proteins within the phagosome and endosomal compartment between HIV-1-infected and HIV-1-infected and nanoATV-treated MDM. Up-regulation of Rab5 and −7 proteins was observed in HIV-1 infected cells; in contrast these same proteins were down-regulated in nanoATV-treated HIV-1 infected cells (pink = increased expression, blue = decreased expression). A similar pattern for LAMP1 was also observed. Moreover, DAVID functional enrichment clustering gave similar enrichment results for lysosomes in HIV-1-infected and HIV-1-infected and nanoATV-treated cells by filtering the data sets at a P value <0.01 (Additional files 4, 5, 6). There was a down-regulation of endosomal and lysosomal proteins in the uninfected cells treated with native ATV (Figure 2A) or nanoATV. The latter showed few down-regulated proteins in parallel endosomal compartments (Figure 2B). HIV-1 and native ATV treatment was similar to HIV-1 alone (i.e. high number of altered proteins). There were less numbers of oppositely regulated proteins compared to HIV and nanoATV (data not shown). A composite of these protein network changes are summarized in Figure 3.Figure 3

Bottom Line: NanoART and native ATV treated uninfected cells showed limited effects.The data was confirmed by Western blot.DAVID and KEGG bioinformatics analyses of proteomic data showed relationships between secretory, mobility and phagocytic cell functions and virus and particle trafficking.

View Article: PubMed Central - PubMed

Affiliation: Departments of Pharmacology and Experimental Neuroscience, University of Nebraska Medical Center, 985880 Nebraska Medical Center, Omaha 68198-5880, NE, USA. hegendel@unmc.edu.

ABSTRACT

Background: Long-acting nanoformulated antiretroviral therapy (nanoART) is designed to improve patient regimen adherence, reduce systemic drug toxicities, and facilitate clearance of human immunodeficiency virus type one (HIV-1) infection. While nanoART establishes drug depots within recycling and late monocyte-macrophage endosomes, whether or not this provides a strategic advantage towards viral elimination has not been elucidated.

Results: We applied quantitative SWATH-MS proteomics and cell profiling to nanoparticle atazanavir (nanoATV)-treated and HIV-1 infected human monocyte-derived macrophages (MDM). Native ATV and uninfected cells served as controls. Both HIV-1 and nanoATV engaged endolysosomal trafficking for assembly and depot formation, respectively. Notably, the pathways were deregulated in opposing manners by the virus and the nanoATV, likely by viral clearance. Paired-sample z-scores, of the proteomic data sets, showed up- and down- regulation of Rab-linked endolysosomal proteins. NanoART and native ATV treated uninfected cells showed limited effects. The data was confirmed by Western blot. DAVID and KEGG bioinformatics analyses of proteomic data showed relationships between secretory, mobility and phagocytic cell functions and virus and particle trafficking.

Conclusions: We posit that modulation of endolysosomal pathways by antiretroviral nanoparticles provides a strategic path to combat HIV infection.

Show MeSH
Related in: MedlinePlus