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Synthetic DNA delivery by electroporation promotes robust in vivo sulfation of broadly neutralizing anti-HIV immunoadhesin eCD4-Ig

BACKGROUND: Despite vigorous and ongoing efforts, active immunizations have yet to induce broadly neutralizing antibodies (bNAbs) against HIV-1. An alternative approach is to achieve prophylaxis with long-term expression of potent biologic HIV-1 inhibitors with Adeno-associated Virus (AAV), which co...

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Autores principales: Xu, Ziyang, Wise, Megan C., Choi, Hyeree, Perales-Puchalt, Alfredo, Patel, Ami, Tello-Ruiz, Edgar, Chu, Jacqueline D., Muthumani, Kar, Weiner, David B.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6161476/
https://www.ncbi.nlm.nih.gov/pubmed/30174283
http://dx.doi.org/10.1016/j.ebiom.2018.08.027
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author Xu, Ziyang
Wise, Megan C.
Choi, Hyeree
Perales-Puchalt, Alfredo
Patel, Ami
Tello-Ruiz, Edgar
Chu, Jacqueline D.
Muthumani, Kar
Weiner, David B.
author_facet Xu, Ziyang
Wise, Megan C.
Choi, Hyeree
Perales-Puchalt, Alfredo
Patel, Ami
Tello-Ruiz, Edgar
Chu, Jacqueline D.
Muthumani, Kar
Weiner, David B.
author_sort Xu, Ziyang
collection PubMed
description BACKGROUND: Despite vigorous and ongoing efforts, active immunizations have yet to induce broadly neutralizing antibodies (bNAbs) against HIV-1. An alternative approach is to achieve prophylaxis with long-term expression of potent biologic HIV-1 inhibitors with Adeno-associated Virus (AAV), which could however be limited by hosts' humoral and cellular responses. An approach that facilitates in vivo production of these complex molecules independent of viral-vectored delivery will be a major advantage. METHODS: We used synthetic DNA and electroporation (DNA/EP) to deliver an anti-HIV-1 immunoadhesin eCD4-Ig in vivo. In addition, we engineered a TPST2 enzyme variant (IgE-TPST2), characterized its intracellular trafficking patterns and determined its ability to post-translationally sulfate eCD4-Ig in vivo. FINDINGS: With a single round of DNA injection, a peak expression level of 80-100μg/mL was observed in mice 14 days post injection (d.p.i). The engineered IgE-TPST2 enzyme trafficked efficiently to the Trans-Golgi Network (TGN). Co-administrating low dose of plasmid IgE-TPST2 with plasmid eCD4-Ig enhanced the potency of eCD4-Ig by three-fold in the ex vivo neutralization assay against the global panel of HIV-1 pseudoviruses. INTERPRETATION: This work provides a proof-of-concept for delivering anti-HIV-1 immunoadhesins by advanced nucleic acid technology and modulating protein functions in vivo with targeted enzyme-mediated post-translational modifications. FUNDING: This work is supported by NIH IPCAVD Grant U19 Al109646-04, Martin Delaney Collaboration for HIV Cure Research and W.W. Smith Charitable Trust.
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spelling pubmed-61614762018-10-01 Synthetic DNA delivery by electroporation promotes robust in vivo sulfation of broadly neutralizing anti-HIV immunoadhesin eCD4-Ig Xu, Ziyang Wise, Megan C. Choi, Hyeree Perales-Puchalt, Alfredo Patel, Ami Tello-Ruiz, Edgar Chu, Jacqueline D. Muthumani, Kar Weiner, David B. EBioMedicine Research paper BACKGROUND: Despite vigorous and ongoing efforts, active immunizations have yet to induce broadly neutralizing antibodies (bNAbs) against HIV-1. An alternative approach is to achieve prophylaxis with long-term expression of potent biologic HIV-1 inhibitors with Adeno-associated Virus (AAV), which could however be limited by hosts' humoral and cellular responses. An approach that facilitates in vivo production of these complex molecules independent of viral-vectored delivery will be a major advantage. METHODS: We used synthetic DNA and electroporation (DNA/EP) to deliver an anti-HIV-1 immunoadhesin eCD4-Ig in vivo. In addition, we engineered a TPST2 enzyme variant (IgE-TPST2), characterized its intracellular trafficking patterns and determined its ability to post-translationally sulfate eCD4-Ig in vivo. FINDINGS: With a single round of DNA injection, a peak expression level of 80-100μg/mL was observed in mice 14 days post injection (d.p.i). The engineered IgE-TPST2 enzyme trafficked efficiently to the Trans-Golgi Network (TGN). Co-administrating low dose of plasmid IgE-TPST2 with plasmid eCD4-Ig enhanced the potency of eCD4-Ig by three-fold in the ex vivo neutralization assay against the global panel of HIV-1 pseudoviruses. INTERPRETATION: This work provides a proof-of-concept for delivering anti-HIV-1 immunoadhesins by advanced nucleic acid technology and modulating protein functions in vivo with targeted enzyme-mediated post-translational modifications. FUNDING: This work is supported by NIH IPCAVD Grant U19 Al109646-04, Martin Delaney Collaboration for HIV Cure Research and W.W. Smith Charitable Trust. Elsevier 2018-08-30 /pmc/articles/PMC6161476/ /pubmed/30174283 http://dx.doi.org/10.1016/j.ebiom.2018.08.027 Text en © 2018 The Authors http://creativecommons.org/licenses/by-nc-nd/4.0/ This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
spellingShingle Research paper
Xu, Ziyang
Wise, Megan C.
Choi, Hyeree
Perales-Puchalt, Alfredo
Patel, Ami
Tello-Ruiz, Edgar
Chu, Jacqueline D.
Muthumani, Kar
Weiner, David B.
Synthetic DNA delivery by electroporation promotes robust in vivo sulfation of broadly neutralizing anti-HIV immunoadhesin eCD4-Ig
title Synthetic DNA delivery by electroporation promotes robust in vivo sulfation of broadly neutralizing anti-HIV immunoadhesin eCD4-Ig
title_full Synthetic DNA delivery by electroporation promotes robust in vivo sulfation of broadly neutralizing anti-HIV immunoadhesin eCD4-Ig
title_fullStr Synthetic DNA delivery by electroporation promotes robust in vivo sulfation of broadly neutralizing anti-HIV immunoadhesin eCD4-Ig
title_full_unstemmed Synthetic DNA delivery by electroporation promotes robust in vivo sulfation of broadly neutralizing anti-HIV immunoadhesin eCD4-Ig
title_short Synthetic DNA delivery by electroporation promotes robust in vivo sulfation of broadly neutralizing anti-HIV immunoadhesin eCD4-Ig
title_sort synthetic dna delivery by electroporation promotes robust in vivo sulfation of broadly neutralizing anti-hiv immunoadhesin ecd4-ig
topic Research paper
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6161476/
https://www.ncbi.nlm.nih.gov/pubmed/30174283
http://dx.doi.org/10.1016/j.ebiom.2018.08.027
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