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Long-Term Systemic Expression of a Novel PD-1 Blocking Nanobody from an AAV Vector Provides Antitumor Activity without Toxicity

Immune checkpoint blockade using monoclonal antibodies (mAbs) able to block programmed death-1 (PD-1)/PD-L1 axis represents a promising treatment for cancer. However, it requires repetitive systemic administration of high mAbs doses, often leading to adverse effects. We generated a novel nanobody ag...

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Autores principales: Silva-Pilipich, Noelia, Martisova, Eva, Ballesteros-Briones, María Cristina, Hervas-Stubbs, Sandra, Casares, Noelia, González-Sapienza, Gualberto, Smerdou, Cristian, Vanrell, Lucia
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7761623/
https://www.ncbi.nlm.nih.gov/pubmed/33276580
http://dx.doi.org/10.3390/biomedicines8120562
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author Silva-Pilipich, Noelia
Martisova, Eva
Ballesteros-Briones, María Cristina
Hervas-Stubbs, Sandra
Casares, Noelia
González-Sapienza, Gualberto
Smerdou, Cristian
Vanrell, Lucia
author_facet Silva-Pilipich, Noelia
Martisova, Eva
Ballesteros-Briones, María Cristina
Hervas-Stubbs, Sandra
Casares, Noelia
González-Sapienza, Gualberto
Smerdou, Cristian
Vanrell, Lucia
author_sort Silva-Pilipich, Noelia
collection PubMed
description Immune checkpoint blockade using monoclonal antibodies (mAbs) able to block programmed death-1 (PD-1)/PD-L1 axis represents a promising treatment for cancer. However, it requires repetitive systemic administration of high mAbs doses, often leading to adverse effects. We generated a novel nanobody against PD-1 (Nb11) able to block PD-1/PD-L1 interaction for both mouse and human molecules. Nb11 was cloned into an adeno-associated virus (AAV) vector downstream of four different promoters (CMV, CAG, EF1α, and SFFV) and its expression was analyzed in cells from rodent (BHK) and human origin (Huh-7). Nb11 was expressed at high levels in vitro reaching 2–20 micrograms/mL with all promoters, except SFFV, which showed lower levels. Nb11 in vivo expression was evaluated in C57BL/6 mice after intravenous administration of AAV8 vectors. Nb11 serum levels increased steadily along time, reaching 1–3 microgram/mL two months post-treatment with the vector having the CAG promoter (AAV-CAG-Nb11), without evidence of toxicity. To test the antitumor potential of this vector, mice that received AAV-CAG-Nb11, or saline as control, were challenged with colon adenocarcinoma cells (MC38). AAV-CAG-Nb11 treatment prevented tumor formation in 30% of mice, significantly increasing survival. These data suggest that continuous expression of immunomodulatory nanobodies from long-term expression vectors could have antitumor effects with low toxicity.
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spelling pubmed-77616232020-12-26 Long-Term Systemic Expression of a Novel PD-1 Blocking Nanobody from an AAV Vector Provides Antitumor Activity without Toxicity Silva-Pilipich, Noelia Martisova, Eva Ballesteros-Briones, María Cristina Hervas-Stubbs, Sandra Casares, Noelia González-Sapienza, Gualberto Smerdou, Cristian Vanrell, Lucia Biomedicines Article Immune checkpoint blockade using monoclonal antibodies (mAbs) able to block programmed death-1 (PD-1)/PD-L1 axis represents a promising treatment for cancer. However, it requires repetitive systemic administration of high mAbs doses, often leading to adverse effects. We generated a novel nanobody against PD-1 (Nb11) able to block PD-1/PD-L1 interaction for both mouse and human molecules. Nb11 was cloned into an adeno-associated virus (AAV) vector downstream of four different promoters (CMV, CAG, EF1α, and SFFV) and its expression was analyzed in cells from rodent (BHK) and human origin (Huh-7). Nb11 was expressed at high levels in vitro reaching 2–20 micrograms/mL with all promoters, except SFFV, which showed lower levels. Nb11 in vivo expression was evaluated in C57BL/6 mice after intravenous administration of AAV8 vectors. Nb11 serum levels increased steadily along time, reaching 1–3 microgram/mL two months post-treatment with the vector having the CAG promoter (AAV-CAG-Nb11), without evidence of toxicity. To test the antitumor potential of this vector, mice that received AAV-CAG-Nb11, or saline as control, were challenged with colon adenocarcinoma cells (MC38). AAV-CAG-Nb11 treatment prevented tumor formation in 30% of mice, significantly increasing survival. These data suggest that continuous expression of immunomodulatory nanobodies from long-term expression vectors could have antitumor effects with low toxicity. MDPI 2020-12-02 /pmc/articles/PMC7761623/ /pubmed/33276580 http://dx.doi.org/10.3390/biomedicines8120562 Text en © 2020 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Silva-Pilipich, Noelia
Martisova, Eva
Ballesteros-Briones, María Cristina
Hervas-Stubbs, Sandra
Casares, Noelia
González-Sapienza, Gualberto
Smerdou, Cristian
Vanrell, Lucia
Long-Term Systemic Expression of a Novel PD-1 Blocking Nanobody from an AAV Vector Provides Antitumor Activity without Toxicity
title Long-Term Systemic Expression of a Novel PD-1 Blocking Nanobody from an AAV Vector Provides Antitumor Activity without Toxicity
title_full Long-Term Systemic Expression of a Novel PD-1 Blocking Nanobody from an AAV Vector Provides Antitumor Activity without Toxicity
title_fullStr Long-Term Systemic Expression of a Novel PD-1 Blocking Nanobody from an AAV Vector Provides Antitumor Activity without Toxicity
title_full_unstemmed Long-Term Systemic Expression of a Novel PD-1 Blocking Nanobody from an AAV Vector Provides Antitumor Activity without Toxicity
title_short Long-Term Systemic Expression of a Novel PD-1 Blocking Nanobody from an AAV Vector Provides Antitumor Activity without Toxicity
title_sort long-term systemic expression of a novel pd-1 blocking nanobody from an aav vector provides antitumor activity without toxicity
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7761623/
https://www.ncbi.nlm.nih.gov/pubmed/33276580
http://dx.doi.org/10.3390/biomedicines8120562
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