A Multispecific Checkpoint Inhibitor Nanofitin with a Fast Tumor Accumulation Property and Anti-Tumor Activity in Immune Competent Mice

Immune checkpoint inhibitors have revolutionized cancer treatment but remain limited by on-target/off-tumor effects that narrow their therapeutic window. Although PD-L1 is mainly expressed by tumor cells, these effects could reduce bloodstream availability and tumor accumulation of PD-L1 inhibitors....

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Main Authors: Perrine Jacquot, Javier Muñoz-Garcia, Antoine Léger, Antoine Babuty, Manon Taupin, Laurie Fradet, Fabio Dupont, Marie-Françoise Heymann, Mathieu Cinier, Dominique Heymann
Format: Article
Language:English
Published: MDPI AG 2025-03-01
Series:Biomolecules
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Online Access:https://www.mdpi.com/2218-273X/15/4/471
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author Perrine Jacquot
Javier Muñoz-Garcia
Antoine Léger
Antoine Babuty
Manon Taupin
Laurie Fradet
Fabio Dupont
Marie-Françoise Heymann
Mathieu Cinier
Dominique Heymann
author_facet Perrine Jacquot
Javier Muñoz-Garcia
Antoine Léger
Antoine Babuty
Manon Taupin
Laurie Fradet
Fabio Dupont
Marie-Françoise Heymann
Mathieu Cinier
Dominique Heymann
author_sort Perrine Jacquot
collection DOAJ
description Immune checkpoint inhibitors have revolutionized cancer treatment but remain limited by on-target/off-tumor effects that narrow their therapeutic window. Although PD-L1 is mainly expressed by tumor cells, these effects could reduce bloodstream availability and tumor accumulation of PD-L1 inhibitors. Enhancing tumor specificity through bispecific proteins targeting two tumor-associated antigens offers a promising strategy. This study evaluated a bispecific Nanofitin, B10–B11, targeting PD-L1 and EGFR. In vitro, B10–B11 efficiently bound to human A431 and murine CT26 cell lines, validating these models for in vivo studies. Nanofitins’ accumulation in tumors and their anti-tumor efficacy were assessed, respectively, in A431 xenograft and CT26 immunocompetent mouse models. In both experiments, B10–B11 was compared with its albumin binding fused counterpart (B10–B11-ABNF). This study showed that the dual-targeting approach with the bispecific Nanofitin enhanced in vitro PD-L1 neutralization compared to the monomeric form and led to in vivo anti-tumor activity evidenced by reduced tumor growth and increased CD3+ T cells and F4/80+ macrophages in tumors. This activity was further correlated with Nanofitin’s tumor accumulation at 7 h post-injection, which was highest for the B10–B11-ABNF. This study highlights the potential of bispecific Nanofitins, particularly with albumin binding to enable rapid and uniform tumor accumulation of effective PD-L1 immunotherapy.
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spelling doaj-art-e0f48f93d57b47d3a75ac535bb157ff02025-08-20T02:24:45ZengMDPI AGBiomolecules2218-273X2025-03-0115447110.3390/biom15040471A Multispecific Checkpoint Inhibitor Nanofitin with a Fast Tumor Accumulation Property and Anti-Tumor Activity in Immune Competent MicePerrine Jacquot0Javier Muñoz-Garcia1Antoine Léger2Antoine Babuty3Manon Taupin4Laurie Fradet5Fabio Dupont6Marie-Françoise Heymann7Mathieu Cinier8Dominique Heymann9Affilogic SAS, 24 rue de la Rainière, 44300 Nantes, FranceUMR6286, US2B, CNRS, Nantes Université, 44322 Nantes, FranceAffilogic SAS, 24 rue de la Rainière, 44300 Nantes, FranceUMR6286, US2B, CNRS, Nantes Université, 44322 Nantes, FranceResearch Pathology Platform, Institut de Cancérologie de l’Ouest, 44805 Saint-Herblain, FranceUMR6286, US2B, CNRS, Nantes Université, 44322 Nantes, FranceAffilogic SAS, 24 rue de la Rainière, 44300 Nantes, FranceUMR6286, US2B, CNRS, Nantes Université, 44322 Nantes, FranceAffilogic SAS, 24 rue de la Rainière, 44300 Nantes, FranceUMR6286, US2B, CNRS, Nantes Université, 44322 Nantes, FranceImmune checkpoint inhibitors have revolutionized cancer treatment but remain limited by on-target/off-tumor effects that narrow their therapeutic window. Although PD-L1 is mainly expressed by tumor cells, these effects could reduce bloodstream availability and tumor accumulation of PD-L1 inhibitors. Enhancing tumor specificity through bispecific proteins targeting two tumor-associated antigens offers a promising strategy. This study evaluated a bispecific Nanofitin, B10–B11, targeting PD-L1 and EGFR. In vitro, B10–B11 efficiently bound to human A431 and murine CT26 cell lines, validating these models for in vivo studies. Nanofitins’ accumulation in tumors and their anti-tumor efficacy were assessed, respectively, in A431 xenograft and CT26 immunocompetent mouse models. In both experiments, B10–B11 was compared with its albumin binding fused counterpart (B10–B11-ABNF). This study showed that the dual-targeting approach with the bispecific Nanofitin enhanced in vitro PD-L1 neutralization compared to the monomeric form and led to in vivo anti-tumor activity evidenced by reduced tumor growth and increased CD3+ T cells and F4/80+ macrophages in tumors. This activity was further correlated with Nanofitin’s tumor accumulation at 7 h post-injection, which was highest for the B10–B11-ABNF. This study highlights the potential of bispecific Nanofitins, particularly with albumin binding to enable rapid and uniform tumor accumulation of effective PD-L1 immunotherapy.https://www.mdpi.com/2218-273X/15/4/471bispecific Nanofitinprogrammed cell death-ligand 1 (PD-L1)epidermal growth factor receptor (EGFR)on-target/off-tumor effecttumoral accumulation
spellingShingle Perrine Jacquot
Javier Muñoz-Garcia
Antoine Léger
Antoine Babuty
Manon Taupin
Laurie Fradet
Fabio Dupont
Marie-Françoise Heymann
Mathieu Cinier
Dominique Heymann
A Multispecific Checkpoint Inhibitor Nanofitin with a Fast Tumor Accumulation Property and Anti-Tumor Activity in Immune Competent Mice
Biomolecules
bispecific Nanofitin
programmed cell death-ligand 1 (PD-L1)
epidermal growth factor receptor (EGFR)
on-target/off-tumor effect
tumoral accumulation
title A Multispecific Checkpoint Inhibitor Nanofitin with a Fast Tumor Accumulation Property and Anti-Tumor Activity in Immune Competent Mice
title_full A Multispecific Checkpoint Inhibitor Nanofitin with a Fast Tumor Accumulation Property and Anti-Tumor Activity in Immune Competent Mice
title_fullStr A Multispecific Checkpoint Inhibitor Nanofitin with a Fast Tumor Accumulation Property and Anti-Tumor Activity in Immune Competent Mice
title_full_unstemmed A Multispecific Checkpoint Inhibitor Nanofitin with a Fast Tumor Accumulation Property and Anti-Tumor Activity in Immune Competent Mice
title_short A Multispecific Checkpoint Inhibitor Nanofitin with a Fast Tumor Accumulation Property and Anti-Tumor Activity in Immune Competent Mice
title_sort multispecific checkpoint inhibitor nanofitin with a fast tumor accumulation property and anti tumor activity in immune competent mice
topic bispecific Nanofitin
programmed cell death-ligand 1 (PD-L1)
epidermal growth factor receptor (EGFR)
on-target/off-tumor effect
tumoral accumulation
url https://www.mdpi.com/2218-273X/15/4/471
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