A novel therapeutic approach targeting PD-L1 in HNSCC and bone marrow-derived mesenchymal stem cells hampers pro-metastatic features in vitro: perspectives for blocking tumor-stroma communication and signaling

Abstract Background Current conventional treatment regimens for head and neck squamous cell carcinoma (HNSCC), are poorly effective because of the emergence of resistance mechanisms. Many studies have reported how the tumor microenvironment influences tumor response to immune checkpoint inhibitors t...

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Main Authors: Ylenia Ferrara, Debora Latino, Angela Costagliola di Polidoro, Angela Oliver, Annachiara Sarnella, Maria Grazia Caprio, Laura Cerchia, Menotti Ruvo, Annamaria Sandomenico, Antonella Zannetti
Format: Article
Language:English
Published: BMC 2025-02-01
Series:Cell Communication and Signaling
Online Access:https://doi.org/10.1186/s12964-025-02073-7
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author Ylenia Ferrara
Debora Latino
Angela Costagliola di Polidoro
Angela Oliver
Annachiara Sarnella
Maria Grazia Caprio
Laura Cerchia
Menotti Ruvo
Annamaria Sandomenico
Antonella Zannetti
author_facet Ylenia Ferrara
Debora Latino
Angela Costagliola di Polidoro
Angela Oliver
Annachiara Sarnella
Maria Grazia Caprio
Laura Cerchia
Menotti Ruvo
Annamaria Sandomenico
Antonella Zannetti
author_sort Ylenia Ferrara
collection DOAJ
description Abstract Background Current conventional treatment regimens for head and neck squamous cell carcinoma (HNSCC), are poorly effective because of the emergence of resistance mechanisms. Many studies have reported how the tumor microenvironment influences tumor response to immune checkpoint inhibitors targeting PD-1/PD-L1. It has been reported that overexpression of PD-L1 correlates with and is involved in cancer progression by promoting epithelial-to-mesenchymal-transition (EMT) program, stemness and tumor cell invasiveness through AKT and MAPK pathways. In this study, we investigated how bone marrow mesenchymal stem cells (BM-MSCs) recruited and educated by HNSCC cells are able to promote tumor cell invasion and EMT program. In addition, we analyzed how the crosstalk between stromal cells and tumor cells can affect PD-L1 expression levels. In this context, we developed and characterized a novel anti-PD-L1 recombinant Fab (rFab’) and tested its ability to potentiate the effect of cisplatin. Methods BM-MSCs and HNSCC cells co-cultures, cell migration and invasion were performed using Boyden chambers. The effect of treatments on cell viability and growth were analyzed by MTT and clonogenic assay, respectively. The anti-PD-L1 rFab’ was prepared in E. Coli and tested for its binding on HNSCC cells and BM-MSCs by FACS analysis and fluorescence microscopy. PD-L1, p-AKT, p-ERK, N-cadherin and β-catenin expression levels were analyzed by western blotting. Results BM-MSCs were induced by tumor cells to migrate, invade and to trans-differentiate in cancer associated fibroblasts (CAFs) as demonstrated by increased expression levels of α-SMA and FAP-α. BM-MSCs contributed to HNSCC invasiveness by increasing p-AKT, p-ERK, N-cadherin and β-catenin expression levels. When BM-MSCs and HNSCC cells were co-cultured the level of PD-L1 expression was enhanced in both cells indicating a reciprocal support in favoring tumor aggressiveness. Tumor cell treatment with rFab’ anti-PD-L1 reduced their viability, growth, migration and invasion and blunted the underlying signaling pathways. In addition, rFab’ anti-PD-L1 was able to potentiate the antitumor effect of cisplatin on HNSCC cells. Conclusions BM-MSCs recruited and educated by HNSCC cells support tumor cell aggressiveness via PD-L1. A novel rFab’ anti-PD-L1 reduces HNSCC proliferation, migration and invasion and potentiates the cisplatin effect suggesting its potential to be conjugated with drugs for immuno-cytotoxic therapy.
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spelling doaj-art-a70ef8bfe71e45038b6a2024519210722025-08-20T02:48:33ZengBMCCell Communication and Signaling1478-811X2025-02-0123112010.1186/s12964-025-02073-7A novel therapeutic approach targeting PD-L1 in HNSCC and bone marrow-derived mesenchymal stem cells hampers pro-metastatic features in vitro: perspectives for blocking tumor-stroma communication and signalingYlenia Ferrara0Debora Latino1Angela Costagliola di Polidoro2Angela Oliver3Annachiara Sarnella4Maria Grazia Caprio5Laura Cerchia6Menotti Ruvo7Annamaria Sandomenico8Antonella Zannetti9Istituto di Biostrutture e Bioimmagini-CNRIstituto di Biostrutture e Bioimmagini-CNRIstituto di Biostrutture e Bioimmagini-CNRIstituto di Biostrutture e Bioimmagini-CNRIstituto di Biostrutture e Bioimmagini-CNRIstituto di Biostrutture e Bioimmagini-CNRIstituto degli Endotipi in Oncologia, Metabolismo e Immunologia “G. Salvatore”, CNRIstituto di Biostrutture e Bioimmagini-CNRIstituto di Biostrutture e Bioimmagini-CNRIstituto di Biostrutture e Bioimmagini-CNRAbstract Background Current conventional treatment regimens for head and neck squamous cell carcinoma (HNSCC), are poorly effective because of the emergence of resistance mechanisms. Many studies have reported how the tumor microenvironment influences tumor response to immune checkpoint inhibitors targeting PD-1/PD-L1. It has been reported that overexpression of PD-L1 correlates with and is involved in cancer progression by promoting epithelial-to-mesenchymal-transition (EMT) program, stemness and tumor cell invasiveness through AKT and MAPK pathways. In this study, we investigated how bone marrow mesenchymal stem cells (BM-MSCs) recruited and educated by HNSCC cells are able to promote tumor cell invasion and EMT program. In addition, we analyzed how the crosstalk between stromal cells and tumor cells can affect PD-L1 expression levels. In this context, we developed and characterized a novel anti-PD-L1 recombinant Fab (rFab’) and tested its ability to potentiate the effect of cisplatin. Methods BM-MSCs and HNSCC cells co-cultures, cell migration and invasion were performed using Boyden chambers. The effect of treatments on cell viability and growth were analyzed by MTT and clonogenic assay, respectively. The anti-PD-L1 rFab’ was prepared in E. Coli and tested for its binding on HNSCC cells and BM-MSCs by FACS analysis and fluorescence microscopy. PD-L1, p-AKT, p-ERK, N-cadherin and β-catenin expression levels were analyzed by western blotting. Results BM-MSCs were induced by tumor cells to migrate, invade and to trans-differentiate in cancer associated fibroblasts (CAFs) as demonstrated by increased expression levels of α-SMA and FAP-α. BM-MSCs contributed to HNSCC invasiveness by increasing p-AKT, p-ERK, N-cadherin and β-catenin expression levels. When BM-MSCs and HNSCC cells were co-cultured the level of PD-L1 expression was enhanced in both cells indicating a reciprocal support in favoring tumor aggressiveness. Tumor cell treatment with rFab’ anti-PD-L1 reduced their viability, growth, migration and invasion and blunted the underlying signaling pathways. In addition, rFab’ anti-PD-L1 was able to potentiate the antitumor effect of cisplatin on HNSCC cells. Conclusions BM-MSCs recruited and educated by HNSCC cells support tumor cell aggressiveness via PD-L1. A novel rFab’ anti-PD-L1 reduces HNSCC proliferation, migration and invasion and potentiates the cisplatin effect suggesting its potential to be conjugated with drugs for immuno-cytotoxic therapy.https://doi.org/10.1186/s12964-025-02073-7
spellingShingle Ylenia Ferrara
Debora Latino
Angela Costagliola di Polidoro
Angela Oliver
Annachiara Sarnella
Maria Grazia Caprio
Laura Cerchia
Menotti Ruvo
Annamaria Sandomenico
Antonella Zannetti
A novel therapeutic approach targeting PD-L1 in HNSCC and bone marrow-derived mesenchymal stem cells hampers pro-metastatic features in vitro: perspectives for blocking tumor-stroma communication and signaling
Cell Communication and Signaling
title A novel therapeutic approach targeting PD-L1 in HNSCC and bone marrow-derived mesenchymal stem cells hampers pro-metastatic features in vitro: perspectives for blocking tumor-stroma communication and signaling
title_full A novel therapeutic approach targeting PD-L1 in HNSCC and bone marrow-derived mesenchymal stem cells hampers pro-metastatic features in vitro: perspectives for blocking tumor-stroma communication and signaling
title_fullStr A novel therapeutic approach targeting PD-L1 in HNSCC and bone marrow-derived mesenchymal stem cells hampers pro-metastatic features in vitro: perspectives for blocking tumor-stroma communication and signaling
title_full_unstemmed A novel therapeutic approach targeting PD-L1 in HNSCC and bone marrow-derived mesenchymal stem cells hampers pro-metastatic features in vitro: perspectives for blocking tumor-stroma communication and signaling
title_short A novel therapeutic approach targeting PD-L1 in HNSCC and bone marrow-derived mesenchymal stem cells hampers pro-metastatic features in vitro: perspectives for blocking tumor-stroma communication and signaling
title_sort novel therapeutic approach targeting pd l1 in hnscc and bone marrow derived mesenchymal stem cells hampers pro metastatic features in vitro perspectives for blocking tumor stroma communication and signaling
url https://doi.org/10.1186/s12964-025-02073-7
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