@article{10902/29989,
year = {2023},
month = {1},
url = {https://hdl.handle.net/10902/29989},
abstract = {Background
Cancer-associated fibroblasts (CAFs) are major players in tumor-stroma communication, and participate in several cancer hallmarks to drive tumor progression and metastatic dissemination. This study investigates the driving effects of tumor-secreted factors on CAF biology, with the ultimate goal of identifying effective therapeutic targets/strategies for head and neck squamous cell carcinomas (HNSCC).

Methods
Functionally, conditioned media (CM) from different HNSCC-derived cell lines and normal keratinocytes (Kc) were tested on the growth and invasion of populations of primary CAFs and normal fibroblasts (NFs) using 3D invasion assays in collagen matrices. The changes in MMPs expression were evaluated by RT-qPCR and kinase enrichment was analyzed using mass spectrometry phosphoproteomics.

Results
Our results consistently demonstrate that HNSCC-secreted factors (but not Kc CM) specifically and robustly promoted pro-invasive properties in both CAFs and NFs, thereby reflecting the plasticity of fibroblast subtypes. Concomitantly, HNSCC-secreted factors massively increased metalloproteinases levels in CAFs and NFs. By contrast, HNSCC CM and Kc CM exhibited comparable growth-promoting effects on stromal fibroblasts. Mechanistically, phosphoproteomic analysis predominantly revealed phosphorylation changes in fibroblasts upon treatment with HNSCC CM, and various promising kinases were identified: MKK7, MKK4, ASK1, RAF1, BRAF, ARAF, COT, PDK1, RSK2 and AKT1. Interestingly, pharmacologic inhibition of RAF1/BRAF using sorafenib emerged as the most effective drug to block tumor-promoted fibroblast invasion without affecting fibroblast viability

Conclusions
Our findings demonstrate that HNSCC-secreted factors specifically fine tune the invasive potential of stromal fibroblasts, thereby generating tumor-driven pro-invasive niches, which in turn to ultimately facilitate cancer cell dissemination. Furthermore, the RAF/BRAF inhibitor sorafenib was identified as a promising candidate to effectively target the onset of pro-invasive clusters of stromal fibroblasts in the HNSCC microenvironment.},
organization = {Funding: This study was supported by the Instituto de Salud Carlos III (ISCIII, Spain) through the project grants PI19/00560, PI22/00167 and CIBERONC (CB16/12/00390) and co-funded by the European Union; by FINBA-ISPA (Spain) through the project grant 2021-047- INTRAMURAL NOV-ALTES (sponsored by Janssen); by the Instituto de Investigación Sanitaria del Principado de Asturias (ISPA, Spain), by Fundación Bancaria Cajastur-IUOPA, and Universidad de Oviedo (Spain). Also through the grant “Ayudas para Grupos de Investigación de Organismos del Principado de Asturias 2021–2023” (IDI/2021/000079), funded by Principado de Asturias (Spain) through FICYT and the FEDER Funding Program from the European Union. SAT is recipient of a Sara Borrell postdoctoral fellowship from ISCIII, Spain (CD20/00006), and FHP recipient of a Maria Zambrano grant at the University of Oviedo, Spain. LPF is recipient of an FPU-PhD fellowship from the Spanish Ministry of Education, Spain (FPU20/01588), and IMJ recipient of a Severo Ochoa predoctoral fellowship from the Principado de Asturias, Spain (BP20- 152).
Acknowledgments: We thank Dr. Reidar Grenman and Dr. Alberto Gandarillas for kindly providing UT-SCC40 and NFs. We want to particularly acknowledge for its collaboration the Principado de Asturias BioBank (PT17/0015/0023 and PT20/00161), financed jointly by Servicio de Salud del Principado de Asturias, Instituto de Salud Carlos III and Fundación Bancaria Cajastur and integrated in the Spanish National Biobanks Network. Images were created using Biorender.com},
publisher = {Editions Scientifiques Elsevier},
publisher = {Biomedicine & Pharmacotherapy, 2023, 158, 114176},
title = {Driving role of head and neck cancer cell secretome on the invasion of stromal fibroblasts: mechanistic insights by phosphoproteomics},
author = {Prieto-Fernández, Llara and Villaronga, María de los Ángeles and Hermida-Prado, Francisco and Hijazi, Maruan and Montoro-Jiménez, Irene and Pevida, Marta and Llames, Sara and Rodrigo, Juan Pablo and Cutillas, Pedro and Calvo González, Fernando and García-Pedrero, Juana María and Álvarez-Teijeiro, Saúl},
}