Cargando…

Organotypic Co-Cultures as a Novel 3D Model for Head and Neck Squamous Cell Carcinoma

Background: Head and neck squamous cell carcinomas (HNSCC) are phenotypically and molecularly heterogeneous and frequently develop therapy resistance. Reliable patient-derived 3D tumor models are urgently needed to further study the complex pathogenesis of these tumors and to overcome treatment fail...

Descripción completa

Detalles Bibliográficos
Autores principales: Engelmann, Luca, Thierauf, Julia, Koerich Laureano, Natalia, Stark, Hans-Juergen, Prigge, Elena-Sophie, Horn, Dominik, Freier, Kolja, Grabe, Niels, Rong, Chao, Federspil, Philippe, Zaoui, Karim, Plinkert, Peter K., Rotter, Nicole, von Knebel Doeberitz, Magnus, Hess, Jochen, Affolter, Annette
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7463661/
https://www.ncbi.nlm.nih.gov/pubmed/32824777
http://dx.doi.org/10.3390/cancers12082330
_version_ 1783577184171982848
author Engelmann, Luca
Thierauf, Julia
Koerich Laureano, Natalia
Stark, Hans-Juergen
Prigge, Elena-Sophie
Horn, Dominik
Freier, Kolja
Grabe, Niels
Rong, Chao
Federspil, Philippe
Zaoui, Karim
Plinkert, Peter K.
Rotter, Nicole
von Knebel Doeberitz, Magnus
Hess, Jochen
Affolter, Annette
author_facet Engelmann, Luca
Thierauf, Julia
Koerich Laureano, Natalia
Stark, Hans-Juergen
Prigge, Elena-Sophie
Horn, Dominik
Freier, Kolja
Grabe, Niels
Rong, Chao
Federspil, Philippe
Zaoui, Karim
Plinkert, Peter K.
Rotter, Nicole
von Knebel Doeberitz, Magnus
Hess, Jochen
Affolter, Annette
author_sort Engelmann, Luca
collection PubMed
description Background: Head and neck squamous cell carcinomas (HNSCC) are phenotypically and molecularly heterogeneous and frequently develop therapy resistance. Reliable patient-derived 3D tumor models are urgently needed to further study the complex pathogenesis of these tumors and to overcome treatment failure. Methods: We developed a three-dimensional organotypic co-culture (3D-OTC) model for HNSCC that maintains the architecture and cell composition of the individual tumor. A dermal equivalent (DE), composed of healthy human-derived fibroblasts and viscose fibers, served as a scaffold for the patient sample. DEs were co-cultivated with 13 vital HNSCC explants (non-human papillomavirus (HPV) driven, n = 7; HPV-driven, n = 6). Fractionated irradiation was applied to 5 samples (non-HPV-driven, n = 2; HPV-driven n = 3). To evaluate expression of ki-67, cleaved caspase-3, pan-cytokeratin, p16(INK4a), CD45, ∝smooth muscle actin and vimentin over time, immunohistochemistry and immunofluorescence staining were performed Patient checkup data were collected for up to 32 months after first diagnosis. Results: All non-HPV-driven 3D-OTCs encompassed proliferative cancer cells during cultivation for up to 21 days. Proliferation indices of primaries and 3D-OTCs were comparable and consistent over time. Overall, tumor explants displayed heterogeneous growth patterns (i.e., invasive, expansive, silent). Cancer-associated fibroblasts and leukocytes could be detected for up to 21 days. HPV DNA was detectable in both primary and 3D-OTCs (day 14) of HPV-driven tumors. However, p16(INK4a) expression levels were varying. Morphological alterations and radioresistant tumor cells were detected in 3D-OTC after fractionated irradiation in HPV-driven and non-driven samples. Conclusions: Our 3D-OTC model for HNSCC supports cancer cell survival and proliferation in their original microenvironment. The model enables investigation of invasive cancer growth and might, in the future, serve as a platform to perform sensitivity testing upon treatment to predict therapy response.
format Online
Article
Text
id pubmed-7463661
institution National Center for Biotechnology Information
language English
publishDate 2020
publisher MDPI
record_format MEDLINE/PubMed
spelling pubmed-74636612020-09-02 Organotypic Co-Cultures as a Novel 3D Model for Head and Neck Squamous Cell Carcinoma Engelmann, Luca Thierauf, Julia Koerich Laureano, Natalia Stark, Hans-Juergen Prigge, Elena-Sophie Horn, Dominik Freier, Kolja Grabe, Niels Rong, Chao Federspil, Philippe Zaoui, Karim Plinkert, Peter K. Rotter, Nicole von Knebel Doeberitz, Magnus Hess, Jochen Affolter, Annette Cancers (Basel) Article Background: Head and neck squamous cell carcinomas (HNSCC) are phenotypically and molecularly heterogeneous and frequently develop therapy resistance. Reliable patient-derived 3D tumor models are urgently needed to further study the complex pathogenesis of these tumors and to overcome treatment failure. Methods: We developed a three-dimensional organotypic co-culture (3D-OTC) model for HNSCC that maintains the architecture and cell composition of the individual tumor. A dermal equivalent (DE), composed of healthy human-derived fibroblasts and viscose fibers, served as a scaffold for the patient sample. DEs were co-cultivated with 13 vital HNSCC explants (non-human papillomavirus (HPV) driven, n = 7; HPV-driven, n = 6). Fractionated irradiation was applied to 5 samples (non-HPV-driven, n = 2; HPV-driven n = 3). To evaluate expression of ki-67, cleaved caspase-3, pan-cytokeratin, p16(INK4a), CD45, ∝smooth muscle actin and vimentin over time, immunohistochemistry and immunofluorescence staining were performed Patient checkup data were collected for up to 32 months after first diagnosis. Results: All non-HPV-driven 3D-OTCs encompassed proliferative cancer cells during cultivation for up to 21 days. Proliferation indices of primaries and 3D-OTCs were comparable and consistent over time. Overall, tumor explants displayed heterogeneous growth patterns (i.e., invasive, expansive, silent). Cancer-associated fibroblasts and leukocytes could be detected for up to 21 days. HPV DNA was detectable in both primary and 3D-OTCs (day 14) of HPV-driven tumors. However, p16(INK4a) expression levels were varying. Morphological alterations and radioresistant tumor cells were detected in 3D-OTC after fractionated irradiation in HPV-driven and non-driven samples. Conclusions: Our 3D-OTC model for HNSCC supports cancer cell survival and proliferation in their original microenvironment. The model enables investigation of invasive cancer growth and might, in the future, serve as a platform to perform sensitivity testing upon treatment to predict therapy response. MDPI 2020-08-18 /pmc/articles/PMC7463661/ /pubmed/32824777 http://dx.doi.org/10.3390/cancers12082330 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
Engelmann, Luca
Thierauf, Julia
Koerich Laureano, Natalia
Stark, Hans-Juergen
Prigge, Elena-Sophie
Horn, Dominik
Freier, Kolja
Grabe, Niels
Rong, Chao
Federspil, Philippe
Zaoui, Karim
Plinkert, Peter K.
Rotter, Nicole
von Knebel Doeberitz, Magnus
Hess, Jochen
Affolter, Annette
Organotypic Co-Cultures as a Novel 3D Model for Head and Neck Squamous Cell Carcinoma
title Organotypic Co-Cultures as a Novel 3D Model for Head and Neck Squamous Cell Carcinoma
title_full Organotypic Co-Cultures as a Novel 3D Model for Head and Neck Squamous Cell Carcinoma
title_fullStr Organotypic Co-Cultures as a Novel 3D Model for Head and Neck Squamous Cell Carcinoma
title_full_unstemmed Organotypic Co-Cultures as a Novel 3D Model for Head and Neck Squamous Cell Carcinoma
title_short Organotypic Co-Cultures as a Novel 3D Model for Head and Neck Squamous Cell Carcinoma
title_sort organotypic co-cultures as a novel 3d model for head and neck squamous cell carcinoma
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7463661/
https://www.ncbi.nlm.nih.gov/pubmed/32824777
http://dx.doi.org/10.3390/cancers12082330
work_keys_str_mv AT engelmannluca organotypiccoculturesasanovel3dmodelforheadandnecksquamouscellcarcinoma
AT thieraufjulia organotypiccoculturesasanovel3dmodelforheadandnecksquamouscellcarcinoma
AT koerichlaureanonatalia organotypiccoculturesasanovel3dmodelforheadandnecksquamouscellcarcinoma
AT starkhansjuergen organotypiccoculturesasanovel3dmodelforheadandnecksquamouscellcarcinoma
AT priggeelenasophie organotypiccoculturesasanovel3dmodelforheadandnecksquamouscellcarcinoma
AT horndominik organotypiccoculturesasanovel3dmodelforheadandnecksquamouscellcarcinoma
AT freierkolja organotypiccoculturesasanovel3dmodelforheadandnecksquamouscellcarcinoma
AT grabeniels organotypiccoculturesasanovel3dmodelforheadandnecksquamouscellcarcinoma
AT rongchao organotypiccoculturesasanovel3dmodelforheadandnecksquamouscellcarcinoma
AT federspilphilippe organotypiccoculturesasanovel3dmodelforheadandnecksquamouscellcarcinoma
AT zaouikarim organotypiccoculturesasanovel3dmodelforheadandnecksquamouscellcarcinoma
AT plinkertpeterk organotypiccoculturesasanovel3dmodelforheadandnecksquamouscellcarcinoma
AT rotternicole organotypiccoculturesasanovel3dmodelforheadandnecksquamouscellcarcinoma
AT vonknebeldoeberitzmagnus organotypiccoculturesasanovel3dmodelforheadandnecksquamouscellcarcinoma
AT hessjochen organotypiccoculturesasanovel3dmodelforheadandnecksquamouscellcarcinoma
AT affolterannette organotypiccoculturesasanovel3dmodelforheadandnecksquamouscellcarcinoma