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Single-cell RNAseq and longitudinal proteomic analysis of a novel semi-spontaneous urothelial cancer model reveals tumor cell heterogeneity and pretumoral urine protein alterations

Bladder cancer, one of the most prevalent malignancies worldwide, remains hard to classify due to a staggering molecular complexity. Despite a plethora of diagnostic tools and therapies, it is hard to outline the key steps leading up to the transition from high-risk non–muscle-invasive bladder cance...

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Autores principales: Kerzeli, Iliana K., Lord, Martin, Doroszko, Milena, Elgendy, Ramy, Chourlia, Aikaterini, Stepanek, Ivan, Larsson, Elinor, van Hooren, Luuk, Nelander, Sven, Malmstrom, Per-Uno, Dragomir, Anca, Segersten, Ulrika, Mangsbo, Sara M.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8262791/
https://www.ncbi.nlm.nih.gov/pubmed/34232958
http://dx.doi.org/10.1371/journal.pone.0253178
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author Kerzeli, Iliana K.
Lord, Martin
Doroszko, Milena
Elgendy, Ramy
Chourlia, Aikaterini
Stepanek, Ivan
Larsson, Elinor
van Hooren, Luuk
Nelander, Sven
Malmstrom, Per-Uno
Dragomir, Anca
Segersten, Ulrika
Mangsbo, Sara M.
author_facet Kerzeli, Iliana K.
Lord, Martin
Doroszko, Milena
Elgendy, Ramy
Chourlia, Aikaterini
Stepanek, Ivan
Larsson, Elinor
van Hooren, Luuk
Nelander, Sven
Malmstrom, Per-Uno
Dragomir, Anca
Segersten, Ulrika
Mangsbo, Sara M.
author_sort Kerzeli, Iliana K.
collection PubMed
description Bladder cancer, one of the most prevalent malignancies worldwide, remains hard to classify due to a staggering molecular complexity. Despite a plethora of diagnostic tools and therapies, it is hard to outline the key steps leading up to the transition from high-risk non–muscle-invasive bladder cancer (NMIBC) to muscle-invasive bladder cancer (MIBC). Carcinogen-induced murine models can recapitulate urothelial carcinogenesis and natural anti-tumor immunity. Herein, we have developed and profiled a novel model of progressive NMIBC based on 10 weeks of OH-BBN exposure in hepatocyte growth factor/cyclin dependent kinase 4 (R24C) (Hgf-Cdk4(R24C)) mice. The profiling of the model was performed by histology grading, single cell transcriptomic and proteomic analysis, while the derivation of a tumorigenic cell line was validated and used to assess in vivo anti-tumor effects in response to immunotherapy. Established NMIBC was present in females at 10 weeks post OH-BBN exposure while neoplasia was not as advanced in male mice, however all mice progressed to MIBC. Single cell RNA sequencing analysis revealed an intratumoral heterogeneity also described in the human disease trajectory. Moreover, although immune activation biomarkers were elevated in urine during carcinogen exposure, anti-programmed cell death protein 1 (anti-PD1) monotherapy did not prevent tumor progression. Furthermore, anti-PD1 immunotherapy did not control the growth of subcutaneous tumors formed by the newly derived urothelial cancer cell line. However, treatment with CpG-oligodeoxynucleotides (ODN) significantly decreased tumor volume, but only in females. In conclusion, the molecular map of this novel preclinical model of bladder cancer provides an opportunity to further investigate pharmacological therapies ahead with regards to both targeted drugs and immunotherapies to improve the strategies of how we should tackle the heterogeneous tumor microenvironment in urothelial bladder cancer to improve responses rates in the clinic.
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spelling pubmed-82627912021-07-19 Single-cell RNAseq and longitudinal proteomic analysis of a novel semi-spontaneous urothelial cancer model reveals tumor cell heterogeneity and pretumoral urine protein alterations Kerzeli, Iliana K. Lord, Martin Doroszko, Milena Elgendy, Ramy Chourlia, Aikaterini Stepanek, Ivan Larsson, Elinor van Hooren, Luuk Nelander, Sven Malmstrom, Per-Uno Dragomir, Anca Segersten, Ulrika Mangsbo, Sara M. PLoS One Research Article Bladder cancer, one of the most prevalent malignancies worldwide, remains hard to classify due to a staggering molecular complexity. Despite a plethora of diagnostic tools and therapies, it is hard to outline the key steps leading up to the transition from high-risk non–muscle-invasive bladder cancer (NMIBC) to muscle-invasive bladder cancer (MIBC). Carcinogen-induced murine models can recapitulate urothelial carcinogenesis and natural anti-tumor immunity. Herein, we have developed and profiled a novel model of progressive NMIBC based on 10 weeks of OH-BBN exposure in hepatocyte growth factor/cyclin dependent kinase 4 (R24C) (Hgf-Cdk4(R24C)) mice. The profiling of the model was performed by histology grading, single cell transcriptomic and proteomic analysis, while the derivation of a tumorigenic cell line was validated and used to assess in vivo anti-tumor effects in response to immunotherapy. Established NMIBC was present in females at 10 weeks post OH-BBN exposure while neoplasia was not as advanced in male mice, however all mice progressed to MIBC. Single cell RNA sequencing analysis revealed an intratumoral heterogeneity also described in the human disease trajectory. Moreover, although immune activation biomarkers were elevated in urine during carcinogen exposure, anti-programmed cell death protein 1 (anti-PD1) monotherapy did not prevent tumor progression. Furthermore, anti-PD1 immunotherapy did not control the growth of subcutaneous tumors formed by the newly derived urothelial cancer cell line. However, treatment with CpG-oligodeoxynucleotides (ODN) significantly decreased tumor volume, but only in females. In conclusion, the molecular map of this novel preclinical model of bladder cancer provides an opportunity to further investigate pharmacological therapies ahead with regards to both targeted drugs and immunotherapies to improve the strategies of how we should tackle the heterogeneous tumor microenvironment in urothelial bladder cancer to improve responses rates in the clinic. Public Library of Science 2021-07-07 /pmc/articles/PMC8262791/ /pubmed/34232958 http://dx.doi.org/10.1371/journal.pone.0253178 Text en © 2021 Kerzeli et al https://creativecommons.org/licenses/by/4.0/This is an open access article distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
spellingShingle Research Article
Kerzeli, Iliana K.
Lord, Martin
Doroszko, Milena
Elgendy, Ramy
Chourlia, Aikaterini
Stepanek, Ivan
Larsson, Elinor
van Hooren, Luuk
Nelander, Sven
Malmstrom, Per-Uno
Dragomir, Anca
Segersten, Ulrika
Mangsbo, Sara M.
Single-cell RNAseq and longitudinal proteomic analysis of a novel semi-spontaneous urothelial cancer model reveals tumor cell heterogeneity and pretumoral urine protein alterations
title Single-cell RNAseq and longitudinal proteomic analysis of a novel semi-spontaneous urothelial cancer model reveals tumor cell heterogeneity and pretumoral urine protein alterations
title_full Single-cell RNAseq and longitudinal proteomic analysis of a novel semi-spontaneous urothelial cancer model reveals tumor cell heterogeneity and pretumoral urine protein alterations
title_fullStr Single-cell RNAseq and longitudinal proteomic analysis of a novel semi-spontaneous urothelial cancer model reveals tumor cell heterogeneity and pretumoral urine protein alterations
title_full_unstemmed Single-cell RNAseq and longitudinal proteomic analysis of a novel semi-spontaneous urothelial cancer model reveals tumor cell heterogeneity and pretumoral urine protein alterations
title_short Single-cell RNAseq and longitudinal proteomic analysis of a novel semi-spontaneous urothelial cancer model reveals tumor cell heterogeneity and pretumoral urine protein alterations
title_sort single-cell rnaseq and longitudinal proteomic analysis of a novel semi-spontaneous urothelial cancer model reveals tumor cell heterogeneity and pretumoral urine protein alterations
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8262791/
https://www.ncbi.nlm.nih.gov/pubmed/34232958
http://dx.doi.org/10.1371/journal.pone.0253178
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