Single-Cell Analyses of a Novel Mouse Urothelial Carcinoma Model Reveal a Role of Tumor-Associated Macrophages in Response to Anti-PD-1 Therapy
Approximately 80% of patients with advanced bladder cancer do not respond to immune checkpoint inhibitor (ICI) immunotherapy. Therefore, there is an urgent unmet need to develop clinically relevant preclinical models so that factors governing immunotherapy responses can be studied in immunocompetent...
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| Format: | Article |
| Language: | English |
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MDPI AG
2022-05-01
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| Series: | Cancers |
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| Online Access: | https://www.mdpi.com/2072-6694/14/10/2511 |
| _version_ | 1797500968832598016 |
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| author | Dongbo Xu Li Wang Kyle Wieczorek Yali Zhang Zinian Wang Jianmin Wang Bo Xu Prashant K. Singh Yanqing Wang Xiaojing Zhang Yue Wu Gary J. Smith Kristopher Attwood Yuesheng Zhang David W. Goodrich Qiang Li |
| author_facet | Dongbo Xu Li Wang Kyle Wieczorek Yali Zhang Zinian Wang Jianmin Wang Bo Xu Prashant K. Singh Yanqing Wang Xiaojing Zhang Yue Wu Gary J. Smith Kristopher Attwood Yuesheng Zhang David W. Goodrich Qiang Li |
| author_sort | Dongbo Xu |
| collection | DOAJ |
| description | Approximately 80% of patients with advanced bladder cancer do not respond to immune checkpoint inhibitor (ICI) immunotherapy. Therefore, there is an urgent unmet need to develop clinically relevant preclinical models so that factors governing immunotherapy responses can be studied in immunocompetent mice. We developed a line of mouse triple knockout (TKO: <i>Trp53</i>, <i>Pten</i>, <i>Rb1</i>) urothelial carcinoma organoids transplanted into immunocompetent mice. These bladder tumors recapitulate the molecular phenotypes and heterogeneous immunotherapy responses observed in human bladder cancers. The TKO organoids were characterized in vivo and in vitro and compared to the widely used MB49 murine bladder cancer model. RNAseq analysis of the TKO tumors demonstrated a basal subtype. The TKO xenografts demonstrated the expression of urothelial markers (CK5, CK7, GATA3, and p63), whereas MB49 subcutaneous xenografts did not express urothelial markers. Anti-PD-1 immunotherapy resulted in a mixed pattern of treatment responses for individual tumors. Eight immune cell types were identified (basophils, B cells, dendritic cells, macrophages, monocytes, neutrophils, NK cells, and T cells) in ICI-treated xenografts. Responder xenografts displayed significantly increased immune cell infiltration (15.3%, 742 immune cells/4861 total cells) compared to the non-responder tumors (10.1%, 452 immune cells/4459 total cells, Fisher Exact Test <i>p</i> < 0.0001). Specifically, there were more T cells (1.0% vs. 0.4%, <i>p</i> = 0.002) and macrophages (8.6% vs. 6.4%, <i>p</i> = 0.0002) in responder xenografts than in non-responder xenografts. In conclusion, we have developed a novel preclinical model that exhibits a mixed pattern of response to anti-PD-1 immunotherapy. The higher percentage of macrophage tumor infiltration in responders suggests a potential role for the innate immune microenvironment in regulating ICI treatment responses. |
| first_indexed | 2024-03-10T03:11:31Z |
| format | Article |
| id | doaj.art-55f5e29ecd1d41c9a28a5c3b4f903e46 |
| institution | Directory Open Access Journal |
| issn | 2072-6694 |
| language | English |
| last_indexed | 2024-03-10T03:11:31Z |
| publishDate | 2022-05-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Cancers |
| spelling | doaj.art-55f5e29ecd1d41c9a28a5c3b4f903e462023-11-23T10:24:13ZengMDPI AGCancers2072-66942022-05-011410251110.3390/cancers14102511Single-Cell Analyses of a Novel Mouse Urothelial Carcinoma Model Reveal a Role of Tumor-Associated Macrophages in Response to Anti-PD-1 TherapyDongbo Xu0Li Wang1Kyle Wieczorek2Yali Zhang3Zinian Wang4Jianmin Wang5Bo Xu6Prashant K. Singh7Yanqing Wang8Xiaojing Zhang9Yue Wu10Gary J. Smith11Kristopher Attwood12Yuesheng Zhang13David W. Goodrich14Qiang Li15Department of Urology, Roswell Park Comprehensive Cancer Center, Buffalo, NY 14203, USADepartment of Urology, Roswell Park Comprehensive Cancer Center, Buffalo, NY 14203, USADepartment of Urology, Roswell Park Comprehensive Cancer Center, Buffalo, NY 14203, USADepartment of Biostatistics & Bioinformatics, Roswell Park Comprehensive Cancer Center, Buffalo, NY 14203, USADepartments of Cancer Prevention and Control, Roswell Park Comprehensive Cancer Center, Buffalo, NY 14203, USADepartment of Biostatistics & Bioinformatics, Roswell Park Comprehensive Cancer Center, Buffalo, NY 14203, USADepartments of Pathology, Roswell Park Comprehensive Cancer Center, Buffalo, NY 14203, USADepartments of Cancer Genetics & Genomics, Roswell Park Comprehensive Cancer Center, Buffalo, NY 14203, USADepartment of Pharmacology & Therapeutics, Roswell Park Comprehensive Cancer Center, Buffalo, NY 14203, USADepartment of Pharmacology & Therapeutics, Roswell Park Comprehensive Cancer Center, Buffalo, NY 14203, USADepartment of Urology, Roswell Park Comprehensive Cancer Center, Buffalo, NY 14203, USADepartment of Urology, Roswell Park Comprehensive Cancer Center, Buffalo, NY 14203, USADepartment of Biostatistics & Bioinformatics, Roswell Park Comprehensive Cancer Center, Buffalo, NY 14203, USADepartment of Pharmacology & Therapeutics, Roswell Park Comprehensive Cancer Center, Buffalo, NY 14203, USADepartment of Pharmacology & Therapeutics, Roswell Park Comprehensive Cancer Center, Buffalo, NY 14203, USADepartment of Urology, Roswell Park Comprehensive Cancer Center, Buffalo, NY 14203, USAApproximately 80% of patients with advanced bladder cancer do not respond to immune checkpoint inhibitor (ICI) immunotherapy. Therefore, there is an urgent unmet need to develop clinically relevant preclinical models so that factors governing immunotherapy responses can be studied in immunocompetent mice. We developed a line of mouse triple knockout (TKO: <i>Trp53</i>, <i>Pten</i>, <i>Rb1</i>) urothelial carcinoma organoids transplanted into immunocompetent mice. These bladder tumors recapitulate the molecular phenotypes and heterogeneous immunotherapy responses observed in human bladder cancers. The TKO organoids were characterized in vivo and in vitro and compared to the widely used MB49 murine bladder cancer model. RNAseq analysis of the TKO tumors demonstrated a basal subtype. The TKO xenografts demonstrated the expression of urothelial markers (CK5, CK7, GATA3, and p63), whereas MB49 subcutaneous xenografts did not express urothelial markers. Anti-PD-1 immunotherapy resulted in a mixed pattern of treatment responses for individual tumors. Eight immune cell types were identified (basophils, B cells, dendritic cells, macrophages, monocytes, neutrophils, NK cells, and T cells) in ICI-treated xenografts. Responder xenografts displayed significantly increased immune cell infiltration (15.3%, 742 immune cells/4861 total cells) compared to the non-responder tumors (10.1%, 452 immune cells/4459 total cells, Fisher Exact Test <i>p</i> < 0.0001). Specifically, there were more T cells (1.0% vs. 0.4%, <i>p</i> = 0.002) and macrophages (8.6% vs. 6.4%, <i>p</i> = 0.0002) in responder xenografts than in non-responder xenografts. In conclusion, we have developed a novel preclinical model that exhibits a mixed pattern of response to anti-PD-1 immunotherapy. The higher percentage of macrophage tumor infiltration in responders suggests a potential role for the innate immune microenvironment in regulating ICI treatment responses.https://www.mdpi.com/2072-6694/14/10/2511single-cell analysesmouse urothelial carcinomamouse modelICI immunotherapyanti-PD-1tumor-associated macrophages |
| spellingShingle | Dongbo Xu Li Wang Kyle Wieczorek Yali Zhang Zinian Wang Jianmin Wang Bo Xu Prashant K. Singh Yanqing Wang Xiaojing Zhang Yue Wu Gary J. Smith Kristopher Attwood Yuesheng Zhang David W. Goodrich Qiang Li Single-Cell Analyses of a Novel Mouse Urothelial Carcinoma Model Reveal a Role of Tumor-Associated Macrophages in Response to Anti-PD-1 Therapy Cancers single-cell analyses mouse urothelial carcinoma mouse model ICI immunotherapy anti-PD-1 tumor-associated macrophages |
| title | Single-Cell Analyses of a Novel Mouse Urothelial Carcinoma Model Reveal a Role of Tumor-Associated Macrophages in Response to Anti-PD-1 Therapy |
| title_full | Single-Cell Analyses of a Novel Mouse Urothelial Carcinoma Model Reveal a Role of Tumor-Associated Macrophages in Response to Anti-PD-1 Therapy |
| title_fullStr | Single-Cell Analyses of a Novel Mouse Urothelial Carcinoma Model Reveal a Role of Tumor-Associated Macrophages in Response to Anti-PD-1 Therapy |
| title_full_unstemmed | Single-Cell Analyses of a Novel Mouse Urothelial Carcinoma Model Reveal a Role of Tumor-Associated Macrophages in Response to Anti-PD-1 Therapy |
| title_short | Single-Cell Analyses of a Novel Mouse Urothelial Carcinoma Model Reveal a Role of Tumor-Associated Macrophages in Response to Anti-PD-1 Therapy |
| title_sort | single cell analyses of a novel mouse urothelial carcinoma model reveal a role of tumor associated macrophages in response to anti pd 1 therapy |
| topic | single-cell analyses mouse urothelial carcinoma mouse model ICI immunotherapy anti-PD-1 tumor-associated macrophages |
| url | https://www.mdpi.com/2072-6694/14/10/2511 |
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