A platform for multisite immune profiling of premetastatic pancreatic cancer at single-cell resolution
Abstract Background Pancreatic ductal adenocarcinoma (PDAC) is characterized by exceedingly high rates of metastatic progression, with the liver representing the most common site of distant spread. Here, we established a platform for multisite immune profiling of human PDAC encompassing the tumor, p...
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Springer
2025-08-01
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| Series: | Cancer Immunology, Immunotherapy |
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| Online Access: | https://doi.org/10.1007/s00262-025-04146-5 |
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| author | Elishama N. Kanu Ashley A. Fletcher Jiayin Bao Ethan S. Agritelley Julia Button Austin M. Eckhoff Karrie Comatas Tao Wang Bin-Jin Hwang Michael E. Lidsky Sabino Zani Dan G. Blazer Peter J. Allen Zhicheng Ji Frank J. Lowery Sri Krishna Nicholas D. Klemen Daniel P. Nussbaum Erika J. Crosby |
| author_facet | Elishama N. Kanu Ashley A. Fletcher Jiayin Bao Ethan S. Agritelley Julia Button Austin M. Eckhoff Karrie Comatas Tao Wang Bin-Jin Hwang Michael E. Lidsky Sabino Zani Dan G. Blazer Peter J. Allen Zhicheng Ji Frank J. Lowery Sri Krishna Nicholas D. Klemen Daniel P. Nussbaum Erika J. Crosby |
| author_sort | Elishama N. Kanu |
| collection | DOAJ |
| description | Abstract Background Pancreatic ductal adenocarcinoma (PDAC) is characterized by exceedingly high rates of metastatic progression, with the liver representing the most common site of distant spread. Here, we established a platform for multisite immune profiling of human PDAC encompassing the tumor, peripheral circulation, and premetastatic liver, to more comprehensively study how various immune subsets might contribute to patient outcomes. Methods Tumor, liver, and blood samples were obtained from patients undergoing resection for non-metastatic PDAC. Derived immune cells underwent paired single-cell RNA and TCR sequencing. Immune composition, cell-type functional profiles, and T cell clonal expansion patterns were evaluated across tissue sites. Results In total, 106,539 immune cells were sequenced, of which 85,748 met criteria for analysis. We identified 32 cell populations, of which seven demonstrated significant enrichment within a particular tissue, highlighting that this workflow possesses the granularity needed for identifying potential future biomarkers. Functional profiling revealed tissue-specific differences in cell phenotypes. This included terminally differentiated exhausted CD8 T cells within the tumor, highly active Tregs within the premetastatic liver and tumor, and M1 versus M2 polarization of liver and tumor macrophage populations, respectively. Within the tumor, expanded Treg clones were uniquely abundant, and while expanded clones could be tracked to the blood and premetastatic liver, many of these mapped back to known viral antigens. Leveraging previously validated gene sets, we show how these can be applied to predict the tumor reactivity of intratumoral T cells using transcriptional signatures. We demonstrated a high degree of concordance between multiple independent signatures and tracked high-priority TCRs within the blood and liver. Conclusion This study demonstrates the feasibility of a platform, which has already been implemented into ongoing clinical protocols, for immune profiling of human PDAC across the sites most relevant to metastatic progression. Future applications of this work can monitor immune populations throughout metastatic progression to build a temporal database of immune phenotypes and track association with clinical outcomes. |
| format | Article |
| id | doaj-art-14c29031d42e428b87cdd01c0436f438 |
| institution | Kabale University |
| issn | 1432-0851 |
| language | English |
| publishDate | 2025-08-01 |
| publisher | Springer |
| record_format | Article |
| series | Cancer Immunology, Immunotherapy |
| spelling | doaj-art-14c29031d42e428b87cdd01c0436f4382025-08-24T11:32:27ZengSpringerCancer Immunology, Immunotherapy1432-08512025-08-0174911610.1007/s00262-025-04146-5A platform for multisite immune profiling of premetastatic pancreatic cancer at single-cell resolutionElishama N. Kanu0Ashley A. Fletcher1Jiayin Bao2Ethan S. Agritelley3Julia Button4Austin M. Eckhoff5Karrie Comatas6Tao Wang7Bin-Jin Hwang8Michael E. Lidsky9Sabino Zani10Dan G. Blazer11Peter J. Allen12Zhicheng Ji13Frank J. Lowery14Sri Krishna15Nicholas D. Klemen16Daniel P. Nussbaum17Erika J. Crosby18Department of Surgery, Duke University Medical CenterDepartment of Surgery, Duke University Medical CenterDepartment of Surgery, Duke University Medical CenterDuke University School of Medicine, Duke UniversityDepartment of Surgery, Duke University Medical CenterDepartment of Surgery, Duke University Medical CenterDepartment of Surgery, Duke University Medical CenterDepartment of Surgery, Duke University Medical CenterDepartment of Surgery, Duke University Medical CenterDepartment of Surgery, Duke University Medical CenterDepartment of Surgery, Duke University Medical CenterDepartment of Surgery, Duke University Medical CenterDepartment of Surgery, Duke University Medical CenterDepartment of Biostatistics and Bioinformatics, Duke University School of Medicine, Duke UniversitySurgery Branch, Center for Cancer Research, National Cancer Institute, National Institutes of HealthSurgery Branch, Center for Cancer Research, National Cancer Institute, National Institutes of HealthSurgery Branch, Center for Cancer Research, National Cancer Institute, National Institutes of HealthDepartment of Surgery, Duke University Medical CenterDepartment of Surgery, Duke University Medical CenterAbstract Background Pancreatic ductal adenocarcinoma (PDAC) is characterized by exceedingly high rates of metastatic progression, with the liver representing the most common site of distant spread. Here, we established a platform for multisite immune profiling of human PDAC encompassing the tumor, peripheral circulation, and premetastatic liver, to more comprehensively study how various immune subsets might contribute to patient outcomes. Methods Tumor, liver, and blood samples were obtained from patients undergoing resection for non-metastatic PDAC. Derived immune cells underwent paired single-cell RNA and TCR sequencing. Immune composition, cell-type functional profiles, and T cell clonal expansion patterns were evaluated across tissue sites. Results In total, 106,539 immune cells were sequenced, of which 85,748 met criteria for analysis. We identified 32 cell populations, of which seven demonstrated significant enrichment within a particular tissue, highlighting that this workflow possesses the granularity needed for identifying potential future biomarkers. Functional profiling revealed tissue-specific differences in cell phenotypes. This included terminally differentiated exhausted CD8 T cells within the tumor, highly active Tregs within the premetastatic liver and tumor, and M1 versus M2 polarization of liver and tumor macrophage populations, respectively. Within the tumor, expanded Treg clones were uniquely abundant, and while expanded clones could be tracked to the blood and premetastatic liver, many of these mapped back to known viral antigens. Leveraging previously validated gene sets, we show how these can be applied to predict the tumor reactivity of intratumoral T cells using transcriptional signatures. We demonstrated a high degree of concordance between multiple independent signatures and tracked high-priority TCRs within the blood and liver. Conclusion This study demonstrates the feasibility of a platform, which has already been implemented into ongoing clinical protocols, for immune profiling of human PDAC across the sites most relevant to metastatic progression. Future applications of this work can monitor immune populations throughout metastatic progression to build a temporal database of immune phenotypes and track association with clinical outcomes.https://doi.org/10.1007/s00262-025-04146-5Single cell RNAseqPDACPre-metastatic liverTCRT cell clonality |
| spellingShingle | Elishama N. Kanu Ashley A. Fletcher Jiayin Bao Ethan S. Agritelley Julia Button Austin M. Eckhoff Karrie Comatas Tao Wang Bin-Jin Hwang Michael E. Lidsky Sabino Zani Dan G. Blazer Peter J. Allen Zhicheng Ji Frank J. Lowery Sri Krishna Nicholas D. Klemen Daniel P. Nussbaum Erika J. Crosby A platform for multisite immune profiling of premetastatic pancreatic cancer at single-cell resolution Cancer Immunology, Immunotherapy Single cell RNAseq PDAC Pre-metastatic liver TCR T cell clonality |
| title | A platform for multisite immune profiling of premetastatic pancreatic cancer at single-cell resolution |
| title_full | A platform for multisite immune profiling of premetastatic pancreatic cancer at single-cell resolution |
| title_fullStr | A platform for multisite immune profiling of premetastatic pancreatic cancer at single-cell resolution |
| title_full_unstemmed | A platform for multisite immune profiling of premetastatic pancreatic cancer at single-cell resolution |
| title_short | A platform for multisite immune profiling of premetastatic pancreatic cancer at single-cell resolution |
| title_sort | platform for multisite immune profiling of premetastatic pancreatic cancer at single cell resolution |
| topic | Single cell RNAseq PDAC Pre-metastatic liver TCR T cell clonality |
| url | https://doi.org/10.1007/s00262-025-04146-5 |
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