CFTR Expression Analysis for Subtyping of Human Pancreatic Cancer Organoids
Background. Organoid cultures of human pancreatic ductal adenocarcinoma (PDAC) have become a promising tool for tumor subtyping and individualized chemosensitivity testing. PDACs have recently been grouped into different molecular subtypes with clinical impact based on cytokeratin-81 (KRT81) and hep...
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| Language: | English |
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Wiley
2019-01-01
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| Series: | Stem Cells International |
| Online Access: | http://dx.doi.org/10.1155/2019/1024614 |
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| author | Alexander Hennig Laura Wolf Beatrix Jahnke Heike Polster Therese Seidlitz Kristin Werner Daniela E. Aust Jochen Hampe Marius Distler Jürgen Weitz Daniel E. Stange Thilo Welsch |
| author_facet | Alexander Hennig Laura Wolf Beatrix Jahnke Heike Polster Therese Seidlitz Kristin Werner Daniela E. Aust Jochen Hampe Marius Distler Jürgen Weitz Daniel E. Stange Thilo Welsch |
| author_sort | Alexander Hennig |
| collection | DOAJ |
| description | Background. Organoid cultures of human pancreatic ductal adenocarcinoma (PDAC) have become a promising tool for tumor subtyping and individualized chemosensitivity testing. PDACs have recently been grouped into different molecular subtypes with clinical impact based on cytokeratin-81 (KRT81) and hepatocyte nuclear factor 1A (HNF1A). However, a suitable antibody for HNF1A is currently unavailable. The present study is aimed at establishing subtyping in PDAC organoids using an alternative marker. Methods. A PDAC organoid biobank was generated from human primary tumor samples containing 22 lines. Immunofluorescence staining was established and done for 10 organoid lines for cystic fibrosis transmembrane conductance regulator (CFTR) and KRT81. Quantitative real-time PCR (qPCR) was performed for CFTR and HNF1A. A chemotherapeutic drug response analysis was done using gemcitabine, 5-FU, oxaliplatin, and irinotecan. Results. A biobank of patient-derived PDAC organoids was established. The efficiency was 71% (22/31) with 68% for surgical resections and 83% for fine needle aspirations. Organoids could be categorized into the established quasimesenchymal, exocrine-like, and classical subtypes based on KRT81 and CFTR immunoreactivity. CFTR protein expression was confirmed on the transcript level. CFTR and HNF1A transcript expression levels positively correlated (n=10; r=0.927; p=0.001). PDAC subtypes of the primary tumors and the corresponding organoid lines were identical for most of the cases analyzed (6/7). Treatment with chemotherapeutic drugs revealed tendencies but no significant differences regarding drug responses. Conclusions. Human PDAC organoids can be classified into known subtypes based on KRT81 and CFTR immunoreactivity. CFTR and HNF1A mRNA levels correlated well. Furthermore, subtype-specific immunoreactivity matched well between PDAC organoids and the respective primary tumor tissue. Subtyping of human PDACs using CFTR might constitute an alternative to HNF1A and should be further investigated. |
| format | Article |
| id | doaj-art-0564d3d4636847e09cc31b2fcf70f1d3 |
| institution | Kabale University |
| issn | 1687-966X 1687-9678 |
| language | English |
| publishDate | 2019-01-01 |
| publisher | Wiley |
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| series | Stem Cells International |
| spelling | doaj-art-0564d3d4636847e09cc31b2fcf70f1d32025-02-03T06:13:08ZengWileyStem Cells International1687-966X1687-96782019-01-01201910.1155/2019/10246141024614CFTR Expression Analysis for Subtyping of Human Pancreatic Cancer OrganoidsAlexander Hennig0Laura Wolf1Beatrix Jahnke2Heike Polster3Therese Seidlitz4Kristin Werner5Daniela E. Aust6Jochen Hampe7Marius Distler8Jürgen Weitz9Daniel E. Stange10Thilo Welsch11Department of Visceral, Thoracic and Vascular Surgery, Medical Faculty and University Hospital Carl Gustav Carus, Technische Universität Dresden, Dresden, GermanyDepartment of Visceral, Thoracic and Vascular Surgery, Medical Faculty and University Hospital Carl Gustav Carus, Technische Universität Dresden, Dresden, GermanyDepartment of Visceral, Thoracic and Vascular Surgery, Medical Faculty and University Hospital Carl Gustav Carus, Technische Universität Dresden, Dresden, GermanyDepartment of Visceral, Thoracic and Vascular Surgery, Medical Faculty and University Hospital Carl Gustav Carus, Technische Universität Dresden, Dresden, GermanyDepartment of Visceral, Thoracic and Vascular Surgery, Medical Faculty and University Hospital Carl Gustav Carus, Technische Universität Dresden, Dresden, GermanyDepartment of Visceral, Thoracic and Vascular Surgery, Medical Faculty and University Hospital Carl Gustav Carus, Technische Universität Dresden, Dresden, GermanyInstitute of Pathology and Tumour- and Normal Tissue Bank of the University Cancer Center (UCC), University Hospital Carl Gustav Carus, Medical Faculty, Technische Universität Dresden, Dresden, GermanyMedical Department I, Medical Faculty and University Hospital Carl Gustav Carus, Technische Universität Dresden, Dresden, GermanyDepartment of Visceral, Thoracic and Vascular Surgery, Medical Faculty and University Hospital Carl Gustav Carus, Technische Universität Dresden, Dresden, GermanyDepartment of Visceral, Thoracic and Vascular Surgery, Medical Faculty and University Hospital Carl Gustav Carus, Technische Universität Dresden, Dresden, GermanyDepartment of Visceral, Thoracic and Vascular Surgery, Medical Faculty and University Hospital Carl Gustav Carus, Technische Universität Dresden, Dresden, GermanyDepartment of Visceral, Thoracic and Vascular Surgery, Medical Faculty and University Hospital Carl Gustav Carus, Technische Universität Dresden, Dresden, GermanyBackground. Organoid cultures of human pancreatic ductal adenocarcinoma (PDAC) have become a promising tool for tumor subtyping and individualized chemosensitivity testing. PDACs have recently been grouped into different molecular subtypes with clinical impact based on cytokeratin-81 (KRT81) and hepatocyte nuclear factor 1A (HNF1A). However, a suitable antibody for HNF1A is currently unavailable. The present study is aimed at establishing subtyping in PDAC organoids using an alternative marker. Methods. A PDAC organoid biobank was generated from human primary tumor samples containing 22 lines. Immunofluorescence staining was established and done for 10 organoid lines for cystic fibrosis transmembrane conductance regulator (CFTR) and KRT81. Quantitative real-time PCR (qPCR) was performed for CFTR and HNF1A. A chemotherapeutic drug response analysis was done using gemcitabine, 5-FU, oxaliplatin, and irinotecan. Results. A biobank of patient-derived PDAC organoids was established. The efficiency was 71% (22/31) with 68% for surgical resections and 83% for fine needle aspirations. Organoids could be categorized into the established quasimesenchymal, exocrine-like, and classical subtypes based on KRT81 and CFTR immunoreactivity. CFTR protein expression was confirmed on the transcript level. CFTR and HNF1A transcript expression levels positively correlated (n=10; r=0.927; p=0.001). PDAC subtypes of the primary tumors and the corresponding organoid lines were identical for most of the cases analyzed (6/7). Treatment with chemotherapeutic drugs revealed tendencies but no significant differences regarding drug responses. Conclusions. Human PDAC organoids can be classified into known subtypes based on KRT81 and CFTR immunoreactivity. CFTR and HNF1A mRNA levels correlated well. Furthermore, subtype-specific immunoreactivity matched well between PDAC organoids and the respective primary tumor tissue. Subtyping of human PDACs using CFTR might constitute an alternative to HNF1A and should be further investigated.http://dx.doi.org/10.1155/2019/1024614 |
| spellingShingle | Alexander Hennig Laura Wolf Beatrix Jahnke Heike Polster Therese Seidlitz Kristin Werner Daniela E. Aust Jochen Hampe Marius Distler Jürgen Weitz Daniel E. Stange Thilo Welsch CFTR Expression Analysis for Subtyping of Human Pancreatic Cancer Organoids Stem Cells International |
| title | CFTR Expression Analysis for Subtyping of Human Pancreatic Cancer Organoids |
| title_full | CFTR Expression Analysis for Subtyping of Human Pancreatic Cancer Organoids |
| title_fullStr | CFTR Expression Analysis for Subtyping of Human Pancreatic Cancer Organoids |
| title_full_unstemmed | CFTR Expression Analysis for Subtyping of Human Pancreatic Cancer Organoids |
| title_short | CFTR Expression Analysis for Subtyping of Human Pancreatic Cancer Organoids |
| title_sort | cftr expression analysis for subtyping of human pancreatic cancer organoids |
| url | http://dx.doi.org/10.1155/2019/1024614 |
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