Generation of a double binary transgenic zebrafish model to study myeloid gene regulation in response to oncogene activation in melanocytes
A complex network of inflammatory genes is closely linked to somatic cell transformation and malignant disease. Immune cells and their associated molecules are responsible for detecting and eliminating cancer cells as they establish themselves as the precursors of a tumour. By the time a patient has...
| Main Authors: | , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
The Company of Biologists
2018-04-01
|
| Series: | Disease Models & Mechanisms |
| Subjects: | |
| Online Access: | http://dmm.biologists.org/content/11/4/dmm030056 |
| _version_ | 1818143412416675840 |
|---|---|
| author | Amy Kenyon Daria Gavriouchkina Jernej Zorman Vanessa Chong-Morrison Giorgio Napolitani Vincenzo Cerundolo Tatjana Sauka-Spengler |
| author_facet | Amy Kenyon Daria Gavriouchkina Jernej Zorman Vanessa Chong-Morrison Giorgio Napolitani Vincenzo Cerundolo Tatjana Sauka-Spengler |
| author_sort | Amy Kenyon |
| collection | DOAJ |
| description | A complex network of inflammatory genes is closely linked to somatic cell transformation and malignant disease. Immune cells and their associated molecules are responsible for detecting and eliminating cancer cells as they establish themselves as the precursors of a tumour. By the time a patient has a detectable solid tumour, cancer cells have escaped the initial immune response mechanisms. Here, we describe the development of a double binary zebrafish model that enables regulatory programming of the myeloid cells as they respond to oncogene-activated melanocytes to be explored, focussing on the initial phase when cells become the precursors of cancer. A hormone-inducible binary system allows for temporal control of expression of different Ras oncogenes (NRasQ61K, HRasG12V and KRasG12V) in melanocytes, leading to proliferation and changes in morphology of the melanocytes. This model was coupled to binary cell-specific biotagging models allowing in vivo biotinylation and subsequent isolation of macrophage or neutrophil nuclei for regulatory profiling of their active transcriptomes. Nuclear transcriptional profiling of neutrophils, performed as they respond to the earliest precursors of melanoma in vivo, revealed an intricate landscape of regulatory factors that may promote progression to melanoma, including Serpinb1l4, Fgf1, Fgf6, Cathepsin H, Galectin 1 and Galectin 3. The model presented here provides a powerful platform to study the myeloid response to the earliest precursors of melanoma. |
| first_indexed | 2024-12-11T11:31:15Z |
| format | Article |
| id | doaj.art-fb7f3c33b46e46eba6f4c95d6c67fff1 |
| institution | Directory Open Access Journal |
| issn | 1754-8403 1754-8411 |
| language | English |
| last_indexed | 2024-12-11T11:31:15Z |
| publishDate | 2018-04-01 |
| publisher | The Company of Biologists |
| record_format | Article |
| series | Disease Models & Mechanisms |
| spelling | doaj.art-fb7f3c33b46e46eba6f4c95d6c67fff12022-12-22T01:08:52ZengThe Company of BiologistsDisease Models & Mechanisms1754-84031754-84112018-04-0111410.1242/dmm.030056030056Generation of a double binary transgenic zebrafish model to study myeloid gene regulation in response to oncogene activation in melanocytesAmy Kenyon0Daria Gavriouchkina1Jernej Zorman2Vanessa Chong-Morrison3Giorgio Napolitani4Vincenzo Cerundolo5Tatjana Sauka-Spengler6 University of Oxford, Weatherall Institute of Molecular Medicine, Radcliffe Department of Medicine, Oxford OX3 9DS, United Kingdom University of Oxford, Weatherall Institute of Molecular Medicine, Radcliffe Department of Medicine, Oxford OX3 9DS, United Kingdom University of Oxford, Weatherall Institute of Molecular Medicine, Radcliffe Department of Medicine, Oxford OX3 9DS, United Kingdom University of Oxford, Weatherall Institute of Molecular Medicine, Radcliffe Department of Medicine, Oxford OX3 9DS, United Kingdom University of Oxford, Weatherall Institute of Molecular Medicine, MRC Human Immunology Unit, Radcliffe Department of Medicine, Oxford OX3 9DS, United Kingdom University of Oxford, Weatherall Institute of Molecular Medicine, MRC Human Immunology Unit, Radcliffe Department of Medicine, Oxford OX3 9DS, United Kingdom University of Oxford, Weatherall Institute of Molecular Medicine, Radcliffe Department of Medicine, Oxford OX3 9DS, United Kingdom A complex network of inflammatory genes is closely linked to somatic cell transformation and malignant disease. Immune cells and their associated molecules are responsible for detecting and eliminating cancer cells as they establish themselves as the precursors of a tumour. By the time a patient has a detectable solid tumour, cancer cells have escaped the initial immune response mechanisms. Here, we describe the development of a double binary zebrafish model that enables regulatory programming of the myeloid cells as they respond to oncogene-activated melanocytes to be explored, focussing on the initial phase when cells become the precursors of cancer. A hormone-inducible binary system allows for temporal control of expression of different Ras oncogenes (NRasQ61K, HRasG12V and KRasG12V) in melanocytes, leading to proliferation and changes in morphology of the melanocytes. This model was coupled to binary cell-specific biotagging models allowing in vivo biotinylation and subsequent isolation of macrophage or neutrophil nuclei for regulatory profiling of their active transcriptomes. Nuclear transcriptional profiling of neutrophils, performed as they respond to the earliest precursors of melanoma in vivo, revealed an intricate landscape of regulatory factors that may promote progression to melanoma, including Serpinb1l4, Fgf1, Fgf6, Cathepsin H, Galectin 1 and Galectin 3. The model presented here provides a powerful platform to study the myeloid response to the earliest precursors of melanoma.http://dmm.biologists.org/content/11/4/dmm030056BiotaggingMacrophageMelanocyteNeutrophilOncogeneZebrafish |
| spellingShingle | Amy Kenyon Daria Gavriouchkina Jernej Zorman Vanessa Chong-Morrison Giorgio Napolitani Vincenzo Cerundolo Tatjana Sauka-Spengler Generation of a double binary transgenic zebrafish model to study myeloid gene regulation in response to oncogene activation in melanocytes Disease Models & Mechanisms Biotagging Macrophage Melanocyte Neutrophil Oncogene Zebrafish |
| title | Generation of a double binary transgenic zebrafish model to study myeloid gene regulation in response to oncogene activation in melanocytes |
| title_full | Generation of a double binary transgenic zebrafish model to study myeloid gene regulation in response to oncogene activation in melanocytes |
| title_fullStr | Generation of a double binary transgenic zebrafish model to study myeloid gene regulation in response to oncogene activation in melanocytes |
| title_full_unstemmed | Generation of a double binary transgenic zebrafish model to study myeloid gene regulation in response to oncogene activation in melanocytes |
| title_short | Generation of a double binary transgenic zebrafish model to study myeloid gene regulation in response to oncogene activation in melanocytes |
| title_sort | generation of a double binary transgenic zebrafish model to study myeloid gene regulation in response to oncogene activation in melanocytes |
| topic | Biotagging Macrophage Melanocyte Neutrophil Oncogene Zebrafish |
| url | http://dmm.biologists.org/content/11/4/dmm030056 |
| work_keys_str_mv | AT amykenyon generationofadoublebinarytransgeniczebrafishmodeltostudymyeloidgeneregulationinresponsetooncogeneactivationinmelanocytes AT dariagavriouchkina generationofadoublebinarytransgeniczebrafishmodeltostudymyeloidgeneregulationinresponsetooncogeneactivationinmelanocytes AT jernejzorman generationofadoublebinarytransgeniczebrafishmodeltostudymyeloidgeneregulationinresponsetooncogeneactivationinmelanocytes AT vanessachongmorrison generationofadoublebinarytransgeniczebrafishmodeltostudymyeloidgeneregulationinresponsetooncogeneactivationinmelanocytes AT giorgionapolitani generationofadoublebinarytransgeniczebrafishmodeltostudymyeloidgeneregulationinresponsetooncogeneactivationinmelanocytes AT vincenzocerundolo generationofadoublebinarytransgeniczebrafishmodeltostudymyeloidgeneregulationinresponsetooncogeneactivationinmelanocytes AT tatjanasaukaspengler generationofadoublebinarytransgeniczebrafishmodeltostudymyeloidgeneregulationinresponsetooncogeneactivationinmelanocytes |