Shugo Sasaki1,2,3,4, Michelle Y Y Lee5, Yuka Wakabayashi6, Luka Suzuki6, Helena Winata5, Miwa Himuro6, Taka-Aki Matsuoka7, Iichiro Shimomura7, Hirotaka Watada6,8, Francis C Lynn9,10, Takeshi Miyatsuka11,12,13. 1. Department of Metabolic Medicine, Osaka University Graduate School of Medicine, Osaka, Japan. shugosasaki@endmet.med.osaka-u.ac.jp. 2. Department of Diabetes Care Medicine, Osaka University Graduate School of Medicine, Osaka, Japan. shugosasaki@endmet.med.osaka-u.ac.jp. 3. Diabetes Research Group, BC Children's Hospital Research Institute, Vancouver, BC, Canada. shugosasaki@endmet.med.osaka-u.ac.jp. 4. Department of Surgery, School of Biomedical Engineering, University of British Columbia, Vancouver, BC, Canada. shugosasaki@endmet.med.osaka-u.ac.jp. 5. Diabetes Research Group, BC Children's Hospital Research Institute, Vancouver, BC, Canada. 6. Department of Metabolism and Endocrinology, Juntendo University Graduate School of Medicine, Tokyo, Japan. 7. Department of Metabolic Medicine, Osaka University Graduate School of Medicine, Osaka, Japan. 8. Center for Identification of Diabetic Therapeutic Targets, Juntendo University Graduate School of Medicine, Tokyo, Japan. 9. Diabetes Research Group, BC Children's Hospital Research Institute, Vancouver, BC, Canada. francis.lynn@ubc.ca. 10. Department of Surgery, School of Biomedical Engineering, University of British Columbia, Vancouver, BC, Canada. francis.lynn@ubc.ca. 11. Department of Metabolism and Endocrinology, Juntendo University Graduate School of Medicine, Tokyo, Japan. miyatsuka-takeshi@umin.net. 12. Center for Identification of Diabetic Therapeutic Targets, Juntendo University Graduate School of Medicine, Tokyo, Japan. miyatsuka-takeshi@umin.net. 13. Department of Endocrinology, Diabetes and Metabolism, Kitasato University School of Medicine, Kanagawa, Japan. miyatsuka-takeshi@umin.net.
Abstract
AIMS/HYPOTHESIS: While pancreatic beta cells have been shown to originate from endocrine progenitors in ductal regions, it remains unclear precisely where beta cells emerge from and which transcripts define newborn beta cells. We therefore investigated characteristics of newborn beta cells extracted by a time-resolved reporter system. METHODS: We established a mouse model, 'Ins1-GFP; Timer', which provides spatial information during beta cell neogenesis with high temporal resolution. Single-cell RNA-sequencing (scRNA-seq) was performed on mouse beta cells sorted by fluorescent reporter to uncover transcriptomic profiles of newborn beta cells. scRNA-seq of human embryonic stem cell (hESC)-derived beta-like cells was also performed to compare newborn beta cell features between mouse and human. RESULTS: Fluorescence imaging of Ins1-GFP; Timer mouse pancreas successfully dissected newly generated beta cells as green fluorescence-dominant cells. This reporter system revealed that, as expected, some newborn beta cells arise close to the ducts (βduct); unexpectedly, the others arise away from the ducts and adjacent to blood vessels (βvessel). Single-cell transcriptomic analyses demonstrated five distinct populations among newborn beta cells, confirming spatial heterogeneity of beta cell neogenesis such as high probability of glucagon-positive βduct, musculoaponeurotic fibrosarcoma oncogene family B (MafB)-positive βduct and musculoaponeurotic fibrosarcoma oncogene family A (MafA)-positive βvessel cells. Comparative analysis with scRNA-seq data of mouse newborn beta cells and hESC-derived beta-like cells uncovered transcriptional similarity between mouse and human beta cell neogenesis including microsomal glutathione S-transferase 1 (MGST1)- and synaptotagmin 13 (SYT13)-highly-expressing state. CONCLUSIONS/ INTERPRETATION: The combination of time-resolved histological imaging with single-cell transcriptional mapping demonstrated novel features of spatial and transcriptional heterogeneity in beta cell neogenesis, which will lead to a better understanding of beta cell differentiation for future cell therapy. DATA AVAILABILITY: Raw and processed single-cell RNA-sequencing data for this study has been deposited in the Gene Expression Omnibus under accession number GSE155742.
AIMS/HYPOTHESIS: While pancreatic beta cells have been shown to originate from endocrine progenitors in ductal regions, it remains unclear precisely where beta cells emerge from and which transcripts define newborn beta cells. We therefore investigated characteristics of newborn beta cells extracted by a time-resolved reporter system. METHODS: We established a mouse model, 'Ins1-GFP; Timer', which provides spatial information during beta cell neogenesis with high temporal resolution. Single-cell RNA-sequencing (scRNA-seq) was performed on mouse beta cells sorted by fluorescent reporter to uncover transcriptomic profiles of newborn beta cells. scRNA-seq of human embryonic stem cell (hESC)-derived beta-like cells was also performed to compare newborn beta cell features between mouse and human. RESULTS: Fluorescence imaging of Ins1-GFP; Timer mouse pancreas successfully dissected newly generated beta cells as green fluorescence-dominant cells. This reporter system revealed that, as expected, some newborn beta cells arise close to the ducts (βduct); unexpectedly, the others arise away from the ducts and adjacent to blood vessels (βvessel). Single-cell transcriptomic analyses demonstrated five distinct populations among newborn beta cells, confirming spatial heterogeneity of beta cell neogenesis such as high probability of glucagon-positive βduct, musculoaponeurotic fibrosarcoma oncogene family B (MafB)-positive βduct and musculoaponeurotic fibrosarcoma oncogene family A (MafA)-positive βvessel cells. Comparative analysis with scRNA-seq data of mouse newborn beta cells and hESC-derived beta-like cells uncovered transcriptional similarity between mouse and human beta cell neogenesis including microsomal glutathione S-transferase 1 (MGST1)- and synaptotagmin 13 (SYT13)-highly-expressing state. CONCLUSIONS/ INTERPRETATION: The combination of time-resolved histological imaging with single-cell transcriptional mapping demonstrated novel features of spatial and transcriptional heterogeneity in beta cell neogenesis, which will lead to a better understanding of beta cell differentiation for future cell therapy. DATA AVAILABILITY: Raw and processed single-cell RNA-sequencing data for this study has been deposited in the Gene Expression Omnibus under accession number GSE155742.
Authors: Manami Hara; Xiaoyu Wang; Toshihiko Kawamura; Vytas P Bindokas; Restituto F Dizon; Sergio Y Alcoser; Mark A Magnuson; Graeme I Bell Journal: Am J Physiol Endocrinol Metab Date: 2002-09-17 Impact factor: 4.310
Authors: Martin Enge; H Efsun Arda; Marco Mignardi; John Beausang; Rita Bottino; Seung K Kim; Stephen R Quake Journal: Cell Date: 2017-09-28 Impact factor: 41.582
Authors: V M Schwitzgebel; D W Scheel; J R Conners; J Kalamaras; J E Lee; D J Anderson; L Sussel; J D Johnson; M S German Journal: Development Date: 2000-08 Impact factor: 6.868