A Pro-Endocrine Pancreatic Islet Transcriptional Program Established During Development Is Retained in Human Gallbladder Epithelial Cells

Mugdha V. Joglekar; Subhshri Sahu; Wilson K.M. Wong; Sarang N. Satoor; Charlotte X. Dong; Ryan J. Farr; Michael D. Williams; Prapti Pandya; Gaurang Jhala; Sundy N.Y. Yang; Yi Vee Chew; Nicola Hetherington; Dhan Thiruchevlam; Sasikala Mitnala; Guduru V. Rao; Duvvuru Nageshwar Reddy; Thomas Loudovaris; Wayne J. Hawthorne; Andrew G. Elefanty; Vinay M. Joglekar; Edouard G. Stanley; David Martin; Helen E. Thomas; David Tosh; Louise T. Dalgaard; Anandwardhan A. Hardikar

doi:10.1016/j.jcmgh.2022.01.008

A Pro-Endocrine Pancreatic Islet Transcriptional Program Established During Development Is Retained in Human Gallbladder Epithelial Cells

Mugdha V. Joglekar, Subhshri Sahu, Wilson K.M. Wong, Sarang N. Satoor, Charlotte X. Dong, Ryan J. Farr, Michael D. Williams, Prapti Pandya, Gaurang Jhala, Sundy N.Y. Yang, Yi Vee Chew, Nicola Hetherington, Dhan Thiruchevlam, Sasikala Mitnala, Guduru V. Rao, Duvvuru Nageshwar Reddy, Thomas Loudovaris, Wayne J. Hawthorne, Andrew G. Elefanty, Vinay M. JoglekarEdouard G. Stanley, David Martin, Helen E. Thomas, David Tosh, Louise T. Dalgaard, Anandwardhan A. Hardikar

Research output: Contribution to journal › Article › peer-review

Abstract

Background & Aims: Pancreatic islet β-cells are factories for insulin production; however, ectopic expression of insulin also is well recognized. The gallbladder is a next-door neighbor to the developing pancreas. Here, we wanted to understand if gallbladders contain functional insulin-producing cells. Methods: We compared developing and adult mouse as well as human gallbladder epithelial cells and islets using immunohistochemistry, flow cytometry, enzyme-linked immunosorbent assays, RNA sequencing, real-time polymerase chain reaction, chromatin immunoprecipitation, and functional studies. Results: We show that the epithelial lining of developing, as well as adult, mouse and human gallbladders naturally contain interspersed cells that retain the capacity to actively transcribe, translate, package, and release insulin. We show that human gallbladders also contain functional insulin-secreting cells with the potential to naturally respond to glucose in vitro and in situ. Notably, in a non-obese diabetic (NOD) mouse model of type 1 diabetes, we observed that insulin-producing cells in the gallbladder are not targeted by autoimmune cells. Interestingly, in human gallbladders, insulin splice variants are absent, although insulin splice forms are observed in human islets. Conclusions: In summary, our biochemical, transcriptomic, and functional data in mouse and human gallbladder epithelial cells collectively show the evolutionary and developmental similarities between gallbladder and the pancreas that allow gallbladder epithelial cells to continue insulin production in adult life. Understanding the mechanisms regulating insulin transcription and translation in gallbladder epithelial cells would help guide future studies in type 1 diabetes therapy.

Original language	English
Pages (from-to)	1530-1553.e4
Number of pages	23
Journal	Cellular and Molecular Gastroenterology and Hepatology
Volume	13
Issue number	5
Early online date	12 Jan 2022
DOIs	https://doi.org/10.1016/j.jcmgh.2022.01.008
Publication status	Published - 31 Dec 2022

Bibliographical note

Funding Information:
Funding Supported mainly by a 2008–2009 British Council (UK–India Educational Research Initiative) exchange program (A.A.H. and D.T.), a 2012–2015 National Health and Medical Research Council project grant (A.A.H.), and the Danish Diabetes Academy visiting professorship (A.A.H. and L.T.D.). Also funded by the Australian Research Council (2012–2016) and the Juvenile Diabetes Research Foundation (JDRF) Australian Type 1 Diabetes Clinical Research Network (grant 4-CDA2016-228-MB; a special initiative of the Australian Research Council) (A.A.H.), the JDRF USA postdoctoral fellowship (2012–2014), advanced post-doctoral fellowship (2015–2018), and the JDRF career transition award (2019–2021) (M.V.J.). The University of Sydney, Australian postgraduate award (University Post-graduate Award or Australian Post-graduate Award), and JDRF Australia PhD top-up awards (W.K.M.W. and R.J.F.), and Council of Scientific and Industrial Research fellowships (Government of India) (S.S.) are acknowledged. W.K.M.W. currently is funded through a grant from the Leona M. and Harry B. Helmsley Charitable Trust (grant 2018PG-T1D009) in collaboration with the JDRF Australian Type 1 Diabetes Clinical Research Network grant 3-SRA-2019-694-M-B (A.A.H.). Also supported by the Summer Scholarship from the National Health and Medical Research Council Clinical Trials Centre, University of Sydney (P.P.).

Funding Information:
The authors acknowledge and are thankful for the support from surgical teams, consenting donors or families of the donors, as well as infrastructure support obtained through the Rebecca L Cooper Foundation (A.A.H.) and provided by the National Centre for Cell Science, India and the Faculty of Medicine and Health, University of Sydney. All authors acknowledge the assistance from Dr Andrew M. Holland and Dr Suzanne J. Micallef in the provision of the Pdx1 GFP/w reporter mice, Professor Manami Hara for the MIP-GFP mice, and the support from Professor Jonathan Slack and Professor Harry Heimberg for the viral vectors used in the study. The authors also acknowledge support from Dr Ramesh Dumbre, Dr Pabitra Sahoo, Ms Fahmida Khan, Ms Sophie Breedveld, Ms Mariah Taleb, RPA Pathology Services, Bosch Facilities (Bosch Mass Spectometry Facility and Live Cell Analysis Facility) at the University of Sydney. A.G.E. and E.G.S were supported by Australian National and Health and Medical Research Council grants (GNT1079004, GNT1117596, GNT1129861, GNT1138717, GNT1123277), and by the Stafford Fox Medical Research Foundation. Additional infrastructure funding to the Murdoch Children's Research Institute was provided by the Australian Government National Health and Medical Research Council Independent Research Institute Infrastructure Support Scheme and the Victorian Government's Operational Infrastructure Support Program.

Funding Information:
The authors acknowledge and are thankful for the support from surgical teams, consenting donors or families of the donors, as well as infrastructure support obtained through the Rebecca L Cooper Foundation (A.A.H.) and provided by the National Centre for Cell Science, India and the Faculty of Medicine and Health, University of Sydney. All authors acknowledge the assistance from Dr Andrew M. Holland and Dr Suzanne J. Micallef in the provision of the Pdx1GFP/w reporter mice, Professor Manami Hara for the MIP-GFP mice, and the support from Professor Jonathan Slack and Professor Harry Heimberg for the viral vectors used in the study. The authors also acknowledge support from Dr Ramesh Dumbre, Dr Pabitra Sahoo, Ms Fahmida Khan, Ms Sophie Breedveld, Ms Mariah Taleb, RPA Pathology Services, Bosch Facilities (Bosch Mass Spectometry Facility and Live Cell Analysis Facility) at the University of Sydney. A.G.E. and E.G.S were supported by Australian National and Health and Medical Research Council grants (GNT1079004, GNT1117596, GNT1129861, GNT1138717, GNT1123277), and by the Stafford Fox Medical Research Foundation. Additional infrastructure funding to the Murdoch Children's Research Institute was provided by the Australian Government National Health and Medical Research Council Independent Research Institute Infrastructure Support Scheme and the Victorian Government's Operational Infrastructure Support Program. Mugdha V. Joglekar (Conceptualization: Supporting; Data curation: Equal; Formal analysis: Equal; Investigation: Lead; Methodology: Equal; Software: Equal; Supervision: Equal; Writing ? original draft: Lead; Writing ? review & editing: Equal), Subhshri Sahu (Data curation: Equal; Formal analysis: Equal; Methodology: Equal; Software: Supporting; Writing ? review & editing: Equal), Wilson KM Wong (Data curation: Equal; Formal analysis: Equal; Methodology: Supporting; Software: Equal; Validation: Equal; Writing ? review & editing: Equal), Sarang N. Satoor (Data curation: Supporting; Formal analysis: Supporting; Methodology: Supporting), Charlotte X. Dong (Software: Supporting; Validation: Supporting), Ryan J Farr (Data curation: Supporting; Methodology: Supporting), Michael D. Williams (Data curation: Supporting), Prapti Pandya (Data curation: Supporting), Gaurang Jhala (Data curation: Supporting; Resources: Supporting), Sundy N.Y. Yang (Data curation: Supporting), Yi Vee Chew (Data curation: Supporting), Nicola Hetherington (Data curation: Supporting), Dhan Thiruchevlam (Resources: Supporting), Sasikala Mitnala (Data curation: Supporting; Resources: Supporting), Guduru V Rao (Resources: Supporting), Duvvuru Nageshwar Reddy (Resources: Supporting), Thomas Loudovaris (Resources: Supporting), Wayne J. Hawthorne (Resources: Supporting), Andrew G. Elefanty (Resources: Supporting), Vinay M. Joglekar (Resources: Supporting), Edouard G. Stanley (Resources: Supporting), David Martin (Resources: Supporting), Helen E. Thomas (Resources: Supporting), David Tosh (Conceptualization: Supporting; Funding acquisition: Supporting; Supervision: Supporting; Writing ? original draft: Supporting; Writing ? review & editing: Supporting), Louise T. Dalgaard (Software: Equal; Validation: Equal; Writing ? review & editing: Supporting), Anandwardhan A. Hardikar (Conceptualization: Lead; Data curation: Supporting; Funding acquisition: Lead; Investigation: Supporting; Project administration: Lead; Supervision: Lead; Visualization: Supporting; Writing ? original draft: Equal; Writing ? review & editing: Equal), Sequencing data are available through GEO data sets cited in text.

Data Availability Statement Sequencing data are available through GEO data sets cited in text.

Keywords

Differentiation
Gallbladder Development
Insulin
Islets
Splice Variants

ASJC Scopus subject areas

Hepatology
Gastroenterology

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

Access to Document

10.1016/j.jcmgh.2022.01.008Licence: CC BY-NC-ND

Cite this

Joglekar, M. V., Sahu, S., Wong, W. K. M., Satoor, S. N., Dong, C. X., Farr, R. J., Williams, M. D., Pandya, P., Jhala, G., Yang, S. N. Y., Chew, Y. V., Hetherington, N., Thiruchevlam, D., Mitnala, S., Rao, G. V., Reddy, D. N., Loudovaris, T., Hawthorne, W. J., Elefanty, A. G., ... Hardikar, A. A. (2022). A Pro-Endocrine Pancreatic Islet Transcriptional Program Established During Development Is Retained in Human Gallbladder Epithelial Cells. Cellular and Molecular Gastroenterology and Hepatology, 13(5), 1530-1553.e4. https://doi.org/10.1016/j.jcmgh.2022.01.008

A Pro-Endocrine Pancreatic Islet Transcriptional Program Established During Development Is Retained in Human Gallbladder Epithelial Cells. / Joglekar, Mugdha V.; Sahu, Subhshri; Wong, Wilson K.M. et al.
In: Cellular and Molecular Gastroenterology and Hepatology, Vol. 13, No. 5, 31.12.2022, p. 1530-1553.e4.

Research output: Contribution to journal › Article › peer-review

Joglekar, MV, Sahu, S, Wong, WKM, Satoor, SN, Dong, CX, Farr, RJ, Williams, MD, Pandya, P, Jhala, G, Yang, SNY, Chew, YV, Hetherington, N, Thiruchevlam, D, Mitnala, S, Rao, GV, Reddy, DN, Loudovaris, T, Hawthorne, WJ, Elefanty, AG, Joglekar, VM, Stanley, EG, Martin, D, Thomas, HE, Tosh, D, Dalgaard, LT & Hardikar, AA 2022, 'A Pro-Endocrine Pancreatic Islet Transcriptional Program Established During Development Is Retained in Human Gallbladder Epithelial Cells', Cellular and Molecular Gastroenterology and Hepatology, vol. 13, no. 5, pp. 1530-1553.e4. https://doi.org/10.1016/j.jcmgh.2022.01.008

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title = "A Pro-Endocrine Pancreatic Islet Transcriptional Program Established During Development Is Retained in Human Gallbladder Epithelial Cells",

abstract = "Background & Aims: Pancreatic islet β-cells are factories for insulin production; however, ectopic expression of insulin also is well recognized. The gallbladder is a next-door neighbor to the developing pancreas. Here, we wanted to understand if gallbladders contain functional insulin-producing cells. Methods: We compared developing and adult mouse as well as human gallbladder epithelial cells and islets using immunohistochemistry, flow cytometry, enzyme-linked immunosorbent assays, RNA sequencing, real-time polymerase chain reaction, chromatin immunoprecipitation, and functional studies. Results: We show that the epithelial lining of developing, as well as adult, mouse and human gallbladders naturally contain interspersed cells that retain the capacity to actively transcribe, translate, package, and release insulin. We show that human gallbladders also contain functional insulin-secreting cells with the potential to naturally respond to glucose in vitro and in situ. Notably, in a non-obese diabetic (NOD) mouse model of type 1 diabetes, we observed that insulin-producing cells in the gallbladder are not targeted by autoimmune cells. Interestingly, in human gallbladders, insulin splice variants are absent, although insulin splice forms are observed in human islets. Conclusions: In summary, our biochemical, transcriptomic, and functional data in mouse and human gallbladder epithelial cells collectively show the evolutionary and developmental similarities between gallbladder and the pancreas that allow gallbladder epithelial cells to continue insulin production in adult life. Understanding the mechanisms regulating insulin transcription and translation in gallbladder epithelial cells would help guide future studies in type 1 diabetes therapy.",

keywords = "Differentiation, Gallbladder Development, Insulin, Islets, Splice Variants",

author = "Joglekar, {Mugdha V.} and Subhshri Sahu and Wong, {Wilson K.M.} and Satoor, {Sarang N.} and Dong, {Charlotte X.} and Farr, {Ryan J.} and Williams, {Michael D.} and Prapti Pandya and Gaurang Jhala and Yang, {Sundy N.Y.} and Chew, {Yi Vee} and Nicola Hetherington and Dhan Thiruchevlam and Sasikala Mitnala and Rao, {Guduru V.} and Reddy, {Duvvuru Nageshwar} and Thomas Loudovaris and Hawthorne, {Wayne J.} and Elefanty, {Andrew G.} and Joglekar, {Vinay M.} and Stanley, {Edouard G.} and David Martin and Thomas, {Helen E.} and David Tosh and Dalgaard, {Louise T.} and Hardikar, {Anandwardhan A.}",

note = "Funding Information: Funding Supported mainly by a 2008–2009 British Council (UK–India Educational Research Initiative) exchange program (A.A.H. and D.T.), a 2012–2015 National Health and Medical Research Council project grant (A.A.H.), and the Danish Diabetes Academy visiting professorship (A.A.H. and L.T.D.). Also funded by the Australian Research Council (2012–2016) and the Juvenile Diabetes Research Foundation (JDRF) Australian Type 1 Diabetes Clinical Research Network (grant 4-CDA2016-228-MB; a special initiative of the Australian Research Council) (A.A.H.), the JDRF USA postdoctoral fellowship (2012–2014), advanced post-doctoral fellowship (2015–2018), and the JDRF career transition award (2019–2021) (M.V.J.). The University of Sydney, Australian postgraduate award (University Post-graduate Award or Australian Post-graduate Award), and JDRF Australia PhD top-up awards (W.K.M.W. and R.J.F.), and Council of Scientific and Industrial Research fellowships (Government of India) (S.S.) are acknowledged. W.K.M.W. currently is funded through a grant from the Leona M. and Harry B. Helmsley Charitable Trust (grant 2018PG-T1D009) in collaboration with the JDRF Australian Type 1 Diabetes Clinical Research Network grant 3-SRA-2019-694-M-B (A.A.H.). Also supported by the Summer Scholarship from the National Health and Medical Research Council Clinical Trials Centre, University of Sydney (P.P.). Funding Information: The authors acknowledge and are thankful for the support from surgical teams, consenting donors or families of the donors, as well as infrastructure support obtained through the Rebecca L Cooper Foundation (A.A.H.) and provided by the National Centre for Cell Science, India and the Faculty of Medicine and Health, University of Sydney. All authors acknowledge the assistance from Dr Andrew M. Holland and Dr Suzanne J. Micallef in the provision of the Pdx1 GFP/w reporter mice, Professor Manami Hara for the MIP-GFP mice, and the support from Professor Jonathan Slack and Professor Harry Heimberg for the viral vectors used in the study. The authors also acknowledge support from Dr Ramesh Dumbre, Dr Pabitra Sahoo, Ms Fahmida Khan, Ms Sophie Breedveld, Ms Mariah Taleb, RPA Pathology Services, Bosch Facilities (Bosch Mass Spectometry Facility and Live Cell Analysis Facility) at the University of Sydney. A.G.E. and E.G.S were supported by Australian National and Health and Medical Research Council grants (GNT1079004, GNT1117596, GNT1129861, GNT1138717, GNT1123277), and by the Stafford Fox Medical Research Foundation. Additional infrastructure funding to the Murdoch Children's Research Institute was provided by the Australian Government National Health and Medical Research Council Independent Research Institute Infrastructure Support Scheme and the Victorian Government's Operational Infrastructure Support Program. Funding Information: The authors acknowledge and are thankful for the support from surgical teams, consenting donors or families of the donors, as well as infrastructure support obtained through the Rebecca L Cooper Foundation (A.A.H.) and provided by the National Centre for Cell Science, India and the Faculty of Medicine and Health, University of Sydney. All authors acknowledge the assistance from Dr Andrew M. Holland and Dr Suzanne J. Micallef in the provision of the Pdx1GFP/w reporter mice, Professor Manami Hara for the MIP-GFP mice, and the support from Professor Jonathan Slack and Professor Harry Heimberg for the viral vectors used in the study. The authors also acknowledge support from Dr Ramesh Dumbre, Dr Pabitra Sahoo, Ms Fahmida Khan, Ms Sophie Breedveld, Ms Mariah Taleb, RPA Pathology Services, Bosch Facilities (Bosch Mass Spectometry Facility and Live Cell Analysis Facility) at the University of Sydney. A.G.E. and E.G.S were supported by Australian National and Health and Medical Research Council grants (GNT1079004, GNT1117596, GNT1129861, GNT1138717, GNT1123277), and by the Stafford Fox Medical Research Foundation. Additional infrastructure funding to the Murdoch Children's Research Institute was provided by the Australian Government National Health and Medical Research Council Independent Research Institute Infrastructure Support Scheme and the Victorian Government's Operational Infrastructure Support Program. Mugdha V. Joglekar (Conceptualization: Supporting; Data curation: Equal; Formal analysis: Equal; Investigation: Lead; Methodology: Equal; Software: Equal; Supervision: Equal; Writing ? original draft: Lead; Writing ? review & editing: Equal), Subhshri Sahu (Data curation: Equal; Formal analysis: Equal; Methodology: Equal; Software: Supporting; Writing ? review & editing: Equal), Wilson KM Wong (Data curation: Equal; Formal analysis: Equal; Methodology: Supporting; Software: Equal; Validation: Equal; Writing ? review & editing: Equal), Sarang N. Satoor (Data curation: Supporting; Formal analysis: Supporting; Methodology: Supporting), Charlotte X. Dong (Software: Supporting; Validation: Supporting), Ryan J Farr (Data curation: Supporting; Methodology: Supporting), Michael D. Williams (Data curation: Supporting), Prapti Pandya (Data curation: Supporting), Gaurang Jhala (Data curation: Supporting; Resources: Supporting), Sundy N.Y. Yang (Data curation: Supporting), Yi Vee Chew (Data curation: Supporting), Nicola Hetherington (Data curation: Supporting), Dhan Thiruchevlam (Resources: Supporting), Sasikala Mitnala (Data curation: Supporting; Resources: Supporting), Guduru V Rao (Resources: Supporting), Duvvuru Nageshwar Reddy (Resources: Supporting), Thomas Loudovaris (Resources: Supporting), Wayne J. Hawthorne (Resources: Supporting), Andrew G. Elefanty (Resources: Supporting), Vinay M. Joglekar (Resources: Supporting), Edouard G. Stanley (Resources: Supporting), David Martin (Resources: Supporting), Helen E. Thomas (Resources: Supporting), David Tosh (Conceptualization: Supporting; Funding acquisition: Supporting; Supervision: Supporting; Writing ? original draft: Supporting; Writing ? review & editing: Supporting), Louise T. Dalgaard (Software: Equal; Validation: Equal; Writing ? review & editing: Supporting), Anandwardhan A. Hardikar (Conceptualization: Lead; Data curation: Supporting; Funding acquisition: Lead; Investigation: Supporting; Project administration: Lead; Supervision: Lead; Visualization: Supporting; Writing ? original draft: Equal; Writing ? review & editing: Equal), Sequencing data are available through GEO data sets cited in text. Data Availability Statement Sequencing data are available through GEO data sets cited in text.",

year = "2022",

month = dec,

day = "31",

doi = "10.1016/j.jcmgh.2022.01.008",

language = "English",

volume = "13",

pages = "1530--1553.e4",

journal = "Cellular and Molecular Gastroenterology and Hepatology",

issn = "2352-345X",

publisher = "Elsevier",

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}

TY - JOUR

T1 - A Pro-Endocrine Pancreatic Islet Transcriptional Program Established During Development Is Retained in Human Gallbladder Epithelial Cells

AU - Joglekar, Mugdha V.

AU - Sahu, Subhshri

AU - Wong, Wilson K.M.

AU - Satoor, Sarang N.

AU - Dong, Charlotte X.

AU - Farr, Ryan J.

AU - Williams, Michael D.

AU - Pandya, Prapti

AU - Jhala, Gaurang

AU - Yang, Sundy N.Y.

AU - Chew, Yi Vee

AU - Hetherington, Nicola

AU - Thiruchevlam, Dhan

AU - Mitnala, Sasikala

AU - Rao, Guduru V.

AU - Reddy, Duvvuru Nageshwar

AU - Loudovaris, Thomas

AU - Hawthorne, Wayne J.

AU - Elefanty, Andrew G.

AU - Joglekar, Vinay M.

AU - Stanley, Edouard G.

AU - Martin, David

AU - Thomas, Helen E.

AU - Tosh, David

AU - Dalgaard, Louise T.

AU - Hardikar, Anandwardhan A.

N1 - Funding Information: Funding Supported mainly by a 2008–2009 British Council (UK–India Educational Research Initiative) exchange program (A.A.H. and D.T.), a 2012–2015 National Health and Medical Research Council project grant (A.A.H.), and the Danish Diabetes Academy visiting professorship (A.A.H. and L.T.D.). Also funded by the Australian Research Council (2012–2016) and the Juvenile Diabetes Research Foundation (JDRF) Australian Type 1 Diabetes Clinical Research Network (grant 4-CDA2016-228-MB; a special initiative of the Australian Research Council) (A.A.H.), the JDRF USA postdoctoral fellowship (2012–2014), advanced post-doctoral fellowship (2015–2018), and the JDRF career transition award (2019–2021) (M.V.J.). The University of Sydney, Australian postgraduate award (University Post-graduate Award or Australian Post-graduate Award), and JDRF Australia PhD top-up awards (W.K.M.W. and R.J.F.), and Council of Scientific and Industrial Research fellowships (Government of India) (S.S.) are acknowledged. W.K.M.W. currently is funded through a grant from the Leona M. and Harry B. Helmsley Charitable Trust (grant 2018PG-T1D009) in collaboration with the JDRF Australian Type 1 Diabetes Clinical Research Network grant 3-SRA-2019-694-M-B (A.A.H.). Also supported by the Summer Scholarship from the National Health and Medical Research Council Clinical Trials Centre, University of Sydney (P.P.). Funding Information: The authors acknowledge and are thankful for the support from surgical teams, consenting donors or families of the donors, as well as infrastructure support obtained through the Rebecca L Cooper Foundation (A.A.H.) and provided by the National Centre for Cell Science, India and the Faculty of Medicine and Health, University of Sydney. All authors acknowledge the assistance from Dr Andrew M. Holland and Dr Suzanne J. Micallef in the provision of the Pdx1 GFP/w reporter mice, Professor Manami Hara for the MIP-GFP mice, and the support from Professor Jonathan Slack and Professor Harry Heimberg for the viral vectors used in the study. The authors also acknowledge support from Dr Ramesh Dumbre, Dr Pabitra Sahoo, Ms Fahmida Khan, Ms Sophie Breedveld, Ms Mariah Taleb, RPA Pathology Services, Bosch Facilities (Bosch Mass Spectometry Facility and Live Cell Analysis Facility) at the University of Sydney. A.G.E. and E.G.S were supported by Australian National and Health and Medical Research Council grants (GNT1079004, GNT1117596, GNT1129861, GNT1138717, GNT1123277), and by the Stafford Fox Medical Research Foundation. Additional infrastructure funding to the Murdoch Children's Research Institute was provided by the Australian Government National Health and Medical Research Council Independent Research Institute Infrastructure Support Scheme and the Victorian Government's Operational Infrastructure Support Program. Funding Information: The authors acknowledge and are thankful for the support from surgical teams, consenting donors or families of the donors, as well as infrastructure support obtained through the Rebecca L Cooper Foundation (A.A.H.) and provided by the National Centre for Cell Science, India and the Faculty of Medicine and Health, University of Sydney. All authors acknowledge the assistance from Dr Andrew M. Holland and Dr Suzanne J. Micallef in the provision of the Pdx1GFP/w reporter mice, Professor Manami Hara for the MIP-GFP mice, and the support from Professor Jonathan Slack and Professor Harry Heimberg for the viral vectors used in the study. The authors also acknowledge support from Dr Ramesh Dumbre, Dr Pabitra Sahoo, Ms Fahmida Khan, Ms Sophie Breedveld, Ms Mariah Taleb, RPA Pathology Services, Bosch Facilities (Bosch Mass Spectometry Facility and Live Cell Analysis Facility) at the University of Sydney. A.G.E. and E.G.S were supported by Australian National and Health and Medical Research Council grants (GNT1079004, GNT1117596, GNT1129861, GNT1138717, GNT1123277), and by the Stafford Fox Medical Research Foundation. Additional infrastructure funding to the Murdoch Children's Research Institute was provided by the Australian Government National Health and Medical Research Council Independent Research Institute Infrastructure Support Scheme and the Victorian Government's Operational Infrastructure Support Program. Mugdha V. Joglekar (Conceptualization: Supporting; Data curation: Equal; Formal analysis: Equal; Investigation: Lead; Methodology: Equal; Software: Equal; Supervision: Equal; Writing ? original draft: Lead; Writing ? review & editing: Equal), Subhshri Sahu (Data curation: Equal; Formal analysis: Equal; Methodology: Equal; Software: Supporting; Writing ? review & editing: Equal), Wilson KM Wong (Data curation: Equal; Formal analysis: Equal; Methodology: Supporting; Software: Equal; Validation: Equal; Writing ? review & editing: Equal), Sarang N. Satoor (Data curation: Supporting; Formal analysis: Supporting; Methodology: Supporting), Charlotte X. Dong (Software: Supporting; Validation: Supporting), Ryan J Farr (Data curation: Supporting; Methodology: Supporting), Michael D. Williams (Data curation: Supporting), Prapti Pandya (Data curation: Supporting), Gaurang Jhala (Data curation: Supporting; Resources: Supporting), Sundy N.Y. Yang (Data curation: Supporting), Yi Vee Chew (Data curation: Supporting), Nicola Hetherington (Data curation: Supporting), Dhan Thiruchevlam (Resources: Supporting), Sasikala Mitnala (Data curation: Supporting; Resources: Supporting), Guduru V Rao (Resources: Supporting), Duvvuru Nageshwar Reddy (Resources: Supporting), Thomas Loudovaris (Resources: Supporting), Wayne J. Hawthorne (Resources: Supporting), Andrew G. Elefanty (Resources: Supporting), Vinay M. Joglekar (Resources: Supporting), Edouard G. Stanley (Resources: Supporting), David Martin (Resources: Supporting), Helen E. Thomas (Resources: Supporting), David Tosh (Conceptualization: Supporting; Funding acquisition: Supporting; Supervision: Supporting; Writing ? original draft: Supporting; Writing ? review & editing: Supporting), Louise T. Dalgaard (Software: Equal; Validation: Equal; Writing ? review & editing: Supporting), Anandwardhan A. Hardikar (Conceptualization: Lead; Data curation: Supporting; Funding acquisition: Lead; Investigation: Supporting; Project administration: Lead; Supervision: Lead; Visualization: Supporting; Writing ? original draft: Equal; Writing ? review & editing: Equal), Sequencing data are available through GEO data sets cited in text. Data Availability Statement Sequencing data are available through GEO data sets cited in text.

PY - 2022/12/31

Y1 - 2022/12/31

N2 - Background & Aims: Pancreatic islet β-cells are factories for insulin production; however, ectopic expression of insulin also is well recognized. The gallbladder is a next-door neighbor to the developing pancreas. Here, we wanted to understand if gallbladders contain functional insulin-producing cells. Methods: We compared developing and adult mouse as well as human gallbladder epithelial cells and islets using immunohistochemistry, flow cytometry, enzyme-linked immunosorbent assays, RNA sequencing, real-time polymerase chain reaction, chromatin immunoprecipitation, and functional studies. Results: We show that the epithelial lining of developing, as well as adult, mouse and human gallbladders naturally contain interspersed cells that retain the capacity to actively transcribe, translate, package, and release insulin. We show that human gallbladders also contain functional insulin-secreting cells with the potential to naturally respond to glucose in vitro and in situ. Notably, in a non-obese diabetic (NOD) mouse model of type 1 diabetes, we observed that insulin-producing cells in the gallbladder are not targeted by autoimmune cells. Interestingly, in human gallbladders, insulin splice variants are absent, although insulin splice forms are observed in human islets. Conclusions: In summary, our biochemical, transcriptomic, and functional data in mouse and human gallbladder epithelial cells collectively show the evolutionary and developmental similarities between gallbladder and the pancreas that allow gallbladder epithelial cells to continue insulin production in adult life. Understanding the mechanisms regulating insulin transcription and translation in gallbladder epithelial cells would help guide future studies in type 1 diabetes therapy.

AB - Background & Aims: Pancreatic islet β-cells are factories for insulin production; however, ectopic expression of insulin also is well recognized. The gallbladder is a next-door neighbor to the developing pancreas. Here, we wanted to understand if gallbladders contain functional insulin-producing cells. Methods: We compared developing and adult mouse as well as human gallbladder epithelial cells and islets using immunohistochemistry, flow cytometry, enzyme-linked immunosorbent assays, RNA sequencing, real-time polymerase chain reaction, chromatin immunoprecipitation, and functional studies. Results: We show that the epithelial lining of developing, as well as adult, mouse and human gallbladders naturally contain interspersed cells that retain the capacity to actively transcribe, translate, package, and release insulin. We show that human gallbladders also contain functional insulin-secreting cells with the potential to naturally respond to glucose in vitro and in situ. Notably, in a non-obese diabetic (NOD) mouse model of type 1 diabetes, we observed that insulin-producing cells in the gallbladder are not targeted by autoimmune cells. Interestingly, in human gallbladders, insulin splice variants are absent, although insulin splice forms are observed in human islets. Conclusions: In summary, our biochemical, transcriptomic, and functional data in mouse and human gallbladder epithelial cells collectively show the evolutionary and developmental similarities between gallbladder and the pancreas that allow gallbladder epithelial cells to continue insulin production in adult life. Understanding the mechanisms regulating insulin transcription and translation in gallbladder epithelial cells would help guide future studies in type 1 diabetes therapy.

KW - Differentiation

KW - Gallbladder Development

KW - Insulin

KW - Islets

KW - Splice Variants

UR - http://www.scopus.com/inward/record.url?scp=85126525918&partnerID=8YFLogxK

U2 - 10.1016/j.jcmgh.2022.01.008

DO - 10.1016/j.jcmgh.2022.01.008

M3 - Article

C2 - 35032693

AN - SCOPUS:85126525918

SN - 2352-345X

VL - 13

SP - 1530-1553.e4

JO - Cellular and Molecular Gastroenterology and Hepatology

JF - Cellular and Molecular Gastroenterology and Hepatology

IS - 5

ER -

A Pro-Endocrine Pancreatic Islet Transcriptional Program Established During Development Is Retained in Human Gallbladder Epithelial Cells

Abstract

Bibliographical note

Keywords

ASJC Scopus subject areas

UN SDGs

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Confocal Laser Scanning Confocal Microscope (LSM)

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A Pro-Endocrine Pancreatic Islet Transcriptional Program Established During Development Is Retained in Human Gallbladder Epithelial Cells

Abstract

Bibliographical note

Keywords

ASJC Scopus subject areas

UN SDGs

Access to Document

Other files and links

Fingerprint

Equipment

Confocal Laser Scanning Confocal Microscope (LSM)

Cite this