Mitchell-Riley syndrome iPSCs exhibit reduced pancreatic endoderm differentiation due to a mutation in RFX6

Jamie Trott; Yunus Alpagu; Ee Kim Tan; Mohammad Shboul; Yousif Dawood; Michael Elsy; Heike Wollmann; Vincent Tano; Carine Bonnard; Shermaine Eng; Gunaseelan Narayanan; Seetanshu Junnarkar; Stephen Wearne; James Strutt; Aakash Kumar; Lucian B. Tomaz; Pierre Alexis Goy; Slim Mzoughi; Rachel Jennings; Jaco Hagoort; Ascia Eskin; Hane Lee; Stanley F. Nelson; Fawaz Al-Kazaleh; Mohammad El-Khateeb; Rajaa Fathallah; Harsha Shah; Jonathan Goeke; Sarah R. Langley; Ernesto Guccione; Neil Hanley; Bernadette S. de Bakker; Bruno Reversade; N. Ray Dunn

doi:10.1242/dev.194878

Mitchell-Riley syndrome iPSCs exhibit reduced pancreatic endoderm differentiation due to a mutation in RFX6

Jamie Trott, Yunus Alpagu, Ee Kim Tan, Mohammad Shboul, Yousif Dawood, Michael Elsy, Heike Wollmann, Vincent Tano, Carine Bonnard, Shermaine Eng, Gunaseelan Narayanan, Seetanshu Junnarkar, Stephen Wearne, James Strutt, Aakash Kumar, Lucian B. Tomaz, Pierre Alexis Goy, Slim Mzoughi, Rachel Jennings, Jaco HagoortAscia Eskin, Hane Lee, Stanley F. Nelson, Fawaz Al-Kazaleh, Mohammad El-Khateeb, Rajaa Fathallah, Harsha Shah, Jonathan Goeke, Sarah R. Langley, Ernesto Guccione, Neil Hanley, Bernadette S. de Bakker, Bruno Reversade, N. Ray Dunn^*

^*Corresponding author for this work

Medicine and Health

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

8 Citations (Scopus)

Abstract

Mitchell-Riley syndrome (MRS) is caused by recessive mutations in the regulatory factor X6 gene (RFX6) and is characterised by pancreatic hypoplasia and neonatal diabetes. To determine why individuals with MRS specifically lack pancreatic endocrine cells, we micro-CT imaged a 12-week-old foetus homozygous for the nonsense mutation RFX6 c.1129C>T, which revealed loss of the pancreas body and tail. From this foetus, we derived iPSCs and show that differentiation of these cells in vitro proceeds normally until generation of pancreatic endoderm, which is significantly reduced. We additionally generated an RFX6^HA reporter allele by gene targeting in wild-type H9 cells to precisely define RFX6 expression and in parallel performed in situ hybridisation for RFX6 in the dorsal pancreatic bud of a Carnegie stage 14 human embryo. Both in vitro and in vivo, we find that RFX6 specifically labels a subset of PDX1-expressing pancreatic endoderm. In summary, RFX6 is essential for efficient differentiation of pancreatic endoderm, and its absence in individuals with MRS specifically impairs formation of endocrine cells of the pancreas head and tail.

Original language	English
Article number	dev194878
Journal	Development (Cambridge)
Volume	147
Issue number	21
DOIs	https://doi.org/10.1242/dev.194878
Publication status	Published - Nov 2020

Bibliographical note

Funding Information:
J.T. was funded by a National Medical Research Council Award Young Investigator Research Grant (OFYIRG18May-0049). Y.A. was awarded a Singapore

Funding Information:
International Graduate Award by the Agency for Science, Technology and Research (A*STAR) Graduate Academy. N.R.D. and B.R. were provided with core funding by the A*STAR Institute of Medical Biology and were additionally supported by an Economic Development Board Singapore ‘Singapore Childhood Undiagnosed Diseases’ program grant (IAF311019) and an Agency for Science, Technology and Research Strategic Positioning Fund ‘Genetic Orphan Diseases Adopted: Fostering Innovation Therapy’ (GODAFIT) grant.

Publisher Copyright:
© 2020. Published by The Company of Biologists Ltd

Keywords

Genetic disease
In vitro differentiation
Mitchell-Riley syndrome
Pancreas development
RFX6

ASJC Scopus subject areas

Molecular Biology
Developmental Biology

UN SDGs

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

Access to Document

10.1242/dev.194878

Cite this

Trott, J., Alpagu, Y., Tan, E. K., Shboul, M., Dawood, Y., Elsy, M., Wollmann, H., Tano, V., Bonnard, C., Eng, S., Narayanan, G., Junnarkar, S., Wearne, S., Strutt, J., Kumar, A., Tomaz, L. B., Goy, P. A., Mzoughi, S., Jennings, R., ... Dunn, N. R. (2020). Mitchell-Riley syndrome iPSCs exhibit reduced pancreatic endoderm differentiation due to a mutation in RFX6. Development (Cambridge), 147(21), Article dev194878. https://doi.org/10.1242/dev.194878

@article{3f2467088db4411dad5f95fa0136bc32,

title = "Mitchell-Riley syndrome iPSCs exhibit reduced pancreatic endoderm differentiation due to a mutation in RFX6",

abstract = "Mitchell-Riley syndrome (MRS) is caused by recessive mutations in the regulatory factor X6 gene (RFX6) and is characterised by pancreatic hypoplasia and neonatal diabetes. To determine why individuals with MRS specifically lack pancreatic endocrine cells, we micro-CT imaged a 12-week-old foetus homozygous for the nonsense mutation RFX6 c.1129C>T, which revealed loss of the pancreas body and tail. From this foetus, we derived iPSCs and show that differentiation of these cells in vitro proceeds normally until generation of pancreatic endoderm, which is significantly reduced. We additionally generated an RFX6HA reporter allele by gene targeting in wild-type H9 cells to precisely define RFX6 expression and in parallel performed in situ hybridisation for RFX6 in the dorsal pancreatic bud of a Carnegie stage 14 human embryo. Both in vitro and in vivo, we find that RFX6 specifically labels a subset of PDX1-expressing pancreatic endoderm. In summary, RFX6 is essential for efficient differentiation of pancreatic endoderm, and its absence in individuals with MRS specifically impairs formation of endocrine cells of the pancreas head and tail.",

keywords = "Genetic disease, In vitro differentiation, Mitchell-Riley syndrome, Pancreas development, RFX6",

author = "Jamie Trott and Yunus Alpagu and Tan, {Ee Kim} and Mohammad Shboul and Yousif Dawood and Michael Elsy and Heike Wollmann and Vincent Tano and Carine Bonnard and Shermaine Eng and Gunaseelan Narayanan and Seetanshu Junnarkar and Stephen Wearne and James Strutt and Aakash Kumar and Tomaz, {Lucian B.} and Goy, {Pierre Alexis} and Slim Mzoughi and Rachel Jennings and Jaco Hagoort and Ascia Eskin and Hane Lee and Nelson, {Stanley F.} and Fawaz Al-Kazaleh and Mohammad El-Khateeb and Rajaa Fathallah and Harsha Shah and Jonathan Goeke and Langley, {Sarah R.} and Ernesto Guccione and Neil Hanley and {de Bakker}, {Bernadette S.} and Bruno Reversade and Dunn, {N. Ray}",

note = "Funding Information: J.T. was funded by a National Medical Research Council Award Young Investigator Research Grant (OFYIRG18May-0049). Y.A. was awarded a Singapore Funding Information: International Graduate Award by the Agency for Science, Technology and Research (A*STAR) Graduate Academy. N.R.D. and B.R. were provided with core funding by the A*STAR Institute of Medical Biology and were additionally supported by an Economic Development Board Singapore {\textquoteleft}Singapore Childhood Undiagnosed Diseases{\textquoteright} program grant (IAF311019) and an Agency for Science, Technology and Research Strategic Positioning Fund {\textquoteleft}Genetic Orphan Diseases Adopted: Fostering Innovation Therapy{\textquoteright} (GODAFIT) grant. Publisher Copyright: {\textcopyright} 2020. Published by The Company of Biologists Ltd",

year = "2020",

month = nov,

doi = "10.1242/dev.194878",

language = "English",

volume = "147",

journal = "Development (Cambridge)",

issn = "0950-1991",

publisher = "The Company of Biologists Ltd.",

number = "21",

}

Trott, J, Alpagu, Y, Tan, EK, Shboul, M, Dawood, Y, Elsy, M, Wollmann, H, Tano, V, Bonnard, C, Eng, S, Narayanan, G, Junnarkar, S, Wearne, S, Strutt, J, Kumar, A, Tomaz, LB, Goy, PA, Mzoughi, S, Jennings, R, Hagoort, J, Eskin, A, Lee, H, Nelson, SF, Al-Kazaleh, F, El-Khateeb, M, Fathallah, R, Shah, H, Goeke, J, Langley, SR, Guccione, E, Hanley, N, de Bakker, BS, Reversade, B & Dunn, NR 2020, 'Mitchell-Riley syndrome iPSCs exhibit reduced pancreatic endoderm differentiation due to a mutation in RFX6', Development (Cambridge), vol. 147, no. 21, dev194878. https://doi.org/10.1242/dev.194878

TY - JOUR

T1 - Mitchell-Riley syndrome iPSCs exhibit reduced pancreatic endoderm differentiation due to a mutation in RFX6

AU - Trott, Jamie

AU - Alpagu, Yunus

AU - Tan, Ee Kim

AU - Shboul, Mohammad

AU - Dawood, Yousif

AU - Elsy, Michael

AU - Wollmann, Heike

AU - Tano, Vincent

AU - Bonnard, Carine

AU - Eng, Shermaine

AU - Narayanan, Gunaseelan

AU - Junnarkar, Seetanshu

AU - Wearne, Stephen

AU - Strutt, James

AU - Kumar, Aakash

AU - Tomaz, Lucian B.

AU - Goy, Pierre Alexis

AU - Mzoughi, Slim

AU - Jennings, Rachel

AU - Hagoort, Jaco

AU - Eskin, Ascia

AU - Lee, Hane

AU - Nelson, Stanley F.

AU - Al-Kazaleh, Fawaz

AU - El-Khateeb, Mohammad

AU - Fathallah, Rajaa

AU - Shah, Harsha

AU - Goeke, Jonathan

AU - Langley, Sarah R.

AU - Guccione, Ernesto

AU - Hanley, Neil

AU - de Bakker, Bernadette S.

AU - Reversade, Bruno

AU - Dunn, N. Ray

N1 - Funding Information: J.T. was funded by a National Medical Research Council Award Young Investigator Research Grant (OFYIRG18May-0049). Y.A. was awarded a Singapore Funding Information: International Graduate Award by the Agency for Science, Technology and Research (A*STAR) Graduate Academy. N.R.D. and B.R. were provided with core funding by the A*STAR Institute of Medical Biology and were additionally supported by an Economic Development Board Singapore ‘Singapore Childhood Undiagnosed Diseases’ program grant (IAF311019) and an Agency for Science, Technology and Research Strategic Positioning Fund ‘Genetic Orphan Diseases Adopted: Fostering Innovation Therapy’ (GODAFIT) grant. Publisher Copyright: © 2020. Published by The Company of Biologists Ltd

PY - 2020/11

Y1 - 2020/11

N2 - Mitchell-Riley syndrome (MRS) is caused by recessive mutations in the regulatory factor X6 gene (RFX6) and is characterised by pancreatic hypoplasia and neonatal diabetes. To determine why individuals with MRS specifically lack pancreatic endocrine cells, we micro-CT imaged a 12-week-old foetus homozygous for the nonsense mutation RFX6 c.1129C>T, which revealed loss of the pancreas body and tail. From this foetus, we derived iPSCs and show that differentiation of these cells in vitro proceeds normally until generation of pancreatic endoderm, which is significantly reduced. We additionally generated an RFX6HA reporter allele by gene targeting in wild-type H9 cells to precisely define RFX6 expression and in parallel performed in situ hybridisation for RFX6 in the dorsal pancreatic bud of a Carnegie stage 14 human embryo. Both in vitro and in vivo, we find that RFX6 specifically labels a subset of PDX1-expressing pancreatic endoderm. In summary, RFX6 is essential for efficient differentiation of pancreatic endoderm, and its absence in individuals with MRS specifically impairs formation of endocrine cells of the pancreas head and tail.

AB - Mitchell-Riley syndrome (MRS) is caused by recessive mutations in the regulatory factor X6 gene (RFX6) and is characterised by pancreatic hypoplasia and neonatal diabetes. To determine why individuals with MRS specifically lack pancreatic endocrine cells, we micro-CT imaged a 12-week-old foetus homozygous for the nonsense mutation RFX6 c.1129C>T, which revealed loss of the pancreas body and tail. From this foetus, we derived iPSCs and show that differentiation of these cells in vitro proceeds normally until generation of pancreatic endoderm, which is significantly reduced. We additionally generated an RFX6HA reporter allele by gene targeting in wild-type H9 cells to precisely define RFX6 expression and in parallel performed in situ hybridisation for RFX6 in the dorsal pancreatic bud of a Carnegie stage 14 human embryo. Both in vitro and in vivo, we find that RFX6 specifically labels a subset of PDX1-expressing pancreatic endoderm. In summary, RFX6 is essential for efficient differentiation of pancreatic endoderm, and its absence in individuals with MRS specifically impairs formation of endocrine cells of the pancreas head and tail.

KW - Genetic disease

KW - In vitro differentiation

KW - Mitchell-Riley syndrome

KW - Pancreas development

KW - RFX6

UR - http://www.scopus.com/inward/record.url?scp=85095861690&partnerID=8YFLogxK

U2 - 10.1242/dev.194878

DO - 10.1242/dev.194878

M3 - Article

C2 - 33033118

AN - SCOPUS:85095861690

SN - 0950-1991

VL - 147

JO - Development (Cambridge)

JF - Development (Cambridge)

IS - 21

M1 - dev194878

ER -

Mitchell-Riley syndrome iPSCs exhibit reduced pancreatic endoderm differentiation due to a mutation in RFX6

Abstract

Bibliographical note

Keywords

ASJC Scopus subject areas

UN SDGs

Access to Document

Fingerprint

Cite this