Generation of glucocorticoid-producing cells derived from human pluripotent stem cells

Gerard Ruiz-Babot; Ariane Eceiza; Fernando Abollo-Jiménez; Maria Malyukov; Diana L Carlone; Kleiton Borges; Alexandra Rodrigues Da Costa; Shamma Qarin; Takuya Matsumoto; Ryuji Morizane; William C Skarnes; Barbara Ludwig; Paul J Chapple; Leonardo Guasti; Helen L Storr; Stefan R Bornstein; David T Breault

doi:10.1016/j.crmeth.2023.100627

Generation of glucocorticoid-producing cells derived from human pluripotent stem cells

Cell Rep Methods. 2023 Nov 20;3(11):100627. doi: 10.1016/j.crmeth.2023.100627. Epub 2023 Nov 3.

Authors

Gerard Ruiz-Babot¹, Ariane Eceiza², Fernando Abollo-Jiménez³, Maria Malyukov⁴, Diana L Carlone², Kleiton Borges², Alexandra Rodrigues Da Costa⁵, Shamma Qarin⁶, Takuya Matsumoto⁷, Ryuji Morizane⁸, William C Skarnes⁹, Barbara Ludwig⁴, Paul J Chapple⁵, Leonardo Guasti⁵, Helen L Storr⁵, Stefan R Bornstein¹⁰, David T Breault¹¹

Affiliations

¹ Division of Endocrinology, Boston Children's Hospital, Harvard Medical School, Boston, MA, USA; Harvard Stem Cell Institute, Cambridge, MA, USA; Department of Medicine, University Hospital Carl Gustav Carus, Dresden, Germany. Electronic address: gerard.ruiz-babot@uniklinikum-dresden.de.
² Division of Endocrinology, Boston Children's Hospital, Harvard Medical School, Boston, MA, USA; Harvard Stem Cell Institute, Cambridge, MA, USA.
³ Maimonides Biomedical Research Institute of Cordoba (IMIBIC), Córdoba, Spain.
⁴ Department of Medicine, University Hospital Carl Gustav Carus, Dresden, Germany.
⁵ Centre for Endocrinology, William Harvey Research Institute, Bart's and the London School of Medicine and Dentistry, Queen Mary University of London, London, UK.
⁶ Wellcome-MRC Cambridge Stem Cell Institute, Cambridge Biomedical Campus, University of Cambridge, Puddicombe Way, Cambridge, UK.
⁷ Wyss Institute for Biologically Inspired Engineering, Harvard University, Cambridge, MA, USA; Nephrology Division, Massachusetts General Hospital, Boston, MA, USA.
⁸ Harvard Stem Cell Institute, Cambridge, MA, USA; Wyss Institute for Biologically Inspired Engineering, Harvard University, Cambridge, MA, USA; Nephrology Division, Massachusetts General Hospital, Boston, MA, USA.
⁹ Cellular Engineering, The Jackson Laboratory for Genomic Medicine, Farmington, CT 06032, USA.
¹⁰ Department of Medicine, University Hospital Carl Gustav Carus, Dresden, Germany; Division of Endocrinology, Diabetes and Nutritional Sciences, Faculty of Life Sciences and Medicine, King's College London, London, UK.
¹¹ Division of Endocrinology, Boston Children's Hospital, Harvard Medical School, Boston, MA, USA; Harvard Stem Cell Institute, Cambridge, MA, USA. Electronic address: david.breault@childrens.harvard.edu.

Abstract

Adrenal insufficiency is a life-threatening condition resulting from the inability to produce adrenal hormones in a dose- and time-dependent manner. Establishing a cell-based therapy would provide a physiologically responsive approach for the treatment of this condition. We report the generation of large numbers of human-induced steroidogenic cells (hiSCs) from human pluripotent stem cells (hPSCs). Directed differentiation of hPSCs into hiSCs recapitulates the initial stages of human adrenal development. Following expression of steroidogenic factor 1, activation of protein kinase A signaling drives a steroidogenic gene expression profile most comparable to human fetal adrenal cells, and leads to dynamic secretion of steroid hormones, in vitro. Moreover, expression of the adrenocorticotrophic hormone (ACTH) receptor/co-receptor (MC2R/MRAP) results in dose-dependent ACTH responsiveness. This protocol recapitulates adrenal insufficiency resulting from loss-of-function mutations in AAAS, which cause the enigmatic triple A syndrome. Our differentiation protocol generates sufficient numbers of hiSCs for cell-based therapy and offers a platform to study disorders causing adrenal insufficiency.

Keywords: ACTH; CP: Stem cell; adrenal insufficiency; adrenal steroid; allgrove syndrome; cell therapy; human pluripotent stem cells; steroidogenesis; triple A syndrome.

MeSH terms

Adrenal Insufficiency* / genetics
Adrenocorticotropic Hormone / pharmacology
Glucocorticoids / pharmacology
Humans
Pluripotent Stem Cells* / metabolism
Receptors, Corticotropin

Substances

Glucocorticoids
Adrenocorticotropic Hormone
Receptors, Corticotropin

Supplementary concepts

Achalasia Addisonianism Alacrimia syndrome

Grants and funding

DP2 DK133821/DK/NIDDK NIH HHS/United States