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

Cell Reports: Methods (Nov 2023)

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

Affiliations

Gerard Ruiz-Babot: 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; Corresponding author
Ariane Eceiza: Division of Endocrinology, Boston Children’s Hospital, Harvard Medical School, Boston, MA, USA; Harvard Stem Cell Institute, Cambridge, MA, USA
Fernando Abollo-Jiménez: Maimonides Biomedical Research Institute of Cordoba (IMIBIC), Córdoba, Spain
Maria Malyukov: Department of Medicine, University Hospital Carl Gustav Carus, Dresden, Germany
Diana L. Carlone: Division of Endocrinology, Boston Children’s Hospital, Harvard Medical School, Boston, MA, USA; Harvard Stem Cell Institute, Cambridge, MA, USA
Kleiton Borges: Division of Endocrinology, Boston Children’s Hospital, Harvard Medical School, Boston, MA, USA; Harvard Stem Cell Institute, Cambridge, MA, USA
Alexandra Rodrigues Da Costa: Centre for Endocrinology, William Harvey Research Institute, Bart’s and the London School of Medicine and Dentistry, Queen Mary University of London, London, UK
Shamma Qarin: Wellcome-MRC Cambridge Stem Cell Institute, Cambridge Biomedical Campus, University of Cambridge, Puddicombe Way, Cambridge, UK
Takuya Matsumoto: Wyss Institute for Biologically Inspired Engineering, Harvard University, Cambridge, MA, USA; Nephrology Division, Massachusetts General Hospital, Boston, MA, USA
Ryuji Morizane: 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
William C. Skarnes: Cellular Engineering, The Jackson Laboratory for Genomic Medicine, Farmington, CT 06032, USA
Barbara Ludwig: Department of Medicine, University Hospital Carl Gustav Carus, Dresden, Germany
Paul J. Chapple: Centre for Endocrinology, William Harvey Research Institute, Bart’s and the London School of Medicine and Dentistry, Queen Mary University of London, London, UK
Leonardo Guasti: Centre for Endocrinology, William Harvey Research Institute, Bart’s and the London School of Medicine and Dentistry, Queen Mary University of London, London, UK
Helen L. Storr: Centre for Endocrinology, William Harvey Research Institute, Bart’s and the London School of Medicine and Dentistry, Queen Mary University of London, London, UK
Stefan R. Bornstein: 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
David T. Breault: Division of Endocrinology, Boston Children’s Hospital, Harvard Medical School, Boston, MA, USA; Harvard Stem Cell Institute, Cambridge, MA, USA; Corresponding author

Journal volume & issue: Vol. 3, no. 11
p. 100627

Abstract

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Summary: 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. Motivation: Adrenal insufficiency is a life-threatening condition resulting from the inability to produce steroid hormones in a dose and time-dependent manner. The ability to generate cells that produce cortisol in response to adrenocorticotrophic hormone (ACTH), the main regulator of glucocorticoid production in humans, combined with the development of encapsulation and immunoprotecting technologies, will open future opportunities to treat this condition using cell-based therapies. Current protocols have demonstrated the generation of fetal-like steroid-producing cells from hPSCs. However, the resulting cells fail to secrete cortisol, the main glucocorticoid in humans. We therefore sought to develop a protocol that enables the generation of billions of cells that produce cortisol and can respond to ACTH stimulation.

CP: Stem cell

Published in Cell Reports: Methods

ISSN: 2667-2375 (Online)
Publisher: Elsevier
Country of publisher: United States
LCC subjects: Technology: Chemical technology: Biotechnology; Science: Chemistry: Organic chemistry: Biochemistry
Website: https://www.cell.com/cell-reports-methods

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