In Vivo CaV3 Channel Inhibition Promotes Maturation of Glucose-Dependent Ca2+ Signaling in Human iPSC-Islets

Kaixuan Zhao, Yue Shi, Jia Yu, Lina Yu, Martin Köhler, Amber Mael, Anthony Kolton, Thomas Joyce, Jon Odorico, Per-olof Berggren, Shao-nian Yang

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Abstract

CaV3 channels are ontogenetically downregulated with the maturation of certain electrically excitable cells, including pancreatic β cells. Abnormally exaggerated CaV3 channels drive the dedifferentiation of mature β cells. This led us to question whether excessive CaV3 channels, retained mistakenly in engineered human-induced pluripotent stem cell-derived islet (hiPSC-islet) cells, act as an obstacle to hiPSC-islet maturation. We addressed this question by using the anterior chamber of the eye (ACE) of immunodeficient mice as a site for recapitulation of in vivo hiPSC-islet maturation in combination with intravitreal drug infusion, intravital microimaging, measurements of cytoplasmic-free Ca2+ concentration ([Ca2+]i) and patch clamp analysis. We observed that the ACE is well suited for recapitulation, observation and intervention of hiPSC-islet maturation. Intriguingly, intraocular hiPSC-islet grafts, retrieved intact following intravitreal infusion of the CaV3 channel blocker NNC55-0396, exhibited decreased basal [Ca2+]i levels and increased glucose-stimulated [Ca2+]i responses. Insulin-expressing cells of these islet grafts indeed expressed the NNC55-0396 target CaV3 channels. Intraocular hiPSC-islets underwent satisfactory engraftment, vascularization and light scattering without being influenced by the intravitreally infused NNC55-0396. These data demonstrate that inhibiting CaV3 channels facilitates the maturation of glucose-activated Ca2+ signaling in hiPSC-islets, supporting the notion that excessive CaV3 channels as a developmental error impede the maturation of engineered hiPSC-islet insulin-expressing cells.
Original languageEnglish
Article number807
Pages (from-to)1-15
Number of pages15
JournalBiomedicines
Volume11
Issue number3
Early online date7 Mar 2023
DOIs
Publication statusPublished online - 7 Mar 2023

Bibliographical note

Funding Information:
This work was supported by grants from Berth von Kantzow’s Foundation, ERC-2018-AdG 834860 EYELETS, the Family Erling-Persson Foundation, Funds at Karolinska Institutet, the Stichting af Jochnick Foundation, the Strategic Research Program in Diabetes at Karolinska Institutet, the Swedish Alzheimer Association, the Swedish Diabetes Association, the Swedish Foundation for Strategic Research, the Swedish Research Council and the Novo Nordisk Foundation. R.M.S. has been supported by NIH (NIDDK) SBIR grants (2R44DKI 04497-02, 1R43DKl04497-01, 1R43DK108441-01Al, 1R43DK112472-01, 5R44DK104497-03 and 1 R43DK109832-01A1).

Funding Information:
P.-O.B. is the founder and CEO of the biotech company BioCrine AB. M.K. and S.-N.Y. are consultants to BioCrine. A.K. and J.O. are co-founders of Regenerative Medical Solutions, Inc. and have equity interest in the company. The research reported here was supported in part by funding provided by Regenerative Medical Solutions, Inc. A.M. and T.J. are employees of Regenerative Medical Solutions, Inc.

Publisher Copyright:
© 2023 by the authors.

Keywords

  • anterior chamber of the eye (ACE)
  • calcium channel
  • cytoplasmic-free Ca2+ concentration ([Ca2+]i)
  • stem cell
  • in vivo confocal microscopy
  • islet
  • cytoplasmic-free Ca concentration ([Ca ] )

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