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Modeling Genetic Risk of β-Cell Dysfunction in Human Induced Pluripotent Stem Cells From Patients Carrying the MTNR1B Risk Variant

Singh, Tania LU ; Kalamajski, Sebastian LU ; Cunha, Joãp P M C M ; Hladkou, Siarhei LU ; Roberts, Fiona LU ; Gheibi, Sevda LU ; Soltanian, Anahita LU ; Yektay Farahmand, Kaveh LU orcid ; Ekström, Ola LU and Mamidi, Anant LU , et al. (2025) In Journal of Pineal Research 77(5). p.1-16
Abstract

Disruptions in circadian rhythm, partly controlled by the hormone melatonin, increase the risk of type 2 diabetes (T2D). Accordingly, a variant of the gene encoding the melatonin receptor 1B (MTNR1B) is robustly associated with increased risk of T2D. This single-nucleotide polymorphism (SNP; rs10830963; G-allele) is an expression quantitative trait locus (eQTL) in human pancreatic islets, conferring increased expression of MTNR1B, which is thought to perturb pancreatic β-cell function. To understand this pathogenic mechanism in detail, we utilized human induced pluripotent stem cells (hiPSC), derived from individuals with T2D carrying the MTNR1B G-allele. Patient-derived fibroblasts were reprogrammed to hiPSC and single-base genome... (More)

Disruptions in circadian rhythm, partly controlled by the hormone melatonin, increase the risk of type 2 diabetes (T2D). Accordingly, a variant of the gene encoding the melatonin receptor 1B (MTNR1B) is robustly associated with increased risk of T2D. This single-nucleotide polymorphism (SNP; rs10830963; G-allele) is an expression quantitative trait locus (eQTL) in human pancreatic islets, conferring increased expression of MTNR1B, which is thought to perturb pancreatic β-cell function. To understand this pathogenic mechanism in detail, we utilized human induced pluripotent stem cells (hiPSC), derived from individuals with T2D carrying the MTNR1B G-allele. Patient-derived fibroblasts were reprogrammed to hiPSC and single-base genome editing by CRISPR/Cas9 was employed to create isogenic lines of either the C/C or G/G genotypes (nonrisk and risk, respectively). In addition, the human embryonic stem cell (hESC) line (HUES4) was subjected to genome editing to create isogenic lines of either the C/C or G/G genotypes. hiPSC and hESC were differentiated into β-like cells, using a 50-day 2D protocol. Single-base genome editing generated cells with the desired genotype at a success rate of > 90%. Expression of stage-specific markers confirmed differentiation of both hiPSC and hESC into β-cells. MTNR1B mRNA levels were consistently low in differentiated β-cells, precluding quantitative analysis of gene expression. Western blot analyses indicated slightly higher levels of the MTNR1B protein in differentiated β-cells carrying the risk allele, which is in accord with the notion that rs10830963 (G-allele) functions as an eQTL in β-cells. Insulin secretion in response to the combination of high glucose and IBMX was comparable between genotypes, whereas the addition of melatonin appeared to reduce insulin secretion more efficiently in cells carrying the G-allele. While our data suggest elevated MTNR1B protein levels in stem cell-derived β-like cells carrying the risk allele, these cells do not appear to be sufficiently mature to establish rs10830963 as an eQTL at the mRNA level. The observed nominal increase in melatonin sensitivity in G-allele-carrying cells is suggestive of a functional contribution of rs10830963 to β-cell dysfunction; however, this interpretation remains tentative and will require further validation in more mature β-cell models.

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publication status
published
subject
keywords
Humans, Induced Pluripotent Stem Cells/metabolism, Receptor, Melatonin, MT2/genetics, Insulin-Secreting Cells/metabolism, Polymorphism, Single Nucleotide, Diabetes Mellitus, Type 2/genetics, Quantitative Trait Loci, Cell Differentiation, Genetic Predisposition to Disease
in
Journal of Pineal Research
volume
77
issue
5
article number
e70073
pages
1 - 16
publisher
Wiley-Blackwell
external identifiers
  • pmid:40898610
  • scopus:105014826030
ISSN
1600-079X
DOI
10.1111/jpi.70073
language
English
LU publication?
yes
additional info
© 2025 The Author(s). Journal of Pineal Research published by John Wiley & Sons Ltd.
id
87ad7713-a3a8-403b-a60c-1a824872b9d5
date added to LUP
2025-09-09 10:57:59
date last changed
2025-09-10 04:05:56
@article{87ad7713-a3a8-403b-a60c-1a824872b9d5,
  abstract     = {{<p>Disruptions in circadian rhythm, partly controlled by the hormone melatonin, increase the risk of type 2 diabetes (T2D). Accordingly, a variant of the gene encoding the melatonin receptor 1B (MTNR1B) is robustly associated with increased risk of T2D. This single-nucleotide polymorphism (SNP; rs10830963; G-allele) is an expression quantitative trait locus (eQTL) in human pancreatic islets, conferring increased expression of MTNR1B, which is thought to perturb pancreatic β-cell function. To understand this pathogenic mechanism in detail, we utilized human induced pluripotent stem cells (hiPSC), derived from individuals with T2D carrying the MTNR1B G-allele. Patient-derived fibroblasts were reprogrammed to hiPSC and single-base genome editing by CRISPR/Cas9 was employed to create isogenic lines of either the C/C or G/G genotypes (nonrisk and risk, respectively). In addition, the human embryonic stem cell (hESC) line (HUES4) was subjected to genome editing to create isogenic lines of either the C/C or G/G genotypes. hiPSC and hESC were differentiated into β-like cells, using a 50-day 2D protocol. Single-base genome editing generated cells with the desired genotype at a success rate of &gt; 90%. Expression of stage-specific markers confirmed differentiation of both hiPSC and hESC into β-cells. MTNR1B mRNA levels were consistently low in differentiated β-cells, precluding quantitative analysis of gene expression. Western blot analyses indicated slightly higher levels of the MTNR1B protein in differentiated β-cells carrying the risk allele, which is in accord with the notion that rs10830963 (G-allele) functions as an eQTL in β-cells. Insulin secretion in response to the combination of high glucose and IBMX was comparable between genotypes, whereas the addition of melatonin appeared to reduce insulin secretion more efficiently in cells carrying the G-allele. While our data suggest elevated MTNR1B protein levels in stem cell-derived β-like cells carrying the risk allele, these cells do not appear to be sufficiently mature to establish rs10830963 as an eQTL at the mRNA level. The observed nominal increase in melatonin sensitivity in G-allele-carrying cells is suggestive of a functional contribution of rs10830963 to β-cell dysfunction; however, this interpretation remains tentative and will require further validation in more mature β-cell models.</p>}},
  author       = {{Singh, Tania and Kalamajski, Sebastian and Cunha, Joãp P M C M and Hladkou, Siarhei and Roberts, Fiona and Gheibi, Sevda and Soltanian, Anahita and Yektay Farahmand, Kaveh and Ekström, Ola and Mamidi, Anant and Franks, Paul W and Rosengren, Anders and Semb, Henrik and Mulder, Hindrik and Fex, Malin}},
  issn         = {{1600-079X}},
  keywords     = {{Humans; Induced Pluripotent Stem Cells/metabolism; Receptor, Melatonin, MT2/genetics; Insulin-Secreting Cells/metabolism; Polymorphism, Single Nucleotide; Diabetes Mellitus, Type 2/genetics; Quantitative Trait Loci; Cell Differentiation; Genetic Predisposition to Disease}},
  language     = {{eng}},
  month        = {{09}},
  number       = {{5}},
  pages        = {{1--16}},
  publisher    = {{Wiley-Blackwell}},
  series       = {{Journal of Pineal Research}},
  title        = {{Modeling Genetic Risk of β-Cell Dysfunction in Human Induced Pluripotent Stem Cells From Patients Carrying the MTNR1B Risk Variant}},
  url          = {{http://dx.doi.org/10.1111/jpi.70073}},
  doi          = {{10.1111/jpi.70073}},
  volume       = {{77}},
  year         = {{2025}},
}