PMID- 37113489
OWN - NLM
STAT- MEDLINE
DCOM- 20230501
LR  - 20230503
IS  - 1664-2392 (Print)
IS  - 1664-2392 (Electronic)
IS  - 1664-2392 (Linking)
VI  - 14
DP  - 2023
TI  - Characterization of the functional and transcriptomic effects of pro-inflammatory 
      cytokines on human EndoC-betaH5 beta cells.
PG  - 1128523
LID - 10.3389/fendo.2023.1128523 [doi]
LID - 1128523
AB  - OBJECTIVE: EndoC-betaH5 is a newly established human beta-cell model which may be 
      superior to previous model systems. Exposure of beta cells to pro-inflammatory 
      cytokines is widely used when studying immune-mediated beta-cell failure in type 
      1 diabetes. We therefore performed an in-depth characterization of the effects of 
      cytokines on EndoC-betaH5 cells. METHODS: The sensitivity profile of EndoC-betaH5 cells 
      to the toxic effects of interleukin-1beta (IL-1beta), interferon gamma (IFNgamma) and tumor 
      necrosis factor-alpha (TNFalpha) was examined in titration and time-course experiments. 
      Cell death was evaluated by caspase-3/7 activity, cytotoxicity, viability, TUNEL 
      assay and immunoblotting. Activation of signaling pathways and major 
      histocompatibility complex (MHC)-I expression were examined by immunoblotting, 
      immunofluorescence, and real-time quantitative PCR (qPCR). Insulin and chemokine 
      secretion were measured by ELISA and Meso Scale Discovery multiplexing 
      electrochemiluminescence, respectively. Mitochondrial function was evaluated by 
      extracellular flux technology. Global gene expression was characterized by 
      stranded RNA sequencing. RESULTS: Cytokines increased caspase-3/7 activity and 
      cytotoxicity in EndoC-betaH5 cells in a time- and dose-dependent manner. The 
      proapoptotic effect of cytokines was primarily driven by IFNgamma signal 
      transduction. Cytokine exposure induced MHC-I expression and chemokine production 
      and secretion. Further, cytokines caused impaired mitochondrial function and 
      diminished glucose-stimulated insulin secretion. Finally, we report significant 
      changes to the EndoC-betaH5 transcriptome including upregulation of the human 
      leukocyte antigen (HLA) genes, endoplasmic reticulum stress markers, and 
      non-coding RNAs, in response to cytokines. Among the differentially expressed 
      genes were several type 1 diabetes risk genes. CONCLUSION: Our study provides 
      detailed insight into the functional and transcriptomic effects of cytokines on 
      EndoC-betaH5 cells. This information should be useful for future studies using this 
      novel beta-cell model.
CI  - Copyright (c) 2023 Frorup, Gerwig, Svane, Mendes Lopes de Melo, Henriksen, Floyel, 
      Pociot, Kaur and Storling.
FAU - Frorup, Caroline
AU  - Frorup C
AD  - Translational Type 1 Diabetes Research, Clinical Research, Steno Diabetes Center 
      Copenhagen, Herlev, Denmark.
FAU - Gerwig, Rebekka
AU  - Gerwig R
AD  - Translational Type 1 Diabetes Research, Clinical Research, Steno Diabetes Center 
      Copenhagen, Herlev, Denmark.
FAU - Svane, Cecilie Amalie Sondergaard
AU  - Svane CAS
AD  - Translational Type 1 Diabetes Research, Clinical Research, Steno Diabetes Center 
      Copenhagen, Herlev, Denmark.
FAU - Mendes Lopes de Melo, Joana
AU  - Mendes Lopes de Melo J
AD  - Translational Type 1 Diabetes Research, Clinical Research, Steno Diabetes Center 
      Copenhagen, Herlev, Denmark.
FAU - Henriksen, Kristine
AU  - Henriksen K
AD  - Translational Type 1 Diabetes Research, Clinical Research, Steno Diabetes Center 
      Copenhagen, Herlev, Denmark.
FAU - Floyel, Tina
AU  - Floyel T
AD  - Translational Type 1 Diabetes Research, Clinical Research, Steno Diabetes Center 
      Copenhagen, Herlev, Denmark.
FAU - Pociot, Flemming
AU  - Pociot F
AD  - Translational Type 1 Diabetes Research, Clinical Research, Steno Diabetes Center 
      Copenhagen, Herlev, Denmark.
AD  - Faculty of Health and Medical Sciences, University of Copenhagen, Copenhagen, 
      Denmark.
FAU - Kaur, Simranjeet
AU  - Kaur S
AD  - Translational Type 1 Diabetes Research, Clinical Research, Steno Diabetes Center 
      Copenhagen, Herlev, Denmark.
FAU - Storling, Joachim
AU  - Storling J
AD  - Translational Type 1 Diabetes Research, Clinical Research, Steno Diabetes Center 
      Copenhagen, Herlev, Denmark.
AD  - Department of Biomedical Sciences, University of Copenhagen, Copenhagen, Denmark.
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20230411
PL  - Switzerland
TA  - Front Endocrinol (Lausanne)
JT  - Frontiers in endocrinology
JID - 101555782
RN  - 0 (Cytokines)
RN  - EC 3.4.22.- (Caspase 3)
RN  - 82115-62-6 (Interferon-gamma)
RN  - 0 (Chemokines)
SB  - IM
MH  - Humans
MH  - *Cytokines
MH  - Transcriptome
MH  - *Diabetes Mellitus, Type 1
MH  - Caspase 3/genetics
MH  - Interferon-gamma/pharmacology
MH  - Chemokines
PMC - PMC10126300
OTO - NOTNLM
OT  - RNA-Seq
OT  - apoptosis
OT  - inflammation
OT  - insulin
OT  - model system
OT  - pancreatic beta cells
OT  - signaling
OT  - type 1 diabetes
COIS- The authors declare that the research was conducted in the absence of any 
      commercial or financial relationships that could be construed as a potential 
      conflict of interest.
EDAT- 2023/04/28 06:42
MHDA- 2023/05/01 06:42
PMCR- 2023/01/01
CRDT- 2023/04/28 02:29
PHST- 2022/12/20 00:00 [received]
PHST- 2023/03/10 00:00 [accepted]
PHST- 2023/05/01 06:42 [medline]
PHST- 2023/04/28 06:42 [pubmed]
PHST- 2023/04/28 02:29 [entrez]
PHST- 2023/01/01 00:00 [pmc-release]
AID - 10.3389/fendo.2023.1128523 [doi]
PST - epublish
SO  - Front Endocrinol (Lausanne). 2023 Apr 11;14:1128523. doi: 
      10.3389/fendo.2023.1128523. eCollection 2023.