Changes in up and down regulated gene expression after transient suppression of Arx gene in pancreatic alphaTC1-6 cell line
2023
Аутори:
Đorđević, MarijaFeuerstein-Akgoz, Clarissa
Arambašić Jovanović, Jelena
Tolić, Anja
Rajić, Jovana
Sarić, Ana
Grdović, Nevena
Dinić, Svetlana
Uskoković, Aleksandra
Mihailović, Mirjana
Gerhauser, Clarissa
Jurkowski, Tomasz
Vidaković, Melita
Остала ауторства
Dobrijević, ZoranaТип документа:
Конференцијски прилог (Објављена верзија)
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© 2023 by the Institute of Molecular Genetics and Genetic Engineering, University of Belgrade
Метаподаци
Приказ свих података о документуАпстракт:
Introduction: The Aristaless-related homeobox (Arx) gene plays a key role in the development and maintaining pancreatic alpha cell phenotype, and as such represents an excellent target for alpha cell identity change towards insulin-producing cells. Therefore, this cell switch and increase in beta cell mass could be of potential use in diabetes management.
Methods: On the fifth day after transient transfection with dCas9-Dnmt3a3L-KRAB construct and four gRNAs targeting Arx promoter, αTC1-6 were sorted to collect GFP-positive (transfected) cells (EpiC). The mRNA-seq libraries were pooled in equimolar amounts and sequenced in a single end-setting on the Illumina NextSeq 500 High output machine with 75 bases long reads. KEGG pathway overrepresentation analysis was performed using an application on all significantly up- or downregulated genes using default settings.
Results: Directed induction of DNA methylation on the Arx gene promoter reduces its expression and causes the up-regulation of 357 genes, while 266 genes were down-regulated in EpiC compared to Mock transfected cells. The KEGG pathways analysis of biological processes confirmed several biological pathways associated with genes differentially expressed in EpiC vs. Mock transfected cells at the 5th posttransfection day (pval ≤ 0.05). As the most significant, up-regulated pathways we found Type II diabetes, Insulin secretion, Longevity regulation pathways. As significant, down-regulated pathways pop-up Fatty acid metabolism and PPAR signaling pathway.
Conclusion: Reduction of ArxmRNA level is sufficient to initiate the transdifferentiation process of alpha cells into insulin-producing cells by triggering several biological pathways tight related to insulin secretion and function.
Кључне речи:
CRISPR/dCas9; alpha cells; methylation; Arx gene; epigenetic editingФинансирање / пројекти:
- European Foundation for the Study of Diabetes (EFSD), European Diabetes Research Programme in Cellular Plasticity Underlying the Pathophysiology of Type 2 Diabetes, research grant from Astra Zeneca
- Министарство науке, технолошког развоја и иновација Републике Србије, институционално финансирање - 200007 (Универзитет у Београду, Институт за биолошка истраживања 'Синиша Станковић') (RS-MESTD-inst-2020-200007)
У:
- Dobrijević Z, editor. Abstract Book: CoMBoS2 - the Second Congress of Molecular Biologists of Serbia; 2023 Oct 6-8; Belgrade, Serbia. Belgrade: Institute of Molecular Genetics and Genetic Engineering, University of Belgrade ; 2023. p. 151. (Trends in Molecular Biology; Special Issue).