Transdifferentiation of pancreatic alpha to beta cells via targeted epigenome editing by Epi-CRISPRs-s directed DNA methylation
2017
Аутори:
Đorđević, MarijaArambašić Jovanović, Jelena
Tolić, Anja
Đorđević, Miloš
Mihailović, Mirjana
Grdović, Nevena
Uskoković, Aleksandra
Rajić, Jovana
Poznanović, Goran
Dinić, Svetlana
Jurkowski, Tomasz
Vidaković, Melita
Остала ауторства
Mourdjeva, MilenaKistanova, Elena
Fazeli, Alireza
Brevini, Tiziana
Тип документа:
Конференцијски прилог (Објављена верзија)
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© 2017 COST Office
Метаподаци
Приказ свих података о документуАпстракт:
We propose to transdifferentiate alpha to beta cells using our recently developed Epi-CRISPRs, a novel synthetic epigenetic tool. Using this methodology we are able to induce straightforward, one-step cell transdifferentiation by targeted DNA methylation and suppression of homeobox gene Arx that is essential for maintaining pancreatic alpha cell identity.
The Epi-CRISPR constructs with and one or four different guide RNAs for specific targeting the promoter region of Arx, were transiently transfected in alphaTCl-6 cells (alpha-cells). The success of a-cells transdifferentiation into insulin-producing cells was evaluated by measuring Arx, glucagon (Glu) and insulin (lns2) mRNA level, amount of secreted insulin and by immunostaining of insulin and glucagon in the cells.
Our study will be valuable for later subsequent Epi-CRISPRs use in mouse in vivo model of diabetes and eventually as a potential therapy for diabetes attenuation in humans.
Финансирање / 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
- Сигнални молекули у дијабетесу: идентификација потенцијалних биолошких маркера укључених у модификацију и интеграцију сигналних путева у циљу предикције и интервенције у дијабетесу (RS-MESTD-Basic Research (BR or ON)-173020)
У:
- Mourdjeva M, Kistanova E, Fazeli A, Brevini T, editors. Proceedings of CellFit meeting: In vitro 3-D total cell guidance and fitness; 2017 Sep 12-14; Albena Resort, Bulgaria. Institute of Biology and Immunology of Reproduction, BAS, COST Action CA16119 and Mouseprint Ltd.; 2017. p. 80.