TY  - CONF
AU  - Uskoković, Aleksandra
PY  - 2023
UR  - http://radar.ibiss.bg.ac.rs/handle/123456789/6308
AB  - Epigenetic editing has a powerful potential to direct reprogramming of cellular phenotype that can be used in disease modeling. The reprogramming of cells from different origin into insulin-producing cells could provide a solution for restoring functional beta cell mass in diabetic patients. We used CRISPR/dCas9-based epigenetic tool for targeted hypermethylation of Arx promoter and its subsequent suppression in mouse pancreatic α cell line. By epigenetic silencing of Arx we successfully triggered a direct, transient switch of pancreatic α- to insulin-producing cells obtaining approximately 1% of transiently transfected cells which were able to produce 35% more insulin than Mock transfected α cells. As a future perspective we intend to address the potential use of epigenetic editing tool as a pre-therapeutic approach in triple-negative breast cancers (TNBCs) with unknown mutational signature of BRCA1. The BRCA1methylation (BRCAness) as a predictor for response to therapeutics such as PARPi would allow direct TNBC treatment without previous screening for BRCA1 mutations. The main objective will be to induce BRCAness by suppressing BRCA1 expression in TNBC cells via targeted DNA methylation of BRCA1 promoter using the synthetic epigenetic editing tool. This approach would enable the faster decision toward the use of newest medicaments to increase cells’ apoptosis and cancer cell diminishment.
PB  - Belgrade: Institute of Molecular Genetics and Genetic Engineering, University of Belgrade
C3  - Abstract Book: CoMBoS2 - the Second Congress of Molecular Biologists of Serbia; 2023 Oct 6-8; Belgrade, Serbia
T1  - Epigenetic editing as a potential therapeutic tool for the treatment of noncommunicable diseases
SP  - 132
EP  - 132
UR  - https://hdl.handle.net/21.15107/rcub_ibiss_6308
ER  - 

@conference{
author = "Uskoković, Aleksandra",
year = "2023",
abstract = "Epigenetic editing has a powerful potential to direct reprogramming of cellular phenotype that can be used in disease modeling. The reprogramming of cells from different origin into insulin-producing cells could provide a solution for restoring functional beta cell mass in diabetic patients. We used CRISPR/dCas9-based epigenetic tool for targeted hypermethylation of Arx promoter and its subsequent suppression in mouse pancreatic α cell line. By epigenetic silencing of Arx we successfully triggered a direct, transient switch of pancreatic α- to insulin-producing cells obtaining approximately 1% of transiently transfected cells which were able to produce 35% more insulin than Mock transfected α cells. As a future perspective we intend to address the potential use of epigenetic editing tool as a pre-therapeutic approach in triple-negative breast cancers (TNBCs) with unknown mutational signature of BRCA1. The BRCA1methylation (BRCAness) as a predictor for response to therapeutics such as PARPi would allow direct TNBC treatment without previous screening for BRCA1 mutations. The main objective will be to induce BRCAness by suppressing BRCA1 expression in TNBC cells via targeted DNA methylation of BRCA1 promoter using the synthetic epigenetic editing tool. This approach would enable the faster decision toward the use of newest medicaments to increase cells’ apoptosis and cancer cell diminishment.",
publisher = "Belgrade: Institute of Molecular Genetics and Genetic Engineering, University of Belgrade",
journal = "Abstract Book: CoMBoS2 - the Second Congress of Molecular Biologists of Serbia; 2023 Oct 6-8; Belgrade, Serbia",
title = "Epigenetic editing as a potential therapeutic tool for the treatment of noncommunicable diseases",
pages = "132-132",
url = "https://hdl.handle.net/21.15107/rcub_ibiss_6308"
}

Uskoković, A.. (2023). Epigenetic editing as a potential therapeutic tool for the treatment of noncommunicable diseases. in 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., 132-132.
https://hdl.handle.net/21.15107/rcub_ibiss_6308

Uskoković A. Epigenetic editing as a potential therapeutic tool for the treatment of noncommunicable diseases. in Abstract Book: CoMBoS2 - the Second Congress of Molecular Biologists of Serbia; 2023 Oct 6-8; Belgrade, Serbia. 2023;:132-132.
https://hdl.handle.net/21.15107/rcub_ibiss_6308 .

Uskoković, Aleksandra, "Epigenetic editing as a potential therapeutic tool for the treatment of noncommunicable diseases" in Abstract Book: CoMBoS2 - the Second Congress of Molecular Biologists of Serbia; 2023 Oct 6-8; Belgrade, Serbia (2023):132-132,
https://hdl.handle.net/21.15107/rcub_ibiss_6308 .

TY  - CONF
AU  - Vidaković, Melita
AU  - Đorđević, Marija
AU  - Arambašić Jovanović, Jelena
AU  - Tolić, Anja
AU  - Đorđević, Miloš
AU  - Mihailović, Mirjana
AU  - Grdović, Nevena
AU  - Uskoković, Aleksandra
AU  - Rajić, Jovana
AU  - Poznanović, Goran
AU  - Dinić, Svetlana
AU  - Jurkowski, Tomasz
PY  - 2017
UR  - http://radar.ibiss.bg.ac.rs/handle/123456789/5816
AB  - Introduction and aim: Diabetes is the perfect candidate for cell replacement therapy since it is caused by either an absolute (type 1 diabetes) or relative (type 2 diabetes) defect of insulin-producing pancreatic beta cells (b-cells}. We focused on applying a novel synthetic epigenetic tool (Epi-CRISPRs) for a straightforward, one­-step transdifferentiation of mouse pancreatic alpha (a-cells) to b-cell by targeted DNA methylation and suppression of genes essential for maintaining pancreatic cell identity (homeobox Arx gene (Arx)). 
Methods: The a-cells were transiently transfected with four different Epi-CRISPR constructs and co-transfected with a single guided RNA (gRNA} or with a mix of different gRNAs all targeting different promoter regions of Arx. After 5, 8 and 12 days post-transfection, DNA and RNA were isolated and the cells were immunostained. The transdifferentiated cells were analysed for key features of bona fide cells, using qPCR to assess Arx expression, and immunostaining of insulin/glucagon and ELISA for measuring secreted insulin. 
Results: We succeeded to transiently transfect a-cells with Epi-CRISPR constructs and 275 gRNA/mix gRNA. The suppression of Arx in a-cells was confirmed on days 5 and 8 post-transfection. The reduction of glucagon synthesis and beginning of insulin production in transfected a-cell was confirmed and visualised by immunostaining. Whether DNA methylation-mediated suppression of Arx in a-cells lead to their transdifferentiation to insulin-producing cells, will be confirmed by bisulfite sequencing. 
Conclusion: We are on the right course of developing a clear-cut technology capable of providing a perfect delivery system for increasing the number of insulin­producing cells in vitro.
PB  - Belgrade: University of Belgrade, Faculty of Biology
C3  - Book of Abstracts: 1st Congress of Molecular Biologists of Serbia: CoMBoS; 2017 Sep 20-21; Belgrade, Serbia.
T1  - Therapeutic genome methylation for cell reprogramming editing: use of Epi-CRISPR-induced targeted DNA
SP  - 10
UR  - https://hdl.handle.net/21.15107/rcub_ibiss_5816
ER  - 

@conference{
author = "Vidaković, Melita and Đorđević, Marija and Arambašić Jovanović, Jelena and Tolić, Anja and Đorđević, Miloš and Mihailović, Mirjana and Grdović, Nevena and Uskoković, Aleksandra and Rajić, Jovana and Poznanović, Goran and Dinić, Svetlana and Jurkowski, Tomasz",
year = "2017",
abstract = "Introduction and aim: Diabetes is the perfect candidate for cell replacement therapy since it is caused by either an absolute (type 1 diabetes) or relative (type 2 diabetes) defect of insulin-producing pancreatic beta cells (b-cells}. We focused on applying a novel synthetic epigenetic tool (Epi-CRISPRs) for a straightforward, one­-step transdifferentiation of mouse pancreatic alpha (a-cells) to b-cell by targeted DNA methylation and suppression of genes essential for maintaining pancreatic cell identity (homeobox Arx gene (Arx)). 
Methods: The a-cells were transiently transfected with four different Epi-CRISPR constructs and co-transfected with a single guided RNA (gRNA} or with a mix of different gRNAs all targeting different promoter regions of Arx. After 5, 8 and 12 days post-transfection, DNA and RNA were isolated and the cells were immunostained. The transdifferentiated cells were analysed for key features of bona fide cells, using qPCR to assess Arx expression, and immunostaining of insulin/glucagon and ELISA for measuring secreted insulin. 
Results: We succeeded to transiently transfect a-cells with Epi-CRISPR constructs and 275 gRNA/mix gRNA. The suppression of Arx in a-cells was confirmed on days 5 and 8 post-transfection. The reduction of glucagon synthesis and beginning of insulin production in transfected a-cell was confirmed and visualised by immunostaining. Whether DNA methylation-mediated suppression of Arx in a-cells lead to their transdifferentiation to insulin-producing cells, will be confirmed by bisulfite sequencing. 
Conclusion: We are on the right course of developing a clear-cut technology capable of providing a perfect delivery system for increasing the number of insulin­producing cells in vitro.",
publisher = "Belgrade: University of Belgrade, Faculty of Biology",
journal = "Book of Abstracts: 1st Congress of Molecular Biologists of Serbia: CoMBoS; 2017 Sep 20-21; Belgrade, Serbia.",
title = "Therapeutic genome methylation for cell reprogramming editing: use of Epi-CRISPR-induced targeted DNA",
pages = "10",
url = "https://hdl.handle.net/21.15107/rcub_ibiss_5816"
}

Vidaković, M., Đorđević, M., Arambašić Jovanović, J., Tolić, A., Đorđević, M., Mihailović, M., Grdović, N., Uskoković, A., Rajić, J., Poznanović, G., Dinić, S.,& Jurkowski, T.. (2017). Therapeutic genome methylation for cell reprogramming editing: use of Epi-CRISPR-induced targeted DNA. in Book of Abstracts: 1st Congress of Molecular Biologists of Serbia: CoMBoS; 2017 Sep 20-21; Belgrade, Serbia.
Belgrade: University of Belgrade, Faculty of Biology., 10.
https://hdl.handle.net/21.15107/rcub_ibiss_5816

Vidaković M, Đorđević M, Arambašić Jovanović J, Tolić A, Đorđević M, Mihailović M, Grdović N, Uskoković A, Rajić J, Poznanović G, Dinić S, Jurkowski T. Therapeutic genome methylation for cell reprogramming editing: use of Epi-CRISPR-induced targeted DNA. in Book of Abstracts: 1st Congress of Molecular Biologists of Serbia: CoMBoS; 2017 Sep 20-21; Belgrade, Serbia.. 2017;:10.
https://hdl.handle.net/21.15107/rcub_ibiss_5816 .

Vidaković, Melita, Đorđević, Marija, Arambašić Jovanović, Jelena, Tolić, Anja, Đorđević, Miloš, Mihailović, Mirjana, Grdović, Nevena, Uskoković, Aleksandra, Rajić, Jovana, Poznanović, Goran, Dinić, Svetlana, Jurkowski, Tomasz, "Therapeutic genome methylation for cell reprogramming editing: use of Epi-CRISPR-induced targeted DNA" in Book of Abstracts: 1st Congress of Molecular Biologists of Serbia: CoMBoS; 2017 Sep 20-21; Belgrade, Serbia. (2017):10,
https://hdl.handle.net/21.15107/rcub_ibiss_5816 .