PHENYLBUTYRIC ACID REDUCES MOLECULAR MARKERS OF ER STRESS-INDUCED APOPTOSIS IN GLYCOGEN STORAGE DISEASE TYPE IB IN VITRO MODEL SYSTEM
Аутори
Parezanović, MarinaStevanović, Nina
Anđelković, Marina
Ugrin, Milena
Pavlović, Sonja
Stojiljković, Maja
Skakić, Anita
Конференцијски прилог (Објављена верзија)
Метаподаци
Приказ свих података о документуАпстракт
Background: The current therapy for glycogen
storage disease Ib (GSD Ib) fails to prevent the
development of renal dysfunction, and hepatocellular/
renal carcinoma in many patients. Therefore,
new therapies for the treatment of life-threatening
complications of GSD Ib are of great interest. Recent
studies revealed that chronic endoplasmic reticulum
(ER) stress and increased apoptosis are
involved in pathogenesis of GSD Ib, whereas small
molecule phenylbutyrate (4-PBA) showed the capability
of reducing ER stress-induced apoptosis.
Methods: To analyze the function of 4-PBA
as ER stress inhibitor, we created a G6PT-deficient
FlpInHEK293 cell line using the CRISPR/Cas9
knockout method and tested if 4-PBA could decrease
chronic metabolic stress and prevent cell
death. We analyzed molecular markers of unfolded
protein response (ATF4, DDIT3, HSPA5, XBP1s),
and apoptosis (BCL2/BAX, CASP3, CASP7) in
G6PT-deficient cells before and upon the treatment
using RT-qPCR method.Resu...lts: Treatment with the most effective
dose of 1 mM 4-PBA reduced the expression of
executioner caspases (CASP3, CASP7) and increased
the BCL2/BAX ratio, indicating a reduced
apoptosis level. Additionally, 4-PBA decreased
UPR marker expression in G6PT-deficient cells.
Our results proved the concept that 4-PBA could
alleviate markers of ER stress detected in the GSD
Ib in vitro model system and prevent cell death.
Conclusion: We demonstrated, for the first
time, the potential of 4-PBA to be repurposed for
patients with GSD Ib and open perspectives for
translational research that could contribute to a
knowledge of GSD Ib treatments and other genetic
diseases where chronic ER stress-induced apoptosis
contribute to the disease pathology.
Кључне речи:
GSD Ib in vitro model system / CRISPR/Cas9 / 4-PBA treatment / ER stress / apoptosis / rare diseasesИзвор:
International Journal of Medical Genetics, 2023, 26, Supplement, 54-54Издавач:
- International Journal of Medical Genetic
Напомена:
- ABSTRACT BOOK: “Genetic Diseases from Diagnostics to Prevention and Therapy” October 05-14th Balkan Congress of Human Genetics & 9th Rare Disease SEE Meeting 2023; Skopje, October 05-07, 2023
Институција/група
Institut za molekularnu genetiku i genetičko inženjerstvoTY - CONF AU - Parezanović, Marina AU - Stevanović, Nina AU - Anđelković, Marina AU - Ugrin, Milena AU - Pavlović, Sonja AU - Stojiljković, Maja AU - Skakić, Anita PY - 2023 UR - https://imagine.imgge.bg.ac.rs/handle/123456789/2176 AB - Background: The current therapy for glycogen storage disease Ib (GSD Ib) fails to prevent the development of renal dysfunction, and hepatocellular/ renal carcinoma in many patients. Therefore, new therapies for the treatment of life-threatening complications of GSD Ib are of great interest. Recent studies revealed that chronic endoplasmic reticulum (ER) stress and increased apoptosis are involved in pathogenesis of GSD Ib, whereas small molecule phenylbutyrate (4-PBA) showed the capability of reducing ER stress-induced apoptosis. Methods: To analyze the function of 4-PBA as ER stress inhibitor, we created a G6PT-deficient FlpInHEK293 cell line using the CRISPR/Cas9 knockout method and tested if 4-PBA could decrease chronic metabolic stress and prevent cell death. We analyzed molecular markers of unfolded protein response (ATF4, DDIT3, HSPA5, XBP1s), and apoptosis (BCL2/BAX, CASP3, CASP7) in G6PT-deficient cells before and upon the treatment using RT-qPCR method.Results: Treatment with the most effective dose of 1 mM 4-PBA reduced the expression of executioner caspases (CASP3, CASP7) and increased the BCL2/BAX ratio, indicating a reduced apoptosis level. Additionally, 4-PBA decreased UPR marker expression in G6PT-deficient cells. Our results proved the concept that 4-PBA could alleviate markers of ER stress detected in the GSD Ib in vitro model system and prevent cell death. Conclusion: We demonstrated, for the first time, the potential of 4-PBA to be repurposed for patients with GSD Ib and open perspectives for translational research that could contribute to a knowledge of GSD Ib treatments and other genetic diseases where chronic ER stress-induced apoptosis contribute to the disease pathology. PB - International Journal of Medical Genetic C3 - International Journal of Medical Genetics T1 - PHENYLBUTYRIC ACID REDUCES MOLECULAR MARKERS OF ER STRESS-INDUCED APOPTOSIS IN GLYCOGEN STORAGE DISEASE TYPE IB IN VITRO MODEL SYSTEM EP - 54 IS - Supplement SP - 54 VL - 26 UR - https://hdl.handle.net/21.15107/rcub_imagine_2176 ER -
@conference{ author = "Parezanović, Marina and Stevanović, Nina and Anđelković, Marina and Ugrin, Milena and Pavlović, Sonja and Stojiljković, Maja and Skakić, Anita", year = "2023", abstract = "Background: The current therapy for glycogen storage disease Ib (GSD Ib) fails to prevent the development of renal dysfunction, and hepatocellular/ renal carcinoma in many patients. Therefore, new therapies for the treatment of life-threatening complications of GSD Ib are of great interest. Recent studies revealed that chronic endoplasmic reticulum (ER) stress and increased apoptosis are involved in pathogenesis of GSD Ib, whereas small molecule phenylbutyrate (4-PBA) showed the capability of reducing ER stress-induced apoptosis. Methods: To analyze the function of 4-PBA as ER stress inhibitor, we created a G6PT-deficient FlpInHEK293 cell line using the CRISPR/Cas9 knockout method and tested if 4-PBA could decrease chronic metabolic stress and prevent cell death. We analyzed molecular markers of unfolded protein response (ATF4, DDIT3, HSPA5, XBP1s), and apoptosis (BCL2/BAX, CASP3, CASP7) in G6PT-deficient cells before and upon the treatment using RT-qPCR method.Results: Treatment with the most effective dose of 1 mM 4-PBA reduced the expression of executioner caspases (CASP3, CASP7) and increased the BCL2/BAX ratio, indicating a reduced apoptosis level. Additionally, 4-PBA decreased UPR marker expression in G6PT-deficient cells. Our results proved the concept that 4-PBA could alleviate markers of ER stress detected in the GSD Ib in vitro model system and prevent cell death. Conclusion: We demonstrated, for the first time, the potential of 4-PBA to be repurposed for patients with GSD Ib and open perspectives for translational research that could contribute to a knowledge of GSD Ib treatments and other genetic diseases where chronic ER stress-induced apoptosis contribute to the disease pathology.", publisher = "International Journal of Medical Genetic", journal = "International Journal of Medical Genetics", title = "PHENYLBUTYRIC ACID REDUCES MOLECULAR MARKERS OF ER STRESS-INDUCED APOPTOSIS IN GLYCOGEN STORAGE DISEASE TYPE IB IN VITRO MODEL SYSTEM", pages = "54-54", number = "Supplement", volume = "26", url = "https://hdl.handle.net/21.15107/rcub_imagine_2176" }
Parezanović, M., Stevanović, N., Anđelković, M., Ugrin, M., Pavlović, S., Stojiljković, M.,& Skakić, A.. (2023). PHENYLBUTYRIC ACID REDUCES MOLECULAR MARKERS OF ER STRESS-INDUCED APOPTOSIS IN GLYCOGEN STORAGE DISEASE TYPE IB IN VITRO MODEL SYSTEM. in International Journal of Medical Genetics International Journal of Medical Genetic., 26(Supplement), 54-54. https://hdl.handle.net/21.15107/rcub_imagine_2176
Parezanović M, Stevanović N, Anđelković M, Ugrin M, Pavlović S, Stojiljković M, Skakić A. PHENYLBUTYRIC ACID REDUCES MOLECULAR MARKERS OF ER STRESS-INDUCED APOPTOSIS IN GLYCOGEN STORAGE DISEASE TYPE IB IN VITRO MODEL SYSTEM. in International Journal of Medical Genetics. 2023;26(Supplement):54-54. https://hdl.handle.net/21.15107/rcub_imagine_2176 .
Parezanović, Marina, Stevanović, Nina, Anđelković, Marina, Ugrin, Milena, Pavlović, Sonja, Stojiljković, Maja, Skakić, Anita, "PHENYLBUTYRIC ACID REDUCES MOLECULAR MARKERS OF ER STRESS-INDUCED APOPTOSIS IN GLYCOGEN STORAGE DISEASE TYPE IB IN VITRO MODEL SYSTEM" in International Journal of Medical Genetics, 26, no. Supplement (2023):54-54, https://hdl.handle.net/21.15107/rcub_imagine_2176 .