CRISPR/Cas9 genome editing of SLC37A4 gene elucidates the role of molecular markers of endoplasmic reticulum stress and apoptosis in renal involvement in glycogen storage disease type Ib
Нема приказа
Аутори
Skakić, AnitaAnđelković, Marina
Tošić, Nataša
Klaassen, Kristel
Đorđević, Maja
Pavlović, Sonja
Stojiljković, Maja
Чланак у часопису (Објављена верзија)
Метаподаци
Приказ свих података о документуАпстракт
Glycogen storage disease type Ib (GSD Ib) is an autosomal recessive disorder, caused by a deficiency of ubiquitously expressed SLC37A4 protein. Deficiency of SLC37A4 leads to abnormal storage of glycogen in the liver and kidneys, resulting in long-term complications of renal disease and hepatocellular adenomas, whose mechanisms are poorly understood. Molecular markers of the adaptive responses to the metabolic stress caused by a deficiency of SLC37A4, such as markers related to the endoplasmic reticulum (ER) stress and unfolded protein response (UPR), have not been extensively studied. The aim of this study was to investigate the expression of molecular markers of the UPR response and apoptosis related to a deficiency of SLC37A4 in kidney cells. For that purpose, we intended to establish a human kidney cell model system for GSD Ib. The novel variant c.248G gt A, found in GSD Ib patients, was introduced into the Flp-In (TM) T-REx (TM)-293 cell line using CRISPR/Cas9-mediated precise g...ene editing method, resulting in significant decrease of SLC37A4 gene expression. In this model system we used RT-qPCR analysis to investigate the expression of molecular markers of the UPR response (ATF4, DDIT3, HSPA5, and XBP1s) and apoptosis (BCL2, BAX). We demonstrated that under chronic metabolic stress conditions caused by SLC37A4 deficiency, the ER stress-induced UPR was triggered, resulting in suppression of the UPR molecular markers and cell survival promotion (decreased expression levels of ATF4, DDIT3, HSPA5, with the exception of XHE1s). However, persistent metabolic stress overrides an adaptation and induces apoptosis through increased expression of pro-apoptotic markers (decreased ratio of BCL2/BAX genes). We established a cellular model system characterized by a deficiency of SLC37A4, which presents pathological manifestations of GSD Ib in the kidney. Expression analysis in a novel model system supports the hypothesis that renal dysfunction in the GSD Ib is partly due to the ER stress and increased apoptosis.
Кључне речи:
Unfolded protein response / SLC37A4 gene / GSD Ib in vitro model system / ER-mediated apoptosis / ER stress / CRISPR/Cas9 gene knockinИзвор:
Gene, 2019, 703, 17-25Издавач:
- Elsevier Science Bv, Amsterdam
Финансирање / пројекти:
- Ретке болести: молекуларна патофизиологија, дијагностички и терапијски модалитети и социјални, етички и правни аспекти (RS-MESTD-Integrated and Interdisciplinary Research (IIR or III)-41004)
DOI: 10.1016/j.gene.2019.04.002
ISSN: 0378-1119
PubMed: 30951856
WoS: 000469904800004
Scopus: 2-s2.0-85063992028
Институција/група
Institut za molekularnu genetiku i genetičko inženjerstvoTY - JOUR AU - Skakić, Anita AU - Anđelković, Marina AU - Tošić, Nataša AU - Klaassen, Kristel AU - Đorđević, Maja AU - Pavlović, Sonja AU - Stojiljković, Maja PY - 2019 UR - https://imagine.imgge.bg.ac.rs/handle/123456789/1254 AB - Glycogen storage disease type Ib (GSD Ib) is an autosomal recessive disorder, caused by a deficiency of ubiquitously expressed SLC37A4 protein. Deficiency of SLC37A4 leads to abnormal storage of glycogen in the liver and kidneys, resulting in long-term complications of renal disease and hepatocellular adenomas, whose mechanisms are poorly understood. Molecular markers of the adaptive responses to the metabolic stress caused by a deficiency of SLC37A4, such as markers related to the endoplasmic reticulum (ER) stress and unfolded protein response (UPR), have not been extensively studied. The aim of this study was to investigate the expression of molecular markers of the UPR response and apoptosis related to a deficiency of SLC37A4 in kidney cells. For that purpose, we intended to establish a human kidney cell model system for GSD Ib. The novel variant c.248G gt A, found in GSD Ib patients, was introduced into the Flp-In (TM) T-REx (TM)-293 cell line using CRISPR/Cas9-mediated precise gene editing method, resulting in significant decrease of SLC37A4 gene expression. In this model system we used RT-qPCR analysis to investigate the expression of molecular markers of the UPR response (ATF4, DDIT3, HSPA5, and XBP1s) and apoptosis (BCL2, BAX). We demonstrated that under chronic metabolic stress conditions caused by SLC37A4 deficiency, the ER stress-induced UPR was triggered, resulting in suppression of the UPR molecular markers and cell survival promotion (decreased expression levels of ATF4, DDIT3, HSPA5, with the exception of XHE1s). However, persistent metabolic stress overrides an adaptation and induces apoptosis through increased expression of pro-apoptotic markers (decreased ratio of BCL2/BAX genes). We established a cellular model system characterized by a deficiency of SLC37A4, which presents pathological manifestations of GSD Ib in the kidney. Expression analysis in a novel model system supports the hypothesis that renal dysfunction in the GSD Ib is partly due to the ER stress and increased apoptosis. PB - Elsevier Science Bv, Amsterdam T2 - Gene T1 - CRISPR/Cas9 genome editing of SLC37A4 gene elucidates the role of molecular markers of endoplasmic reticulum stress and apoptosis in renal involvement in glycogen storage disease type Ib EP - 25 SP - 17 VL - 703 DO - 10.1016/j.gene.2019.04.002 ER -
@article{ author = "Skakić, Anita and Anđelković, Marina and Tošić, Nataša and Klaassen, Kristel and Đorđević, Maja and Pavlović, Sonja and Stojiljković, Maja", year = "2019", abstract = "Glycogen storage disease type Ib (GSD Ib) is an autosomal recessive disorder, caused by a deficiency of ubiquitously expressed SLC37A4 protein. Deficiency of SLC37A4 leads to abnormal storage of glycogen in the liver and kidneys, resulting in long-term complications of renal disease and hepatocellular adenomas, whose mechanisms are poorly understood. Molecular markers of the adaptive responses to the metabolic stress caused by a deficiency of SLC37A4, such as markers related to the endoplasmic reticulum (ER) stress and unfolded protein response (UPR), have not been extensively studied. The aim of this study was to investigate the expression of molecular markers of the UPR response and apoptosis related to a deficiency of SLC37A4 in kidney cells. For that purpose, we intended to establish a human kidney cell model system for GSD Ib. The novel variant c.248G gt A, found in GSD Ib patients, was introduced into the Flp-In (TM) T-REx (TM)-293 cell line using CRISPR/Cas9-mediated precise gene editing method, resulting in significant decrease of SLC37A4 gene expression. In this model system we used RT-qPCR analysis to investigate the expression of molecular markers of the UPR response (ATF4, DDIT3, HSPA5, and XBP1s) and apoptosis (BCL2, BAX). We demonstrated that under chronic metabolic stress conditions caused by SLC37A4 deficiency, the ER stress-induced UPR was triggered, resulting in suppression of the UPR molecular markers and cell survival promotion (decreased expression levels of ATF4, DDIT3, HSPA5, with the exception of XHE1s). However, persistent metabolic stress overrides an adaptation and induces apoptosis through increased expression of pro-apoptotic markers (decreased ratio of BCL2/BAX genes). We established a cellular model system characterized by a deficiency of SLC37A4, which presents pathological manifestations of GSD Ib in the kidney. Expression analysis in a novel model system supports the hypothesis that renal dysfunction in the GSD Ib is partly due to the ER stress and increased apoptosis.", publisher = "Elsevier Science Bv, Amsterdam", journal = "Gene", title = "CRISPR/Cas9 genome editing of SLC37A4 gene elucidates the role of molecular markers of endoplasmic reticulum stress and apoptosis in renal involvement in glycogen storage disease type Ib", pages = "25-17", volume = "703", doi = "10.1016/j.gene.2019.04.002" }
Skakić, A., Anđelković, M., Tošić, N., Klaassen, K., Đorđević, M., Pavlović, S.,& Stojiljković, M.. (2019). CRISPR/Cas9 genome editing of SLC37A4 gene elucidates the role of molecular markers of endoplasmic reticulum stress and apoptosis in renal involvement in glycogen storage disease type Ib. in Gene Elsevier Science Bv, Amsterdam., 703, 17-25. https://doi.org/10.1016/j.gene.2019.04.002
Skakić A, Anđelković M, Tošić N, Klaassen K, Đorđević M, Pavlović S, Stojiljković M. CRISPR/Cas9 genome editing of SLC37A4 gene elucidates the role of molecular markers of endoplasmic reticulum stress and apoptosis in renal involvement in glycogen storage disease type Ib. in Gene. 2019;703:17-25. doi:10.1016/j.gene.2019.04.002 .
Skakić, Anita, Anđelković, Marina, Tošić, Nataša, Klaassen, Kristel, Đorđević, Maja, Pavlović, Sonja, Stojiljković, Maja, "CRISPR/Cas9 genome editing of SLC37A4 gene elucidates the role of molecular markers of endoplasmic reticulum stress and apoptosis in renal involvement in glycogen storage disease type Ib" in Gene, 703 (2019):17-25, https://doi.org/10.1016/j.gene.2019.04.002 . .