title: Expression of heterologous oxalate decarboxylase in HEK293 cells confers protection against oxalate induced oxidative stress as a therapeutic approach for calcium oxalate stone disease.
abstract: Oxalates stimulate alterations in renal epithelial cells and thereby induce calcium oxalate (CaOx) stone formation. Bacillus subtilis YvrK gene encodes for oxalate decarboxylase (OxdC) which degrades oxalate to formate and CO2. The present work is aimed to clone the oxdC gene in a mammalian expression vector pcDNA and transfect into Human Embryonic Kidney 293 (HEK293) cells and evaluate the oxdC expression, cell survival rate and oxalate degrading efficiency. The results indicate cell survival rate of HEK293/pcDNAOXDC cells pre-incubated with oxalate was enhanced by 28%. HEK293/pcDNAOXDC cells expressing OxdC treated with oxalate, significantly restored antioxidant activity, mitochondrial membrane potential and intracellular reactive oxygen species (ROS) generation compared with HEK293/pcDNA. Apoptotic marker caspase 3 downregulation illustrates HEK293/pcDNAOXDC cells were able to survive under oxalate-mediated oxidative stress. The findings suggest HEK293 cells expressing oxdC capable of degrading oxalate protect cells from oxidative damage and thus serve as a therapeutic option for prevention of CaOx stone disease. [Formula: see text].
authors.affiliation: a Department of Biochemistry, Centre for Excellence in Genomics Science , School of Biological Sciences, Madurai Kamaraj University , Madurai , India.
authors.forename: Abhishek
authors.initials: A
authors.lastname: Albert
authors.affiliation: a Department of Biochemistry, Centre for Excellence in Genomics Science , School of Biological Sciences, Madurai Kamaraj University , Madurai , India.
authors.forename: Vidhi
authors.initials: V
authors.lastname: Tiwari
authors.affiliation: a Department of Biochemistry, Centre for Excellence in Genomics Science , School of Biological Sciences, Madurai Kamaraj University , Madurai , India.
authors.forename: Eldho
authors.initials: E
authors.lastname: Paul
authors.affiliation: a Department of Biochemistry, Centre for Excellence in Genomics Science , School of Biological Sciences, Madurai Kamaraj University , Madurai , India.
authors.forename: Divya
authors.initials: D
authors.lastname: Ganesan
authors.affiliation: b DBT-IPLS Programme, School of Biological Science , Madurai Kamaraj University , Madurai , India.
authors.forename: Mahesh
authors.initials: M
authors.lastname: Ayyavu
authors.affiliation: a Department of Biochemistry, Centre for Excellence in Genomics Science , School of Biological Sciences, Madurai Kamaraj University , Madurai , India.
authors.forename: Ritu
authors.initials: R
authors.lastname: Kujur
authors.affiliation: c Department of Oral Biology, School of Dental Medicine , University at Buffalo , Buffalo , NY , USA.
authors.forename: Sasikumar
authors.initials: S
authors.lastname: Ponnusamy
authors.affiliation: d Department of Molecular Biology , Centre for Excellence in Genomics Science, School of Biological Sciences, Madurai Kamaraj University , Madurai , India.
authors.forename: Kathiresan
authors.initials: K
authors.lastname: Shanmugam
authors.affiliation: e Department of Physiology and Pharmacology "Vittorio Erspamer" , Sapienza University of Rome , Rome , Italy.
authors.forename: Luciano
authors.initials: L
authors.lastname: Saso
authors.affiliation: a Department of Biochemistry, Centre for Excellence in Genomics Science , School of Biological Sciences, Madurai Kamaraj University , Madurai , India.
authors.forename: Selvam
authors.initials: S
authors.lastname: Govindan Sadasivam
mesh_terms: D002129:Calcium Oxalate / Q000737:chemistry*; D057809:HEK293 Cells; D006801:Humans; D007669:Kidney Calculi / Q000378:metabolism / Q000517:prevention & control*; D053078:Membrane Potential, Mitochondrial; D010070:Oxalates / Q000494:pharmacology*; D018384:Oxidative Stress / Q000187:drug effects*; D017382:Reactive Oxygen Species / Q000378:metabolism
chemical_list: D010070:Oxalates; D017382:Reactive Oxygen Species; D002129:Calcium Oxalate
keywords: Calcium oxalate; gene therapy; hyperoxaluria; oxalate decarboxylase; urolithiasis