The use of urinary and kidney SILAM proteomics to monitor kidney response to high dose morpholino oligonucleotides in the mdx mouse. (2015)

• Record Type: Journal Article
• Authors: Zhang, Aiping; Uaesoontrachoon, Kitipong; Shaughnessy, Conner; Das, Jharna R.; Rayavarapu, Sree; Brown, Kristy J.; Ray, Patricio E.; Nagaraju, Kanneboyina; van den Anker, John N.; Hoffman, Eric P.; Hathout, Yetrib
• Is Part Of: Toxicology reports. Volume 2(2015)
• Page Start: 838
• Page End: 849
• Publication Date: 2015
• Subjects: Phosphorodiamidate morpholino (PubChem CID: 22140692) -- Isoflurane (PubChem CID: 3763) -- Formic acid (PubChem CID: 284) -- Acetonitrile (PubChem CID: 6342) -- Acetone (PubChem CID: 180) -- Methanol (PubChem CID: 887); 5-OPase 5-oxoprolinase -- Aass alpha-aminoadipic semialdehyde synthase -- AKI acute kidney injury -- AMY2 pancreatic amylase α2 -- Anx2 annexin 2 -- AP-A glutamyl aminopeptidase -- AP-N aminopeptidase N -- CI-B22 NADH dehydrogenase [ubiquinone] 1 beta subcomplex subunit 9 -- Cytcox cytochrome c oxidase subunit 2 -- DMD Duchenne muscular dystrophy -- EGF pro-epidermal growth factor -- FABPH fatty acid binding protein heart type -- PK pharmacokinetics -- GGT1 gamma glutamytransferase 1 -- H&E hematoxylin and eosin -- IP2 integrated proteomics pipeline -- KIM-1 kidney injury molecule-1 -- mAspAT mitochondrial aspartate aminotransferase -- Mep-1 Meprin A subunit alpha -- NGAL neutrophil gelatinase-associated lipocalin -- OPN osteopontin -- PAS periodic acid shift -- PCB mitochondrial pyruvate carboxylase -- PMO phosphorodiamidate morpholino oligonucleotide -- PSs phosphorothioates -- SILAM stable isotope labeling in mammals -- SUn serum urea nitrogen; PMO -- Urinary biomarkers -- mdx-23 -- Duchenne muscular dystrophy -- Clusterin -- GGT1; Toxicology -- Periodicals; Clinical toxicology -- Periodicals; Drug-Related Side Effects and Adverse Reactions; Hazardous Substances; Poisoning; Toxicology; Electronic journals; Periodicals; Periodicals; 571.9505
• DOI: 10.1016/j.toxrep.2015.05.008
• Languages: English
• ISSNs: 2214-7500

Abstract

Abstract: Phosphorodiamidate morpholino oligonucleotides (PMO) are used as a promising exon-skipping gene therapy for Duchenne muscular dystrophy (DMD). One potential complication of high dose PMO therapy is its transient accumulation in the kidneys. Therefore new urinary biomarkers are needed to monitor this treatment. Here, we carried out a pilot proteomic profiling study using stable isotope labeling in mammals (SILAM) strategy to identify new biomarkers to monitor the effect of PMO on the kidneys of the dystrophin deficient mouse model for DMD (mdx-23). We first assessed the baseline renal status of the mdx-23 mouse compared to the wild type (C57BL10) mouse, and then followed the renal outcome of mdx-23 mouse treated with a single high dose intravenous PMO injection (800 mg/kg). Surprisingly, untreated mdx-23 mice showed evidence of renal injury at baseline, which was manifested by albuminuria, increased urine output, and changes in established urinary biomarker of acute kidney injury (AKI). The PMO treatment induced further transient renal injury, which peaked at 7 days, and returned to almost the baseline status at 30 days post-treatment. In the kidney, the SILAM approach followed by western blot validation identified changes in Meprin A subunit alpha at day 2, then returned to normal levels at days 7 and 30 after PMO injection. In the urine, SILAM approach identified an increase in Clusterin and γ-glutamyl transpeptidase 1 as potential candidates to monitor the transient renal accumulation of PMO. These results, which were confirmed by Western blots or ELISA, demonstrate the value of the SILAM approach to identify new candidate biomarkers of renal injury in mdx-23 mice treated with high dose PMO.