Profiling the Skeletal Muscle Proteome in Patients on Atypical Antipsychotics and Mood Stabilizers

Abstract

Atypical antipsychotics (AAP) are used in the treatment of severe mental illness. They are associated with several metabolic side effects including insulin resistance. The skeletal muscle is the primary tissue responsible for insulin-stimulated glucose uptake. Dysfunction of protein regulation within the skeletal muscle following treatment with AAPs may play a role in the associated metabolic side effects. The objective of this study was to measure protein abundance in the skeletal muscle of patients on long-term AAP or mood stabilizer treatment. Cross-sectional muscle biopsies were obtained from patients with bipolar disorder and global protein abundance was measured using stable isotope labeling by amino acid (SILAC) combined with high-performance liquid chromatography-electrospray ionization tandem mass spectrometry (HPLC-ESI-MS/MS). Sixteen patients completed muscle biopsies and were included in the proteomic analyses. A total of 40 proteins were significantly different between the AAP group and the mood stabilizer group. In-silico pathway analysis identified significant enrichment in several pathways including glucose metabolism, cell cycle, apoptosis, and folate metabolism. Proteome abundance changes also differed based on protein biological processes and function. In summary, significant differences in proteomic profiles were identified in the skeletal muscle between patients on AAPs and mood stabilizers. Future work is needed to validate these findings in prospectively sampled populations.

Effect of Insulin and Pioglitazone on Protein Phosphatase 2A Interaction Partners in Primary Human Skeletal Muscle Cells Derived from Obese Insulin-Resistant Participants

“…For each lane, three slices (250–75, 75–37, and 37–10 kDa) were excised, destained, and exposed to 50 mM DTT, followed by iodoacetamide (IAA) treatment, in-gel trypsin digestion, peptide purification by ZipTip Pipette tips, and UPLC-ESI-MS/MS analysis using an Orbitrap Fusion Lumos as described. 50 , 53 …”

“…Subsequently, the beads were boiled in 30 μL of 2 × SDS buffer containing 5% mercaptoethanol at 95 °C for 5 min, Bead eluates were resolved by 10% 1D-SDS-PAGE. For each lane, three slices (250–75, 75–37, and 37–10 kDa) were excised, destained, and exposed to 50 mM DTT, followed by iodoacetamide (IAA) treatment, in-gel trypsin digestion, peptide purification by ZipTip Pipette tips, and UPLC-ESI-MS/MS analysis using an Orbitrap Fusion Lumos as described. , …”

“…For each lane, three slices (250−75, 75−37, and 37−10 kDa) were excised, destained, and exposed to 50 mM DTT, followed by iodoacetamide (IAA) treatment, in-gel trypsin digestion, peptide purification by ZipTip Pipette tips, and UPLC-ESI-MS/MS analysis using an Orbitrap Fusion Lumos as described. 50,53 The peptide mixture was separated with a gradient of 2− 35% buffer B (100% ACN and 0.1% FA) in 70 min, starting with 2% B and increase to 5% in 1 min, to 25% B in 60 min, to 35% B in 10 min, to 90% B in 1 min, hold at 90% B for 5 min, and then 2% buffer B at a flow rate of 400 nL/min on a C 18reversed phase column (75 μm ID, 45 cm length) packed inhouse with ReproSil-Pur C18-AQ μm resin (Dr. Maisch GmbH) in buffer A (0.1% FA). MS spectra were generated from Orbitrap Fusion Lumos (ThermoFisher) equipped with a nano-LC-based electrospray ionization source and coupled online to a Dionex Ultimate 3000 UPLC system (Thermo Fisher Scientific).…”