Adaptation to hypothermia is important for skeletal muscle cells under physiological stress and is used for therapeutic hypothermia (mild hypothermia at 32 ⁰C). We show that hypothermic preconditioning at 32 ⁰C for 72 hours improves the differentiation of skeletal muscle myoblasts using both C2C12 and primary myoblasts isolated from 3 month and 18-month-old mice. The proteomic analysis of myoblasts exposed to hypothermia revealed that levels of the cold-shock protein RBM3, an RNA-binding protein, increases with both acute and chronic exposure to hypothermic stress, and is necessary for the enhanced differentiation and maintenance of mitochondrial metabolism. We also show that overexpression of RBM3 at 37 ⁰C is sufficient to promote mitochondrial metabolism, cellular proliferation, and differentiation of C2C12 and primary myoblasts. Proteomic analysis of C2C12 myoblasts overexpressing RBM3 show significant enrichment of pathways involved in fatty acid metabolism, RNA metabolism and the electron transport chain. Overall, we show that the cold-shock protein RBM3 is a critical factor that can be used for controlling the metabolic network of myoblasts.

This suggests that delivering RBM3 to muscles in-vivo could be an important approach to promote muscle metabolism. Therefore, we are developing a lipid nanoparticle-based mRNA delivery for efficient transfection into i) myotubes in-vitro ii) skeletal muscle 3D organoids and iii) in-vivo mouse model.

Protein domains are subunits of a protein that are classified according to three aspects of the subunits: Structural, Functional, and folding. Classification and identification of domains can provide insights into the structural and functional activities of a protein. In this presentation, I will be talking about domain classification with the updates I have made to DIAL, an in-house algorithm developed by (Sowdhamini and Blundell,1995). I will also discuss identifying domains from proteomic sequences by talking about my work in identifying Prolyl Oligopeptidases (POPs) from the proteomic assemblies of the human gut microbiota.  

Not disclosed yet!