Upregulation of Mitochondrial Biogenesis in Neuronal Cells via CREB-Mediated Transcription of TFAM (S20.007)

Objective: Mitochondrial dysfunction has severe cellular consequences and is linked to aging and neurological disorders including Parkinson’s disease. Therefore, upregulation of mitochondrial biogenesis is an important area of research. Here, we have undertaken a novel approach to upregulate mitochondrial biogenesis in cultured neuronal cells. Background: RNS60 is normal saline modified by a process involving Taylor-Couette-Poiseuille (TCP) flow under elevated oxygen pressure. RNS60 is proposed to contain charge-stabilized nanostructures consisting of an oxygen nanobubble core surrounded by an electrical double-layer at the liquid/gas interface. RNS60 has been reported to be neuro-protective. Here, we tested the effects of RNS60 on mitochondrial biogenesis in dopaminergic neurons. Methods: MN9D dopaminergic neurons and mouse primary dopaminergic neurons were treated with RNS60, ONS60 (saline containing a comparable level of dissolved oxygen without the TCP modification), NS (normal saline), or RNS10.3 (TCP-modified normal saline without added oxygen) followed by monitoring mitochondrial biogenesis. Results: RNS60, but not NS, ONS60 or RNS10.3, increased the expression of genes associated with mitochondrial biogenesis and upregulated mitochondrial biogenesis in dopaminergic neurons. On the other hand, RNS60 had no effect on lysosomal biogenesis and genes associated with lysosomal biogenesis. Interestingly, we found that RNS60 upregulated mitochondrial transcription factor A (TFAM), and siRNA knockdown of TFAM abrogated the ability of RNS60 to increase mitochondrial biogenesis. Furthermore, we delineated that RNS60 increased the transcription of TFAM via type IA phosphatidylinositol (PI) 3-kinase-mediated activation of CREB. Accordingly, knockdown of the PI-3 kinase - CREB pathway suppressed RNS60-mediated mitochondrial biogenesis. Conclusions: These results describe a novel property of RNS60 of stimulating mitochondrial biogenesis via PI 3-kinase-CREB-mediated up-regulation of TFAM, which may be of therapeutic benefit in different neurodegenerative disorders. This study has been supported by Revalesio Corporation, Tacoma, WA. Disclosure: Dr. Kundu has nothing to disclose. Dr. Chandra has nothing to disclose. Dr. Rangasamy has nothing to disclose. Dr. Ghosh has nothing to disclose. Dr. Watson has received personal compensation for activities with Revalesio Corporation as an employee. Dr. Pahan has nothing to disclose.