The work in Dr. Narayan’s laboratory focuses on mitigating reactive oxygen species (ROS)-related oxidative damage to protein disulfide isomerase (PDI), the chief oxidoreductase chaperone in the endoplasmic reticulum responsible for preventing neurotoxicity and accumulation of misfolded cellular debris. Reactive oxygen species (ROS) insult has been convincingly implicated in a host of pathological and behavioral changes in both neuronal (cellular) and animal models, eventually resulting in compromised or increased morbidity and mortality. Particularly, ROS-mediated accumulation of ?-synuclein is known to affect synaptic plasticity in the dentate gyrus and conversely, corrective transformation of ROS attenuates the deleterious role of superoxide in modulating synaptic plasticity, learning and memory.
Significance of the work: Our long-term goal is to bring to clinical trials therapeutics that address neurodegeneration not only for rescuing behavioral plasticity, but also prevention of related neuropathies. It is our vision that someday potent ethnopharma-based neurodegeneration preventatives can be incorporated into processed foods and “vaccinate” populations from early childhood. The benefits of our approach are: 1) The research thrust is preventative and can be used to target populations over a wide age, gender and ethnic spectrum 2) Curcumin has FDA clearance 3) Curcumin lacks adverse side-effects associated with chemotherapeutics.
Methods to be learned: Dr Narayan’s laboratory uses biochemical methods in their approach. Students will develop and characterize select bisphenols as potent NOx scavengers, use molecular recognition and then target validation using in vivo assays.
Participants will initiate work to establish the ROS-scavenging ability of select curcumin and other polyphenolic-analogs previously extracted or available in the laboratory. The anti-ROS activity will be determined using model NOx generators routinely employed in Dr. Narayan’s laboratory. The students will apply reversed-phase HPLC to fractionate the reactions products of the polyphenols post exposure to the NOx generators; mass spectrometric analysis will aid in determining the identity of the radical-scavenged products, if any. Successful radical scavengers will then be introduced into a human derived neuroblastoma cell line (SH-SY5Y), prior to oxidative insult of the cells using rotenone. Neuronal integrity against debris accumulation, a key biomarker for eventual compromised neuronal plasticity, will be monitored using green fluorescent protein tagged synphilin-1 through fluorescence microscopy.