Presentation on theme: "1 Mitigating Effects of Oxidation in Aging and Diseases M. Shchepinov, R.J. Molinari *, V. Shmanai, C.Clarke, A. Manning-Bog March 5, 2010 ASENT Pipeline."— Presentation transcript:
1 Mitigating Effects of Oxidation in Aging and Diseases M. Shchepinov, R.J. Molinari *, V. Shmanai, C.Clarke, A. Manning-Bog March 5, 2010 ASENT Pipeline Session * Presentor
Background & Hypothesis Many CNS disease etiologies have recently been associated with oxidation damage of mitochondrial membranes, Such oxidation occurs at a very few, weak-link chemical bonds in polyunsaturated fatty acids, Isotope effect can stabilize target bonds in well-understood ways, with little, if any, toxicity. Supplementing isotope-‘fortified’ components into essential fatty acids will increase resistance to oxidation, and may mitigate disease.
3 Oxidation by ROS and Simplest Isotope Protection ROS abstract hydrogens from PUFAs Some specific stabilization schema.
4 Preliminary Data:. D-4 Linolenic Acid Co-enzyme Q10 deficient yeast oxidative stress models. Complete replacement of dietary PUFA with d-PUFA, Treatment of MPTP mice with dietary PUFAs.
5 Yeast Co-Q10 minus Oxidative Stress Model Linolenic Acid vs. D-4 Linolenic Acid * Clarke Lab, UCLA
MPTP Mouse Study Primary goal: Does dietary D-PUFA protect nigrostriatal dopamine from MPTP-induced depletion? Secondary goals: -Do dietary D-PUFAs incorporate into the brain ? -Are they toxic ? -Does dietary D-PUFA decrease nigrostriatal lipid hydroperoxide formation (HNE) due to MPTP challenge ?
Dietary PUFA dosing: Fat-free chow (5-6g/mouse) supplemented with saturated fats, oleic acid, and H-PUFA or D-PUFA (30 mg linolenic and 30 mg linoleic coated on pellets) for 6 days + 6 days post MPTP Toxicant challenge: Single injection of MPTP (40 mg/kg) or saline vehicle, i.p. Cohorts (4) were: H-PUFA-saline; H-PUFA-MPTP; D- PUFA-saline; D-PUFA-MPTP Readouts: Striatal dopamine levels 4-HNE immunohistochemistry/nigral stereology Remaining brain for total deuterium levels Study Design:
* p = 0.0077 Nigrostriatal dopamine levels Striatal DA (ng/mg protein) n=3 n=4 Manning-Bog Lab, Stanford Research Institute
Brain tissues in treated, saline samples were highly enriched in deuterium. No observed mortality or side effects from dietary dosing, normal weight gain in all mice except as noted below. MPTP produced marked dopamine depletion of >75% with abnormally high mortality in the untreated, MPTP cohort (3/7) vs. 1/7 in D-PUFA cohort. Study Notations: Less than 1 in 7 were removed due to dissected protein amounts in excess of two std. deviations (n=2), or failure to thrive (n=1) Additional Observations
10 Further Strategies Focus on diseases in which oxidation products of PUFAs (e.g. 4-HNE) are directly implicated (PD, AMD, ALS, ALZ, CHF) Expand POC efficacy and toxicity of PUFA reinforcement in predictive cellular models & mammals Retrotope supplies materials/expertise for trials, work done by disease model experts Triage and accelerate disease programs with new funding / partners when POC established Develop early regulatory Tox Approach
Acknowledgments InvestigatorInstitutionWork Co-PI: Mikhail Shchepinov, CSO Robert Molinari *, CEO Retrotope, Inc.All studies Co-PI: Amy Manning-Bog RA, Vivian Chou Advisor: J. Wm. Langston Stanford Research Institute/ The Parkinson’s Institute Preclinical in vivo MPTP modeling studies Collaborator: Catherine Clarke UCLA Department of Chemistry Co-Q10 Yeast Studies Collaborator: Vadim Shmanai Belarussian Academy of Sciences, Institute for Physical Organic Chemistry Chemical Synthesis of deuterated PUFAs Contractor: Eric Pollock Univ. of Arkansas Stable Isotopes Laboratory Mass Spec for deuterated PUFA incorporation This work was funded by the Michael J. Fox Foundation for Parkinson’s Research