1 mitigating effects of oxidation in aging and diseases m. shchepinov, r.j. molinari *, v. shmanai,...
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Mitigating Effects of Oxidation in Mitigating Effects of Oxidation in Aging and DiseasesAging and Diseases
M. Shchepinov, R.J. Molinari *, V. Shmanai, M. Shchepinov, R.J. Molinari *, V. Shmanai, C.Clarke, A. Manning-Bog C.Clarke, A. Manning-Bog
March 5, 2010March 5, 2010ASENT Pipeline SessionASENT 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.
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Oxidation by ROS and Simplest Isotope Protection
ROS abstract hydrogens from PUFAs
Some specific stabilization schema.
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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.
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Yeast Co-Q10 minus Oxidative Stress ModelLinolenic 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 stereologyRemaining brain for total deuterium levels
Study Design:
* p = 0.0077
Nigrostriatal dopamine levels
Str
iata
l DA
(n
g/m
g p
rote
in)
n=3 n=3 n=4 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
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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 Investigator Institution Work
Co-PI: Mikhail Shchepinov, CSORobert Molinari *, CEO
Retrotope, Inc. All studies
Co-PI: Amy Manning-BogRA, Vivian Chou
Advisor: J. Wm. Langston
Stanford Research Institute/The Parkinson’s Institute
Preclinical in vivo MPTP modeling studies
Collaborator:Catherine Clarke
UCLADepartment 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 ArkansasStable Isotopes Laboratory
Mass Spec for deuterated PUFA incorporation
This work was funded by the Michael J. Fox Foundation for Parkinson’s Research