Dietary Ketosis: Fatty Acids Activate AMPK Energy Circuits Modulating Global Methylation

Nutritional epigenetics denotes gene-diet interactions and highlights the modulatory role of
nutrition in aging and age-related diseases such as cancer, CVD (cardiovascular disease),
diabetes and neurodegenerative disorders. Nutrients are a source of epigenetic modification;
they are able to regulate the placement of histone modifiers distinguishing phenotype from
genotype. The energy status of the cells (fed or fasted) modulates the regulation of global
DNA methylation via the S-adenosylmethionine (SAM), the methyltransferase inhibitor,
S-adenosylhomocysteine (SAH) axis and whole blood histamine levels. Insulin resistance and
hyperinsulinemia dysregulate cellular signals leading to metabolic inflexibility. Chronic
elevations in insulin with long-standing impairments in glucose delivery are associated with
profound changes in epigenetic patterns due to over-activation of the mTOR kinase pathway and
repression of AMPK (adenosine monophosphate-activated protein kinase). Dietary ketosis is
known to change the metabolic status of the cells by increasing the AMP/ATP (adenosine
monophosphate/adenosine tri-phosphate) ratio. AMPK activation adapts rRNA (ribosomal
ribonucleic acid) synthesis away from growth/biosynthesis and toward ATP availability and
utilization, thereby, attenuating the progression of hypo-metabolic diseases in the body and
the brain at the level of the genome.

The study will explore whether early stage memory loss (SMC & MCI) and the comorbidity of
Metabolic Syndrome are symptomatic of peripheral and cerebral cellular hypo-metabolism
induced by chronic insulin resistance. We will attempt to show that consequential to systemic
hyperinsulinemia, aberrant crosstalk between the mitochondria and nuclear genome results in
the dysregulation of the regulatory kinases mediating metabolic state and
intracellular/extracellular signaling: mTOR and AMPK. The suppression of AMPK signals with
chronic overexpression of mTOR signaling will adapt rRNA synthesis away from nutrient
availability and toward ATP consuming processes: the biosynthesis of cholesterol,
triglycerides, glycogen with inhibition of fatty acid oxidation, histone acetylation with a
down-regulation of NAD+ (nicotinamide adenine dinucleotide coenzyme) and SAHH
(S-adenosylhomocysteine hydrolase) cofactors leading to global DNA hypo-methylation and local
hyper-methylation, suppression of the SAM/SAH ratio, the inhibition of SIRT (sirtuin)
expression and normalized whole blood histamine levels. These epigenetic shifts mediate
global metabolic inflexibility by channeling fuel substrates toward cytosolic, substrate
level phosphorylation (SLP) via over expression of the glycolytic enzymes including PDK
(pyruvate dehydrogenase kinase) and away from mitochondrial oxidation mediated by the
suppression of PDC (pyruvate dehydrogenase complex), the major regulatory gateway of
metabolism between glycolysis and citric acid cycle. We will attempt to show that activation
of the AMPK pathway via induced and controlled dietary ketosis will inhibit mTOR signaling
away from the biosynthesis of energy and SLP toward the generation of ATP by increasing the
cellular AMP/ATP ratio, thus regulating oxidative metabolic signals and attenuating global,
cellular hypo-metabolism evidenced by marked reductions in lipid synthesis and LP-IR score
(particle concentration and size), HgA1c (hemoglobin A1c), fasting insulin/HOMA-IR
(homeostatic model assessment of insulin resistance), blood ketones, fasting triglycerides
together with epigenetic regulation of DNA methylation status including normalized whole
blood histamine levels and homocysteine regulation. Improvement in cerebral glucose
metabolism and corresponding diagnosis of SMC/MCI will be assessed by the objective changes
in the outcome measures of MoCA (Montreal Cognitive Assessment), MMSE (Mini-Mental State
Exam) and BVMT-R (Brief Visuospatial Memory Test-Revised) administered at baseline and week
12.

Inclusion Criteria:

- Male or Female (age 35-80)

- Previously diagnosed with MetS and/or T2DM as measured by possessing at least 2 of the
following physiological measures: type II diabetes, BMI >30, HgA1c > 5.7, waist/height
ratio >.6, fasting glucose > 125

- Subjective Memory Complaints (SCM) - Subjects score > 3 'yes' answers on the
Subjective Memory --Complaints Questionnaire

- Previously diagnosed with Mild Cognitive Impairment (MCI)

Exclusion Criteria:

- Previously diagnosed with Alzheimer's disease (AD), dementia or Parkinson's disease