Mitochondrial Methylation in Type 2 Diabetes
Mitochondrial Methylation in Type 2 Diabetes
The overarching goal of this proposal is to determine whether DNA methylation of the
 mitochondrial DNA impairs mitochondrial function in insulin resistant states such as
 overweight/obesity and type 2 diabetes.
To determine whether differences in human skeletal muscle DNA methylation patterns in the
 mitochondrial and nuclear genome can explain the lower abundance of ETC and OXPHOS mRNA and
 protein observed in insulin resistant skeletal muscle of overweight/obese and type 2 diabetic
 participants. To determine whether patterns of human skeletal muscle DNA methylation in the
 mitochondrial and nuclear genome are predictive of ETC function. We will isolate skeletal
 muscle mitochondria from metabolically well-characterized lean insulin sensitive,
 overweight/obese insulin resistant nondiabetic and obese insulin resistant type 2 diabetic
 volunteers, and functionally evaluate each ETC complex (I - IV) and complex V (ATP synthase).
Inclusion Criteria:

 1. Subjects must be 21-55 years old

 2. Body Mass Index:

 Lean, healthy BMI ≤25; Overweight,non-diabetic BMI 25-29.9; Obese, non-diabetic BMI
 30-50; Type 2 Diabetes (per the American Diabetes Association criteria)

 3. Subjects must be able to communicate meaningfully with the investigator and must be
 legally competent to provide written informed consent.

 4. Female subjects must be non-lactating and will be eligible only if they have a
 negative pregnancy test throughout the study period, and must agree to use acceptable
 birth control (hormonal contraceptives, barrier methods, have an intrauterine device,
 or surgical sterilization)

 5. Subjects must have the following laboratory values:

 -  Hematocrit ≥ 35 vol%

 -  Serum creatinine ≤ 1.6 mg/dl

 -  AST (SGOT) < 2 times upper limit of normal

 -  ALT (SGPT) < 2 times upper limit of normal

 -  Alkaline phosphatase < 2 times upper limit of normal

 -  Triglycerides < 150 mg/dl for nondiabetics

 -  Triglycerides <300 for diabetics

 -  INR ≤ 1.3

 -  HbA1c ≤ 10

 Exclusion Criteria:

 1. Subjects must not be receiving any of the following medications: thiazide or
 furosemide diuretics, beta-blockers, or other chronic medications with known adverse
 effects on glucose tolerance levels unless the patient has been on a stable dose of
 such agents for the past three months before entry into the study. Subjects must not
 be taking estrogens or other hormonal replacement therapy unless the subject has been
 on these agents on a stable dose for the prior three months. Subjects taking systemic
 glucocorticoids are excluded. Patients with type 2 diabetes will be excluded if they
 are taking thiazolidinediones.

 2. Subjects receiving Gemfibrozil must not also be receiving a statin.

 3. Subjects with a history of clinically significant heart disease (New York Heart
 Classification greater than grade II; more than non-specific ST-T wave changes on the
 EKG), peripheral vascular disease (history of claudication), or pulmonary disease
 (dyspnea on exertion of one flight or less; abnormal breath sounds on auscultation)
 will not be studied.

 4. Recent systemic or pulmonary embolus, untreated high-risk proliferative retinopathy,
 recent retinal hemorrhage, uncontrolled hypertension, systolic BP>160, diastolic
 BP>95, autonomic neuropathy, resting heart rate >100, electrolyte abnormalities.