Treatment for Non-Alcoholic Fatty Liver With Different Doses of Vitamin E

Non-alcoholic fatty liver disease (NAFLD) is the most common cause for liver test
abnormalities in the western world, and an increasingly rising cause for liver-related
morbidity and mortality. Vitamin E, a fat-soluble anti-oxidant was recently found to be an
effective treatment for NAFLD; however, its mechanism of action is unclear. In a controlled
clinical trial vitamin E treatment was shown to significantly reduce the hepatic fat burden,
suggesting mechanisms other than reducing oxidative stress are involved. Furthermore, the
optimal dose of vitamin E to treat NAFLD is unknown.

We propose a phase IIa study to determine the optimal dose of vitamin E and its mechanism and
site of action. In this study we aim to enroll up to 90 patients with NAFLD. Initially, all
patients will undergo 12 weeks of intensive lifestyle modification. Following that, all
patients will be randomized to treatment with 3 different doses of natural vitamin E (rrr-
-tocopherol at 200, 400 or 800 IU/d) for 24 weeks. The primary end points for efficacy are
normalization of liver enzymes and reduction in liver fat contents by magnetic resonance
spectroscopy. Patients will undergo liver and adipose tissue biopsies before vitamin E
treatment and after 4 weeks of therapy, and the biopsy samples will be used to measure
changes in gene expression and markers of oxidative stress. This will be coupled with
extensive phenotyping before and after treatment using serological, radiological and dynamic
endocrine testing and is aimed at finding the dose-response characteristics of vitamin E in
NAFLD, and allowing us to understand the mechanism of its action.

After 24 weeks of randomized treatment, all patients will be switched to a dose of 800 IU/ml
and will continue treatment for up to 30 months, at the end of which another liver biopsy
will be performed. From this phase we will assess the effects of dose increase of vitamin E
on liver enzymes and fat content, and will determine the effect of long-term treatment on
histological outcome.

- INCLUSION CRITERIA:

Clinical suspicion of NAFLD, defined by the presence of at least two of the following
criteria:

- Suggestion of liver fat by an imaging study (ultrasound, CT scan, MRI or MR
spectroscopy) performed in the 6 months prior to enrollment.

- Elevated aminotransferase levels (ALT > 31 U/L for men or > 19 U/L for women, or AST >
30 U/L) on at least two occasions in the 6 months preceding enrollment.

- Presence of the metabolic syndrome, defined according to the modified AHA/NCEP
criteria as the presence of at least three of:

- Abdominal obesity, defined as waist circumference > 102 cm for men or > 88 cm for
women

- Elevated triglycerides (> 150 mg/dL) or the use of medication to lower
triglycerides

- Reduced HDL cholesterol (< 40 mg/DL for men or < 50 mg/dL for women)

- Elevated blood pressure (> 135/80 mmHg) or use of medication for hypertension

- Elevated fasting glucose levels (> 100 mg/dL) or use of anti-diabetic medication

- For the purpose of inclusion, the presence of overt diabetes mellitus type 2
will be considered equivalent to the presence of the metabolic syndrome,
even if the other criteria are absent.

- Estimated average alcohol consumption < 30 g/d for men or < 20 g/d for women in the 6
months prior to enrollment and no binge-drinking behavior.

- Age > 18 years at enrollment

- Willingness to participate in the study

EXCLUSION CRITERIA:

- Chronic infection with hepatitis C virus (HCV) or hepatitis B virus (HBV). Patients
who were treated successfully for HCV and achieved sustained virological response can
be eligible for enrollment > 18 months after treatment cessation. Patients who are
inactive carriers of HBV (HBV DNA < 1000 copies/mL, HBeAg negative, Anti HDV negative)
for at least 12 months prior to enrollment are also eligible. Patients receiving
antiviral therapy are ineligible.

- Concomitant liver disease such as autoimmune hepatitis, primary biliary cirrhosis,
primary sclerosing cholangitis, Wilson s disease, alpha-1 antitrypsin deficiency.

- Presence of definite or probable drug-induced liver injury. In the case of
lipid-lowering, anti-hypertensive or anti-diabetic medications that are suspected to
cause elevation of aminotransferases, patients will be eligible if treatment is
associated with stable enzyme levels for at least 6 months and inclusion criteria 1a.
and 1c. are both present.

- Treatment with medications known to cause fatty liver disease such as atypical
neuroleptics, tetracycline, methotrexate or tamoxifen

- Uncontrolled hypo- or hyperthyroidism.

- Decompensated advanced liver disease, defined as direct bilirubin > 0.5 g/dL, PT > 18,
albumin < 3 g/dL, or history of ascites, encephalopathy, variceal bleeding,
spontaneous bacterial peritonitis or liver transplant.

- Active coronary artery disease, defined as persistent angina pectoris, reversible
ischemia on cardiac stress test or imaging, or the presence of significant coronary
artery disease on imaging or catheterization. Patients with coronary artery disease
that was treated by angioplasty or bypass surgery may be eligible if they have no
evidence of active disease >= 1 year after intervention, can safely stop antiplatelet
and anticoagulant medications before the performance of invasive procedures, and have
adequate ventricular function as assessed by echocardiography or cardiology
consultation. These patients will require cardiology consultation and clearance prior
to enrollment.

- Congestive heart failure.

- Chronic kidney disease, with creatinine clearance < 60 ml/h.

- Uncontrolled diabetes mellitus. Patients may be enrolled if they have been on stable
therapy with any anti-diabetic agent for at least 3 months prior to enrollment, are
not foreseen by the physician treating their diabetes to require antidiabetic
medication or dose changes during the trial and have an HbA1c <= 7.5% on enrollment.

- Treatment with vitamin E. Patients who are currently taking vitamin E as a supplement
will be requested to stop for at least 3 months before becoming eligible for
enrollment. Patients who are taking vitamin E for a medical indication other than
NAFLD will not be eligible.

- Contraindication to or inability to undergo a liver biopsy.

- Patients who had a liver biopsy performed <= 2 years before enrollment, unless they
are willing to undergo all of the trial biopsies, knowing that these biopsies are
purely for research and are not clinically indicated. This will be clearly documented
in the patients charts prior to enrollment.

- Patients with coagulopathy (PT/PTT values that are prolonged >= 3 seconds from the
upper limit of the normal, including treatment with oral and parenteral
anticoagulants), thrombocytopenia (<70,000), or platelet dysfunction will not be
enrolled because of potential increase in risk of bleeding with vitamin E treatment.
Antiplatelet agents taken for cardiovascular prevention will not exclude patients,
unless they cannot be stopped safely for the performance of a liver biopsy.

- Maldigestion or malabsorption that can interfere with absorption of vitamin E
including: steatorrhea of all causes, chronic pancreatitis, cystic fibrosis, short
bowel syndrome, severe cholestasis, orlistat treatment and similar conditions

- Inability to swallow vitamin E capsules

- Allergy to vitamin E

- Alcohol or substance abuse within the past 12 months. Patients will be required to
have an AUDIT score of 7 or less18, and drink no more than 14 drinks/week (for men) or
7 drinks/week (for women).

- For women of childbearing age, pregnancy or inability (or unwillingness) to practice
contraception for the duration of the study or breast feeding.

- Inability to understand and give informed consent for participation.