Dried Blood Spot- Statin Pilot Study

Despite reductions in cardiovascular disease (CVD) mortality in the past 20 years, CVD
remains the leading cause of mortality in the United States, accounting for ~1 of every 3
deaths. CVD costs the U.S. over $100 billion annually and is an important field for targeted
primary and secondary prevention. Dyslipidemia, particularly a high low-density-lipoprotein
cholesterol (LDL-C) level, is a well-established cardiovascular risk factor, and fasting
lipid panels (including total cholesterol, LDL-C, triglycerides, and high-density-lipoprotein
cholesterol [HDL-C]) are included in the recent 2013 ACC/AHA Risk Assessment Guidelines.
Clinical trials have shown that low density lipoprotein cholesterol (LDL-C) reduction with
statin therapy predicts cardiovascular event reduction. About a quarter of all adults >45
years old were on statin therapy in 2008, and this will likely increase due to the recent
2013 ACC/AHA Cholesterol Management guidelines. These guidelines recommend statin therapy in
all patients with clinical atherosclerotic CVD, all patients with LDL-C = 190 mg/dL, patients
age 40-75 years with diabetes and LDL-C 70-189 mg/dL, and patients with an estimated 10-year
atherosclerotic CVD risk = 7.5%. For all of these patients, a fasting lipid panel should be
drawn prior to statin initiation as well as during follow-up to assess medication and
lifestyle adherence. These fasting lipid panels are obtained via conventional phlebotomy via
venopuncture in an office-based or hospital laboratory setting.

Research protein quantitation with mass spectrometry and enzyme-linked immunosorbent assay
(ELISA) are technologies that allow for sensitive quantitation of protein biomarkers and
targets, including lipoproteins. Most importantly, multiple reaction monitoring (MRM) mass
spectrometry is able to assess samples from a dried blood spot (DBS), whose advantages
include minimal volume requirements, ease of sample attainment by finger stick with minimal
training required, ease of transport, and sample stability. DBS sampling has been used for
clinical and pre-clinical studies, simplifying sample collection and handling. However,
research protein quantitation with mass spectrometry and ELISA has not been compared against
conventional blood sample for evaluating changes in cholesterol levels during statin therapy.

While LDL-C is well-established in predicting CVD event reduction, other laboratory lipid,
thrombotic and inflammatory biomarkers have been shown to be associated with atherosclerotic
plaque development or CVD risk. These include Apo A-I, Apo B, Apo E, IgM, plasminogen,
TIMP-1, Von Willebrand factor, antithrombin III, cystatin C, mesothelin, C-reactive protein,
SAA, LPS-binding protein, mannose-binding lectin, myeloperoxidase, fibrinogen, alpha-1-acid
glycoprotein, soluble transferrin receptor, haptoglobin. All of these CVD protein biomarkers
can be measured by research protein quantitation techniques.

In a pilot study of 20 subjects, we aim to:

1. measure changes in a panel of CVD protein biomarkers before and after initiation of
statin therapy.

2. compare clinical laboratory measurements of LDL-C and HDL-C in conventional phlebotomy
blood samples with research protein quantitation mass spectrometry and ELISA
measurements of LDL-C (Apo B) and HDL-C (Apo A-I) proteins in DBS and plasma.

Inclusion criteria:

1. Men and women age> 18 years who are initiated on statin therapy in the clinical setting.

Exclusion criteria:

1. History of rhabdomyolysis

2. History of prior allergic reaction to statins

3. History of liver failure

4. Contraindications to antecubital phlebotomy or finger stick (including those with
bilateral dialysis AV fistulae).