Circulating Adenosine Levels Before and After Intravenous (IV) Persantine

Objectives:

The overall goal of this proposal is to develop methods to achieve heart and vascular
protection from ischemia and thus improve soldier's performance in adverse environment. The
major hypothesis is that new approach and method can be developed to enhance resistance to
stress-induced circulatory insufficiency and myocardial ischemia. The goals here are to
determine whether a decreased adenosine transporter function is associated with a reduced
physiological responsiveness to the vasculo-protective drug persantine using two in vitro
endpoints: the ability of persantine 1) to inhibit platelet aggregation and 2) to inhibit
[3H] uridine uptake. Both are endpoints that indicate physiological responsiveness. Both
relate directly to the cardiovascular protective effects of , that is, persantine the
availability of extracellular adenosine level and the anti-platelet property. Specifically,
the relationship between circulating adenosine increase to persantine in vivo and blockade
of radio-labeled uridine uptake by erythrocytes and of platelet aggregation by the drug in
vitro will be determined. These investigations will recruit subjects undergoing persantine
stress testing in the Nuclear Cardiology Laboratory.

Scientific Background and Significance:

Development of methods to protect from skeletal and cardiovascular insufficiency is the main
objective of the current research. Adenosine is a potent cyto-protective hormone released
during ischemia. The goal of this clinical research project are to test the presence of
genetic polymorphisms in the adenosine transporter gene and to determine whether it is
associated with an altered persantine responsiveness. The hypothesis is that some or all of
these polymorphisms are associated with a decreased responsiveness to persantine and that
increasing the dose of persantine will overcome the relative non-response.

Specific Evaluations:

-Persantine Administration: Persantine will be administered after baseline evaluation with
intravenous dose (0.56 mg/kg) over 5 minutes.

-Adenosine levels: Adenosine levels will be measured at baseline, and 2 minutes after the
persantine dose. We have recently modified and adapted a method to measure nanomolar
concentration of adenosine in human blood.

-Adenosine transporter function: The transporter function will be determined by the ability
of persantine to inhibit erythrocyte uptake of radio-labeled uridine in vitro.

- Anti-platelet effect of persantine: The platelet aggregation response to persantine will
be determined via whole blood aggregometry in vitro where the ex vivo platelet response to
the drug can be quantified.

Preliminary Studies:

Preliminary data showed that two non-synonymous single nucleotide polymorphisms occurring at
a frequency of 3-4% exist in the persantine-binding regions of the adenosine transporter
gene. The goal of the present study is to determine the functional significance of these
polymorphisms by testing the association of these polymorphisms with the ability of
persantine to inhibit uridine (uridine uses the same transporter) uptake and platelet
aggregation. These represent the in vitro functional endpoints in the subjects with the
polymorphism. The investigators will also study the association of these polymorphisms with
any clinical characteristics such as the incidence of MI, acute coronary syndrome, coronary
bypass or stenting procedures. Since this is NOT an interventional study, these are not
considered clinical outcomes or endpoints in the traditional sense when an intervention is
carried out. These are considered clinical characteristics or phenotypes that associate
with the genotype of polymorphisms. Primary and secondary outcomes and endpoints: The goal
of the present study is to determine the functional significance of these polymorphisms by
testing the association of these polymorphisms with the ability of persantine to inhibit
uridine (uridine uses the same transporter) uptake and platelet aggregation. These
represent the in vitro functional endpoints in the subjects with the polymorphism. The
investigators will also study the association of these polymorphisms with any clinical
characteristics such as the incidence of MI, acute coronary syndrome, coronary bypass or
stenting procedures. Since this is NOT an interventional study, these are not considered
clinical outcomes or endpoints in the traditional sense when an intervention is carried out.
These are considered clinical characteristics or phenotypes that associate with the
genotype of polymorphisms.

Inclusion Criteria:

- Subjects with or without coronary artery disease undergoing a Persantine nuclear
stress test

Exclusion Criteria:

- Oral persantine use within 24 hours

- Second or third degree AV block, or sick sinus syndrome without a functioning
pacemaker

- Active asthma or bronchospasm

- Those with end-stage liver disease such as cirrhosis or active hepatitis such as > 5
fold liver enzyme elevation will not be included

- Anemia (Hct < 30)

- Myocardial infarction within 30 days

- Severe left ventricular dysfunction (EF < 30%)