Search for Genomic and
Proteomic Risk / Protective
Factors Associated with
Coronary Heart Disease
Xiaohong Wu MD, Mehran Haidari MD, Silvio Litovsky
MD, Ward Casscells MD, James T Willerson MD, and
Morteza Naghavi MD.
1. We hypothesize that genes expressed on peripheral blood
inflammatory cells, monocytes, are distinct among following
1)first time heart attack patients 40-70y(with or without traditional
2)people who over 70 with 2 or more risk factors but with no
history of heart attack;
3)apparently healthy people without cardio vascular disease or risk
2. We hypothesize that systemic proteomics study of blood serum from
different group of patients will identify additional candidate
markers, hence result in a much greater capacity to determine
individual risk profiles.
This study will be an analytic case-
control study and either sex, 40 to 80
years old patients will be recruited in
Design– Patient group
1- Heart attack: Blood Samples will be collected 2 weeks after the first time
Myocardial infarction, acute coronary syndromes from
outpatients at Memorial Herman and St. Lukes hospital
2- Suvivors: elderly people w/ 2 or more risk factors but no history of
heart attack. We will recruited this group of patients
houston nursing home.
3- Normal: apparently healthy people without cardio vascular disease or
risk factors from outpatients at Memorial Herman and St.
hospital with other specialty problem.
We classify the key risk factors include:
1. Gender and Age;
5. Diabetes mellitus;
6. Family History;
50 patients from each group will take
part in the study.
Questionnaire will be filled out for
acquiring baseline information
5ml blood with
Freeze -80 0
Rest of the inflammatory
Cells store with RNA
(Ambion) @-80 0
Monocytes Isolation and mRNA
at 260nm and 280n
( 260/280 ratio 1.8-2.0
as high quality)
20ug of RNA for
RNeasy Mini kit(Qiagen)
The Process of Gene expression assay
L L L
10% Biotin-labeled cRNA
Fragment (heat, Mg2+
Hybridize Wash/stain Scan
(Transcript labeling Kit: ENZo Diag)
HuSNP Mapping Process
hybridization scanningWash and staining
(120ng sample DNA) (1:1000 dilution)
HuSNP reagent kit
Hybridization and staining
Cost for gene expression
U133A $12000/30 Arrays
Choice System for
$56 /patient $144 /patient
Total Cost ~ $1300/patient
The expressed genes that have more
than 2 fold changes detected by
Microarray will be further verified by
With the 2-D-gel system proteins are separated by
molecular weight and isoelectric point.
The gel is silver stained and comparison of 2-D gels
of different materials will indicate protein spots which
differ in identity and/or quantity.
Interesting proteins ("differentials") can be
characterized further after analyzing protein digests
by mass spectrometry.
Comparison with public databanks can be done to
reveal the identity and functions of these proteins.
Process of Proteomics
Blood serum protein
We will cooperate with Dr.Fofanov and Dr.
Christoph F. Eick from UH Department of
Computer Science for developing software
and database for further analyzing the gene
expression profile from different group of
patients in order to gain further genetic
information that would help us to be able to
develop new diagnostic and therapies for
Coronary Heart Disease.