Correlation between NOD2/CARD15 and TNF-
gene mutations
and the clinical manifestations of Crohnís
disease
Inflammatory Bowel Diseases (such as
Crohnís disease, ulcerative
colitis, etc.) cause chronic inflammation of the digestive
tract. Abdominal pain, persistent diarrhea, rectal
bleeding, fever, etc. are the most common symptoms.
It is not completely clear what causes Crohnís
disease, but the most accepted theory is that the
bodyís immune system reacts to a virus or a
bacterium by causing an ongoing inflammation in the
intestine. It is also believed that the disease has
multifactorial genetic background. NOD2/CARD15 gene
was identified within chromosome 16, and its three
alleles have shown a statistical association with
Chronís disease in different European Caucasian
populations. Recent studies show that TNF-
(Tumor Necrosis Factor ñalpha) gene polymorphisms
have an important role in phenotype modulation in
patients with Crohnís disease. It has also
been established that the -308 polymorphism in the
TNF- promoter is responsible for certain
extra-intestinal manifestations of the IBD, which
is very important in the prognosis and further treatment
of such patients.The manifestations of mutations in
NOD2/CARD15 gene and the -308 polymorphism in the
TNF- promoter have not been sufficiently
studied in Serbian patients with Crohnís disease.
Thus, it is of scientific and clinical importance
to establish the genotype of the patients at these
loci and stratify the patients by the disease progress
and treatment response.
Research Phases:
Week 1:
1. DNA isolation from the blood samples of patients
with Crohnís disease
and healthy control subjects (phenol/chlorophorm)
Weeks 2-3:
2. Optimization of the PCR-RFLP (Polymerase Chain
Reaction- Restriction
Fragment Length Polymorphism) method for detecting
the R702W mutation in
exon 4 of the NOD2/CARD15 gene
3. Optimization of the PCR-RFLP method
for detecting G908R mutation in
exon 8 of the NOD2/CARD15 gene
4. Optimization of the PCR-RFLP method
for detecting L1007fs mutation in
exon 11 of the NOD2/CARD15 gene
5. Optimization of the PCR-RFLP method
for detecting -308 mutation in
TNF- gene promoter
Weeks 4-9:
6. DNA analysis of patients with Crohnís disease
for the presence of three mutations in NOD2/CARD15
gene
7. DNA analysis of healthy controls
for the presence of three mutations in NOD2/CARD15
gene
8. DNA analysis of patients with Crohnís
disease for the presence of the -308 mutation
9. DNA analysis of healthy controls
for the presence of the -308 mutation
Week 10:
10. Statistical analysis of results
Number of blood samples that will be
analyzed: 70 (out of which, 50 with Crohnís
disease, 20 healthy controls)
Estimated duration of project: ten weeks
This internship proposal was developed
in conjunction with Sonja
Pavlovi, Ph.D., Institute for Molecular Genetics and
Genetic
Engineering, Belgrade, Serbia and Montenegro, who
has agreed to be my
internship supervisor and mentor.
Activities
Associated with Award
Poster Presentation:
Presented at the Monthly
Seminar of the Institute of Molecular Genetics and
Genetic Engineering, Belgrade, Serbia on August 8th,
2006.
Presented at Bryn
Mawr College Summer Science Internship and Fellowship
Poster Session Sept. 7th, 2006.
