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1. CHRISTIAN DOPPLER LABORATORY
for Molecular Cancer Chemoprevention:
Colorectal
cancer (CRC) is the second leading cause of cancer death in the Western
world. Progress has been made in understanding the molecular mechanisms
of colon carcinogenesis (Wnt pathway, microsatellite instability,
CpG-island-methylator phenotype) and in the variety of therapeutic
modalities. However, advanced (metastatic) disease is still associated
with short life expectancy and excessive health care costs. Prevention
of cancer is a promising alternative strategy to safe lives and health
care resources.
Epidemiological
and interventional studies showed that salicylate-derivatives (aspirin,
mesalazine), some non-steroidal anti-inflammatory drugs (NSAIDs), COX-2
inhibitors, or the naturally occurring phenolic acid derivative
curcumin, protect from adenoma or CRC development. From a mechanistic
point of view, these compounds seem to interfere with different
pathways of colon carcinogenesis (e.g. aspirin is active in preventing
sporadic CRC and adenoma recurrence, whereas mesalazine is particularly
efficient in preventing inflammation-associated CRC). In vitro,
mesalazine decreases the proliferation of colorectal epithelial cells,
activates replication checkpoints, and reduces the rate of replication
errors. In contrast, aspirin restrains cell proliferation by inducing a
p21Waf1/Cip1- and ATM-dependent G1-arrest and p53-dependent apoptosis.
In this
Christian Doppler laboratory we plan to study the molecular basis of
cancer chemoprevention using mesalazine, aspirin, and curcumin as model
compounds,
to design and characterize novel molecules based on structure
similarities, to test novel molecules in animal models, and to evaluate
surrogate
markers of early carcinogenesis in biopsy material for testing the
effect of a novel mesalazine preparation (MMX-mesalazine) on CRC
prevention in humans.
By identifying
the mode of action of these key compounds, we expect to improve our
molecular understanding of colon carcinogenesis and our ability to
interfere with such. The designed novel compounds may have better
efficacy and less toxicity, and thus improve cancer prevention,
specifically in high-risk populations such as patients with inflammatory bowel disease or inherited cancer syndromes.
2. Chronic Inflammation and Colorectal Cancer:
Cancer is a disease that develops slowly. For most
solid human tumors there is a 20 year interval between the carcinogen
exposure and clinical detection of cancer. In ulcerative colitis, an
idiopathic chronic inflammatory diseaseof the large intestine,
colorectal cancer (CRC) development occurs at a higher rate and
speed.The basic premise for the development of colorectal cancer is
that cells accumulate millions of DNA mutations. The normal mutation
rate, however, is insufficient to account for the multiple mutations
observed in cancer cells. Therefore, changes that increase mutation
rates are essential for tumor development. This so called "mutator
phenotype" has been hypothesized for many years. It was first
identified in tumors from patients with Lynch-syndrome (i.e. HNPCC or
hereditary nonpolyposis colorectal cancer syndrome) in which multiple
mutations within repetitive DNA-sequences (so called microsatellites)
had been found. These multiple mutations in microsatellites are called
microsatellite instability or MSI. In HNPCC, MSI is caused by germline
mutations in DNA repair genes, the so called mismatch repair system
(MMR). Indeed, MSI is regarded as fingerprint of inactivation of this
DNA repair system that ensures DNA fidelity after each cell division.
In ulcerative colitis, MSI is found in a minor form already in
chronically inflamed mucosa suggesting that impairment of DNA fidelity
during cell division is a key mechanism in the development of
colitis-associated CRC. The reason for this is still unknown. Inhere we
hypothesize that chronic inflammation in the colon creates an
environment, which meets the criteria of a "mutator phenotype".
We intend to simulate such an environment in cell culture and we will
test its effect on the DNA mutation rate in a recently developed
system. Thereby, we transfer the clinical situation into the laboratory
and may proof the existence of an inflammation-associated "mutator
phenotype". The model will be useful for studying molecular mechanisms
in the development of cancer. In particular, we also intend to study
the effects of certain drugs and natural compounds for its ability to
improve DNA fidelity during cell division. The results of our tests can
be transferred into the clinical situations and may be used for
prevention of CRC in ulcerative colitis and eventually also for other
inflammation-associated cancers within the digestive tract.
3) Iron and Megakaryopoiesis:
Inflammatory bowel diseases (IBD) are associated
with high platelet counts (thrombocytosis), platelet activation and an
increased risk in thromboembolism, which is believed to be also caused
by hypercoagulopathy. Thromboembolism and thrombocytosis is not an IBD
specific but more a general event that was considered to be due to
elevated levels of proinflammatory cytokines such as IL-1, TNF, or
IL-6, which may stimulate (directly or indirectly) megakaryopoiesis
and/or increase platelet activity. In this regard, a correlation of
IL-6 (or IL-11) and platelet counts in Crohn's disease has been
demonstrated. However, other intestinal diseases with chronic
iron deficiency anemia without such a systemic inflammatory reactions
may also experience thrombocytosis (e.g. celiac disease). Since
iron deficiency anemia is associated with elevated EPO levels, it was
thought that EPO induced stimulation of megakaryopoiesis may be
responsible for the increase in platelets counts during iron deficiency
anemia. This proposal deals with an inflammation- and
EPO-independent hypothesis of thrombocytosis in IBD.
4) Variant IL-10R1 Signaling:
Interleukin-10 (IL-10) is a regulatory protein that
is secreted from immune cells during active inflammation. It is thought
to counteract inappropriate inflammation by inducing a series of
anti-inflammatory effects in the body. Animal models with IL-10
deficiency exhibit chronic inflammation of the bowel similar to human
Crohn's disease, indicating that IL-10 regulates the immune response in
the gut. All IL-10 induced signals are conducted over interaction of
IL-10 with IL-10 receptor 1 (IL-10R1). We previously identified genetic
variants of the IL-10R1, so called single nucleotide polymorphisms
(SNP), SNP3 and SNP4. The worldwide distribution of these variants had
been studied in 51 native populations. Both were most common in Semites
and Caucasians (up to half of the population) and absent in Asians or
Native Americans. These receptor variants changed the cellular response
to IL-10. We also found that these variants protect individuals from
development of some diseases but confer susceptibility to others
supporting the notion that these genetic variations in the IL-10R1 are
biologically relevant.
Inhere we propose to study the molecular mechanisms behind our
observations. By using standard biochemical methods, we investigate the
differential interaction of IL-10 and the IL-10R1 variants SNP3 and
SNP4. Our research will help to better understand the impact of genetic
variations on the cause of chronic inflammatory disease.
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