Clinical Trial Details
— Status: Completed
Administrative data
| NCT number |
NCT02786602 |
| Other study ID # |
PA16046 |
| Secondary ID |
|
| Status |
Completed |
| Phase |
N/A
|
| First received |
May 26, 2016 |
| Last updated |
March 2, 2017 |
| Start date |
May 2016 |
| Est. completion date |
March 2017 |
Study information
| Verified date |
March 2017 |
| Source |
CHU de Reims |
| Contact |
n/a |
| Is FDA regulated |
No |
| Health authority |
|
| Study type |
Observational
|
Clinical Trial Summary
Colorectal cancer (CRC) is a major public health problem in France and worldwide. CRC is the
third most common cancer in incidence and mortality in France. The vast majority of these
cancers are adenocarcinomas that arise sporadically and develop from precursor lesions:
adenoma. All CRC with the same disease stage do not have the same prognosis. Various
parameters have been identified as factors influencing the prognosis and allows adjustment
of the treatment. The poor histoprognostic factors are vessels and nerves invasion by the
tumor or the mucinous adenocarcinoma subtype. At the molecular level, the presence of
microsatellite instability (MSI) improves the prognosis, while the presence of a BRAF
mutation is an independent poor prognostic factor.
The different molecular pathways of colonic carcinogenesis are the chromosomal instability
pathway, the microsatellite instability pathway inducing errors in DNA mismatch repair and
the CpG Island Methylator Phenotype (CIMP). The hypermethylation of CpG islands of genes
promoters leads to an over or most frequently under gene expression. CIMP is observed in
near 15% of CRC and is associated with specific clinical and pathological features: older
patients, female predominance, right colonic involvement, poorly differentiated or mucinous
adenocarcinomas. From a molecular point of view, the high CIMP phenotype is strongly
associated with the presence of BRAFV600E mutation, the absence of RAS mutation and the
presence of microsatellite instability. The prognostic value of CIMP is actually
controversial. A recent meta-analysis found that the CIMP phenotype was associated with a
poor prognosis. Methylation of some genes promoters as CDKN2A is associated with a poor
prognosis.
LRP-1 (low density lipoprotein receptor-related protein 1) is a multifunctional endocytic
receptor that belongs to the LDL receptors the family. It mediates the clearance of many
extracellular enzymes involved in the spread of cancer cells: metalloproteinases and serine
proteinases. Decrease of LRP-1 activity or loss of LRP-1 expression correlates with
increased aggressiveness of cancer cells in certain types of cancer. The expression of LRP-1
has almost never been studied in CRC. Only one immunohistochemical study of LRP-1 protein
expression in colonic adenocarcinoma has been published to date. This study shows that tumor
cells express LRP-1, but in nearly half the cases, weaker than in normal cells colic. The
mechanisms involved in the decrease of expression are not known.
An epigenetic mechanism might be involved as hypermethylation of the of LRP-1 gene promoter,
especially as the promoter of this gene is rich in CpG islands (methylation targets).
Clinical and prognostic significance of the LRP-1 gene expression and promoter methylation
is actually unknown.
Description:
Colorectal cancer (CRC) is a major public health problem in France and worldwide. CRC is the
third most common cancer in incidence and mortality in France. The vast majority of these
cancers are adenocarcinomas that arise sporadically and develop from precursor lesions:
adenoma. All CRC with the same disease stage do not have the same prognosis. Various
parameters have been identified as factors influencing the prognosis and allows adjustment
of the treatment. The poor histoprognostic factors are vessels and nerves invasion by the
tumor or the mucinous adenocarcinoma subtype. At the molecular level, the presence of
microsatellite instability (MSI) improves the prognosis, while the presence of a BRAF
mutation is an independent poor prognostic factor.
The different molecular pathways of colonic carcinogenesis are the chromosomal instability
pathway, the microsatellite instability pathway inducing errors in DNA mismatch repair and
the CpG Island Methylator Phenotype (CIMP). The hypermethylation of CpG islands of genes
promoters leads to an over or most frequently under gene expression. CIMP is observed in
near 15% of CRC and is associated with specific clinical and pathological features: older
patients, female predominance, right colonic involvement, poorly differentiated or mucinous
adenocarcinomas. From a molecular point of view, the high CIMP phenotype is strongly
associated with the presence of BRAFV600E mutation, the absence of RAS mutation and the
presence of microsatellite instability. The prognostic value of CIMP is actually
controversial. A recent meta-analysis found that the CIMP phenotype was associated with a
poor prognosis. Methylation of some genes promoters as CDKN2A is associated with a poor
prognosis.
LRP-1 (low density lipoprotein receptor-related protein 1) is a multifunctional endocytic
receptor that belongs to the LDL receptors the family. It mediates the clearance of many
extracellular enzymes involved in the spread of cancer cells: metalloproteinases and serine
proteinases. Decrease of LRP-1 activity or loss of LRP-1 expression correlates with
increased aggressiveness of cancer cells in certain types of cancer. The expression of LRP-1
has almost never been studied in CRC. Only one immunohistochemical study of LRP-1 protein
expression in colonic adenocarcinoma has been published to date. This study shows that tumor
cells express LRP-1, but in nearly half the cases, weaker than in normal cells colic. The
mechanisms involved in the decrease of expression are not known.
An epigenetic mechanism might be involved as hypermethylation of the of LRP-1 gene promoter,
especially as the promoter of this gene is rich in CpG islands (methylation targets).
The clinical impact of the level of LRP-1 expression on overall survival of cancer patients
has been assessed in two studies: one on patients suffering from hepatocellular carcinoma
and one on patients suffering from primitive lung adenocarcinomas. In both studies, the
decrease of immunohistochemical and gene expression of LRP-1 correlated with overall
survival decrease. The clinical and prognostic impact of LRP-1 expression in CRC and its
association with a particular molecular or morphological profile has not been studied to
date.
In this work, the investigators will study the methylation pattern of the LRP-1 gene
promoter in a well-characterized series of CRC.
The main objective of this study is to study by methylation analysis the role LRP-1 promoter
gene methylation in the prognosis of CRC.
The secondary objective of this study is to evaluate the prognostic impact of CIMP.
This study will be the first to explore the LRP-1 gene promoter methylation in in CRC. This
study will help to increase knowledge of the prognostic role of LRP-1 gene promoter
methylation in CRC and its possible association with clinical, morphological or molecular
parameters.
This study is a single center retrospective cohort study.
The investigators will first design the list of all eligible patients by using a request in
the software DIAMIC of th pathology department with component code "colon", damage code
"adenocarcinoma " and period "between 2006 and 2012". Patients suffering from Lynch syndrome
or other familial cancer syndrome will be excluded of the study. Informed consent will be
send to all the selected patients.
Once the patients list will be completed:
- The patients' medical records will be reviewed for: sociodemographic and clinical data
colonic tumor location, TNM stage, MSI, RAS and BRAF status, presence of synchronous
metastases, treatment type and duration, patient follow up to the date of 01/06/2016
point (metastases occurrence, disease recurrence, patient death, ...).
- A central review of all pathological slides will be performed for these criteria:
adenocarcinoma subtype according to WHO 2010 classification, differentiation,
association with polyps and their types, pTNM stage, invasion front and budding, stroma
type, presence and amount of tumor necrosis).
- Methylation analysis:
- LRP1 gene promoter methylation analysis by pyrosequencing.
- CIMP status determination by High Resolution Melting analysis of MLH1, MINT1,
MINT2, MINT31 and CDKN2A genes promoters.
Statistical analysis:
- Data description: mean and standard deviation for quantitative variables; number and
percentage for categorical variables.
- Comparison of tumors having a high methylation of the LRP-1 gene promoter and tumors
with not or little methylation of the LRP-1 gene promoter by univariate analysis
(Student's test and the Wilcoxon test, Chi2 or Fisher's exact) and multivariate
(logistic regression).
- Search factors associated with patient outcomes, including the methylation of LRP1
promoter and CIMP status, by univariate analysis (log-rank test) and multivariate (Cox
model).
