Clinical Trial Details
— Status: Completed
Administrative data
| NCT number |
NCT03872622 |
| Other study ID # |
5484-18-SMC |
| Secondary ID |
|
| Status |
Completed |
| Phase |
N/A
|
| First received |
|
| Last updated |
|
| Start date |
November 27, 2018 |
| Est. completion date |
August 29, 2023 |
Study information
| Verified date |
April 2024 |
| Source |
Sheba Medical Center |
| Contact |
n/a |
| Is FDA regulated |
No |
| Health authority |
|
| Study type |
Interventional
|
Clinical Trial Summary
A cohort of CD patients (total n=300) and a cohort of controls (healthy individuals and
family members of CD patients, total n=200) will be recruited.
We will analyze gut host pattern of tissue immune system and epithelial responses to putative
exposome and microbial triggers, expression data in the gut using mRNA expression studies and
genetic analysis of the resident bacterial flora are necessary to identify specific molecules
and signal pathways as well as specific bacterial taxa involved in aberrant response and
instigation of inflammation that is the driver of Crohn's disease. Therefore, We will obtain
stool & blood samples and intestinal biopsy and/or resection specimens from CD patients, and
blood, stool and intestinal biopsy samples from healthy individuals and a-symptomatic family
relatives undergoing colonoscopy for reasons un-related to the study (e.g screening
colonoscopy).
Patients from both cohorts will also undergo environmental and dietary exposure survey. For
environmental exposure, we will use the questionnaire developed by the International
Organization of IBD (IOIBD), with some modification. Questions relate to five main different
areas: (i) Childhood factors up to 20 years; (ii) food habits including daily, weekly or less
frequent consumption; (iii) smoking habits; (iv) sanitary conditions such as the availability
of in-house water tap, hot water tap or flush toilet; and (v) others factors including
physical activity, oral contraceptive pill and stressful events before diagnosis.
For mapping dietary habits we will employ an interview conducted by a trained dietician and
using the validated structured FFQ (Food frequency Questionnaire).
Description:
Only adults>18YO will be included, Two cohorts will be included: A cohort of CD patients
(including specific sub-cohorts of recruited known & new-onset CD and travelling CD patients
and archived pathological specimens of post-operative CD, total n=300) and a cohort of
controls (healthy individuals and family members of CD patients, total n=200). We will obtain
stool & blood samples and intestinal biopsy and/or resection specimens from CD patients, and
blood, stool and intestinal biopsy samples from healthy individuals and a-symptomatic family
relatives undergoing colonoscopy for reasons un-related to the study (e.g screening
colonoscopy).
After signing informed consent, Stool sample and blood sample (20cc) will be obtained.
Enteric infections will be excluded by stool cultures, parasites including entameba, and C.
difficile assay. Inflammatory markers (CRP in blood, calprotectin in stool) will be tested.
Blood samples will undergo immune characterization including in vitro testing by immune cell
activation, cytokine secretion measurements and cell-surface markers analysis by FACS. Two
fecal samples per donor, (≥3g or 3 mL each) will be collected in clean containers at room
temperature, fresh frozen at -20°C and then stored at -80°C within 72h. We will also obtain
at colonoscopy four biopsies from inflamed and non-inflamed ileal and colonic mucosa (total 8
biopsies). From each individual, samples will be divided for analyses at Sheba and for
transfer. Stool will be transferred to Prof. Elinav in Weizmann Institute to perform shotgun
metagenomics of the microbiome. Intestinal biopsy and/or archived resection specimens
(histology) will be transferred to Prof Stappenbeck at Washington University, St. Louis USA,
for histological analysis of innate immune cells and paneth cell morphology analysis by H&E
and immunohistochemistry.
Ileal and rectal biopsies will be collected and stored immediately in RNAlater. Biopsies will
be obtained from each location (either inflamed or not inflamed, total 8 biopsies) and used
for RNA and for microbial DNA extraction. RNA will be used for mRNAseq of tissue, and DNA for
microbial characterization. mRNAseq will be done at the NIH supported digestive health center
(DHC) core at Cincinnati Children Hospital Medical Center (CCHMC), where >700 mRNAseq of
different IBD cohorts including RISK CD and PROTECT UC cohorts were already run with good
results. Microbial 16S amplicon sequencing of stool & biopsy extracted DNA will be done as
previously described.
Patients from both cohorts will also undergo environmental and dietary exposure survey. For
environmental exposure, we will use the questionnaire developed by the International
Organization of IBD (IOIBD), with some modification. The questionnaire consists of 87
questions covering 25 different topics proposed to be environmental risk factors for CD
and/or UC. This questionnaire has been previously used in epidemiological studies
investigating triggers of IBD, including one conducted in South-East Asia and China.
Questions relate to five main different areas: (i) Childhood factors up to 20 years including
breast feeding, appendectomy, tonsillectomy, eczema, vaccinations (tuberculosis, pertussis,
measles, rubella, diphtheria, tetanus, polio), childhood infections (measles, pertussis,
rubella, chickenpox, mumps, scarlet fever) and pet ownership; (ii) food habits including
daily, weekly or less frequent consumption of fruit, vegetables, egg, cereal, bread, cereal,
coffee, tea, juice, sugar and fast food; (iii) smoking habits (current smoker, non-smoker,
ex-smoker); (iv) sanitary conditions such as the availability of in-house water tap, hot
water tap or flush toilet; and (v) others factors including daily physical activity, oral
contraceptive pill and stressful events before diagnosis. We will also add to the IOIBD
questionnaire items pertaining to antibiotic use before and after the age of 15 years, use of
toothpaste and presence of amalgam teeth filling during childhood or later in life.
For mapping dietary habits we will employ an interview conducted by a trained dietician and
using the validated structured FFQ (Food frequency Questionnaire). This tool captures >600
food items, prioritized by their pre-defined frequency in regular diet. Personnel will be
trained in the dietary interview method. FFQ results will be further imported into
computerized FFQ version, which automatically derives and normalizes the nutrients content
from an individual patient diet based on known Israel food-items nutrients' database from the
National Institute of Statistics. As shown in the screen-shot figure from this responsive
software for an exemplary patient, each color bar denotes a specific nutrient consumption by
the individual (e.g protein, oligosaccharides, etc), along with its divergence from WHO
normalized consumption (green colored range). The FFQ methodology will be complemented by a
validation prospective three-day dietary itinerary capture by SmartPhone. Participants will
take electronic pictures of all food items consumed during these three day. Photos will be
uploaded to electronic visual data base, and food items will be identified, recorded and
verified against the retrospective FFQ.
In order to prioritize among the multiple environmental/dietary factors, we will perform a
systematic review and meta-analysis of all publications pertaining to environmental
risk-factors for CD. We will compute pooled odds ratio and point estimates for each variable,
and define the weighted effects between variables using random effect model. This will enable
to dissect and identify factors which have been most reproducibly and robustly found to be
associated with occurrence of CD. We will then prioritize these variables in a step-wise
manner, for the analysis of environmental factors association with microbiome, transcriptome
and PCP perturbations.
