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1. SAMPLE COLLECTION:
NORMAL COLORECTAL MUCOSA FROM ENDOSCOPIC
PROCEDURES AT THE JOHN RADCLIFFE HOSPITAL.
-
PATIENTS GIVEN INFORMED CONSENT
COLLECTED 10ML BLOOD AND/OR COLORECTAL BIOPSIES
BIOPSIES FRESHLY USED TO EXTRACT DNA + RNA
REMAINING TISSUE SNAP FROZEN
2. ASSAY OPTIMIZATION:
BLOOD
- NUCLEATED CELL SEPARATION
- DNA EXTRACTION AND QC
- RNA EXTRACTION AND QC
COLORECTAL BIOPSIES
- TISSUE FRAGMENTATION &
HOMOGENISATION
- DNA EXTRACTION AND QC
- RNA EXTRACTION AND QC
DNA
RNA
- EXTRACTION AND QC
- EXTRACTION AND QC
- 23 RISK SNPS  POLYGENIC
- RETROTRANSCRIPTION AND DNASE
RISK SCORE(PRS)
TREATMENT
- TELOMERE LENGTH (TL) - PATHWAY EXPRESSION ANALYSIS (PEA)
3. BOWEL ASSAYS & CRC RISK:
COMPARED LEVELS OF 3 ASSAYS IN A GROUP OF
HEALTHY CONTROLS VS. PATIENTS WITH
POLYPS/CRC
- 2-STAGE DESIGN: PILOT50 (50 CASES & 50 CONTROLS)
PHASE2 (182 CASES & 57 CONTROLS)
- CASES SHOWED LONGER TL THAN CONTROLS
(OR=1.106;
P=0.025)
- CASES PRESENTED WITH HIGHER NUMBER OF RISK
ALLELES (P=0.007; OR=1.086)
4. BLOOD ASSAYS:
ANALYZE CORRELATION BETWEEN BOWEL MARKERS
AND THEIR BLOOD COUNTERPARTS
- BLOOD TL WAS SIGNIFICANTLY CORRELATED WITH BOWEL TL
- BLOOD TL COULD BE USED AS A PROXY FOR RISK
ASSESSMENT IN A QUICK AND LESS INVASIVE WAY