Validation of Colon Biomarkers for the Early Detection of Colorectal Adenocarcinoma

The goal of this trial is to estimate the sensitivity and specificity of stool vimentin
methylation, serum galectin-3 ligand, and fecal immunochemical testing for colorectal
adenocarcinoma, or 2) screen relevant neoplasms (high-grade dysplasia or adenoma with ≥25%
villous histologic features or adenoma measuring ≥1 cm in the greatest dimension or sessile
serrated polyps measuring 1 cm or more in diameter) as single markers and in combination.
Asymptomatic subjects undergoing a colonoscopic procedure for screening for colorectal cancer
are eligible. Patients who have a first or second positive fecal immunochemical test, a
positive stool guaiac test or a positive Cologuard test are eligible. Up to 2,500 stool blood
or Cologuard positive subjects will be recruited on this protocol. Up to an additional 1,000
subjects who have not had previous FIT tests will be recruited. Subjects with a negative
stool blood or Cologuard test are not eligible for enrollment. Subjects will meet with
research staff prior to initiation of any colonoscopic preparative procedure. After
completing informed consent, they will complete Early Detection Research Network (EDRN) data
element forms. Blood and urine will be obtained following EDRN standard operating procedures
(SOPs). Subjects will be provided with kits to collect stool samples for fecal immunochemical
test (FIT) and processing for stool based biomarkers. The collected samples will be shipped
to the Central Laboratory at the University of Michigan or German Cancer Research Center
(Deutsches Krebsforschungszentrum, DKFZ), Heidelberg, Germany where the stool will be
homogenized, aliquoted, and stored at the Umiversity of Michigan CLASS laboratories . The FIT
tests will be sent to the Central Laboratory at the University of Michigan or to DKFZ for
quantitative analysis following standard operating procedures provided by Eiken Chemical
Company. Data from the screening colonoscopy will be obtained. One year after colonoscopy,
subjects will be contacted to determine if they have had a neoplastic colorectal diagnosis or
other neoplastic events. Data management and protocol coordination will be performed by the
Data Management and Coordinating Center (DMCC) of the EDRN along with the GLNE Prevention
Research Base at the University of Michigan and will include a Web-based front end and
relational database backend, with biosample tracking (VSIMS). Biosamples will be managed in a
high quality repository facility at the University of Michigan.

We will estimate sensitivities and specificities and the corresponding confidence intervals
of the stool DNA tests and serum/plasma tests for detection of invasive colorectal neoplasms
and for screen relevant neoplasias (Aim 1). We will then test the primary hypothesis to
confirm the clinical accuracy of a particular biomarker test or panel (Aim 2). The specific
primary hypothesis will be defined prior to data analysis based on state of the art
information available at that time about candidate biomarkers and tests. Several specific
examples of potential primary hypotheses are given to justify study sample size. Finally,
several alternative tests and multi-marker panels will be evaluated. (Aim 3). In secondary
analysis, we will (a) provide measures of diagnostic accuracy standardized to the age and
gender distribution of US population and (b) assess the effect of subject heterogeneity on
the marker performance. A primary objective is to establish an archive of appropriately
preserved stool, serum, plasma and DNA human biospecimens to be used by EDRN-approved
investigators for future validation and biomarker discovery research (Aim 4).

Inclusion Criteria:

- Adults 40*+ undergoing a first time colonoscopy for screening OR

- Positive guaiac-based occult blood or fecal immunochemical test (e.g. FOBT, FIT) in
the past 12 months (365 days)

- Willing to sign informed consent

- Able to physically tolerate removal of 50 ml of blood

- Willing to collect 2 stool samples

(*age 60 and up in U.S., 50 and up outside US)

Exclusion Criteria:

- Inability to provide informed consent

- History of Inflammatory Bowel Disease

- Overt rectal bleeding within 1 month (30 days) (including due to suspected
hemorrhoids)

- Undergone resection of the colon for any indication

- Subjects with known HIV or chronic viral hepatitis (Hepatitis B and C)

- Subjects with known or suspected HNPCC (Lynch Syndrome) or FAP

- Any cancer within 5 years of enrollment except any of the following:

- Squamous cell carcinoma of the skin or Basal cell carcinoma of the skin

- Carcinoma in situ of the cervix, Stages Ia or Ib invasive squamous cell carcinoma
of the cervix treated by surgery only. (Excluded if had pelvic radiation)

- Stage , 0, I or Ia Grade 1 adenocarcinoma of the endometrium treated with surgery