SBIR-STTR Award

Colorectal carcinoma is the most common gastrointestinal malignancy affecting about 160,000 new cases per year in the United States. Survival rates are closely related to the stage of cancer at the time of diagnosis. Novel approaches for the detection of colon cancer in the early stages by non-invasive methods are critically needed for secondary prevention by mass screening of high risk populations. The new emerging science of COPROCYTOBIOLOGY, involving the isolation of colonic epithelial cells from stool and screening these cell populations for tumor cells carrying cancer-specific markers is an exciting development. This new technology is still a research tool, although a number of publications have appeared showing its usefulness in early cancer diagnosis. The objective of this SBIR, is to standardize the technology, develop prototype kits for the collection of samples and introduce in kit form a simplified high throughput procedure which can be readily adopted by clinical laboratories in a cost effective manner. In optimizing the components of the kits, the outcome variables will be cell yield, viability, and retention of cell surface markers. A standard reference marker will be designated for quality control and data reduction when comparing results from several centers. The transport medium for the sample collection kit will be a solution containing the appropriate preservatives and chemical constituents to keep the colonic cells intact and functional. The high throughput procedure will be modifications of published methodologies, eliminating time consuming steps, and providing carefully pre prepared media for size/density based separations involving readily available benchtop laboratory centrifuges. PROPOSED COMMERCIAL APPLICATIONS: If a standardized kit becomes available for isolating colonic cells noninvasively, it would be an attractive and cost effective approach to the early detection of colon cancer and other colonic diseases. Diagnostic laboratories, research centers, biology labs and oncology research centers will be potential users.

Thesaurus Terms:
cell sorting, colon, colon neoplasm, cytodiagnosis, feces analysis, neoplasm /cancer diagnosis, technology /technique development biomarker, diagnosis design /evaluation, diagnostic test, growth media, tissue /cell culture human tissue

Colorectal carcinoma is the most common gastrointestinal malignancy affecting about 160,000 new cases per year in the United States. Survival rates are closely correlated with the stage of cancer at the time of diagnosis. This Phase II (revised) proposal is based on the fact that it is possible to recover colonic cells from human stool and examine them for biomarkers associated with malignant transformation. Our objective is to develop a noninvasive screening test for colon cancer by demonstrating the expression of tumor associated biomarkers on exfoliated colonocytes isolated from stools of patients with colon cancer. This is a cross-sectional observational study of patients undergoing diagnostic colonoscopy at two medical centers: Sinai Hospital in Baltimore and the Walter Reed Army Medical Center in Washington, D.C. The outcome measures are PCR amplicons of CD 44 splice variants and tumor specific variants of carcinoembryonic antigen (CEA). In addition, cell surface carbohydrate motifs linked to tumorigenesis will be examined by flow cytometry using fluorescently labeled plant lectins that are now to be specific ligands. From previous studies we expect to show a high degree of sensitivity and specificity for detection of colon cancer and its precursor polyps using a panel of these biomarkers. As a reference marker Cytokeratin 19 will be amplified, semiquantitatively, by PCR using cDNA generated from mRNA extracted from the cells. Our objective is to develop a standardized kit for the detection of colon cancer, non-invasively, as a cost-effective screening tool.

Thesaurus Terms:
cell sorting, colon, colon neoplasm, cytodiagnosis, diagnosis design /evaluation, feces analysis, neoplasm /cancer diagnosis biomarker, diagnostic test, growth media, tissue /cell culture clinical research, human subject