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244 High Throughput Assays for Isolation and Characterization of Cancer Stem Cells

Number of anticipated awards: 3-5
(Fast-Track proposals will not be accepted.)
Budget (total costs): Phase I: $150,000;
Phase II: $1,000,000

The deadline for receipt of all contract proposals submitted in response to this solicitation is: 5:00 p.m. Eastern Standard Time Monday, November 5, 2007

Cancer Stem Cells (CSC), represent a small population of cells within a tumor, which drives growth and metastasis, and has the phenotypic properties of stem cells. While chemotherapy and radiation therapy are able to eradicate most of the tumor cells, especially the fast dividing cells, these treatments frequently fail in eradicating the CSC, which might be dormant for years. Eradication of the CSC is thought to be essential for cure.

Currently the identification and isolation of CSC is based on markers that are shared by the same tissue stem cells and occasionally by other cells as well. This is followed by functional assays such as spheroid formation by cancerous cells in vitro for some cancer cells such as breast, prostate and brain, and by the recapitulation of the original tumor by the cancer stem cells in vivo in NOD SCID mice. There is an urgent need for the development of high-throughput assays for the isolation and characterization of cancer stem cells, particularly from premalignant and early stage cancers, as well as from exfoliated cells (e.g., urothelial cells in urine sediment) to improve early detection, diagnosis and prognosis.

Project goals:

  • Select a panel of differentially expressed markers for CSC.
  • Develop affinity reagents to target the differentially expressed markers in the form of monoclonal antibodies or recombinant antibodies, RNA or DNA aptamers, or small chemical molecules.
  • Develop high-throughput assays and methods for reproducible analysis of CSC.
  • Perform analytical validation, e.g., test the reproducibility, sensitivity, specificity in comparison with the current assays.
  • Test the utility of the developed assay in identification and isolation of CSC from premalignant lesions (e.g., DCIS, or HGPIN), early stage cancers and exfoliated cells of cancer patients (e.g., colon, bladder, breast, lung pancreas, and head and neck).

Phase I activities and expected deliverables:

  • Establish the proof of principle: Demonstrate that the developed assay, based on a panel of differentially expressed markers, can detect cancer stem cells at least as well as the current methodologies and functional assays or better.
  • Demonstrate that the same cancer stem cells can be isolated from the original cancer, the premalignant lesion and the exfoliated cells (e.g., for colon cancer).

Phase II activities and expected deliverables:

  • Validation of the assay: High-throughput isolation of cancer stems cells from: (1) 50 cancer patients with premalignant lesions (e.g., DCIS, HPIN); (2) 50 cancer patients with exfoliated (e.g., urine sediment of bladder cancers) cells;
  • Testing the utility of the developed cancer stem markers for early detection, diagnosis and prognosis.

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