Compositions and methods for treating and diagnosing cancer
View Patent ↗The present invention relates to compositions and methods for characterizing, treating and diagnosing cancer. In particular, the present invention provides a cancer stem cell profile, as well as novel stem cell cancer markers useful for the diagnosis, characterization, prognosis and treatment of cancer and in particular the targeting of solid tumor stem cells.
1. A method for obtaining from a colon tumor a cellular composition comprising colon cancer stem cells and non-tumorigenic colon tumor cells, wherein at least 75% of the tumor cells in the composition are tumorigenic colon cancer stem cells and 25% or less of the tumor cells in the composition are non-tumorigenic colon tumor cells, said method comprising:
(a) obtaining a dissociated mixture of colon tumor cells; and
(b) separating the mixture of colon tumor cells into a first fraction comprising at least 75% colon cancer stem cells and 25% or less non-tumorigenic colon tumor cells and a second fraction of colon tumor cells depleted of colon cancer stem cells wherein the separating comprises contacting the mixture with reagents against CD44 and epithelial-specific antigen (ESA); and
(c) collecting said first fraction, wherein said colon cancer stem cells in said first fraction are CD44 + and ESA + .
2. The method of claim 1 , wherein the separating is performed by flow cytometry, fluorescence activated cell sorting (FACS), panning, affinity chromatography or magnetic selection.
3. The method of claim 1 , wherein the separating is performed by fluorescence activated cell sorters (FACS) analysis.
4. The method of claim 1 , wherein the separating further comprises contacting the mixture with reagents against at least one additional reagent against a protein selected from the group consisting of: HES6, CD166, CEACAM6, CD59, and CD49f.
5. A method for preparing a population of colon tumor cells enriched for colon cancer stem cells, wherein the enriched population comprises 75% or more colon cancer stem cells and 25% or less non-tumorigenic colon tumor cells, said method comprising:
(a) obtaining a dissociated mixture of colon tumor cells;
(b) contacting the mixture of colon tumor cells with reagents against CD44, CD166, and ESA;
(c) selecting a first fraction enriched for colon cancer stem cells by their binding to the reagents and a second fraction of colon tumor cells depleted of colon cancer stem cells; and
(d) collecting said first fraction, wherein said colon cancer stem cells in said first fraction are CD44 + and ESA + .
6. The method of claim 5 , wherein the selecting is performed by flow cytometry, fluorescence activated cell sorting (FACS), panning, affinity chromatography, or magnetic selection.
7. The method of claim 5 , wherein the selecting is performed by fluorescence activated cell sorters (FACS) analysis.
8. The method of claim 5 , wherein the reagents with which the mixture of colon tumor cells are contacted further comprise a reagent against HES6.
9. The method of claim 5 , wherein the mixture of colon tumor cells are further contacted with a reagent against CD59.
10. The method of claim 5 , wherein the mixture of colon tumor cells are further contacted with a reagent against CD49f.
11. The method of claim 5 , wherein the mixture of colon tumor cells are further contacted with a reagent against CEACAM6.
12. A method for obtaining from a colon tumor a cellular composition comprising colon cancer stem cells and non-tumorigenic colon tumor cells, wherein at least 75% of the tumor cells in the composition are tumorigenic colon cancer stem cells and 25% or less of the tumor cells in the composition are non-tumorigenic colon cancer tumor cells, said method comprising:
(a) obtaining a dissociated mixture of colon tumor cells;
(b) separating the mixture of colon cancer tumor cells into a first fraction comprising at least 75% colon cancer stem cells and 25% or less non-tumorigenic colon tumor cells and a second fraction of colon tumor cells depleted of colon cancer stem cells, wherein the separating comprises contacting the mixture with reagents against CD44, ESA, and CD49f; and
(c) collecting said first fraction, wherein said colon cancer stem cells in said first fraction are CD44 + , ESA + , and CD49f hi .
13. The method of claim 12 , wherein the separating is performed by flow cytometry, fluorescence activated cell sorting (FACS), panning, affinity chromatography or magnetic selection.
14. The method of claim 12 , wherein the separating is performed by fluorescence activated cell sorters (FACS) analysis.
15. The method of claim 12 , wherein the separating further comprises contacting the mixture with reagents against HES6.
16. The method of claim 12 , wherein the separating further comprises contacting the mixture with reagents against CD166.
17. The method of claim 12 , wherein the separating further comprises contacting the mixture with reagents against CEACAM6.
18. The method of claim 12 , wherein the separating further comprises contacting the mixture with reagents against CD59.