CANCER STEM CELLS
Cancer stem cell populations characterized by expression of CD44 hi , ABCG2, β-catenin, CD117, CD133, ALDH, VLA-2, CD166, CD201, IGFR, and/or EGF1R, and methods of isolating and using the same.
1 . An isolated cancer stem cell population comprising at least 90% cancer stem cells, wherein the cancer stem cells (i) express ABCG2 or express CD44 at a level that is at least 5-fold greater than non-tumorigenic cells of the same origin, (ii) are tumorigenic, (iii) are capable of self-renewal, and (iv) generate tumors comprising non-tumorigenic cells.
2 . The isolated cancer stem cell population of claim 1 , which comprises at least 95% cancer stem cells.
3 . The isolated cancer stem cell population of claim 1 , wherein the cancer stem cells comprise less than about 5% of the origin tumor cell population.
4 . The isolated cancer stem cell population of claim 3 , wherein the cancer stem cells comprise less than about 2% of the origin tumor cell population.
5 . The isolated cancer stem cell population of claim 4 , wherein the cancer stem cells comprise less than about 1% of the origin tumor cell population.
6 . The isolated cancer stem cell population of claim 1 , wherein the cancer stem cells expressing CD44 at a level that is at least 5-fold greater than non-tumorigenic cells of the same origin comprise less than about 50% of the origin tumor cell population.
7 . The isolated cancer stem cell population of claim 6 , wherein the cancer stem cells expressing CD44 at a level that is at least 5-fold greater than non-tumorigenic cells of the same origin comprise less than about 33% of the origin tumor cell population.
8 . The isolated cancer stem cell population of claim 7 , wherein the cancer stem cells expressing CD44 at a level that is at least 5-fold greater than non-tumorigenic cells of the same origin comprise less than about 25% of the origin tumor cell population.
9 . The isolated cancer stem cell population of claim 8 , wherein the cancer stem cells expressing CD44 at a level that is at least 5-fold greater than non-tumorigenic cells of the same origin comprise less than about 15% of the origin tumor cell population.
10 . The isolated cancer stem cell population of claim 9 , wherein the cancer stem cells expressing CD44 at a level that is at least 5-fold greater than non-tumorigenic cells of the same origin comprise less than about 10% of the origin tumor cell population.
11 . The isolated cancer stem cell population of claim 1 , wherein the cancer stem cells additionally express β-catenin, CD117, CD133, ALDH, VLA-2, CD166, CD201, IGFR, EGF1R, or a combination thereof.
12 . The isolated cancer stem cell population of claim 1 , wherein the cancer stem cells do not express differentiation markers.
13 . The isolated cancer stem cell population of claim 12 , wherein the cancer stem cells are depleted of cells expressing CD26, Muc-1, Muc-2, villin, CD24, CEA, or CK20.
14 . The isolated cancer stem cell population of claim 1 , which is derived from colon.
15 . The isolated cancer stem cell population of claim 1 , wherein a subpopulation of about 10 cells has the capacity to form a palpable tumor.
16 . An enriched cancer stem cell population derived from a tumor cell population comprising cancer stem cells and non-tumorigenic cells, wherein the cancer stem cells (i) express ABCG2 or express CD44 at a level that is at least 5-fold greater than non-tumorigenic cells of the same origin, (ii) are tumorigenic, (iii) are capable of self-renewal, (iv) generate tumors comprising non-tumorigenic cells, and (iv) are enriched at least 2-fold compared to the tumor cell population.
17 . The enriched cancer stem cell population of claim 16 , wherein the cancer stem cells are enriched at least 5-fold compared to tumor-derived cell population.
18 . The enriched cancer stem cell population of claim 17 , wherein the cancer stem cells are enriched at least 10-fold compared to tumor-derived cell population.
19 . The enriched cancer stem cell population of claim 18 , wherein the cancer stem cells are enriched at least 50-fold compared to tumor-derived cell population.
20 . The enriched cancer stem cell population of claim 19 , wherein the cancer stem cells are enriched at least 100-fold compared to tumor-derived cell population.
21 . The enriched cancer stem cell population of claim 16 , wherein the cancer stem cells additionally express β-catenin, CD117, CD133, ALDH, VLA-2, CD166, CD201, IGFR, EGF1R, or a combination thereof.
22 . The enriched cancer stem cell population of claim 16 , wherein the cancer stem cells do not express differentiation markers of the tumor cell population.
23 . The enriched cancer stem cell population of claim 22 , wherein the cancer stem cells are depleted of cells expressing CD26, Muc-1, Muc-2, villin, CD24, CEA, or CK20.
24 . The enriched cancer stem cell population of claim 16 , which is derived from colon.
25 . The enriched cancer stem cell population of claim 16 , wherein a subpopulation of about 10 cells has the capacity to form a palpable tumor.
26 . A method of isolating a cancer stem cell population comprising:
(a) providing dissociated tumor cells, wherein a majority of the cells express CD44 at a low level, and wherein a minority of the cells express CD44 at a high level that is at least about 5-fold greater than the low level;
(b) contacting the dissociated tumor cells with an agent that specifically binds to CD44;
(c) selecting cells that specifically bind to the agent of (b) to an extent that shows a high level of CD44 expression that is at least about 5-fold greater than the low level;
whereby a cancer stem cell population is isolated.
27 . The method of claim 26 , wherein the cancer stem cell population comprises at least 90% cancer stem cells.
28 . The method of claim 27 , wherein the cancer stem cell population comprises at least 95% cancer stem cells.
29 . The method of claim 26 , wherein the cancer stem cell population is enriched in cancer stem cells at least 2-fold when compared to the dissociated tumor cells.
30 . The method of claim 29 , wherein the cancer stem cell population is enriched in cancer stem cells at least 5-fold when compared to the dissociated tumor cells.
31 . The method of claim 30 , wherein the cancer stem cell population is enriched in cancer stem cells at least 10-fold when compared to the dissociated tumor cells.
32 . The method of claim 26 , further comprising:
(d) contacting the dissociated tumor cells with an agent that specifically binds to ABCG2; and
(e) selecting cells that specifically bind to the agent of (d).
33 . The method of claim 26 , further comprising:
(d) contacting the dissociated tumor cells with one or more agents that specifically bind to ABCG2, CD117, CD133, ALDH, CD166, CD201, IGFR, EGF1R, or a combination thereof; and
(e) selecting cells that specifically bind to an agent or combination of agents of (d).
34 . The method of claim 26 , further comprising:
(d) contacting the dissociated tumor cells with one or more agents that specifically binds to a differentiation marker expressed by the tumor cells; and
(e) depleting the cancer stem cell population of cells that specifically bind to the one or more agents of (d).
35 . The method of claim 34 , wherein the differentiation marker is CD26.
36 . The method of claim 26 , wherein the agent that specifically binds CD44 is an anti-CD44 antibody.
37 . The method of claim 26 , wherein the selecting cells is performed by flow cytometry, fluorescence activated cell sorting, panning, affinity column separation, or magnetic selection.
38 . The method of claim 26 , wherein the dissociated tumor cells are colon cancer cells.
39 . A cancer stem cell population isolated according to the method of claim 26 .
40 . A method of isolating a cancer stem cell population comprising:
(a) providing dissociated tumor cells;
(b) contacting the dissociated tumor cells with an agent that specifically binds to ABCG2;
(c) selecting cells that specifically bind to the agent of (b);
whereby a cancer stem cell population is isolated.
41 . The method of claim 40 , wherein the cancer stem cell population comprises at least 90% cancer stem cells.
42 . The method of claim 41 , wherein the cancer stem cell population comprises at least 95% cancer stem cells.
43 . The method of claim 40 , wherein the cancer stem cell population is enriched in cancer stem cells at least 10-fold when compared to the dissociated tumor cells.
44 . The method of claim 43 , wherein the cancer stem cell population is enriched in cancer stem cells at least 50-fold when compared to the dissociated tumor cells.
45 . The method of claim 44 , wherein the cancer stem cell population is enriched in cancer stem cells at least 100-fold when compared to the dissociated tumor cells.
46 . The method of claim 40 , further comprising:
(d) contacting the dissociated tumor cells with one or more agents that specifically bind to CD44, CD117, CD133, ALDH, CD166, CD201, IGFR, EGF1R, or a combination thereof; and
(e) selecting cells that specifically bind to the one or more agents of (d).
47 . The method of claim 40 , further comprising:
(d) contacting the dissociated tumor cells with one or more agents that specifically binds to a differentiation marker expressed by the tumor cells; and
(e) depleting the cancer stem cell population of cells that specifically bind to the one or more agents of (d).
48 . The method of claim 40 , wherein the differentiation marker is CD26.
49 . The method of claim 40 , wherein the dissociated tumor cells comprise a majority of cells expressing CD44 at a low level and a minority of cells expressing CD44 at a high level that is at least about 5-fold greater than the low level; and wherein the method further comprises:
(d) contacting the dissociated tumor cells with an agent that specifically binds to CD44; and
(e) selecting cells that bind to the agent of (d) to an extent that shows a high level of CD44 expression that is at least about 5-fold greater than the low level.
50 . The method of claim 40 , wherein the agent that specifically binds ABCG2 is an anti-ABCG2 antibody.
51 . The method of claim 40 , wherein the selecting cells is performed by flow cytometry, fluorescence activated cell sorting, panning, affinity column separation, or magnetic selection.
52 . The method of claim 40 , wherein the dissociated tumor cells are colon cancer cells.
53 . A cancer stem cell population isolated according to the method of claim 40 .
54 . A method of testing efficacy of a cancer drug or candidate cancer drug comprising:
(a) providing an isolated cancer stem cell population according to claim 1 ;
(b) contacting the cancer stem cells with a cancer drug or a candidate cancer drug;
(c) observing a change in tumorigenic potential of the cancer stem cells following contacting the cancer stem cells with the cancer drug or candidate cancer drug.
55 . A method of testing efficacy of a cancer drug or candidate cancer drug comprising:
(a) providing an enriched cancer stem cell population according to claim 16 ;
(b) contacting the cancer stem cells with a cancer drug or a candidate cancer drug;
(c) observing a change in tumorigenic potential of the cancer stem cells following contacting the cancer stem cells with the cancer drug or candidate cancer drug.
56 . A method of testing efficacy of a cancer drug or candidate cancer drug comprising:
(a) providing a cancer stem cell population according to claim 39 ;
(b) contacting the cancer stem cells with a cancer drug or a candidate cancer drug;
(c) observing a change in tumorigenic potential of the cancer stem cells following contacting the cancer stem cells with the cancer drug or candidate cancer drug.
57 . A method of testing efficacy of a cancer drug or candidate cancer drug comprising:
(a) providing a cancer stem cell population according to claim 53 ;
(b) contacting the cancer stem cells with a cancer drug or a candidate cancer drug;
(c) observing a change in tumorigenic potential of the cancer stem cells following contacting the cancer stem cells with the cancer drug or candidate cancer drug.