Isolation, gene expression, and chemotherapeutic resistance of motile cancer cells
View Patent ↗Methods of isolating motile cells of interest from an animal tissue is provided. Also provided are methods of determining mRNA or protein expression of a gene in motile cells of interest from an animal tissue. Additionally, methods of determining whether a cancer in a tissue of a mammal is likely to metastasize are provided. Methods are also provided for inhibiting metastasis of a cancer in a tissue of a mammal. Further provided are methods of determining resistance of a motile cancer cell population in an animal tissue to a chemotherapeutic agent.
1. A method of determining whether a cancer in a tissue of a mammal is likely to metastasize, the method comprising
obtaining a microneedle or capillary filled with a porous matrix comprising a chemotactic factor;
inserting the microneedle or capillary into the tissue for a time sufficient for motile cells to migrate into the porous matrix;
expelling the porous matrix with motile cells from the microneedle or capillary;
combining the porous matrix with microbeads, where the microbeads comprise a binding partner to a surface marker present on macrophages from the tissue;
removing the microbeads and bound macrophages; and
obtaining a gene expression profile of motile tumor cells remaining after removal of the microbeads,
wherein the gene that is profiled includes one or more of collagen type III α1, G-protein coupled receptor 26, Zip code binding protein 1, fibroblast growth factor receptor 1, Arp2/3 p16 subunit, tight junction protein 2, member Ras oncogene family, and epidermal growth factor receptor, and
wherein downregulation of expression of one or more of collagen type III α1, G-protein coupled receptor 26, Zip code binding protein 1, or fibroblast growth factor receptor 1 compared to expression in non-metastatic tumor cells, and/or upregulation of expression of one or more of Arp2/3 p16 subunit, tight junction protein 2, member Ras oncogene family or epidermal growth factor receptor compared to expression in non-metastatic tumor cells indicates that the profiled tumor cells are metastatic tumor cells and that the cancer is likely to metastasize.
2. The method of claim 1 , wherein the tissue is mammary tissue.
3. The method of claim 1 , wherein the porous matrix comprises matrigel.
4. The method of claim 1 , wherein the microneedle or capillary is a microneedle.
5. The method of claim 1 , wherein the binding partner is an antibody.
6. The method of claim 1 , wherein the chemotactic factor is an epidermal growth factor.
7. The method of claim 1 , wherein the binding partner is an antibody is specific for CD11b.
8. The method of claim 1 , wherein the gene expression profile is determined using mRNA expression.
9. The method of claim 1 , wherein the gene expression profile is determined using protein expression.
10. The method of claim 1 , wherein the genes that are profiled include all of collagen type III α1, G-protein coupled receptor 26, Zip code binding protein 1, fibroblast growth factor receptor 1, Arp2/3 p16 subunit, tight junction protein 2, member Ras oncogene family, and epidermal growth factor receptor.
11. A method of identifying metastatic tumor cells from a tumor, the method comprising:
isolating motile tumor cells from the tumor;
obtaining a gene expression profile of the motile tumor cells, wherein the genes that are profiled include all of collagen type III α1, G-protein coupled receptor 26, Zip code binding protein 1, fibroblast growth factor receptor 1, Arp2/3 p16 subunit, tight junction protein 2, member Ras oncogene family, and epidermal growth factor receptor; and
comparing the expression of collagen type III α1, G-protein coupled receptor 26, Zip code binding protein 1, fibroblast growth factor receptor 1, Arp2/3 p16 subunit, tight junction protein 2, member Ras oncogene family and epidermal growth factor receptor in the profiled tumor cells to their expression in non-metastatic tumor cells, wherein downregulation of expression of one or more of collagen type III α1, G-protein coupled receptor 26, Zip code binding protein 1, or fibroblast growth factor receptor 1 compared to expression in non-metastatic tumor cells, and/or upregulation of expression of one or more of Arp2/3 p16 subunit, tight junction protein 2, member Ras oncogene family or epidermal growth factor receptor compared to expression in non-metastatic tumor cells indicates that the profiled tumor cells are metastatic tumor cells, thereby identifying said profiled tumor cells as metastatic tumor cells.
12. The method of claim 11 , wherein the metastatic tumor cells are breast tumor cells.
13. The method of claim 11 , wherein the gene expression profile is determined using mRNA expression.
14. The method of claim 11 , wherein the gene expression profile is determined using protein expression.