Cell culture model for acquired chemoresistance of chronic myelogenous leukemia and related methods for identifying agents to overcome resistance
A method of generating a chronic myelogenic leukemia (CML) acquired chemoresistant culture model is provided. Such a method may comprise providing a naïve blast crisis CML cell line; administering/contacting the cell line with a mutation-inducing dose of imatinib; maintaining a culture of the treated cell line for a period of time until the treated cell line relapses and repopulates the culture; and determining the repopulated cell culture is a CML acquired chemoresistant cell line by detecting a BCR-ABL mutation, wherein the acquired chemoresistance is achieved by a BCR-ABL mutation.
1. A culture model of chronic myelogenic leukemia (CML) acquired chemoresistant cells, the model generated by:
contacting a blast crisis CML cell line, which has previously not been exposed to imatinab before, with a dose of imatinib sufficient to induce a BCR-ABL mutation, wherein the blast crisis CML cell line is a KCL-22 cell line;
maintaining a culture of the imatinib-treated cell line for about two to three weeks until the cell line relapses and repopulates the culture; and
determining that the repopulated cell culture is a CML acquired chemoresistant cell line by detecting a BCR-ABL mutation.
2. The model of claim 1 , wherein the mutation-inducing dose of imatinib is about 2.5 μM or higher.
3. The model of claim 1 , wherein the mutation-inducing dose of imatinib is between about 2.5 μM and about 10 μM.
4. The model of claim 1 , wherein the mutation-inducing dose of imatinib is selected from about 2.5 μM, about 5 μM and about 10 μM.
5. The model of claim 1 , wherein the BCR-ABL mutation is a BCR-ABL kinase domain mutation.
6. The model of claim 5 , wherein the BCR-ABL kinase domain mutation is a T315I BCR-ABL mutation.
7. The model of claim 6 , wherein the BCR-ABL kinase domain mutation is a E255K or a Y253H BCR-ABL mutation or a combination thereof.
8. The model of claim 1 , wherein the BCR-ABL kinase domain mutation is a E255K or a Y253H BCR-ABL mutation or a combination thereof.
9. The model of claim 1 , wherein the BCR-ABL mutation is stable.
10. The model of claim 1 , wherein the mutation-inducing dose of imatinib is a single dose.
11. A culture model of chronic myelogenic leukemia (CML) acquired chemoresistant cells, the model generated by:
contacting a blast crisis CML cell line, which has previously not been exposed to imatinab before, with a single dose of imatinib sufficient to induce a BCR-ABL mutation, wherein the blast crisis CML cell line is a KCL-22 cell line;
maintaining a culture of the imatinib-treated cell line for a period of time until the cell line relapses and repopulates the culture; and
determining that the repopulated cell culture is a CML acquired chemoresistant cell line by detecting a BCR-ABL mutation.
12. The model of claim 11 , wherein the mutation-inducing dose of imatinib is between about 2.5 μM and about 10 μM.
13. The model of claim 11 , wherein the BCR-ABL mutation is a BCR-ABL kinase domain mutation.
14. The model of claim 13 , wherein the BCR-ABL kinase domain mutation is a T315I BCR-ABL mutation.
15. The model of claim 14 , wherein the BCR-ABL kinase domain mutation is a E255K or a Y253H BCR-ABL mutation or a combination thereof.
16. The model of claim 11 , wherein the BCR-ABL kinase domain mutation is a E255K or a Y253H BCR-ABL mutation or a combination thereof.
17. The model of claim 11 , wherein the BCR-ABL mutation is stable.
18. The model of claim 11 , wherein the period of time is about two to three weeks.