Honors College Thesis
 

Role of MEK Mutations in Trametinib Resistant Acute Myeloid Leukemia Cells

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https://ir.library.oregonstate.edu/concern/honors_college_theses/cf95jk383

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  • Acute myeloid leukemia (AML) is a blood malignancy characterized by a differentiation and proliferation defect of myeloid progenitors leading to accumulation of myeloid blasts. It is one of the most common leukemias in adults. Trametinib (Mekinist) is a small molecule inhibitor that targets MEK1 (MAP2K1) and MEK2 (MAPK2K2). It has been approved by the Food and Drug Administration (FDA) to treat advanced melanoma. The benefit of trametinib is limited by the development of resistance. In AML, a combination of trametinib with tyrosine kinase inhibitors (TKIs) has been shown to improve the response in AML cell lines. Moreover, a phase 1/2 study demonstrated trametinib’s benefit in RAS-mutated myeloid malignancies including AML2. This work aims to study trametinib resistance mechanisms in AML. MOLM13, MV4;11, and OCI-AML2 AML cell lines were made resistant to trametinib. Exome sequencing as well as Sanger sequencing revealed MEK1K57N and MEK1L215P mutations in these resistant cells. Therefore, site directed mutagenesis as well as gateway cloning were used to generate MEK1K57N and MEK1L215P open reading frame (ORF) harboring lentivirus vectors. Then, MOLM13 AML cells were transduced with a lentivirus containing wild-type and mutant MEK1 to establish AML cell lines expressing wild type or mutant MEK1. A drug sensitivity assay was used to analyze trametinib sensitivity in these cells. Moreover, western blot analysis was used to study alteration of signaling pathways in acquired trametinib resistant AML cell lines (MOLM13 and OCI-AML2). The drug sensitivity assay demonstrated that MOLM13 cells harboring MEK1K57N has lower sensitivity to trametinib in comparison to MOLM13 cells transduced with wild type MEK1. This indicated that MEK1K57N confers resistance to trametinib in AML cells. The finding confirmed the hypothesis that MEK1K57N plays a role in the drug resistance of trametinib in AML cells. This information can be used to further investigate ways to possible circumvent this mutation and find ways to use trametinib to treat AML. This can potentially be done through using combination therapy, using this information as a treatment selection tool, and finding ways to block the mutated protein from activating.
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