Chromosomal translocation t(9;22) is responsible for appearance of an oncogene encoding BCR/ABL fusion tyrosine kinase, which induce chronic myelogenous leukemia (CML) and a cohort of acute lymphocytic leukemia (ALL). CML usually starts as a relatively benign chronic phase (CML-CP), which progresses to an aggressive disease - blast crisis (CML-BC). Malignant transformation of the disease is associated with accumulation of additional genetic errors. Imatinib mesylate (IM), a small molecule inhibitor of BCR/ABL kinase, revolutionized the treatment of CML-CP. Unfortunately patients may develop resistance to the drug caused by point mutations encoding amino acid substitutions in the BCR/ABL kinase domain. In conclusion, CML cells display genomic instability leading to resistance to IM and malignant progression of the disease. This proposal is focused on determination the mechanisms responsible for these phenomena and subsequently on prevention/inhibition of the development of IM resistance and CML-BC. BCR/ABL kinase stimulates numerous signaling pathways to induce and maintain transformation of hematopoietic cells. We, and others found that phosphatidylinositol-3 kinase (PI-3k) play an essential role in growth factor independent proliferation and protection from apoptosis in CML. Here we propose to study the role of PI-3k and its downstream effectors Akt and Rac in genomic instability in CML stem and progenitor cell populations. Using genetic approach (dominant-negative mutants, siRNA, antisense cDNA, knockout mice) and small molecule inhibitors (for example perifosine, NSC23766) we will determine how PI-3k and its downstream effectors generate the reactive oxygen species (ROS), which in turn may cause oxidative DNA damage and facilitate genomic instability. Long-term in vitro culture and mouse models of CML will be applied here. IM resistance will be detected in clonogenic assays and by sequencing BCR/ABL kinase domain, and chromosomal aberrations will be detected by SNPs and SKY. If successfully accomplished, these experiments will determine if PI-3k pathway inhibitors should be used to improve therapeutic effect of IM and prevent/delay CML progression toward lethal blast crisis.

Public Health Relevance

Although tyrosine kinase inhibitors (TKIs) revolutionized the treatment of chronic myelogenous leukemia (CML), patients can develop resistance to the drug and progress toward fatal malignant blast crisis of the disease. Both phenomena are associated with genetic instability caused by oncogenic BCR/ABL kinase. We will investigate the mechanisms responsible for mutations in BCR/ABL kinase causing TKI-resistance and for chromosomal aberrations leading to blast crisis. Molecular mechanisms of BCR/ABL-dependent phosphatidylinositol (PI-3k)-mediated production of reactive oxygen species (ROS) causing oxidative DNA damage, mutations, and chromosomal aberrations will be determined.

National Institute of Health (NIH)
National Cancer Institute (NCI)
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Basic Mechanisms of Cancer Therapeutics Study Section (BMCT)
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Mufson, R Allan
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Temple University
Schools of Medicine
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Maifrede, Silvia; Nieborowska-Skorska, Margaret; Sullivan-Reed, Katherine et al. (2018) Tyrosine kinase inhibitor-induced defects in DNA repair sensitize FLT3(ITD)-positive leukemia cells to PARP1 inhibitors. Blood 132:67-77
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