Cell Type-Specific Effects of Crizotinib in Human Acute Myeloid Leukemia with TP53 Alterations

Abstract

Introduction: In acute myeloid leukemia (AML) standard therapies induce complete remission in 50-70% of patients, but overall two-year survival is less than 20-30% because of high relapse rates. AML with TP53 alteration is largely insensitive to chemotherapy, modern targeted agents, and hematopoietic stem cell transplantation. Mutations in TP53 are seen in approximately 20% of AML and confer a particularly poor prognosis decreasing the 1-2 year survival rates to 0-10%. Leukemia stem cells (LSCs) are defined as the cells that recapitulate the leukemia and cause relapse. Recent large-scale drug-screening efforts to map treatment vulnerabilities in AML have revealed that crizotinib can reduce the in vitro viability of unselected primary human AML bone marrow samples with TP53 alterations (Tyner et al. Nature 2018). Crizotinib is a tyrosine kinase inhibitor that targets ALK, MET, and RON and is FDA approved for treatment of lung cancer. Since LSCs are largely responsible for AML mortality, we investigated whether crizotinib targets LSCs in primary, diagnostic human AML samples with TP53 alterations. We used mass cytometry (CyTOF2) to determine the cell-type specific effects of this drug on intracellular signaling states. We profiled treated samples with a panel that includes immunophenotypic markers (including LSC markers) and intracellular signaling molecules implicated in AML pathogenesis and aggressiveness. Results: Primary human AML samples with TP53 alterations were treated with crizotinib (10 mM, n=3). There was a reduction of viability by 48 hours of in vitro treatment with crizotinib, relative to vehicle (mean 64% reduction in viability). Crizotinib abolished all colony formation in semi-solid media. Mass cytometry analysis revealed that crizotinib treatment led to a loss of cells bearing LSC-specific markers such as CD123 and TIM3. Crizotinib treatment also induced a consistent loss of phosphorylated-signaling intermediates of the STAT and p38/MAPAPKII pathways in all samples tested. Notably, crizotinib treatment also induced significant reductions in total and phosphorylated NFKB in all samples tested. Conclusions: These data reveal cell-type specific effects of crizotinib in human AML with TP53 alterations. Crizotinib induced a specific loss of LSCs and inhibition of the STAT, p38/MAPAPKII and NFKB pathways. These data suggest that crizotinib inhibition may target LSCs in AML with TP53 alterations. Figure Disclosures No relevant conflicts of interest to declare.

Explainable artificial intelligence for precision medicine in acute myeloid leukemia

“…It has not been enrolled in clinical trials for AML. Nevertheless, it has been used in studies of high-risk AML patients, with TP53 Mut and obtained very promising results ( 34 ).…”

“…It also includes patients who have TP53 Mut genotype, which are biomarkers associated with poor prognosis ( 14 ). MOM recommends treating these patients with Crizotinib, a drug used in other studies with TP53 Mut AML patients which in fact showed very promising results ( 34 ). In addition, this subgroup shows a deficiency in amino acid metabolism which may lead to alternative treatment therapies based on metabolomics.…”

Interpretable precision medicine for acute myeloid leukemia

“…It also includes patients who have TP53 Mut genotype, which are biomarkers associated with poor prognosis [134]. MOM recommends treating these patients with Crizotinib, a drug used in other studies with TP53 Mut AML patients which in fact showed very promising results [107]. In addition, this subgroup shows a deficiency in amino acid metabolism which may lead to alternative treatment therapies based on metabolomics.…”

“…It has not been enrolled in clinical trials for AML. Nevertheless, it has been used in studies of high-risk AML patients, with TP53 Mut and obtained very promising results [107].…”