Dynamic Single-Cell Network Profiles in Acute Myelogenous Leukemia Are Associated with Patient Response to Standard Induction Therapy

Abstract

Purpose Complete response to induction chemotherapy is observed in ~60% of patients with newly diagnosed non-M3 acute myelogenous leukemia (AML). However, no methods exist to predict with high accuracy at the individual patient level the response to standard AML induction therapy. Experimental Design We applied single-cell network profiling (SCNP) using flow cytometry, a tool that allows a comprehensive functional assessment of intracellular signaling pathways in heterogeneous tissues, to two training cohorts of AML samples (n = 34 and 88) to predict the likelihood of response to induction chemotherapy. Results In the first study, univariate analysis identified multiple signaling “nodes” (readouts of modulated intracellular signaling proteins) that correlated with response (i.e., AUCROC ≥ 0.66; P ≤ 0.05) at a level greater than age. After accounting for age, similar findings were observed in the second study. For patients <60 years old, complete response was associated with the presence of intact apoptotic pathways. In patients ≥60 years old, nonresponse was associated with FLT3 ligand–mediated increase in phosphorylated Akt and phosphorylated extracellular signal-regulated kinase. Results were independent of cytogenetics, FLT3 mutational status, and diagnosis of secondary AML. Conclusions These data emphasize the value of performing quantitative SCNP under modulated conditions as a basis for the development of tests highly predictive for response to induction chemotherapy. SCNP provides information distinct from other known prognostic factors such as age, secondary AML, cytogenetics, and molecular alterations and is potentially combinable with the latter to improve clinical decision making. Independent validation studies are warranted.

FACS analysis of Stat3/5 signaling reveals sensitivity to G-CSF and IL-6 as a significant prognostic factor in pediatric AML: a Children's Oncology Group report

“…Recent phospho-flow studies using adult AML samples have found an association between increased basal and induced pY-Stat3 and decreased likelihood of achieving remission. 13,15 We did not find an association between the magnitude of basal or induced pStats and clinical outcome. Our data instead revealed a significant association between the sensitivity of the ligand response and outcome, with increased sensitivity being significantly associated with better survival.…”

“…11,12 More recently, we and others have measured phosphorylated Stat proteins in AML samples by intracellular flow cytometry (phospho-flow). [13][14][15][16][17] This highly sensitive method yields quantitative information at the single-cell level.…”

Multiplexed mAbs: a new strategy in preclinical time-domain imaging of acute myeloid leukemia

“…26,27 They are attractive agents for oncologic imaging, 28,29 but several difficulties must be overcome to achieve the necessary specificity and avidity for in vivo contrast. 30 The number of target antigens per cell is a defining factor in the successful application of mAbs for imaging applications.…”

Association between B-cell receptor responsiveness and disease progression in B-cell chronic lymphocytic leukemia: results from single cell network profiling studies

“…[14][15][16] The association of SCNP data with in vivo acute myeloid leukemia (AML) chemosensitivity/chemoresistance 17 as well as FLT3R signaling deregulation 18 and applications in immunology 19,20 have been previously reported. Here the SCNP assay was first applied in an exploratory study to functionally characterize elements of the BCR signaling network and to assess their association with clinical and prognostic parameters with the goal of ultimately developing a clinically useful tool for the prediction of B-CLL progression and early (<3 years) TTFT in patients with early stage B-CLL.…”