American Association for Cancer Research
21598290cd150298-sup-146420_2_supp_3021087_nq13c5.pdf (4.52 MB)

Supplementary Figures 1 - 8 from Mass Cytometric Functional Profiling of Acute Myeloid Leukemia Defines Cell-Cycle and Immunophenotypic Properties That Correlate with Known Responses to Therapy

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journal contribution
posted on 2023-04-03, 21:03 authored by Gregory K. Behbehani, Nikolay Samusik, Zach B. Bjornson, Wendy J. Fantl, Bruno C. Medeiros, Garry P. Nolan

Supplementary Figure 1. SPADE plots of normal bone marrow sample no. 6. Supplementary Figure 2. The frequency of cells in the indicated (manually-gated) populations. Supplementary Figure 3. Multi-dimensional binning analysis of the CD34+CD38 low subset confirmed that distinct karyotype and genotype-specific immunophenotypic patterns are observed in high-dimensional space. Supplementary Figure 4. AML subtypes are characterized by distinct phosphoprotein signaling patterns. Supplementary Figure 5. viSNE analysis of CD34+CD38 low subset demonstrates signaling aberrancy is most pronounced in cells outside the viSNE space occupied by normal cells. Supplementary Figure 6. Median basal intracellular signaling of CD34+CD38low cells from AML samples of each subtype. Supplementary Figure 7. HU treatment does not increase the fraction of apoptotic cells measured by cPARP levels. Supplementary Figure 8. Histone 3 lysine 9 acetylation decreases with immunophenotypic differentiation in both normal and AML patient samples.



Acute myeloid leukemia (AML) is characterized by a high relapse rate that has been attributed to the quiescence of leukemia stem cells (LSC), which renders them resistant to chemotherapy. However, this hypothesis is largely supported by indirect evidence and fails to explain the large differences in relapse rates across AML subtypes. To address this, bone marrow aspirates from 41 AML patients and five healthy donors were analyzed by high-dimensional mass cytometry. All patients displayed immunophenotypic and intracellular signaling abnormalities within CD34+CD38lo populations, and several karyotype- and genotype-specific surface marker patterns were identified. The immunophenotypic stem and early progenitor cell populations from patients with clinically favorable core-binding factor AML demonstrated a 5-fold higher fraction of cells in S-phase compared with other AML samples. Conversely, LSCs in less clinically favorable FLT3-ITD AML exhibited dramatic reductions in S-phase fraction. Mass cytometry also allowed direct observation of the in vivo effects of cytotoxic chemotherapy.Significance: The mechanisms underlying differences in relapse rates across AML subtypes are poorly understood. This study suggests that known chemotherapy sensitivities of common AML subsets are mediated by cell-cycle differences among LSCs and provides a basis for using in vivo functional characterization of AML cells to inform therapy selection. Cancer Discov; 5(9); 988–1003. ©2015 AACR.See related commentary by Do and Byrd, p. 912.This article is highlighted in the In This Issue feature, p. 893