title: AKT signaling as a novel factor associated with in vitro resistance of human AML to gemtuzumab ozogamicin.
abstract: Gemtuzumab ozogamicin (GO), an immunoconjugate between an anti-CD33 antibody and a calicheamicin-γ(1) derivative, induces remissions and improves survival in a subset of patients with acute myeloid leukemia (AML). As the mechanisms underlying GO and calicheamicin-γ(1) resistance are incompletely understood, we herein used flow cytometry-based single cell network profiling (SCNP) assays to study cellular responses of primary human AML cells to GO. Our data indicate that the extent of DNA damage is quantitatively impacted by CD33 expression and drug efflux activity. However, although DNA damage is required for GO-induced cytotoxicity, it is not sufficient for effective cell kill, suggesting that downstream anti-apoptotic pathways may function as relevant resistance mechanisms. Supporting this notion, we found activated PI3K/AKT signaling to be associated with GO resistance in vitro in primary AML cells. Consistently, the investigational AKT inhibitor MK-2206 significantly sensitized various human AML cells to GO or free calicheamicin-γ(1) with particularly pronounced effects in otherwise GO or free calicheamicin-γ(1)-resistant cells. Likewise, MK-2206 also sensitized primary AML cells to calicheamicin-γ(1). Together, our findings illustrate the capacity of SCNP assays to discover chemotherapy-related biological pathways and signaling networks relevant to GO-induced genotoxic stress. The identification of AKT signaling as being associated with GO resistance in vitro may provide a novel approach to improve the in vivo efficacy of GO/calicheamicin-γ(1) and, by extrapolation, other DNA damage-based therapeutics.
authors: Nodality Inc., South San Francisco, California, United States of America.
authors: David B
authors: DB
authors: Rosen
authors: Kimberly H
authors: KH
authors: Harrington
authors: James A
authors: JA
authors: Cordeiro
authors: Ling Y
authors: LY
authors: Leung
authors: Santosh
authors: S
authors: Putta
authors: Norman
authors: N
authors: Lacayo
authors: George S
authors: GS
authors: Laszlo
authors: Chelsea J
authors: CJ
authors: Gudgeon
authors: Donna E
authors: DE
authors: Hogge
authors: Rachael E
authors: RE
authors: Hawtin
authors: Alessandra
authors: A
authors: Cesano
authors: Roland B
authors: RB
authors: Walter
mesh_terms: D000617:Aminoglycosides / Q000494:pharmacology* / Q000627:therapeutic use; D061067:Antibodies, Monoclonal, Humanized / Q000494:pharmacology* / Q000627:therapeutic use; D000970:Antineoplastic Agents / Q000494:pharmacology*; D045744:Cell Line, Tumor; D004249:DNA Damage; D019008:Drug Resistance, Neoplasm; D053281:Enediynes / Q000494:pharmacology; D000079982:Gemtuzumab; D006575:Heterocyclic Compounds, 3-Ring / Q000494:pharmacology; D006801:Humans; D015470:Leukemia, Myeloid, Acute / Q000378:metabolism* / Q000628:therapy*; D019869:Phosphatidylinositol 3-Kinases / Q000378:metabolism; D047428:Protein Kinase Inhibitors / Q000494:pharmacology; D051057:Proto-Oncogene Proteins c-akt / Q000037:antagonists & inhibitors / Q000502:physiology*; D063268:Sialic Acid Binding Ig-like Lectin 3 / Q000378:metabolism; D015398:Signal Transduction / Q000187:drug effects / Q000502:physiology; D059010:Single-Cell Analysis; D014407:Tumor Cells, Cultured
chemical_list: D000617:Aminoglycosides; D061067:Antibodies, Monoclonal, Humanized; D000970:Antineoplastic Agents; D053281:Enediynes; D000079982:Gemtuzumab; D006575:Heterocyclic Compounds, 3-Ring; D019869:Phosphatidylinositol 3-Kinases; D047428:Protein Kinase Inhibitors; D051057:Proto-Oncogene Proteins c-akt; D063268:Sialic Acid Binding Ig-like Lectin 3; C571163:CD33 protein, human; C548887:MK 2206; C055955:calicheamicin gamma(1)I