SIRPα-antibody fusion proteins stimulate phagocytosis and promote elimination of acute myeloid leukemia cells

Oncotarget. 2017 Feb 14;8(7):11284-11301. doi: 10.18632/oncotarget.14500.

Abstract

CD47, expressed on a variety of tumor cells, confers immune resistance by delivering an inhibitory "don't eat me" signal to phagocytic cells via its myeloid-specific receptor SIRPα. Recent studies have shown that blocking the CD47-SIRPα axis with CD47-directed antibodies or antibody-derivatives enhances phagocytosis and increases antitumor immune effects. However, CD47 expression on healthy cells creates an antigen sink and potential sites of toxicity, limiting the efficacy of CD47-directed therapies. In this study, we first characterized CD47 expression in Acute Myeloid Leukemia (AML) patients (n = 213) and found that CD47 is highly expressed on both AML bulk and stem cells irrespective of the disease state. Furthermore, to inhibit the CD47-SIRPα signaling pathway at the tumor site, we developed a so-called local inhibitory checkpoint monoclonal antibody (licMAB) by grafting the endogenous SIRPα domain to the N-terminus of the light chain of an antibody targeting CD33, a surface antigen expressed in AML. LicMABs selectively bind CD33-expressing cells even in the presence of a large CD33-negative CD47-positive antigen sink, stimulate phagocytosis of AML cells and eliminate AML cell lines and primary, patient-derived AML cells. Our findings qualify licMABs as a promising therapeutic approach to confine the benefit of disrupting the CD47-SIRPα axis to tumor antigen-expressing cells.

Keywords: CD47; SIRPα; acute myeloid leukemia; immunotherapy; therapeutic antibody.

MeSH terms

  • Antibodies, Blocking / pharmacology*
  • Antibodies, Monoclonal / pharmacology
  • Antibody-Dependent Cell Cytotoxicity / immunology
  • Antigens, Differentiation / pharmacology*
  • CD47 Antigen / biosynthesis
  • CD47 Antigen / immunology
  • Cell Separation
  • Flow Cytometry
  • Humans
  • Immunotherapy / methods*
  • Leukemia, Myeloid, Acute / immunology*
  • Microscopy, Confocal
  • Phagocytosis / drug effects*
  • Receptors, Immunologic
  • Sialic Acid Binding Ig-like Lectin 3 / immunology*

Substances

  • Antibodies, Blocking
  • Antibodies, Monoclonal
  • Antigens, Differentiation
  • CD33 protein, human
  • CD47 Antigen
  • CD47 protein, human
  • Receptors, Immunologic
  • SIRPA protein, human
  • Sialic Acid Binding Ig-like Lectin 3