Targeting malignant B cells with an immunotoxin against ROR1

MAbs. May-Jun 2012;4(3):349-61. doi: 10.4161/mabs.19870. Epub 2012 Apr 26.

Abstract

The selective cell surface expression of receptor tyrosine kinase-like orphan receptor 1 (ROR1) in chronic lymphocytic leukemia (CLL) and mantle cell lymphoma (MCL) has made ROR1 a novel and promising target for therapeutic monoclonal antibodies (mAbs). Four mouse mAbs generated by hybridoma technology exhibited specific binding to human ROR1. Epitope mapping studies showed that two mAbs (2A2 and 2D11) recognized N-terminal epitopes in the extracellular region of ROR1 and the other two (1A1 and 1A7) recognized C-terminal epitopes. A ROR1- immunotoxin (BT-1) consisting of truncated Pseudomonas exotoxin A (PE38) and the VH and VL fragments of 2A2-IgG was made recombinantly. Both 2A2-IgG and BT-1 showed dose-dependent and selective binding to primary CLL and MCL cells and MCL cell lines. Kinetic analyses revealed 0.12-nM (2A2-IgG) to 65-nM (BT-1) avidity/affinity to hROR1, depicting bivalent and monovalent interactions, respectively. After binding to cell surface ROR1, 2A2-IgG and BT-1 were partially internalized by primary CLL cells and MCL cell lines, and BT-1 induced profound apoptosis of ROR1-expressing MCL cell lines in vitro (EC 50 = 16 pM-16 nM), but did not affect ROR1-negative cell lines. Our data suggest that ROR1-immunotoxins such as BT-1 could serve as targeted therapeutic agents for ROR1-expressing B cell malignancies and other cancers.

Publication types

  • Research Support, N.I.H., Intramural

MeSH terms

  • ADP Ribose Transferases / therapeutic use*
  • Animals
  • Apoptosis
  • Bacterial Toxins / therapeutic use*
  • Biomarkers, Tumor / metabolism
  • Cell Line, Tumor
  • Exotoxins / therapeutic use*
  • Humans
  • Immunoglobulin Variable Region / immunology
  • Immunotoxins / therapeutic use*
  • Leukemia, Lymphocytic, Chronic, B-Cell / therapy*
  • Lymphoma, Mantle-Cell / therapy*
  • Mice
  • Molecular Targeted Therapy / methods*
  • Receptor Tyrosine Kinase-like Orphan Receptors / antagonists & inhibitors*
  • Receptor Tyrosine Kinase-like Orphan Receptors / immunology
  • Recombinant Fusion Proteins / therapeutic use
  • Virulence Factors / therapeutic use*

Substances

  • Bacterial Toxins
  • Biomarkers, Tumor
  • Exotoxins
  • Immunoglobulin Variable Region
  • Immunotoxins
  • Recombinant Fusion Proteins
  • Virulence Factors
  • ADP Ribose Transferases
  • toxA protein, Pseudomonas aeruginosa
  • ROR1 protein, human
  • Receptor Tyrosine Kinase-like Orphan Receptors