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Apoptosis

Monoclonal antibodies against ROR1 induce apoptosis of chronic lymphocytic leukemia (CLL) cells

Leukemia volume 26, pages 1348–1355 (2012)Cite this article

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Abstract

ROR1 is a receptor tyrosine kinase (RTK) recently identified to be overexpressed at the gene and protein levels in chronic lymphocytic leukemia (CLL). Monoclonal antibodies (MAbs) against RTKs have been successfully applied for therapy of solid tumors. We generated five MAbs against the Ig (n=1), cysteine-rich (CRD) (n=2) and kringle (KNG) (n=2) domains, respectively, of the extracellular part of ROR1. All CLL patients (n=20) expressed ROR1 on the surface of the leukemic cells. A significantly higher frequency of ROR1 expression was found in patients with progressive versus non-progressive disease, and in those with unmutated versus mutated IgVH genes. All five MAbs alone induced apoptosis in the absence of complement or added effector cells (Annexin-V and MTT, as well as cleavage of poly-(ADP ribose)-polymerase, caspase-8 and caspase-9) of CLL cells but not of normal B cells. Most effective were MAbs against CRD and KNG, significantly superior to rituximab (P<0.005). Cross-linking of anti-ROR1 MAbs using the F(ab′)2 fragments of anti-Fc antibodies significantly augmented apoptosis. Two of the MAbs induced complement-dependent cytotoxicity (CDC) similar to that of rituximab and one anti-ROR1 MAb (KNG) (IgG1) showed killing activity by antibody-dependent cellular cytotoxicity. The identified ROR1 epitopes may provide a basis for generating human ROR1 MAbs for therapy.

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Acknowledgements

This study was supported by grants from the CLL Global Research Foundation, the EUCAAD 200755 project funded under the auspices of the EU Seventh Framework Programme, the Cancer and Allergy Foundation, the Swedish Research Council/SIDA/SAREC, the Iranian Ministry of Health and Medical Education, the Swedish Cancer Society, the Cancer Society in Stockholm, the King Gustaf Vth Jubilee Fund, Vinnova, the Karolinska Institutet Foundations and the Stockholm County Council.

Author contributions

AHDM and MHF performed the experiments, analyzed data and wrote the paper. ASK performed experiments and reviewed the manuscript. MMA, AAB, RG, ARM and RH produced MAbs. MJH and FS designed and produced MAbs and reviewed the manuscript. AÖ provided clinical material, analyzed data and wrote the paper. HR designed the study, performed experiments, analyzed data and wrote the paper. HM designed and supervised the study, provided clinical material, analyzed data and wrote the paper.

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Author notes

  1. A H Daneshmanesh and M Hojjat-Farsangi: These authors contributed equally to this work.

Authors and Affiliations

  1. Immune and Gene Therapy Lab, CCK, Karolinska University Hospital Solna, Karolinska Institutet, Stockholm, Sweden

    A H Daneshmanesh, M Hojjat-Farsangi, A S Khan, M Jeddi-Tehrani, A Österborg, H Rabbani & H Mellstedt

  2. Monoclonal Antibody Research Center, Avicenna Research Institute, ACECR, Tehran, Iran

    M Jeddi-Tehrani, A A Bayat, R Ghods, A-R Mahmoudi, R Hadavi, F Shokri & H Rabbani

  3. Reproductive Biology, Biotechnology and Infertility Research Center, Avicenna Research Institute, ACECR, Tehran, Iran

    M Jeddi-Tehrani & M M Akhondi

  4. Departments of Oncology (Radiumhemmet) and Hematology, Karolinska University Hospital Solna, Stockholm, Sweden

    A Österborg & H Mellstedt

  5. Department of Immunology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran

    F Shokri

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  1. A H Daneshmanesh
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Correspondence to H Rabbani.

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Daneshmanesh, A., Hojjat-Farsangi, M., Khan, A. et al. Monoclonal antibodies against ROR1 induce apoptosis of chronic lymphocytic leukemia (CLL) cells. Leukemia 26, 1348–1355 (2012). https://doi.org/10.1038/leu.2011.362

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