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Applied Microbiology and Biotechnology

, Volume 100, Issue 1, pp 263–277 | Cite as

Targeting c-kit receptor in neuroblastomas and colorectal cancers using stem cell factor (SCF)-based recombinant bacterial toxins

  • Swati Choudhary
  • Alessa Pardo
  • Reinhard Rosinke
  • Janendra K. Batra
  • Stefan Barth
  • Rama S. Verma
Applied genetics and molecular biotechnology

Abstract

Autocrine activation of c-kit (KIT receptor tyrosine kinase) has been postulated to be a potent oncogenic driver in small cell lung cancer, neuroblastoma (NB), and poorly differentiated colorectal carcinoma (CRC). Although targeted therapy involving tyrosine kinase inhibitors (TKIs) such as imatinib mesylate is highly effective for gastrointestinal stromal tumor carrying V560G c-kit mutation, it does not show much potential for targeting wild-type KIT (WT-KIT). Our study demonstrates the role of stem cell factor (SCF)-based toxin conjugates for targeting WT-KIT-overexpressing malignancies such as NBs and CRCs. We constructed SCF-based recombinant bacterial toxins by genetically fusing mutated form of natural ligand SCF to receptor binding deficient forms of Diphtheria toxin (DT) or Pseudomonas exotoxin A (ETA') and evaluated their efficacy in vitro. Efficient targeting was achieved in all receptor-positive neuroblastoma (IMR-32 and SHSY5Y) and colon cancer cell lines (COLO 320DM, HCT 116, and DLD-1) but not in receptor-negative breast carcinoma cell line (MCF-7) thereby proving specificity. While dose- and time-dependent cytotoxicity was observed in both neuroblastoma cell lines, COLO 320DM and HCT 116 cells, only an anti-proliferative effect was observed in DLD-1 cells. We prove that these novel targeting agents have promising potential as KIT receptor tyrosine kinase targeting system.

Keywords

Protein-fusion toxins Stem cell factor c-kit receptor Targeting 

Notes

Acknowledgments

We are grateful to Aslam Basha for helping with fermentation, DAAD and MHRD for providing fellowships to SC, and the Department of Biotechnology (DBT), Government of India for providing grant (BT/PR13463/PID/06/488/2010) to support this work.

Conflict of interest

The authors declare that they have no competing interests.

Ethical Statement

This article does not contain any studies with human participants or animals performed by any of the authors.

Supplementary material

253_2015_6978_MOESM1_ESM.pdf (1.4 mb)
ESM 1 (PDF 1,462 kb)

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Copyright information

© Springer-Verlag Berlin Heidelberg 2015

Authors and Affiliations

  • Swati Choudhary
    • 1
  • Alessa Pardo
    • 2
  • Reinhard Rosinke
    • 3
  • Janendra K. Batra
    • 4
  • Stefan Barth
    • 2
    • 3
    • 5
  • Rama S. Verma
    • 1
  1. 1.Department of Biotechnology, Bhupat and Jyoti Mehta School of BiosciencesIndian Institute of Technology MadrasChennaiIndia
  2. 2.Department of Experimental Medicine and Immunotherapy, Institute of Applied Medical EngineeringUniversity Hospital RWTH AachenAachenGermany
  3. 3.Department of Pharmaceutical Product DevelopmentFraunhofer Institute for Molecular Biology and Applied Ecology IMEAachenGermany
  4. 4.National Institute of ImmunologyNew DelhiIndia
  5. 5.UCT Faculty of Health SciencesSouth African Research Chair in Cancer BiotechnologyCape TownSouth Africa

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