Immune infiltration is implicated in the development of acquired resistance to anti-angiogenic cancer therapy. We therefore investigated the correlation between neutrophil infiltration in metastasis of colorectal cancer (CRC) patients and survival after treatment with bevacizumab. Our study identifies CD177+ tumour neutrophil infiltration as an adverse prognostic factor for bevacizumab treatment. We further demonstrate that a novel anti-VEGF/anti-Ang2 compound (BI-880) can overcome resistance to VEGF inhibition in experimental tumour models.


A total of 85 metastatic CRC patients were stratified into cohorts that had either received chemotherapy alone (n = 39) or combined with bevacizumab (n = 46). Tumour CD177+ neutrophil infiltration was correlated to clinical outcome. The impact of neutrophil infiltration on anti-VEGF or anti-VEGF/anti-Ang2 therapy was studied in both xenograft and syngeneic tumour models by immunohistochemistry.


The survival of bevacizumab-treated CRC patients in the presence of CD177+ infiltrates was significantly reduced compared to patients harbouring CD177− metastases. BI-880 treatment reduced the development of hypoxia associated with bevacizumab treatment and improved vascular normalisation in xenografts. Furthermore, neutrophil depletion or BI-880 treatment restored treatment sensitivity in a syngeneic tumour model of anti-VEGF resistance.


Our findings implicate CD177 as a biomarker for bevacizumab and suggest VEGF/Ang2 inhibition as a strategy to overcome neutrophil associated resistance to anti-angiogenic treatment.

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We thank Ali Manav for great technical assistance.

Author contributions

Study design: L.M.S., H.K., O.C. providing of study materials or samples: L.M.S., H.K., O.C., F.G., G.D., M.B., H.A., H.G., A.Q., T.Z., F.H., C.J.B., J.M.S. data generation, analysis and interpretation: L.M.S., H.K., O.C., M.F., F.G., S.D.G., N.R.S., F.T., A.Q., H.G. manuscript writing: all authors.

Author information

Author notes

  1. These authors contributed equally: Hamid Kashkar, Oliver Coutelle.


  1. Cologne Excellence Cluster on Cellular Stress Responses in Aging-Associated Diseases (CECAD), Center for Molecular Medicine Cologne (CMMC) and Institute for Medical Microbiology, Immunology and Hygiene, University of Cologne, CECAD Research Center, Joseph-Stelzmann-Str. 26, 50931, Cologne, Germany

    • Lars Mortimer Schiffmann
    • , Melanie Fritsch
    • , Saskia Diana Günther
    • , Neil Richard Stair
    • , Jens Michael Seeger
    • , Hamid Kashkar
    •  & Oliver Coutelle
  2. Department of General, Visceral and Cancer Surgery, University of Cologne, Kerpener Str. 62, 50937, Cologne, Germany

    • Lars Mortimer Schiffmann
    • , Florian Gebauer
    • , Georg Dieplinger
    • , Marc Bludau
    • , Hakan Alakus
    •  & Christiane Josephine Bruns
  3. Center for Integrated Oncology (CIO) Cologne Bonn, Gastrointestinal Cancer Group Cologne (GCGC), Kerpener Str. 62, 50924, Cologne, Germany

    • Lars Mortimer Schiffmann
    • , Florian Gebauer
    • , Georg Dieplinger
    • , Marc Bludau
    • , Hakan Alakus
    • , Heike Göbel
    • , Alexander Quaas
    • , Thomas Zander
    • , Christiane Josephine Bruns
    •  & Hamid Kashkar
  4. Department of Gynaecology and Obstetrics, University of Cologne, Kerpener Str. 34, 50931, Cologne, Germany

    • Fabinshy Thangarajah
  5. Institute for Pathology, University of Cologne, Kerpener Str. 62, 50937, Cologne, Germany

    • Heike Göbel
    •  & Alexander Quaas
  6. Department I of Internal Medicine, University of Cologne, Kerpener Str. 62, 50924, Cologne, Germany

    • Thomas Zander
  7. Boehringer Ingelheim RCV, Doktor-Boehringer-Gasse 5-11, 1120, Vienna, Austria

    • Frank Hilberg


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H.K. and L.M.S. acknowledge funding from the Deutsche Forschungsgemeinschaft (CRU 286 and CRC1218) and the Deutsche Krebshilfe (70112113). This work was in part financially supported by a collaborative research agreement between the University of Cologne and Boehringer Ingelheim.

Ethical approval:

The study was performed in agreement with the local research ethics guidelines (ethics committee medical faculty, University of Cologne). Human samples were collected at the University Hospital of Cologne with written informed consent from patients with colorectal cancer concurring with the Declaration of Helsinki. All animal experiments were performed in accordance with the German animal protection law as approved by local government authorities.

Competing interests:

The authors declare no competing interests.


This work is published under the standard license to publish agreement. After 12 months the work will become freely available and the license terms will switch to a Creative Commons Attribution 4.0 International (CC BY 4.0).

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Datasets generated and analysed for this work are not publicly available and but are accessible upon request.

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Correspondence to Lars Mortimer Schiffmann.

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