Anti-progranulin/GP88 antibody AG01 inhibits triple negative breast cancer cell proliferation and migration

Abstract

Background

Triple negative breast cancer (TNBC) is characterized by invasiveness and short survival. Identifying novel TNBC-targeted therapies, to potentiate standard of care (SOC) therapy, is an unmet need. Progranulin (PGRN/GP88) is a biological driver of tumorigenesis, survival, and drug resistance in several cancers including breast cancer (BC). PGRN/GP88 tissue expression is an independent prognostic factor of recurrence while elevated serum PGRN/GP88 level is associated with poor outcomes. Since PGRN/GP88 expression is elevated in 30% TNBC, we investigated the involvement of progranulin on TNBC.

Methods

The effect of inhibiting PGRN/GP88 expression in TNBC cells by siRNA was investigated. The effects of a neutralizing anti-human PGRN/GP88 monoclonal antibody AG01 on the proliferation and migration of two TNBC cell lines expressing PGRN/GP88 were then examined in vitro and in vivo.

Results

Inhibition of GP88 expression by siRNA and AG01 treatment to block PGRN/GP88 action reduced proliferation and migration in a dose-dependent fashion in MDA-MB-231 and HS578-T cells. Western blot analysis showed decreased expression of phosphorylated protein kinases p-Src, p-AKT, and p-ERK upon AG01 treatment, as well as inhibition of tumor growth and Ki67 expression in vivo.

Conclusion

PGRN/GP88 represents a therapeutic target with companion diagnostics. Blocking PGRN/GP88 with antibody treatment may provide novel-targeted solutions in TNBC treatment which could eventually address the issue of toxicity and unresponsiveness associated with SOC.

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Data availability

The data used and/or analyzed during the current study are available from the corresponding author on reasonable request.

Abbreviations

AG01:

Anti-progranulin antibody

Akt:

Protein kinase B

BC:

Breast cancer

DAB:

3,3′-Diaminobenzidine

DMEM:

Dulbecco’s Modified Eagle Medium

ECL:

Enhanced chemiluminescence

E-Cadherin:

Epithelial cadherin

EDTA:

Ethylenediaminetetraacetic acid

ERK1/2:

Extracellular signal-regulated protein kinases 1 and 2

FAK:

Focal adhesion kinase

FBS:

Fetal bovine serum

GRN:

Granulin

H&E:

Hematoxylin eosin

HGFR/c-Met-:

Hepatocyte growth factor receptor

HIF-1α:

Hypoxia-inducible factors-1α

HO-1/HMOX1:

Heme oxygenase

HRP:

Horse radish peroxide

IACUC:

Institutional Animal Care and Use Committee

ICAM-1/CD54:

Intercellular Adhesion Molecule 1

IgG-Fc:

Immunoglobulin Fc region

MAPK:

Mitogen-activated protein kinase

MMP-2:

Metalloprotease-2

PBS:

Phosphate buffer saline

PGRN/GP88:

Progranulin, GP88

PVDF:

Polyvinylidene fluoride

R.T.:

Room temperature

SD:

Standard deviation

SEM:

Standard error of the mean

SDS:

Sodium dodecyl sulfate

SDS-PAGE:

Sodium dodecyl sulfate polyacrylamide gel electrophoresis

SiRNA:

Silencing RNA

SOC:

Standard of care

TBST:

Tris-buffered saline

TNBC:

Triple negative breast cancer

TBST:

Tris-buffered saline with Tween-20

TMB:

3,3′,5,5′-Tetramethylbenzidine

TNFR2:

Tumor necrosis factor receptor-2

V t/V o :

Tumor volume on Day t over tumor volume on Day 0

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Acknowledgements

We thank Dr. William S. Twaddell (School of Medicine, University of Maryland, Baltimore) for assisting in animal tumor section evaluation and Dr. Young Suk Lee (University of Maryland, Baltimore) for her review and suggestions on animal data. We also thank Drs. Rena Lapidus, Antonino Passaniti and Amy Fulton (School of Medicine, University of Maryland, Baltimore) for their suggestions and kind review of the manuscript. This work was supported by grant R44 CA 224718 from the National Cancer Institute to GS.

Funding

This work was supported by grant R44 CA 224718 from the National Cancer Institute to Ginette Serrero’s laboratory.

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Contributions

Ginette Serrero and Rupa Guha planned, performed, and analyzed the experimental results, perform data analysis and were primarily responsible for writing and revising the manuscript. Binbin Yue prepared the reagents and carried out the progranulin analysis by ELISA and assisted in performing the animal studies. Jianping Dong assisted in performing the animal studies and the analysis of the in vivo data. Aditi Baneerjee assisted in analyzing the Ki67 stained and HE stained tumor sections for mitotic index, microvessel count. All authors agreed to the submission of the manuscript and its revision.

Corresponding author

Correspondence to Ginette Serrero.

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Conflict of interest

Binbin Yue, Jianping Dong, and Ginette Serrero are employees of A&G Pharmaceutical. Rupa Guha and Aditi Banerjee have no conflicts to declare.

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Guha, R., Yue, B., Dong, J. et al. Anti-progranulin/GP88 antibody AG01 inhibits triple negative breast cancer cell proliferation and migration. Breast Cancer Res Treat (2021). https://doi.org/10.1007/s10549-021-06120-y

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Keywords

  • Progranulin/GP88
  • Triple negative breast cancer
  • Proliferation
  • Migration
  • Ki67
  • Anti-progranulin antibody
  • Progranulin SiRNA