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. 2021 Jan 23;23(1):11.
doi: 10.1186/s13058-021-01391-1.

Leronlimab, a humanized monoclonal antibody to CCR5, blocks breast cancer cellular metastasis and enhances cell death induced by DNA damaging chemotherapy

Xuanmao Jiao  1 , Min Wang  2 , Zhao Zhang  2 , Zhiping Li  2 , Dong Ni  2 , Anthony W Ashton  2   3   4 , Hsin-Yao Tang  5 , David W Speicher  5 , Richard G Pestell  6   7   8
Affiliations

Affiliations

  • 1 Pennsylvania Cancer and Regenerative Medicine Research Center, Baruch S. Blumberg Institute, Pennsylvania Biotechnology Center, 100 East Lancaster Avenue, LIMR R234, Wynnewood, PA, 19096, USA. xuanmao.jiao@bblumberg.org.
  • 2 Pennsylvania Cancer and Regenerative Medicine Research Center, Baruch S. Blumberg Institute, Pennsylvania Biotechnology Center, 100 East Lancaster Avenue, LIMR R234, Wynnewood, PA, 19096, USA.
  • 3 Division of Perinatal Research, Kolling Institute, Northern Sydney Local Health District, St Leonards, NSW, 2065, Australia.
  • 4 Sydney Medical School Northern, University of Sydney, Sydney, NSW, 2006, Australia.
  • 5 Wistar Institute, Philadelphia, PA, 19107, USA.
  • 6 Pennsylvania Cancer and Regenerative Medicine Research Center, Baruch S. Blumberg Institute, Pennsylvania Biotechnology Center, 100 East Lancaster Avenue, LIMR R234, Wynnewood, PA, 19096, USA. richard.pestell@bblumberg.org.
  • 7 Wistar Institute, Philadelphia, PA, 19107, USA. richard.pestell@bblumberg.org.
  • 8 Xavier University School of Medicine, 1000 Woodbury Rd, Suite 109, Woodbury, NY, 11797, USA. richard.pestell@bblumberg.org.

Leronlimab, a humanized monoclonal antibody to CCR5, blocks breast cancer cellular metastasis and enhances cell death induced by DNA damaging chemotherapy

Xuanmao Jiao et al. Breast Cancer Res. .
. 2021 Jan 23;23(1):11.
doi: 10.1186/s13058-021-01391-1.

Authors

Xuanmao Jiao  1 , Min Wang  2 , Zhao Zhang  2 , Zhiping Li  2 , Dong Ni  2 , Anthony W Ashton  2   3   4 , Hsin-Yao Tang  5 , David W Speicher  5 , Richard G Pestell  6   7   8

Affiliations

  • 1 Pennsylvania Cancer and Regenerative Medicine Research Center, Baruch S. Blumberg Institute, Pennsylvania Biotechnology Center, 100 East Lancaster Avenue, LIMR R234, Wynnewood, PA, 19096, USA. xuanmao.jiao@bblumberg.org.
  • 2 Pennsylvania Cancer and Regenerative Medicine Research Center, Baruch S. Blumberg Institute, Pennsylvania Biotechnology Center, 100 East Lancaster Avenue, LIMR R234, Wynnewood, PA, 19096, USA.
  • 3 Division of Perinatal Research, Kolling Institute, Northern Sydney Local Health District, St Leonards, NSW, 2065, Australia.
  • 4 Sydney Medical School Northern, University of Sydney, Sydney, NSW, 2006, Australia.
  • 5 Wistar Institute, Philadelphia, PA, 19107, USA.
  • 6 Pennsylvania Cancer and Regenerative Medicine Research Center, Baruch S. Blumberg Institute, Pennsylvania Biotechnology Center, 100 East Lancaster Avenue, LIMR R234, Wynnewood, PA, 19096, USA. richard.pestell@bblumberg.org.
  • 7 Wistar Institute, Philadelphia, PA, 19107, USA. richard.pestell@bblumberg.org.
  • 8 Xavier University School of Medicine, 1000 Woodbury Rd, Suite 109, Woodbury, NY, 11797, USA. richard.pestell@bblumberg.org.

Abstract

Background: Triple-negative breast cancer (BCa) (TNBC) is a deadly form of human BCa with limited treatment options and poor prognosis. In our prior analysis of over 2200 breast cancer samples, the G protein-coupled receptor CCR5 was expressed in > 95% of TNBC samples. A humanized monoclonal antibody to CCR5 (leronlimab), used in the treatment of HIV-infected patients, has shown minimal side effects in large patient populations.

Methods: A humanized monoclonal antibody to CCR5, leronlimab, was used for the first time in tissue culture and in mice to determine binding characteristics to human breast cancer cells, intracellular signaling, and impact on (i) metastasis prevention and (ii) impact on established metastasis.

Results: Herein, leronlimab was shown to bind CCR5 in multiple breast cancer cell lines. Binding of leronlimab to CCR5 reduced ligand-induced Ca+ 2 signaling, invasion of TNBC into Matrigel, and transwell migration. Leronlimab enhanced the BCa cell killing of the BCa chemotherapy reagent, doxorubicin. In xenografts conducted with Nu/Nu mice, leronlimab reduced lung metastasis of the TNBC cell line, MB-MDA-231, by > 98% at 6 weeks. Treatment with leronlimab reduced the metastatic tumor burden of established TNBC lung metastasis.

Conclusions: The safety profile of leronlimab, together with strong preclinical evidence to both prevent and reduce established breast cancer metastasis herein, suggests studies of clinical efficacy may be warranted.

Keywords: Breast cancer; CCR5; Leronlimab; Metastasis.

PubMed Disclaimer

Conflict of interest statement

R.G.P. holds ownership interests in, CytoDyn Inc. and LightSeed, Inc., receives income from the provision of testimony as an expert witness, serves on the advisory boards of Azure Health, and the MD Anderson Cancer Center Breast Cancer SPORE, and holds ownership interests (value unknown) for several patents and submitted patent applications. The other authors declare that they have no conflict of interest.

Figures

Fig. 1

Fig. 1

Leronlimab binds CCR5 in human…

Fig. 1

Leronlimab binds CCR5 in human breast cancer cells. a In order to determine…

Fig. 1
Leronlimab binds CCR5 in human breast cancer cells. a In order to determine the binding of leronlimab to human CCR5 in breast cancer cells, we used an MDA-MB-231 human breast cancer cell line transfected with a human CCR5 expression vector as a model system (MDA-MB-231-CCR5 cells). A commercial APC conjugated mouse anti-human/mouse/rat CCR5 antibody from R&D (FAB1802A) (APC-αCCR5), was used as a positive control to assess CCR5 positive cells. MDA-MB-231-CCR5 cells were stained with both APC-αCCR5 and leronlimab using the concentration from 1 to 140 mg/ml. Alexa Fluor 488-conjugated mouse anti-human IgG was used as a secondary antibody to measure leronlimab binding cells. Analysis of leronlimab binding with CCR5 by FACS is shown in a. Leronlimab binding with human CCR5 was validated. b The efficiency of leronlimab binding to CCR5-positive cells was up to 98% compared with CCR5 antibody (FAB1802A)
Fig. 2

Fig. 2

Leronlimab blocks human CCL5-CCR5-mediated signaling…

Fig. 2

Leronlimab blocks human CCL5-CCR5-mediated signaling in human breast cancer cells. a MDA-MB-231-CCR5 cells…

Fig. 2
Leronlimab blocks human CCL5-CCR5-mediated signaling in human breast cancer cells. a MDA-MB-231-CCR5 cells were assessed for Ca+ 2 fluxes using Fluo-4 as a calcium concentration indicator. Fluorescence was measured at time points after the addition of the CCR5 ligand, CCL5, in the presence of either CCR5 inhibitors (leronlimab, vicriviroc) or control IgG. b–e Quantitation of the time course of Ca+ 2-induced fluorescence was obtained from living cell imaging (a–d). The CCR5 antagonist, vicriviroc, was used as a positive control (a, e). Leronlimab reduced CCL5-induced calcium responses in MDA-MB-231-CCR5 cells (1.23 ± 0.10, N = 10 for control cells and 0.54 ± 0.13 N = 12 for leronlimab-treated cells. P < 0.001 at calcium peak induce by CCL5)
Fig. 3

Fig. 3

Leronlimab blocks human CCR5-mediated signaling…

Fig. 3

Leronlimab blocks human CCR5-mediated signaling by CCL3 and CCL4 in human breast cancer…

Fig. 3
Leronlimab blocks human CCR5-mediated signaling by CCL3 and CCL4 in human breast cancer cells. a MDA-MB-231-CCR5 cells were assessed for Ca+ 2 fluxes using Fluo-4 as a calcium concentration indicator. Fluorescence was measured at time points after the addition of either CCL3 (a, b) or CCL4 (c, d) in the presence of either CCR5 inhibitors (leronlimab, vicriviroc) or control IgG. b, d Quantitation of the time course of Ca+ 2-induced fluorescence was obtained from living cell imaging (a, c)
Fig. 4

Fig. 4

Leronlimab blocks CCR5-mediated invasion of…

Fig. 4

Leronlimab blocks CCR5-mediated invasion of human breast cancer cells into the extracellular matrix.…

Fig. 4
Leronlimab blocks CCR5-mediated invasion of human breast cancer cells into the extracellular matrix. To test the ability of leronlimab to block cell invasion in 3D Matrigel invasion assay, MDA-MB-231 cells were used. CCL5 was used as chemoattractant to induce invasion. The small-molecule inhibitor of CCR5, vicriviroc, was used as a form of positive control. Leronlimab reduced CCL5-induced MDA-MB-231 breast cancer cell invasion with similar efficacy as vicriviroc (a, b) (855 ± 8.7, N = 8 for control vs. 520 ± 9.1, N = 9 for leronlimab, P < 0.001). We also tested the effects of leronlimab doses on breast cancer cell invasion, and the results showed that both 175 and 350 μg/ml of leronlimab can effectively block MDA-MB-231 cell invasion (c, d)
Fig. 5

Fig. 5

Leronlimab blocks breast cancer metastasis…

Fig. 5

Leronlimab blocks breast cancer metastasis in mice ( a ). The mice were…

Fig. 5
Leronlimab blocks breast cancer metastasis in mice (a). The mice were divided randomly into 4 groups (control, leronlimab, maraviroc, and vicriviroc). MDA-MB-231 cells stably transfected with Luc2-GFP were injected into the mice via the tail vein. The mice in each group were pretreated 1 day before injection with breast cancer cells. The volume of the metastatic tumor formed in the lung was determined by bioluminescence imaging. The bioluminescence images of representative mice from the control, leronlimab, and maraviroc groups are shown in b. The quantitative analysis of tumor size in each group is shown in c. The size of tumors is defined by photon flux (× 109 p/s/cm2/sr). The data was show as mean ± SE. Leronlimab pretreatment decreased breast cancer tumor metastasis to the lung
Fig. 6

Fig. 6

Leronlimab enhances the cell death…

Fig. 6

Leronlimab enhances the cell death induced by doxorubicin, a DNA damage-inducing chemotherapy agent. …

Fig. 6
Leronlimab enhances the cell death induced by doxorubicin, a DNA damage-inducing chemotherapy agent. a MDA-MB-231 cells were treated with 10 μg/ml of leronlimab combined with different dose of doxorubicin for 3 days. The MTT assay was used to determine the relative cell number. The relative absorbance is shown as a fraction of the untreated control. The normalization of leronlimab-treated cells was to leronlimab with no doxorubicin. In b, the cells were treated with maraviroc (100 mM) combined with different doses of doxorubicin, used as a positive control. Data are shown as mean ± SEM for N = 8
Fig. 7

Fig. 7

Leronlimab significantly increases survival in…

Fig. 7

Leronlimab significantly increases survival in mice with established breast cancer lung metastasis. a …

Fig. 7
Leronlimab significantly increases survival in mice with established breast cancer lung metastasis. a Schematic representation of the study design. The mice were injected with MDA-MB-231-pFULG cells via the tail vein. After 7 weeks, when breast cancer lung metastasis was established, the mice were randomly assigned into two cohorts. One cohort was treated with leronlimab (2 mg/mouse, twice a week, 8 mice), and the other was as untreated control. b The survival of mice is plotted with time after the addition of treatment from week 7. c Representative examples of the tumor volume for mice treated with leronlimab, plotted with time after the addition of treatment from week 7. In five mice, the tumor volume decreased as shown (#1, #2, #3, #7, and #8. Mouse #3 was dead between 13 and 14 weeks due to fighting). The decrease in tumor size in mouse #1 was 3-fold to 0.1-fold; #2, 1.2-fold to 0.01-fold; #3, 14.4-fold to 3.7-fold; #7, 7-fold to 0.9-fold; and #8 35.9-fold to 4.7-fold)
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References

    1. Siegel RL, Miller KD, Jemal A. Cancer statistics, 2017. CA Cancer J Clin. 2017;67(1):7–30. - PubMed
    1. Gilabert M, Bertucci F, Esterni B, Madroszyk A, Tarpin C, Jacquemier J, et al. Capecitabine after anthracycline and taxane exposure in HER2-negative metastatic breast cancer patients: response, survival and prognostic factors. Anticancer Res. 2011;31(3):1079–1086. - PubMed
    1. Di Leva G, Cheung DG, Croce CM. miRNA clusters as therapeutic targets for hormone-resistant breast cancer. Expert Rev Endocrinol Metab. 2015;10(6):607–617. - PMC - PubMed
    1. Yu Z, Pestell RG. Small non-coding RNAs govern mammary gland tumorigenesis. J Mammary Gland Biol Neoplasia. 2012;17(1):59–64. - PMC - PubMed
    1. Comprehensive molecular portraits of human breast tumours. Nature. 2012;490(7418):61–70. - PMC - PubMed

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