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. 2015 Jan;9(1):270-81.
doi: 10.1016/j.molonc.2014.08.008. Epub 2014 Aug 30.

Dopamine D2 receptor agonists inhibit lung cancer progression by reducing angiogenesis and tumor infiltrating myeloid derived suppressor cells

Luke H Hoeppner  1 , Ying Wang  1 , Anil Sharma  1 , Naureen Javeed  1 , Virginia P Van Keulen  2 , Enfeng Wang  1 , Ping Yang  3 , Anja C Roden  4 , Tobias Peikert  5 , Julian R Molina  6 , Debabrata Mukhopadhyay  7
Affiliations

Dopamine D2 receptor agonists inhibit lung cancer progression by reducing angiogenesis and tumor infiltrating myeloid derived suppressor cells

Luke H Hoeppner et al. Mol Oncol. 2015 Jan.

Abstract

We sought to determine whether Dopamine D2 Receptor (D2R) agonists inhibit lung tumor progression and identify subpopulations of lung cancer patients that benefit most from D2R agonist therapy. We demonstrate D2R agonists abrogate lung tumor progression in syngeneic (LLC1) and human xenograft (A549) orthotopic murine models through inhibition of tumor angiogenesis and reduction of tumor infiltrating myeloid derived suppressor cells. Pathological examination of human lung cancer tissue revealed a positive correlation between endothelial D2R expression and tumor stage. Lung cancer patients with a smoking history exhibited greater levels of D2R in lung endothelium. Our results suggest D2R agonists may represent a promising individualized therapy for lung cancer patients with high levels of endothelial D2R expression and a smoking history.

Keywords: Angiogenesis; Cabergoline; Dopamine; Dopamine D2 receptor agonists; Lung cancer; VEGF.

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Figures

Figure 1
Figure 1
Dopamine D2 Receptor is upregulated in human lung cancer tissue. A–D: Immunohistochemistry was performed using a monoclonal D2R antibody on formalin‐fixed, paraffin‐embedded tissue as follows: A: mouse brain tissue with no primary D2R antibody (negative control), B: mouse brain tissue (positive control), C: human lung tissue from a healthy individual, D: human lung tissue from a lung adenocarcinoma patient. E–F: Immunohistochemistry of formalin‐fixed, paraffin‐embedded human normal lung (E) and lung adenocarcinoma (F) tissue was performed for Fli‐1 (brown staining) and D2R (red staining). Pink arrows indicate Fli+, D2R‐ endothelial cells (E), green arrows indicate Fli+, D2R+ endothelial cells (F) and black arrows indicate D2R+ myeloid cells.
Figure 2
Figure 2
DA and D2R agonist treatment decreases tumor progression in an LLC1 murine lung cancer model. A–E: C57BL/6 wildtype (B–D) and D2R knockout (B–C) mice were orthotopically injected with 1 × 105 luciferase‐labeled murine LLC1 cells suspended in 80 μl PBS and Matrigel. After establishment of the lung tumor, mice were xenogen imaged four days post‐injection of LLC1 cells. Mice received daily intraperitoneal injections of PBS vehicle (control groups) or 50 mg/kg dopamine (B), 10 mg/kg quinpirole (C), or 5 mg/kg cabergoline (D) (treatment groups) for seven days. Mice were xenogen imaged following treatment. A: Experimental timeline. B–D: Quantitation of LLC1 tumor growth by determining the difference in relative luciferase units (RLU) before and after treatment. E: Pre‐ and post‐treatment xenogen images of vehicle‐ and cabergoline treated mice.
Figure 3
Figure 3
D2R agonist treatment decreases angiogenesis and promotes apoptosis in the endothelium of LLC1 tumor bearing mice. A: Immunohistochemistry was performed using a monoclonal CD31 antibody on formalin‐fixed, paraffin‐embedded tissue harvested from LLC1 tumor bearing mice treated with vehicle (left) or 10 mg/kg quinpirole (right) daily for seven days. B: The amount of CD31‐positive endothelial cells in lung tissue harvested from control and quinpirole‐treated mice (n = 2 for each group) was quantitated by counting the number of CD31‐positive endothelial cells per visual field (10 visual fields were counted for each tissue). C: Co‐immunofluorescence staining for TUNEL (green; bottom left single stain) and CD31 (red; bottom right single stain) was performed on tissue harvested from LLC1 tumor bearing mice treated with vehicle (left) or 10 mg/kg quinpirole (right) daily for seven days. Merged images are shown in the top panel (green: TUNEL, red: CD31, blue: DAPI). D: Colocalization of TUNEL and CD31 staining was quantitated in the immunofluorescent images of lung tissue harvested from control and quinpirole‐treated mice (n = 2 for each group) by counting the number of double positive TUNEL and CD31 endothelial cells per visual field (10 visual fields were counted for each tissue).
Figure 4
Figure 4
D2R agonist treatment decreases tumor progression in a human lung tumor xenograft model. A–C: SCID mice were orthotopically injected with 2 × 106 luciferase‐labeled human A549 lung cancer cells suspended in 80 μl PBS and Matrigel. After establishment of the lung tumor, mice were xenogen imaged eight days post‐injection of A549 cells. Mice received intraperitoneal injections of PBS vehicle (control group, n = 6) or 10 mg/kg quinpirole (treatment group, n = 6) every other day for 17 days. Mice were xenogen imaged following treatment. A: Experimental timeline. B: Quantitation of A549 tumor growth by determining the difference in relative luciferase units (RLU) before and after treatment. C: Pre‐ and post‐treatment xenogen images of vehicle‐ and quinpirole‐treated mice.
Figure 5
Figure 5
D2R is expressed by endothelial cells in human lung tumor tissue. A: Immunohistochemistry was performed using a monoclonal D2R antibody on a human lung cancer tissue microarray, and each tissue was scored for the percentage of endothelial cells that were D2R‐positive. Each point on the graph represents an individual tissue. B: Representative staining from tissue microarray (top: 10× H&E staining, middle: 10× D2R staining, bottom: 20× D2R staining). Red arrows indicate D2R‐positive endothelial cells.
Figure 6
Figure 6
Tumor infiltrating myeloid derived suppressor cells decreased in vivo by D2R agonists and endothelial D2R expression increased in NSCLC patients with a smoking history. A: Representative image of D2R immunohistochemistry of human lung cancer tissue microarray depicting D2R‐positive myeloid/macrophage lineage cells (red arrows). B–C: C57BL/6 wildtype mice were orthotopically injected with 1 × 105 LLC1 cells. Mice were intraperitoneally administered vehicle or cabergoline daily for seven days starting on day five post‐injection of LLC1 cells. After 12 days, the lungs were harvested from euthanized mice and cells were extracted through collagenase digestion. Flow cytometry was used to determine the number (B) and percentage (C) of murine myeloid derived suppressor cells (MDSCs), which were defined as LIN‐, GR1+, and CD11b+. n = 16 mice/treatment group. D: Immunohistochemistry was performed using a monoclonal D2R antibody on a human lung cancer tissue microarray. The graph shows the percentage of endothelial cells stained positive for D2R among lung cancer tissue obtained from smokers (n = 13) compared to those from never smokers (n = 3).
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References

    1. Alberg, A.J. , Shopland, D.R. , Cummings, K.M. , 2014. The 2014 Surgeon General's report: commemorating the 50th Anniversary of the 1964 Report of the Advisory Committee to the US Surgeon General and updating the evidence on the health consequences of cigarette smoking. Am. J. Epidemiol. 179, 403–412. - PMC - PubMed
    1. Basu, S. , Nagy, J.A. , Pal, S. , Vasile, E. , Eckelhoefer, I.A. , Bliss, V.S. , Manseau, E.J. , Dasgupta, P.S. , Dvorak, H.F. , Mukhopadhyay, D. , 2001. The neurotransmitter dopamine inhibits angiogenesis induced by vascular permeability factor/vascular endothelial growth factor. Nat. Med. 7, 569–574. - PubMed
    1. Basu, S. , Sarkar, C. , Chakroborty, D. , Nagy, J. , Mitra, R.B. , Dasgupta, P.S. , Mukhopadhyay, D. , 2004. Ablation of peripheral dopaminergic nerves stimulates malignant tumor growth by inducing vascular permeability factor/vascular endothelial growth factor-mediated angiogenesis. Cancer Res. 64, 5551–5555. - PubMed
    1. Benowitz, N.L. , 2009. Pharmacology of nicotine: addiction, smoking-induced disease, and therapeutics. Annu. Rev. Pharmacol. Toxicol. 49, 57–71. - PMC - PubMed
    1. Bhandarkar, S.S. , Jaconi, M. , Fried, L.E. , Bonner, M.Y. , Lefkove, B. , Govindarajan, B. , Perry, B.N. , Parhar, R. , Mackelfresh, J. , Sohn, A. , Stouffs, M. , Knaus, U. , Yancopoulos, G. , Reiss, Y. , Benest, A.V. , Augustin, H.G. , Arbiser, J.L. , 2009. Fulvene-5 potently inhibits NADPH oxidase 4 and blocks the growth of endothelial tumors in mice. J. Clin. Invest. 119, 2359–2365. - PMC - PubMed

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