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. 2003 Nov 11;100(23):13513-8.
doi: 10.1073/pnas.2235846100. Epub 2003 Oct 31.

A small-molecule antagonist of CXCR4 inhibits intracranial growth of primary brain tumors

Joshua B Rubin  1 , Andrew L Kung, Robyn S Klein, Jennifer A Chan, YanPing Sun, Karl Schmidt, Mark W Kieran, Andrew D Luster, Rosalind A Segal
Affiliations

A small-molecule antagonist of CXCR4 inhibits intracranial growth of primary brain tumors

Joshua B Rubin et al. Proc Natl Acad Sci U S A. .

Abstract

The vast majority of brain tumors in adults exhibit glial characteristics. Brain tumors in children are diverse: Many have neuronal characteristics, whereas others have glial features. Here we show that activation of the Gi protein-coupled receptor CXCR4 is critical for the growth of both malignant neuronal and glial tumors. Systemic administration of CXCR4 antagonist AMD 3100 inhibits growth of intracranial glioblastoma and medulloblastoma xenografts by increasing apoptosis and decreasing the proliferation of tumor cells. This reflects the ability of AMD 3100 to reduce the activation of extracellular signal-regulated kinases 1 and 2 and Akt, all of which are pathways downstream of CXCR4 that promote survival, proliferation, and migration. These studies (i) demonstrate that CXCR4 is critical to the progression of diverse brain malignances and (ii) provide a scientific rationale for clinical evaluation of AMD 3100 in treating both adults and children with malignant brain tumors.

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Figures

Fig. 1.
Fig. 1.
CXCR4 and CXCL12 are expressed in human and mouse brain tumors. (a) CXCR4 immunoreactivity (brown) is in human medulloblastoma cells (Left) but not in cerebellar granule cells (Center). The low-magnification view (Left) shows a densely cellular tumor; the high-power view (Inset) shows that tumor cells are immunoreactive for CXCR4. The low-power view of the cerebellum (CBL) (Center) reveals a normal layered structure, internal granule cell layer (IGL), Purkinje cell layer (PCL), and molecular layer (ML); the high-power view (Inset) shows that granule cells of the IGL exhibit little to no CXCR4 immunoreactivity. Endothelial cell staining for CXCL12 (brown) is seen in medulloblastoma-associated blood vessels (Right, arrow). (b) CXCR4 immunoreactivity (brown) is in GBM cells (Left) but not in an astrocyte of normal cortex (CTX) (Center, blue arrowhead). The low-power view of GBM (Left) reveals a densely cellular tumor composed of cells with fibrillary processes; the high-power view (Inset) shows immunoreactivity for CXCR4 in these cells. (Center) Low-power view of layers II and III of normal cerebral CTX; the high-power view (Inset) reveals a pyramidal neuron on the right, with weak CXCR4 immunostaining, and a cell with features of a nonreactive astrocyte (blue arrowhead) that is negative for CXCR4. Endothelial cell staining for CXCL12 (brown) is seen in GBM-associated blood vessel (Right). (c) CXCR4 (red) staining in tumor cells of a spontaneously arising medulloblastoma in a ptc +/- mouse (Left). CXCR4 immunoreactivity is not present in granule cells of the IGL. Nuclei are counterstained with DAPI. CXCL12 is localized to vascular endothelium of tumor-associated blood vessels. CXCL12 is in red, PECAM is in green (Center), and the merged image is in yellow (Right). An asterisk indicates the same blood vessel in each panel.
Fig. 2.
Fig. 2.
Daoy and U87 cell responses to CXCL12 are blocked by AMD 3100 in vitro. Results with Daoy and U87 cells are shown in Left and Right, respectively. (a) CXCR4 expression in human tumor cell lines. U87 and Daoy cells exhibit punctate staining for CXCR4 (red). (b) CXCL12 is a chemoattractant for Daoy and U87 cells in a Boyden chamber assay. Peak CXCL12 chemotactic responses are blocked by AMD 3100 (solid bars). (c) CXCL12 and Shh are proliferative factors for Daoy cells (open bars), and CXCL12 and PDGF are proliferative factors for U87 cells (open bars). Proliferation in response to CXCL12 is inhibited by AMD 3100 in both cell lines, and AMD 3100 decreases Shh-induced proliferation in Daoy cells (solid bars). (d) The combination of SFM and CXCL12 is equivalent to serum-supplemented media in supporting cell survival (open bars). AMD 3100 blocked the survival effects of CXCL12 (solid bars). Each point represents the mean of three experiments done in triplicate. Data in b and c are mean percent changes relative to control ± SEM. Data in d are percentages TUNEL-positive cells ± SEM. *, P < 0.05; **, P < 0.005 (for the difference between SFM with and without CXCL12). #, P < 0.05; ##, P < 0.005 (for the effect of AMD 3100 on cells exposed to CXCL12). (Scale bars, 100 μm.)
Fig. 3.
Fig. 3.
AMD 3100 inhibits growth of intracranial U87 (a and c) and Daoy (b) xenografts. (a and b) Bioluminescent images from control and treated animals at the start and end of a 3-week treatment. Growth curves were derived from serial measurements in three experiments (four or five animals per treatment group per experiment). Data are mean values ± SEM. (c) Volumetric reconstructions of U87 intracranial xenografts (blue) after 3-week treatment. Tumor reconstructions are overlaid on coronal sections of the mouse brain. Volumes are mean values ± SEM for PBS and AMD 3100 treatment groups (n = 4). *, P < 0.05.
Fig. 4.
Fig. 4.
AMD 3100 increases apoptosis and decreases activation of Erk 1/2 and Akt in tumor cells. (a) Apoptosis is increased in U87 xenografts within 24 h of AMD 3100 treatment compared to PBS control. Serial sections from AMD 3100-treated tumor were stained for PECAM (green) and TUNEL (red). Apoptosis was limited to tumor cells. Increases in apoptosis were also seen in U87 tumors analyzed at 48 h. (b) One week of AMD 3100 treatment increased U87 apoptosis without any change in proliferation. AMD 3100 treatment decreased proliferation and increased apoptosis in Daoy xenografts. Apoptosis was also increased in U87 tumors at 2 and 3 weeks of treatment (data not shown). Data presented are means ± SEM in two tumors per condition (1,000-5,000 cells per tumor). *, P < 0.05; **, P < 0.005. (c) U87 and Daoy xenografts immunostained for pErk 1/2 and pAkt. Immunopositivity of pErk 1/2 and pAkt was decreased in tumors treated for 3 weeks with AMD 3100 compared to those treated with PBS. (Scale bars, 100 μM.)
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References

    1. Ries, L. A. G., Eisner, M. P., Kosary, C. L., Hankey, B. F., Miller, B. A., Clegg, L. & Edwards, B. K., eds. (2001) SEER Cancer Statistics Review (Natl. Cancer Inst., Bethesda).
    1. Kleihues, P., Burger, P. C., Collins, V. P., Newcomb, E. W., Ohgaki, H. & Cavenee, W. K. (2000) in World Health Organization Classification of Tumours: Pathology and Genetics of Tumours of the Central Nervous System, eds. Kleihues, P. C. & Cavenee, W. K. (Int. Agency for Res. on Cancer, Lyon, France), pp. 29-39.
    1. Giangaspero, F., Bigner, S. H., Kleihues, P., Pietsch, T. & Trojanowski, J. Q. (2000) in World Health Organization Classification of Tumours: Pathology and Genetics of Tumours of the Central Nervous System, eds. Kleihues, P. C. & Cavenee, W. K. (Int. Agency for Res. on Cancer, Lyon, France), pp. 129-137.
    1. Luster, A. D. (1998) N. Engl. J. Med. 338, 436-445. - PubMed
    1. Ma, Q., Jones, D., Borghesani, P. R., Segal, R. A., Nagasawa, T., Kishimoto, T., Bronson, R. T. & Springer, T. A. (1998) Proc. Natl. Acad. Sci. USA 95, 9448-9453. - PMC - PubMed

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