An amino-bisphosphonate targets MMP-9–expressing macrophages and angiogenesis to impair cervical carcinogenesis
Enrico Giraudo, … , Masahiro Inoue, Douglas Hanahan
Enrico Giraudo, … , Masahiro Inoue, Douglas Hanahan
Published September 1, 2004
Citation Information: J Clin Invest. 2004;114(5):623-633. https://doi.org/10.1172/JCI22087.
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Article Oncology

An amino-bisphosphonate targets MMP-9–expressing macrophages and angiogenesis to impair cervical carcinogenesis

Abstract

A mouse model involving the human papillomavirus type-16 oncogenes develops cervical cancers by lesional stages analogous to those in humans. In this study the angiogenic phenotype was characterized, revealing intense angiogenesis in high-grade cervical intraepithelial neoplasias (CIN-3) and carcinomas. MMP-9, a proangiogenic protease implicated in mobilization of VEGF, appeared in the stroma concomitant with the angiogenic switch, expressed by infiltrating macrophages, similar to what has been observed in humans. Preclinical trials sought to target MMP-9 and angiogenesis with a prototypical MMP inhibitor and with a bisphosphonate, zoledronic acid (ZA), revealing both to be antiangiogenic, producing effects comparable to a Mmp9 gene KO in impairing angiogenic switching, progression of premalignant lesions, and tumor growth. ZA therapy increased neoplastic epithelial and endothelial cell apoptosis without affecting hyperproliferation, indicating that ZA was not antimitotic. The analyses implicated cellular and molecular targets of ZA’s actions: ZA suppressed MMP-9 expression by infiltrating macrophages and inhibited metalloprotease activity, reducing association of VEGF with its receptor on angiogenic endothelial cells. Given its track record in clinical use with limited toxicity, ZA holds promise as an “unconventional” MMP-9 inhibitor for antiangiogenic therapy of cervical cancer and potentially for additional cancers and other diseases where MMP-9 expression by infiltrating macrophages is evident.

Authors

Enrico Giraudo, Masahiro Inoue, Douglas Hanahan

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Figure 6

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ZA inhibits MMP-9 expression and activation in macrophages. (A) Reduced ...
ZA inhibits MMP-9 expression and activation in macrophages. (A) Reduced MMP-9 expression was detected in macrophages in the stroma adjacent to CIN-3 in ZA-treated mice compared with controls as revealed by colocalization of MMP-9 (green) and CD-68 (red). Arrows show MMP-9–expressing macrophages; arrowheads indicate macrophages that do not express MMP-9. (B) Quantification of double-labeled MMP-9+/CD-68+ cells (5 fields per mouse) revealed a 71% reduction in the PT (n = 8 control, n = 6 ZA-treated) and a 68% reduction in the RT (n = 10 control, n = 8 ZA-treated). Similar results were obtained with double-labeled MMP-9+/F4/80+ cells (not shown). #P < 0.001. (C) Gelatinase activity in tissue extracts was measured by incubation with fluorescin-conjugated gelatin in the absence or presence of the MMP inhibitor 1,10 Phe (4 mM). Gelatinase activity was lower in ZA-treated compared with control cervixes in both PT and RT. **P < 0.01 versus control. N/E2 indicates estrogen-treated normal cervix. (D) Zymography showing the pro- and active forms of gelatinases in tissue lysates from both control and ZA-treated mice. Pro- and active forms of MMP-9 and MMP-2 are indicated by arrows. Scale bar: 25 μm. Values are mean ± SEM. P values were calculated using the Wilcoxon test.