Therapeutic potential of hyaluronan oligosaccharides for bone metastasis of breast cancer

Abstract

Hyaluronan (HA) oligosaccharides were reported to have suppressive effects on various malignant tumors via disruption of receptor HA interactions. However, no studies have focused on the effects of HA oligosaccharides on bone metastasis of breast cancer. In this study, we clarified the effective size of HA oligosaccharides required to inhibit cell growth in the highly invasive breast cancer cell line, MDA-MB-231 cells. Based on the results of cell growth assay, we subsequently analyzed the effects of HA tetrasaccharides, HA decasaccharides, and high molecular weight HA on the other breast cancer cell behaviors in vitro and breast cancer bone metastasis in vivo. HA decasaccharides significantly inhibited cell growth, motility, and invasion, whereas tetrasaccharides did not. HAS2 mRNA expression was altered after the treatment with both tetrasaccharides and decasaccharides. Phosphorylation of Akt was suppressed after 1 h treatment with HA decasaccharides, and the effect was partially reversed by anti-CD44 monoclonal antibody. In vivo, local application of HA decasaccharides inhibited the expansion of osteolytic lesions in tibia on soft X-rays using mouse bone metastasis model of breast cancer. Histological analysis revealed HA accumulation in bone metastatic lesions was perturbed by decasaccharides. These results suggest that HA oligosaccharides suppressed progression of bone metastasis in breast cancer via interruption of endogenous HA-CD44 interaction, and as such, can be a novel therapeutic candidate to limit bone metastasis of breast cancer.

Targeting Hyaluronic Acid Family for Cancer Chemoprevention and Therapy

“…In addition, 4-MU treatment inhibits a variety of HA signaling events, including downregulation of phospho-ErbB2, phospho-Akt, and downstream effectors MMP-2/MMP-9 and IL-8 expression (Fig. 2.1; Lokeshwar et al, 2010; Okuda et al, 2012; Twarock et al, 2011; Urakawa, Nishida, Wasa, et al, 2012). The efficacy of 4-MU as a glucuronate scavenger could contribute to its antitumor effects.…”

“…At concentrations of 0.2–1 mM (i.e., ~35–180 μg/ml), 4-MU inhibits proliferation, motility, and invasion, and causes loss of filopodia and focal adhesions, in a variety of cultured tumor cells (Arai et al, 2011; Hiraga, Ito, & Nakamura, 2013; Lokeshwar et al, 2010; Okuda et al, 2012; Piccioni et al, 2012; Twarock et al, 2011; Uchakina, Ban, & McKallip, 2013; Urakawa et al, 2012). 4-MU inhibits tumor spheroids and osteoclast-like cell formation (Hiraga et al, 2013) and downregulates the expression of both CD44 and RHAMM, suggesting a feedback loop between HA synthesis and HA receptor expression (Lokeshwar et al, 2010).…”

“…Nevertheless, at the concentrations used in cancer cells and tumor models and also the doses at which it is consumed as a dietary supplement, 4-MU is demonstrated to have antitumor activities. For example, in xenograft studies, oral administration of 4-MU has been shown to inhibit tumor growth and metastasis in prostate, B16 melanoma, skin, liver, osteosarcoma, breast, and esophageal cancer model systems (Bhattacharyya et al, 2009; Kudo et al, 2004; Lokeshwar et al, 2010; Nakazawa et al, 2006; Piccioni et al, 2012; Twarock et al, 2011; Urakawa, Nishida, Wasa, et al, 2012; Yoshihara et al, 2005). Recently, 4-MU was shown to inhibit bone metastasis in a breast cancer model (Hiraga et al, 2013).…”

“…Recently, 4-MU was shown to inhibit bone metastasis in a breast cancer model (Hiraga et al, 2013). As in the case of in vitro studies, some mouse xenograft studies have used 4-MU orally at doses as high as 1–3 g/kg; however, in other studies, 4-MU has shown remarkable efficacy at 200–400 mg/kg doses (Arai et al, 2011; Bhattacharyya et al, 2009; Hiraga et al, 2013; Kudo et al, 2004; Nakazawa et al, 2006; Okuda et al, 2012; Piccioni et al, 2012; Twarock et al, 2011; Urakawa, Nishida, Wasa, et al, 2012; Yoshihara et al, 2005). Based on the FDA’s formula of mouse-to-human dose conversion, 200–400 mg/kg doses in mice equates to 1.1–2.2 g/day doses in humans; these are doses at which 4-MU is consumed for improving liver health (Abate et al, 2001; Camarri & Marchettini, 1988; Garrett et al, 1993; Hoffmann et al, 2005; Quaranta et al, 1984; U.S.…”

“…Angiogenic HA fragments have been detected in the urine of patients with bladder cancer and Wilm’s tumor, in the saliva of patients with head and neck cancer, and in bladder and prostate tumor tissues (Franzmann et al, 2003; Kumar, West, Ponting, & Gattamaneni, 1989; Lokeshwar, Obek, Soloway, & Block, 1997; Lokeshwar et al, 2001). In contrast to the angiogenic HA fragments, HA oligosaccharides consisting of 2–3 disaccharide units have been shown to have antitumor activity, presumably because they inhibit HA-induced signaling (Ghatak, Misra, & Toole, 2002; Hosono et al, 2007; Toole, Ghatak, & Misra, 2008; Urakawa et al, 2012). …”