Antitumor activity of sulfated hyaluronic acid fragments in pre-clinical models of bladder cancer

Abstract

// Andre R. Jordan 1 , Soum D. Lokeshwar 2 , Luis E. Lopez 3 , Martin Hennig 3, 4 , Juan Chipollini 5 , Travis Yates 1, + , Marie C. Hupe 3, 4 , Axel S. Merseburger 4 , Aviva Shiedlin 6, * , Wolfgang H. Cerwinka 5, ♠ , Kebin Liu 3 , Vinata B. Lokeshwar 3 1 Sheila and David Fuente Graduate Program in Cancer Biology, Sylvester Comprehensive Cancer Center, University of Miami-Miller School of Medicine, Miami, FL, USA 2 Honors Program in Medical Education, University of Miami-Miller School of Medicine, Miami, FL, USA 3 Department of Biochemistry and Molecular Biology, Medical College of Georgia, Augusta University, Augusta, GA, USA 4 Department of Urology, University of Lübeck, Lübeck, Germany 5 Department of Urology, University of Miami-Miller School of Medicine, Miami, FL, USA 6 Drug and Biomaterial R&D, Genzyme Corporation, Cambridge, MA, USA + Present address: University of Pennsylvania- Department of Cancer Biology, Philadelphia, PA, USA * Present address: PoroGen Corporation, Woburn, MA, USA ♠ Present address: Georgia Pediatric Urology, Children’s Healthcare of Atlanta, Emory University, Atlanta, GA, USA Correspondence to: Vinata B. Lokeshwar, email: vlokeshwar@augusta.edu Keywords: bladder cancer, hyaluronic acid, hyaluronidase, sulfated-HA, targeted therapy Received: February 22, 2016      Accepted: May 16, 2016      Published: July 11, 2016 ABSTRACT Tumor cell-derived hyaluronidase HYAL-1 degrades hyaluronic acid (HA) into angiogenic fragments (AGF: 10-12 disaccharides). AGF support tumor growth and progression. Urine and tissue HAase/HYAL-1 levels are sensitive markers for high-grade bladder cancer (BCa) and its metastasis. In preclinical models of BCa, we evaluated whether o-sulfated AGF (sHA-F) inhibits HAase activity and has antitumor activity. At IC 50 for HAase activity inhibition (5-20 μg/ml [0.4-1.7 μM]), sHA-F significantly inhibited proliferation, motility and invasion of HYAL-1 expressing BCa cells (253J-Lung, HT1376, UMUC-3), P<0.001. sHA-F did not affect the growth of HYAL-1 non-expressing BCa (5637, RT4, T24, TCCSUP) and normal urothelial (Urotsa, SV-HUC1) cells. sHA-F treatment induced apoptosis by death receptor pathway. sHA-F downregulated transcript and/or protein levels of HA receptors (CD44, RHAMM), p-AKT, β-catenin, pβ-Catenin(S552), Snail and Twist but increased levels of pβ-Catenin(T41/S45), pGSK-3α/β(S21/S9) and E-cadherin. sHA-F also inhibited CD44/Phosphoinositide 3-kinase (PI-3K) complex formation and PI-3K activity. AGF addition or myristoylated-AKT overexpression attenuated sHA-F effects. Contrarily, HYAL-1 expression sensitized RT4 cells to sHA-F treatment. In the 253J-L and HT1376 xenograft models, sHA-F treatment significantly inhibited tumor growth (P<0.001), plausibly by inhibiting angiogenesis and HA receptor-PI-3K/AKT signaling. This study delineates that sHA-F targets tumor-associated HA-HAase system and could be potentially useful in BCa treatment.