Heparan-6- O-Endosulfatase 2 Promotes Invasiveness of Head and Neck Squamous Carcinoma Cell Lines in Co-Cultures with Cancer-Associated Fibroblasts

Pritha Mukherjee; Xin Zhou; Julius Benicky; Aswini Panigrahi; Reem Aljuhani; Jian Liu; Laurie Ailles; Vitor H Pomin; Zhangjie Wang; Radoslav Goldman

doi:10.3390/cancers15215168

Heparan-6- O-Endosulfatase 2 Promotes Invasiveness of Head and Neck Squamous Carcinoma Cell Lines in Co-Cultures with Cancer-Associated Fibroblasts

Cancers (Basel). 2023 Oct 27;15(21):5168. doi: 10.3390/cancers15215168.

Authors

Pritha Mukherjee¹, Xin Zhou^{1

2}, Julius Benicky^{1

3}, Aswini Panigrahi^{1

3}, Reem Aljuhani^{3

4}, Jian Liu⁵, Laurie Ailles^{6

7}, Vitor H Pomin^{8

9}, Zhangjie Wang¹⁰, Radoslav Goldman^{1

3

4}

Affiliations

¹ Department of Oncology, Lombardi Comprehensive Cancer Center, Georgetown University, Washington, DC 20057, USA.
² Biotechnology Program, Northern Virginia Community College, Manassas, VA 20109, USA.
³ Clinical and Translational Glycoscience Research Center, Georgetown University, Washington, DC 20057, USA.
⁴ Department of Biochemistry and Molecular & Cellular Biology, Georgetown University, Washington, DC 20057, USA.
⁵ Division of Chemical Biology and Medicinal Chemistry, Eshelman School of Pharmacy, University of North Carolina, Chapel Hill, NC 27599, USA.
⁶ Department of Medical Biophysics, University of Toronto, Toronto, ON M5G 1L7, Canada.
⁷ Princess Margaret Cancer Centre, University Health Network, Toronto, ON M5G 1L7, Canada.
⁸ Department of BioMolecular Sciences, University of Mississippi, Oxford, MS 38677, USA.
⁹ Research Institute of Pharmaceutical Sciences, School of Pharmacy, University of Mississippi, Oxford, MS 38677, USA.
¹⁰ Glycan Therapeutics, LLC, 617 Hutton Street, Raleigh, NC 27606, USA.

Abstract

Local invasiveness of head and neck squamous cell carcinoma (HNSCC) is a complex phenomenon supported by interaction of the cancer cells with the tumor microenvironment (TME). We and others have shown that cancer-associated fibroblasts (CAFs) are a component of the TME that can promote local invasion in HNSCC and other cancers. Here we report that the secretory enzyme heparan-6-O-endosulfatase 2 (Sulf-2) directly affects the CAF-supported invasion of the HNSCC cell lines SCC35 and Cal33 into Matrigel. The Sulf-2 knockout (KO) cells differ from their wild type counterparts in their spheroid growth and formation, and the Sulf-2-KO leads to decreased invasion in a spheroid co-culture model with the CAF. Next, we investigated whether a fucosylated chondroitin sulfate isolated from the sea cucumber Holothuria floridana (HfFucCS) affects the activity of the Sulf-2 enzyme. Our results show that HfFucCS not only efficiently inhibits the Sulf-2 enzymatic activity but, like the Sulf-2 knockout, inhibits Matrigel invasion of SCC35 and Cal33 cells co-cultured with primary HNSCC CAF. These findings suggest that the heparan-6-O-endosulfatases regulate local invasion and could be therapeutically targeted with the inhibitory activity of a marine glycosaminoglycan.

Keywords: Sulf-2 CRISPRCas9 knockout; Sulf-2 inhibitor; cancer invasion; cancer-associated fibroblast; head and neck squamous cell carcinoma; heparan-6-O-endosulfatase.

Abstract

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