Formulation of CRISPR Edited Cells to Evaluate Glypican‐Targeting in Glioblastoma

Heparan sulfate proteoglycans (HSPGs) are ubiquitous protein/carbohydrate complexes that are found on all cell surfaces. They play a role in various biological pathways such as cellular growth and vascularization, processes that are exploited by cancer. Heparanase is an enzyme that modifies HSPGs which can alter the structure of the proteoglycan and change its bioactivity. In glioblastoma, these enzymes are overexpressed and are thought to contribute to the rapid and aggressive growth of cancer. HSPGs, particularly glypicans, are known to be overexpressed in glioblastoma. Here, we generated double and single knockouts of glypican‐3 (GPC3) and glypican‐1 (GPC1) in glioblastoma cells using CRISPR editing with specific sgRNA. Sanger sequencing and western blot determination were utilized to verify the knockout. We then compared these knockouts to unedited glioblastoma cells to understand the importance of glypicans in glioblastoma as well as investigate their sensitivity to platinum‐based chemotherapeutics. The further exploration of targeting HSPGs in general could allow for tailored chemotherapeutic treatments to patients with glioblastoma. This may pave the way for new molecular targets and more effective treatments for glial tumors in the future. Support or Funding Information The President's Commission for the Status of Women for funding my research. My collaborates Dr. Karin Forsberg Nilsson at Uppsala University, Sweden for donating the GL261 cells. Dr. Nicholas P. Farrell at the Massey Cancer Center, Virginia Commonwealth University, Richmond, VA for collaborating with me. Dr. Samantha Katner for her continuous support, encouragement, guidance and great mentorship