Targeted Detection of Soluble Collagen Fragments in U87 Glioblastoma Spheroids
Faculty Mentor Information
Dr. Jonathon Reeck, Boise State University
Presentation Date
7-16-2026
Abstract
Glioblastoma (GBM) is the most aggressive primary brain tumor in adults, characterized by rapid growth, extensive invasion into surrounding brain tissue, high therapy resistance, and nearly universal recurrence despite aggressive treatment. The median survival remains approximately 15 months, with a 5 year survival rate below. Glioblastoma progression is heavily influenced by the remodeling of the tumor's microenvironment and extracellular matrix (ECM). While changes in the matrices drive tumor behavior, specific sub types of collagen roles in growth and development are poorly understood. Collagens work as a paradox, both enabling cell migration yet also providing support and stiffness in tumors. Understanding how these specific subtypes facilitate tumor growth is crucial for therapeutic advancements. We hypothesize that temporal shifts in the ratio of soluble collagen neo-epitopes (cleavage fragments) to intact structural collagens can distinguish early stage tumor progression from benign tissue remodeling and active growth phases. Therefore, the goals of this project are to test the feasibility of targeted detection of soluble fragments using antibodies. Specifically, this study aims to profile collagen expression and its impact on the growth and development in U87 GBM cells across differing chronological stages and growth conditions. The experimental workflow utilized a cell culture of U87 spheroids alongside wound healing assays. Protein collection was performed on both cell lysate and conditioned media obtained from U87 cells at differing times. The workflow started with antibody characterization to ensure reliable protein detection, followed by Western Blot to screen for collagen peptides against a library of collagen antibodies to identify the presence and quantity of collagen over time. Our results demonstrate that specific collagen fragments were detected as soluble fragments. Ultimately, this defines when and under what conditions collagen is being produced in U87 GBM cells, providing crucial insight into the complexities of the tumor matrix. This will allow for greater insight into tumor progression, adding to the impact that we can use collagen dynamics as biomarkers and potentially develop better strategies to interfere with cancer cell migration and target specific cancer subtypes.
Targeted Detection of Soluble Collagen Fragments in U87 Glioblastoma Spheroids
Glioblastoma (GBM) is the most aggressive primary brain tumor in adults, characterized by rapid growth, extensive invasion into surrounding brain tissue, high therapy resistance, and nearly universal recurrence despite aggressive treatment. The median survival remains approximately 15 months, with a 5 year survival rate below. Glioblastoma progression is heavily influenced by the remodeling of the tumor's microenvironment and extracellular matrix (ECM). While changes in the matrices drive tumor behavior, specific sub types of collagen roles in growth and development are poorly understood. Collagens work as a paradox, both enabling cell migration yet also providing support and stiffness in tumors. Understanding how these specific subtypes facilitate tumor growth is crucial for therapeutic advancements. We hypothesize that temporal shifts in the ratio of soluble collagen neo-epitopes (cleavage fragments) to intact structural collagens can distinguish early stage tumor progression from benign tissue remodeling and active growth phases. Therefore, the goals of this project are to test the feasibility of targeted detection of soluble fragments using antibodies. Specifically, this study aims to profile collagen expression and its impact on the growth and development in U87 GBM cells across differing chronological stages and growth conditions. The experimental workflow utilized a cell culture of U87 spheroids alongside wound healing assays. Protein collection was performed on both cell lysate and conditioned media obtained from U87 cells at differing times. The workflow started with antibody characterization to ensure reliable protein detection, followed by Western Blot to screen for collagen peptides against a library of collagen antibodies to identify the presence and quantity of collagen over time. Our results demonstrate that specific collagen fragments were detected as soluble fragments. Ultimately, this defines when and under what conditions collagen is being produced in U87 GBM cells, providing crucial insight into the complexities of the tumor matrix. This will allow for greater insight into tumor progression, adding to the impact that we can use collagen dynamics as biomarkers and potentially develop better strategies to interfere with cancer cell migration and target specific cancer subtypes.