Investigating Internalization, Intracellular Tracking, and Localization of NGR-Tagged Rubredoxin in HT-1080 Fibrosarcoma Cells
Location
Suwanee, GA
Start Date
6-5-2025 1:00 PM
End Date
6-5-2025 4:00 PM
Description
Introduction Fibrosarcoma is a highly aggressive soft tissue sarcoma with limited treatment options and poor response to conventional therapies, necessitating the development of more targeted treatment strategies. CD13, a transmembrane metallopeptidase overexpressed in tumor vasculature, represents a promising target for tumor-selective drug delivery. This study investigates the internalization and subcellular trafficking of an NGR-tagged Rubredoxin holoprotein (NGR-Rb) in HT-1080 fibrosarcoma cells as a potential targeted therapeutic strategy. Rubredoxin, a small iron-containing redox protein derived from Pyrococcus furiosus, is particularly well-suited for this approach due to its extreme stability and ability to function in harsh environments, making it an ideal candidate for tumor-targeted protein therapeutics.
Methods A time-course internalization experiment will be conducted to assess the uptake of NGR-Rb at defined time points (0, 15 min, 45 min, 2 h, and 4 h). Internalization will be confirmed using a dynamin inhibitor, Dynasore, and a 4°C control to differentiate between receptor-mediated and passive uptake. Subcellular localization will be determined through fluorescence microscopy and co-localization studies with nuclear (DAPI), mitochondrial (MitoTracker), and lysosomal (LysoTracker) markers.
Results This study aims to provide insight into the mechanistic pathways governing NGR-Rb internalization and trafficking, offering a foundation for targeted protein-based drug delivery approaches in fibrosarcoma and other CD13-expressing malignancies. By elucidating these mechanisms, this work contributes to the development of innovative, tumor-selective therapeutics designed to improve treatment efficacy while minimizing off-target toxicity.
Discussion The findings of this research will inform the rational design of NGR-conjugated therapeutics, facilitating their integration with chemotherapeutics or immunotherapies for enhanced cancer treatment. Future studies will expand into in vivo models to confirm clinical viability, paving the way for the development of protein-based targeted therapies with reduced systemic toxicity in fibrosarcoma and other CD13-expressing malignancies. As this project advances, it is expected to contribute to the broader field of tumor-targeted therapy, offering a novel approach to drug delivery in fibrosarcoma.
Embargo Period
5-19-2025
Investigating Internalization, Intracellular Tracking, and Localization of NGR-Tagged Rubredoxin in HT-1080 Fibrosarcoma Cells
Suwanee, GA
Introduction Fibrosarcoma is a highly aggressive soft tissue sarcoma with limited treatment options and poor response to conventional therapies, necessitating the development of more targeted treatment strategies. CD13, a transmembrane metallopeptidase overexpressed in tumor vasculature, represents a promising target for tumor-selective drug delivery. This study investigates the internalization and subcellular trafficking of an NGR-tagged Rubredoxin holoprotein (NGR-Rb) in HT-1080 fibrosarcoma cells as a potential targeted therapeutic strategy. Rubredoxin, a small iron-containing redox protein derived from Pyrococcus furiosus, is particularly well-suited for this approach due to its extreme stability and ability to function in harsh environments, making it an ideal candidate for tumor-targeted protein therapeutics.
Methods A time-course internalization experiment will be conducted to assess the uptake of NGR-Rb at defined time points (0, 15 min, 45 min, 2 h, and 4 h). Internalization will be confirmed using a dynamin inhibitor, Dynasore, and a 4°C control to differentiate between receptor-mediated and passive uptake. Subcellular localization will be determined through fluorescence microscopy and co-localization studies with nuclear (DAPI), mitochondrial (MitoTracker), and lysosomal (LysoTracker) markers.
Results This study aims to provide insight into the mechanistic pathways governing NGR-Rb internalization and trafficking, offering a foundation for targeted protein-based drug delivery approaches in fibrosarcoma and other CD13-expressing malignancies. By elucidating these mechanisms, this work contributes to the development of innovative, tumor-selective therapeutics designed to improve treatment efficacy while minimizing off-target toxicity.
Discussion The findings of this research will inform the rational design of NGR-conjugated therapeutics, facilitating their integration with chemotherapeutics or immunotherapies for enhanced cancer treatment. Future studies will expand into in vivo models to confirm clinical viability, paving the way for the development of protein-based targeted therapies with reduced systemic toxicity in fibrosarcoma and other CD13-expressing malignancies. As this project advances, it is expected to contribute to the broader field of tumor-targeted therapy, offering a novel approach to drug delivery in fibrosarcoma.