Date of Award
Fall 11-22-2024
Document Type
Masters Thesis
Degree Name
M.S.
Organizational Unit
College of Natural Science and Mathematics, Chemistry and Biochemistry
First Advisor
Michelle K. Knowles
Second Advisor
Scott Horowitz
Third Advisor
Brian Michel
Fourth Advisor
Mark Siemens
Copyright Statement / License for Reuse
All Rights Reserved.
Keywords
Exocytosis, Exosomes, Interferon-alpha, Lung cancer cells, Multivesicular bodies tracking, Tetherin
Abstract
Cells rely on the secretion of exosomes to communicate with one another and this process is potentially mediated by a protein called tetherin, which anchors exosomes to the cell surface to limit their spread. Tetherin protein has attracted a lot of attention because of its capacity to hinder the propagation of viruses by attaching virions to the surface of host cells. Treatment with interferon-alpha (IFNα) increases the production of tetherin, although it is not known how this affects tetherin localization and exosome-mediated communication. This thesis looks at how the expression of tetherin, which IFNα boosts, affects the dynamics of intercellular communication through exocytosis or the exosome release in A549 (lung cancer) cells. Another poorly understood point is that the tetherin protein is predominately found in human cells though multiple studies proposed on cell membranes. The study uses a multidisciplinary methodology to visualize tetherin’s location both on membranes and in cytosol and colocalization with multivesicular bodies (MVBs) by combining immunofluorescence and Confocal microscope imaging. A nanoparticle (tracking) counter measures secreted extracellular vesicles in different conditions, while Total Internal Reflection Fluorescence (TIRF) microscopy makes examining MVBs fusion events at the cell membrane easier. MATLAB-based quantitative analysis clarifies how tetherin expression affects the kinetics of exosome release. The results of this study provide insights into possible treatment approaches that target tetherin expression and exosome mechanisms governing cellular defense and communication dynamics in diseases or infections spreading.
Copyright Date
11-2024
Publication Statement
Copyright is held by the author. User is responsible for all copyright compliance.
Rights Holder
Md. Rakib Miah
Provenance
Received from author
File Format
application/pdf
Language
English (eng)
Extent
96 pgs
File Size
9.2 MB
Recommended Citation
Miah, Md. Rakib, "Investigating the Role of Tetherin Protein in Multivesicular Bodies Fusions in Lung Cancer Cells" (2024). Electronic Theses and Dissertations. 2502.
https://digitalcommons.du.edu/etd/2502