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Principal Investigator: Ariana Kathryn von Lersner
Organization: VANDERBILT UNIVERSITY
Fiscal Year: 2021
Award: $30,754
Funding agency: National Cancer Institute
PROJECT SUMMARY
Extracellular vesicles (EVs) are a heterogenous group of lipid bilayer enclosed particles secreted from cells and are
found in most, if not all, biological fluids. The term EV encompasses a wide range of vesicle classes, including exosomes
(50-150 nm) and microvesicles (150 nm - 1000+ µm), derived from the endosomal pathway after fusion of multivesicular
bodies with the plasma membrane or from direct budding from the plasma membrane, respectively. EVs are further
characterized by membrane and luminal cargoes that give rise to distinct functional properties. An emerging body of
literature suggests that EVs are major contributors in cell-cell communication in normal physiology and pathological events,
such as cancer. EVs have been shown to affect cancer cell chemotaxis and metastatic organotropism. Notably, a specific
class of EV is dependent upon syntenin-1, an adaptor protein that contains two PSD95/Dlg/zonula occludens 1 (PDZ)
domains, for biogenesis and cargo loading through the endosomal pathway. This suggests that upstream effectors which
bind to syntenin-1 could modulate its availability, and in turn, regulate exosome biogenesis and cargo loading. Activated
Leukocyte Cell Adhesion Molecule (ALCAM) is an adhesion protein in the immunoglobulin superfamily found at sites of
cell-cell junctions. ALCAM is a dynamic regulator of cell adhesion through differential shedding of its ectodomain. Our
laboratory demonstrated that the full-length ALCAM Isoform (Iso1) resists shedding, promotes adhesion and limits
metastasis. Conversely, the alternatively spliced isoform 2 (Iso2) is susceptible to shedding, facilitates motility and
metastasis. ALCAM shedding and elevated expression of ALCAM Iso2 correlates with bladder cancer (BCa) disease
progression. ALCAM can be linked to EV biogenesis through its association with the cytoplasmic scaffolding protein
syntenin-1. Sequestering syntenin-1 via the cytoplasmic tail of ALCAM abrogates motility and metastasis, further
underscoring the functional relevance of the regulatory mechanism. Additional preliminary studies revealed that the
expression of Iso1 suppresses large EV biogenesis while Iso2-facilitates the biogenesis of large EVs. Based on these findings
we hypothesize that ALCAM shedding controls syntenin-dependent EV biogenesis in tumor cells. In addition, we
hypothesize that ALCAM-mediated adhesion can control the pro-migratory function of EVs by regulating their cargo. To
test this hypothesis, I will utilize cell lines expressing different isoforms of ALCAM and BCa patient-derived plasma to
determine the mechanism in which ALCAM-mediated cell adhesion affects EV biology. Experiments proposed in Aim 1
will investigate how modulation of a cells' adhesive state through ALCAM expression alters EV biogenesis and function.
EV biogenesis will be assessed through complementary techniques of nanoparticle tracking analysis, western blotting, and
microflow cytometry. EV function will be determined with the avian embryo model of metastasis and a 3D organotypic
bladder model. Aim 2 will characterize the changes in EV cargos upon ALCAM shedding and disease progression in BCa
through palmitoylated-mass spectrometry. Collectively, these data will define a relationship between cell adhesion EV
biology, both mechanistically and functionally. Additionally, these findings will identify informative molecular markers to
aid in the monitoring of disease state through a non-invasive means.
Terms: <3-D><3-Dimensional><3D><ALCAM><ALCAM gene><Activated-Leukocyte Cell Adhesion Molecule><Adaptor Protein><Adaptor Protein Gene><Adaptor Signaling Protein><Adaptor Signaling Protein Gene><Adhesions><Adhesives><Affect><Alternate Splicing><Alternative RNA Splicing><Alternative Splicing><Aves><Avian><Binding><Biogenesis><Biological><Biology><Birds><Bladder><Bladder Cancer><Bladder Urinary System><Bladder papilloma><Blood Plasma><CD166><CD166 Antigens><CD6 Ligand><Cancer Patient><Cancer Relapse><Cancers><Carcinoma><Cell Adhesion><Cell Body><Cell Communication><Cell Interaction><Cell Junctions><Cell Line><Cell Locomotion><Cell Migration><Cell Movement><Cell membrane><Cell-to-Cell Interaction><CellLine><Cells><Cellular Adhesion><Cellular Migration><Cellular Motility><Chemotaxis><Cytometry><Cytoplasmic Domain><Cytoplasmic Membrane><Cytoplasmic Tail><DLG4><DLG4 gene><Data><Detection><Development><Disease><Disease Progression><Disorder><Electron Microscopy><Embryo><Embryonic><Environment><Epithelial cancer><Event><Goals><Immune Globulins><Immunoblotting><Immunoglobulin Domain><Immunoglobulin-Like Domain><Immunoglobulins><In Vitro><Intercellular Junctions><Intervention><Intervention Strategies><Isoforms><KG-CAM><Laboratories><Length><Leu-CAM Receptor Family><Leukocyte Adhesion Molecules><Leukocyte-Adhesion Receptor Family><Leukocyte-Adhesion Receptors><Link><Lipid Bilayers><Liquid substance><Literature><MEMD><Malignant Bladder Neoplasm><Malignant Cell><Malignant Epithelial Neoplasms><Malignant Epithelial Tumors><Malignant Neoplasms><Malignant Tumor><Malignant Tumor of the Bladder><Malignant neoplasm of urinary bladder><Mass Photometry/Spectrum Analysis><Mass Spectrometry><Mass Spectroscopy><Mass Spectrum><Mass Spectrum Analyses><Mass Spectrum Analysis><Mediating><Mediator><Mediator of Activation><Mediator of activation protein><Membrane><Metastasis><Metastasize><Metastatic Lesion><Metastatic Mass><Metastatic Neoplasm><Metastatic Tumor><Modeling><Molecular><Molecular Interaction><Monitor><Motility><Multivesicular Body><Neoplasm Metastasis><Occluding Junctions><Origin of Life><PSD95><Pathologic><Pathway interactions><Physiology><Plasma><Plasma Membrane><Plasma Serum><Production><Property><Protein Analysis><Protein Isoforms><Proteins><Publishing><Regulation><Research><Reticuloendothelial System, Serum, Plasma><Risk><SAP90><Scaffolding Protein><Secondary Neoplasm><Secondary Tumor><Site><Strains Cell Lines><Techniques><Testing><Tight Junctions><Tumor Cell><Tumor Cell Migration><Tumor-Derived><Urinary Bladder Cancer><Urinary Bladder Malignant Tumor><Urine><Urine Urinary System><Urothelial Papilloma><Vesicle><Western Blotting><Western Immunoblotting><Work><Zonula Occludens><adapter protein><base><cancer cell><cancer metastasis><cancer progression><cell motility><clinical relevance><clinically relevant><cultured cell line><developmental><epithelial carcinoma><exosome><experiment><experimental research><experimental study><extracellular><extracellular vesicles><fluid><in vivo Model><insight><interventional strategy><lipid bilayer membrane><liquid><liquid biopsy><malignancy><member><membrane structure><microvesicles><migration><molecular biomarker><molecular marker><mortality><nano particle><nano-sized particle><nanoparticle><nanosized particle><neoplasm progression><neoplasm/cancer><neoplastic cell><neoplastic progression><novel><particle><pathway><plasmalemma><prevent><preventing><protein blotting><response><response to treatment><synergism><targeted drug therapy><targeted drug treatments><targeted therapeutic><targeted therapeutic agents><targeted therapy><targeted treatment><therapeutic response><three dimensional><treatment response><treatment strategy><tumor cell metastasis><tumor progression><urinary bladder>