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Using a super-resolution microscope,
researchers in Jessica Tanis' lab can image
extracellular vesicles, made visible by a green fluorescent
protein. "To someone who has never looked at EVs before, they look like
stars in the sky," Tanis says. On the right is a portion of the worm
model she uses.
have known for decades that our cells constantly shed tiny pieces known
as extracellular vesicles (EVs), but until relatively recently they
believed that this process was merely a way for cells to rid themselves
of material that was no longer needed.
“For a long time, EVs were thought of as a nonspecific garbage
disposal mechanism,” said Jessica Tanis, assistant professor in the Department of Biological Sciences at the University of Delaware.
As it turns out, nothing could be further from the truth.
Scientists have learned that EVs, despite their extremely small size,
have enormous signaling potential and play a critical role in
communication between cells. They carry unique “cargos” of proteins,
nucleic acids and other materials from the cell that released them. The
transfer of these bioactive molecules by EVs is essential for the
development, and contributes to the progression, of conditions such as
cancer and neurodegenerative diseases.
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“My lab is working on increasing our understanding of these tiny
vesicles that can give you a glimpse into what’s going on in a
particular cell,” said Tanis, whose team was recently awarded a $1.8
million National Institutes of Health (NIH) grant for this research.
“Remarkably, an individual cell can release multiple populations of
EVs, each containing different cargos. We hope that our findings will
provide new insights into how a cell packages and releases these
distinct subsets of EVs. What are the cargos in each particular EV
subtype? What determines which EV a cargo is loaded into? What are the
functions of the different EVs?”
Those are among the questions her research group, which includes
graduate and undergraduate students and a postdoctoral researcher, is
seeking to answer. The NIH grant, “Elucidating biogenesis and cargo
sorting mechanisms for discrete extracellular vesicle subpopulations,”
will support the project for five years.
Tanis works with a worm model, C. elegans, using fluorescently labeled proteins and powerful microscopes housed at UD’s Bioimaging Center
to identify individual EVs. Doctoral student Michael Clupper said the
research is possible only because of the highly sophisticated imaging
equipment available at the center, which is part of the Delaware
Doctoral student Michael Clupper uses UD’s Andor Dragonfly
super-resolution microscope to look for fluorescent extracellular
Clupper came to UD for graduate school and began working on a project
in Tanis’ lab involving a protein potentially linked to Alzheimer’s
disease. When he discovered the protein packaged into EVs, his research
shifted its focus to this new area — and a new skill he’s developed in
working with the Bioimaging Center’s instruments.
“Few research institutions have the means to image EVs,” Clupper
said. “We’re very fortunate at UD. The generalized term for these scopes
is ‘super-resolution microscopes’ because they let you image down to a
single molecule level.”
He and Tanis praised the assistance they’ve received from Jeffrey
Caplan, the imaging center director, and his staff as they learned and
fine-tuned their work with the instruments.
“We’ve had so much support and help,” Tanis said. “And this project
wouldn’t be possible without Mike’s persistence in optimizing the
complex imaging and analysis techniques.”
The biggest technical obstacle lies in the size of the EVs being studied.
“Imaging EVs is so challenging because of how small they are and how
difficult it can be to reliably label them [with fluorescence] in a way
that makes them visible to microscopes,” said Clupper, who now helps
train other members of the research team in the imaging techniques he
uses. “One of the challenges with pioneering work like this is that
there isn’t really a manual or protocol; you have to figure it out
Ultimately, he said, he hopes the work will pave the way for other EV
researchers to use some of the tools and methods that the lab has
developed as they explore new avenues of investigation.
And a great deal of research is being conducted on EVs, Tanis said.
Scientists see potential for using them as biomarkers to diagnose
disease and for engineering them to carry genetic cargo and drugs to
targets within the body as an innovative therapeutic approach.
Article by Ann Manser; photo by Timothy Chaya
Published Feb. 3, 2021