biology

Just What the Doctor Ordered

+
medicine

Facilitation of
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Bu y El
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Wound Healing
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From the Lab Bench to the Clinic
Although the preclinical work with engineered versions of
EGF now occurszat Stanford, Dr. Cochran began creating the
mutant EGF proteins several years earlier. As a postdoctoral
fellow at the Massachusetts Institute of Technology,
Cochran used “directed evolution” to modify the DNA that
codes for EGF by replicating it tens of millions of times
under a mutation-producing process. Then, in order to find
modified EGF
that would be

R
esearch findings from a bioengineer’s lab bench have an most helpful
exciting new application: wound healing. In collaboration in wound

Photo Credit: www.johntodd.com

with the Department of Surgery at the Stanford University healing, she
School of Medicine, Jennifer Cochran, Assistant Professor of sorted through
Bioengineering, is working on expediting the process that the the mutants
body uses to heal skin wounds by delivering a mutated version of with a high-
a naturally occurring protein. throughput
assay to
Natural Wound Healing select ones
Epidermal growth factor (EGF), a naturally occurring that would
protein, is crucial to the process of healing cuts as healthy associate
skin cells gather and multiply at the affected area. By most strongly Cochran shows surgery Pro-
attracting fibroblasts (connective tissue with EGF receptors. Since her fessor Michael Longaker a
cells that produce collagen) to the area initial work at MIT, Cochran has beaker in which yeast are
of the wound, EGF helps to build new found dozens of EGF mutants with making EGF.
skin tissue. Dr. Michael Longaker, approximately 4 - 30 times more
Cochran’s collaborator, explains receptor-binding strength than the naturally occurring
the everyday importance of EGF in growth factor. Experiments are on-going to determine if this
pointing out that “animals lick their increase in receptor binding affinity will cause the engineered
Photo Credit: Dr. Jennifer Cochran

Since her initial work at MIT, Cochran has found dozens of EGF
mutants with approximately four to thirty times more receptor-
binding strength than the naturally occurring growth factor.

EGF (red) binding to wounds because their saliva includes
a portion of the EGF a high concentration of EGF” (News
receptor (grey). Service). After initially being attracted
by EGF, fibroblasts produce their own
EGF, causing keratinocytes (cells forming the outer layer of
skin) to multiply and close the wound. While EGF is one
of nature’s powerful own medicines, it only exists at the
wound site for a short time. Therefore, the Cochran lab has
engineered mutated EGF that hopefully will last longer at
the wound site to promote enhanced healing.
EGF to stay at a wound site for longer periods of time.
While Dr. Cochran created these enhanced proteins at
MIT, by coming to Stanford she has been able to collaborate
with physicians in the Department of Surgery. Cochran
explains, “The clinical collaboration here with Drs. Michael
Longaker and George Yang in the Department of Surgery
has now allowed us to take this project to the next level.
We can start looking at the therapeutic efficacy of these
proteins” (Science News). This past April, the Bioengineering
Department granted $100,000 to Cochran, Longaker, Yang

18 stanford scientific
 biology
and research fellows Dr. Daphne Ly, Dr. Stayce Beck, and +
medicine
Stephen Lee to develop therapeutic applications using these
engineered EGF proteins.

Potential applications of EGF include

“accelerate[d] healing in gastric and
oral ulcers, skin graft donor sites,
corneal epithelial wounds, and ear
drum perforations.”
Mutated Epidermal Growth Factor
The engineered EGF proteins were recombinantly
expressed in yeast cells for preclinical testing. The group
has utilized cell-based “scratch assays” in which a wound is
simulated by scratching a 2-millimeter line through a thin
layer of tissue in a Petri dish and engineered EGF proteins are
added in a saline solution. Preliminary tests have indicated
that mutant EGF causes cells to fill this artificial “wound”

Cochran’s findings offer special

promise to diabetic patients who
suffer from chronic skin lesions.
faster than with natural EGF, indicating a potential for in
vivo wound healing applications. After the scratch assay
tests are complete, the group will test the most promising
mutated proteins on mice.

Looking to the Future

While Cochran’s findings about the mutated form of EGF
give hope to anyone who experiences a wound, they also offer
special promise to diabetic patients who suffer from chronic
skin lesions due to their neutrophils remaining in the wound
site for too long and starting to clear out new tissue instead
of damaged tissue. Cochran claims that other potential
applications of EGF include “accelerate[d] healing in gastric
and oral ulcers, skin graft donor sites, corneal epithelial
wounds, and ear drum perforations.” Additionally, EGF may
be used to regulate nerve injury response and regeneration.
Cochran’s team strives to develop a form of EGF that will
treat chronic wounds that are incurable by natural healing
and believes that this new approach will be more effective,
cheaper and easier to use than alternative solutions for
Scratch assay results after addition of EGF show that Cochran’s mutated version
wound healing. S only takes 48 hours to heal the wound (middle) while it takes 72 hours for
natural EGF to do its job (bottom).

ELIZABETH BURSTEIN is a freshman from Cupertino, California. She is To Learn More:

thinking of majoring in Biomechanical Engineering, with possible
minors in Psychology or French. She also enjoys assisting research, For more information, read Dr. Cochran’s research abstract
dancing, soccer, volleyball, running, playing the piano, and event at http://peds.oxfordjournals.org/cgi/content/abstract/
planning with the Frosh Council. 19/6/245u

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