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James Belknap, DVM, PhD

Award Name CCTS Pilot 2009
Award Date 04/01/2009

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Laser Capture Microdissection in Equine Laminitis

The CCTS Pilot and Collaborative Translational and Clinical Science Program has awarded James Belknap, DVM, PhD, a pilot grant to be used for his research of equine laminitis.

Laminitis is a crippling disease that affects horses. Similar to organ injury in humans with sepsis, it entails an inflammatory injury of the laminae, a soft tissue (of the same cell types that make up our skin) which interdigitates to support the horses’ third phalanx (coffin bone).

This life threatening disease is extremely painful and can be onset by colic, enterocolitis, pleuropneumonia, and metritis.

“It is a devastating disease,” Belknap said. As an equine surgeon, he commonly treats clinical cases of the disease at the OSU Veterinary Teaching Hospital.

Belknap believes that the first step to fighting laminitis is to find out what signaling events in the foot prompt the disease to occur.

“There is a layer of cells attached to the hoof wall called the basal epithelial cells that lose their attachment to the underlying connective attached to the third phalanx,” Belknap said.

This detachment results in the third phalanx tearing loose from the hoof wall and displacing toward the ground.

“It can actually perforate the sole. We need to find out what signaling is going on in these cells that causes them to lose their adherence,” Belknap said.

In order to find out what the signaling events are, the team is using laser capture microdissection to collect and isolate RNA samples from laminar epithelial cells.

“At this point it’s sort of a black box of what’s going on with these cells,” said Belknap.

By using laser capture microdissection to collect the specific layer of cells at the point of failure, a better understanding of the role that the laminar epithelial cells play in this disease will become clear.

Graduate student Britta Leise, MS, DVM, is working in the Belknap laboratory to collect the RNA samples. Once the cells have been collected by Leise, they will be sent to Texas for microarray analysis and gene networking. The cells afflicted with laminitis will be compared to those of clinically normal horses.

“The analysis of these data will focus in on the cellular dysregulation that result in structural failure of the digit in the disease,” Belknap said.

Once the points of dysregulation are identified, Belknap believes that it would be more likely to find an effective combination of medical treatments to combat the crippling disease.

By Becky King, Monday, August 23, 2010

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