Researchers create cell lines to help treat childhood mitochondrial diseases

Mitochondria are well-known for their role as "cell power source"

Mitochondrial diseases are a group of debilitating genetic diseases.

But recent research published in the journal Mitochondrial and BMC Molecular and Cell Biology Aloka Obey Bandara, an associate professor in the Department of Research Biomedical Sciences and the Virginia-Maryland School of Veterinary Medicine and his team provides a glimmer of hope for patients with mitochondrial diseases and their parents

Bandara and Virginia Tech researchers from Blacksburg and Roanoke successfully created a living cell model that mimics mitochondrial disease cells

"Our cell model will allow us to see what happens to the cell and its processes when a child develops a mitochondrial disease

Our body produces life-sustaining energy from the food we eat and the air we breathe

Through a series of reactions, the electron transport chain can remove electrons from nutrients and push them through the mitochondrial membrane, thereby forming a proton gradient

Bandara said: "Sometimes, you can see damage or mutations in the protein of the electron transport chain

The electron transport chain is composed of five protein complexes or groups of proteins

Patients with mitochondrial diseases may have complex I or complex II defects

Although researchers can pinpoint the location of the defect, creating treatments for these mitochondrial diseases has always been a challenge

Bandara hopes that his cell line will not only support the upcoming research, but also support patients and their families who are experiencing mitochondrial disease and all its direct effects

Bandara said: "Parents are often helpless because they can't go to the pharmacy to buy medicine

In order to test drug candidates, researchers must first create cell models as artificial "sick" cells

In order to create cells that mimic mitochondrial diseases, Bandara must use CRISPR/Cas9 technology to "knock out" the parts of the genome that generate complex I and complex II

First, the researchers identified the parts of the genome that needed to be deleted
.
Then, they designed a piece of RNA as their "base"
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Then, the RNA "guides" an enzyme called Cas9 to the gene's home base
.
Cas9 can bind to that point and "cut" it
.

After this process was completed, Bandara performed genome sequencing to confirm that the part was successfully deleted from the genome
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After months of hard work, Bandara and his team created two mutant cell lines, one without complex I and the other without complex II
.

Bandara is one of the few researchers on Virginia Tech's Blacksburg campus to use CRISPR/Cas9 technology to treat mitochondrial diseases
.

After cultivating the mutant cell lines, Bandara placed them in a disease model in which he tested the function of the "diseased" cell line and the "parental" cell line composed of healthy cells
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Through careful analysis, Bandara confirmed that diseased cells consume much less oxygen, grow very slowly, and cannot produce enough ATP to make the cells work normally-the three signs of mitochondrial disease cells
.

Once they confirmed that the knockout cell line correctly mimics the cellular dysfunction of mitochondrial disease, they were able to test a newly developed drug called idefenone
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Bandara research shows that this treatment can restore cell growth and oxygen consumption to a certain extent
.

These cell lines are the result of a fruitful collaboration between experts from the Department of Human Nutrition, Food and Exercise and the Carilion School of Medicine at Virginia Tech
.

The construction of the mutant cell line was directed and supported by David Brown, a former associate professor in the Department of Human Nutrition, Food and Exercise in the School of Agriculture and Life Sciences at Virginia Tech, and is now the Senior Director of Technological Innovation at Stealth BioTherapeutics
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A biotech company in Boston
.

Through this work, the team has obtained two provisional patents for their cells
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One of the cell lines has been patented and licensed to a pharmaceutical company that will develop new therapies for patients with mitochondrial diseases
.

These cells have been provided to interested researchers and pharmaceutical companies around the world through Ximbio.
Ximbio is the world's largest non-profit organization specializing in all types of life science research tools
.

"The cell model with mitochondrial complex I and II defects has a high social and economic impact and serves as a model for testing candidate drugs for mitochondrial dysfunction treatment in a cost- and time-effective manner," said Justin Perry, a graduate of Virginia Tech Sheng, now the business development manager of Ximbio
.

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