DNA 'barcoding' allows rapid testing of nanoparticles for therapeutic delivery

February 07, 2017

Using tiny snippets of DNA as "barcodes," researchers have developed a new technique for rapidly screening the ability of nanoparticles to selectively deliver therapeutic genes to specific organs of the body. The technique could accelerate the development and use of gene therapies for such killers as heart disease, cancer and Parkinson's disease.

Genetic therapies, such as those made from DNA or RNA, are hard to deliver into the right cells in the body. For the past 20 years, scientists have been developing nanoparticles made from a broad range of materials and adding compounds such as cholesterol to help carry these therapeutic agents into cells. But the rapid development of nanoparticle carriers has run into a major bottleneck: the nanoparticles have to be tested, first in cell culture, before a very small number of nanoparticles is tested in animals. With millions of possible combinations, identifying the optimal nanoparticle to target each organ was highly inefficient.

Using DNA strands just 58 nucleotides long, researchers from the University of Florida, Georgia Institute of Technology and Massachusetts Institute of Technology have developed a new testing technique that skips the cell culture testing altogether -- and could allow hundreds of different types of nanoparticles to be tested simultaneously in just a handful of animals.

The original research was done in the laboratories of Robert Langer, the David H. Koch Institute Professor, and Daniel Anderson, the Samuel A. Goldsmith Professor of Applied Biology, at MIT. Supported by the National Institutes of Health, the research was reported February 6 in the journal Proceedings of the National Academy of Sciences.

"We want to understand at a very high level what factors affecting nanoparticle delivery are important," said James Dahlman, an assistant professor in the Wallace H. Coulter Department of Biomedical Engineering at Georgia Tech and Emory University, one of Langer's former graduate students, lead author on the study, and one of the paper's corresponding authors. "This new technique not only allows us to understand what factors are important, but also how disease factors affect the process."

To prepare nanoparticles for testing, the researchers insert a snippet of DNA that is assigned to each type of nanoparticle. The nanoparticles are then injected into mice, whose organs are then examined for presence of the barcodes. By using the same technologies scientists use to sequence the genome, many nanoparticles can be tested simultaneously, each identified by its unique DNA barcode.

Researchers are interested not only in which nanoparticles deliver the therapeutics most effectively, but also which can deliver them selectively to specific organs. Therapeutics targeted to tumors, for example, should be delivered only to the tumor and not to surrounding tissues. Therapeutics for heart disease likewise should selectively accumulate in the heart.

While much of the study was devoted to demonstrating control strategies, the researchers did test how 30 different particles were distributed in eight different tissues of an animal model. This nanoparticle targeting 'heat map' showed that some particles were not taken up at all, while others entered multiple organs. The testing included nanoparticles previously shown to selectivity enter the lungs and liver, and the results of the new technique were consistent with what was already known about those nanoparticles.

The single-strand DNA barcode sequences are about the same size as antisense oligonucleotides, microRNA and siRNA being developed for possible therapeutic uses. Other gene-based therapeutics are larger, and additional research would be needed to determine if the technique could be used with them. In the research reported this week, the nanoparticles were not used to deliver active therapeutics, though that would be a near-term next step.

"In future work, we are hoping to make a thousand particles and instead of evaluating them three at a time, we would hope to test a few hundred simultaneously," Dahlman said. "Nanoparticles can be very complicated because for every biomaterial available, you could make several hundred nanoparticles of different sizes and with different components added."

Once promising nanoparticles are identified with the screening, they would be subjected to additional testing to verify their ability to deliver therapeutics. In addition to accelerating the screening, the new technique may require fewer animals -- perhaps no more than three for each set of nanoparticles tested.

There are a few caveats with the technique. To avoid the possibility of nanoparticles merging, only structures that are stable in aqueous environments can be tested. Only nontoxic nanoparticles can be screened, and researchers must control for potential inflammation generated by the inserted DNA.

In Langer and Anderson's laboratory, Dahlman worked with Kevin Kauffman, who remains at MIT, and Eric Wang, now an assistant professor the University of Florida. Other co-authors of the paper included Yiping Xing, Taylor Shaw, Faryal Mir and Chloe Dlott, all of whom are at MIT.

"Nucleic acid therapies hold considerable promise for treating a range of serious diseases," said Dahlman. "We hope this technique will be used widely in the field, and that it will ultimately bring more clarity to how these drugs affect cells -- and how we can get them to the right locations in the body."

Georgia Institute of Technology

Related Heart Disease Articles from Brightsurf:

Cellular pathway of genetic heart disease similar to neurodegenerative disease
Research on a genetic heart disease has uncovered a new and unexpected mechanism for heart failure.

Mechanism linking gum disease to heart disease, other inflammatory conditions discovered
The link between periodontal (gum) disease and other inflammatory conditions such as heart disease and diabetes has long been established, but the mechanism behind that association has, until now, remained a mystery.

New 'atlas' of human heart cells first step toward precision treatments for heart disease
Scientists have for the first time documented all of the different cell types and genes expressed in the healthy human heart, in research published in the journal Nature.

With a heavy heart: How men and women develop heart disease differently
A new study by researchers from McGill University has uncovered that minerals causing aortic heart valve blockage in men and women are different, a discovery that could change how heart disease is diagnosed and treated.

Heart-healthy diets are naturally low in dietary cholesterol and can help to reduce the risk of heart disease and stroke
Eating a heart-healthy dietary pattern rich in vegetables, fruits, whole grains, low-fat dairy products, poultry, fish, legumes, vegetable oils and nuts, which is also limits salt, red and processed meats, refined-carbohydrates and added sugars, is relatively low in dietary cholesterol and supports healthy levels of artery-clogging LDL cholesterol.

Pacemakers can improve heart function in patients with chemotherapy-induced heart disease
Research has shown that treating chemotherapy-induced cardiomyopathy with commercially available cardiac resynchronization therapy (CRT) delivered through a surgically implanted defibrillator or pacemaker can significantly improve patient outcomes.

Arsenic in drinking water may change heart structure raising risk of heart disease
Drinking water that is contaminated with arsenic may lead to thickening of the heart's main pumping chamber in young adults, according to a new study by researchers at Columbia University Mailman School of Public Health.

New health calculator can help predict heart disease risk, estimate heart age
A new online health calculator can help people determine their risk of heart disease, as well as their heart age, accounting for sociodemographic factors such as ethnicity, sense of belonging and education, as well as health status and lifestyle behaviors.

Wide variation in rate of death between VA hospitals for patients with heart disease, heart failure
Death rates for veterans with ischemic heart disease and chronic heart failure varied widely across the Veterans Affairs (VA) health care system from 2010 to 2014, which could suggest differences in the quality of cardiovascular health care provided by VA medical centers.

Heart failure: The Alzheimer's disease of the heart?
Similar to how protein clumps build up in the brain in people with some neurodegenerative diseases such as Alzheimer's and Parkinson's diseases, protein clumps appear to accumulate in the diseased hearts of mice and people with heart failure, according to a team led by Johns Hopkins University researchers.

Read More: Heart Disease News and Heart Disease Current Events
Brightsurf.com is a participant in the Amazon Services LLC Associates Program, an affiliate advertising program designed to provide a means for sites to earn advertising fees by advertising and linking to Amazon.com.