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Applied Biosystems and Dr. Ronald P. Hart

A partnership for the advancement of science

Ronald P. Hart, Ph.D.

Professor and Associate Director of Cell Biology & Neuroscience at Rutgers, The State University of New Jersey

W.M. Keck Center for Collaborative Neuroscience
Stem Cell Research Center

Dr. Ronald P. Hart is recognized as a leading expert in microarray technologies in models of trauma to the central nervous system. Since establishing functional genomics technologies at the W. M. Keck Center for Collaborative Neuroscience in 2000, Dr. Hart has trained over 75 national and international scientists in the use of microarrays and actively works in partnership with dozens of researchers throughout the world.

We recently spoke with Dr. Hart about his work at the Keck Center and the newly unveiled New Jersey Stem Cell Research Center at Rutgers, and how Applied Biosystems is helping his group accelerate the pace of research in spinal cord injury, neurotrauma, and other neurological conditions through worldwide collaboration.

Tell us a little about your area of expertise, and what you're working on now.

I'm primarily a molecular biologist, but I've been working in the field of neuroscience since I joined the Rutgers faculty in 1985 and became interested in finding new ways to apply molecular biology to neuroscience and neuroimmunology.

In 2000, I was asked to join the Keck Center to provide functional genomics support for the work they were doing on treatments for spinal cord injury. The idea was that the gene expression side of the story could support some of the activities in therapy development and cell transplant work.

Most recently, we have turned our attention to therapeutic cellular transplants and how gene expression changes and molecular mechanisms can either be monitored or harnessed to control cells prior to a transplant for neurotrauma. I work with many laboratories around the world doing functional genomics and interpreting data in areas including immune response to spinal cord injury and regenerative growth of cells.

What made the Keck Center for Collaborative Neuroscience an appealing choice for you?

I felt that the application of functional genomics was best made in the service of many other areas of biology, so it was an exciting opportunity to make the best use of the things that I know—the technologies and the molecular biology. I also liked the idea of working in a small center. We have only four faculty, but it's very interactive with people outside of the group—"collaboration" is right in its name, and that is the key to the Center's success.

Can you give us some examples of those collaborative efforts?

We provided microarray-based functional genomics data and bioinformatics for Phil Popovich at Ohio State University. Our work triggered studies that found Toll-like receptors, which are fundamental cellular signaling mechanisms that are known to work in the immune system, are also triggered after spinal cord injury.

Another important collaboration was with Marie Filbin at Hunter College in New York, who specializes in the inhibitory molecules that prevent neuronal regeneration after spinal cord injury and how cells might be "tricked" to overcome this inhibition. We provided gene expression profiles for cells under various conditions that allowed her lab to identify several new targets for developing regenerative therapies.

These kinds of interactions have proved to be absolutely crucial in keeping things moving and adding value to the work of other researchers.

You've been using Applied Biosystems technologies for years. How have they contributed to the success of your research?

Yes, our use of Applied Biosystems technologies spans several generations of research. A lot of it is very mundane, day-to-day usage that is so important for the progress of any molecular biology lab. For example, it wasn't so long ago that we were doing routine DNA sequencing in our own labs, and now we rely on Applied Biosystems standardized reagents and equipment in core facilities. We've also done a lot of high throughput real-time quantitative PCR over the years using Applied Biosystems technologies.

About two years ago, we started using some of the gene expression tools like the 1700 array series, which I don't think many other people in academics were using at the time. The high-quality data we have derived from those experiments has been instrumental to our understanding of neurogenesis mechanisms. We've even gone so far as to take the array design that was originally constructed and re-annotate it using updated bioinformatics. We just had an article accepted that presented some of our new bioinformatics based on the original designed array that Applied Biosystems produced.

Why do you continue to choose Applied Biosystems over other technologies?

There are several sources for some of the technologies we're using, but it's a no-brainer to turn to the technologies that we know we can depend on to get the job done. In my lab, we have a lot of experience with Applied Biosystems for things like real-time PCR, and adapting them for genomics testing takes so little effort on our part because the tools are already available and proven. So when a vendor tells us they're going to sell us cheaper reagents, I absolutely resist the urge because I know what it would cost me in terms of loss of productivity. At one point, I was using some other quantitative PCR kits for micro RNAs and very quickly switched back to the Applied Biosystems TaqMan® kit version as soon as it came out because of the immediate success and reliability of it.

Let's talk about the new Stem Cell Research Center in New Jersey. What are the goals of the Center, both in the near term and down the road?

The near-term goal is to get the use of embryonic stem cells up and running and available to a large number of basic researchers throughout New Jersey. The key is the validation and documentation of those cells using molecular techniques, and having a core facility to do that is absolutely crucial in getting the work off the ground.

The ultimate goal is to develop human therapies for neurotrauma and other neurological conditions by feeding promising cells and protocols into the Stem Cell Institute of New Jersey, which is envisioned as focusing on translational medicine. Based on an enormous amount of work from many, many research sites, it is clear that transplanted stem cells in one form or another have the potential to restore more function than any other single treatment. Combining stem cells with other strategies is likely to be the first round of truly successful therapies for spinal cord injury and neurotrama as well as some of the degenerative diseases including Parkinson's, Alzheimer's, and MS.

How will Applied Biosystems help you in your work there?

One component of the core Stem Cell Research Center facility at Rutgers will be to derive new, genetically defined cells discarded or donated from IVF usage. The second component will be to expand and validate those cultures prior to distribution to research groups. That validation process will use a number of Applied Biosystems products, including the genomics assays and gene expression assays we will need to show a) that these cells are what we say they are, b) how they relate to other stem cells from other sources, and c) how they can be reproducibly provided to other researchers. As basic scientists, we don't normally think about FDA approval, but that is what is down the road and Applied Biosystems will help us prepare for that eventuality.

When it is fully operational, who will be using the Center?

I think it's going to be a large and very dynamic cast of characters coming through that space. The new lab will be very flexible because it is a state-funded facility with no Federal National Institutes of Health (NIH) dollars involved, so we will be allowed to work with non-NIH-approvable cell lines.

In the first phase, the Center will allow researchers from throughout New Jersey to come and spend time in the space to do their embryonic stem cell work without contaminating their NIH-funded research laboratories. The second phase will include stem cell scientists from other states and around the world. We've already got agreements with groups in Scotland and Spain and we are currently working with a group of Canadian stem cell researchers to expand this network.

Do you expect to take advantage of the new Applied Biosystems Application Support Center (ASC) in Foster City?

The opening of the new Support Center is timed perfectly for where we are with the Stem Cell Research Center. I can see using the ASC to explore how to incorporate the range of Applied Biosystems technologies that we've used in the past as well as some that are just becoming available now. We're also talking with Applied Biosystems about the variations and extensions to existing technologies that we will likely need in the future. For example, we're looking forward to using some of the existing fingerprinting technologies for making sure that we know the source of every cell line that we're studying. In parallel, we will need pathogen identification and mouse contamination tools that are under development now at Applied Biosystems—and we will be happy to work with Applied Biosystems scientists to help validate those new technologies. In the longer term, things like the next-generation SOLiD™ sequencing technology will be very important to us.

What excites you most about the future of the industry and your future work with Applied Biosystems?

One of the tools that I think will be incredibly important down the road is the next generation of DNA sequencing, including the SOLiD™ System technology. To get a stem cell to the point where we can transplant it into a human being, we're going to have to sequence that genome every time for every cell line. The availability of a reliable, accurate, and inexpensive high-throughput deep-sequencing technique is going to be absolutely essential for biological materials that are eventually going to go into humans.

We're really excited about where the field is going in this area, where the technologies are going, and clearly Applied Biosystems is helping to lead the advances. I'm quite anxious to see it in action and talk to the people who are developing and using it at Applied Biosystems.

Do you have any plans in the works to conduct collaborative research with Applied Biosystems scientists at the Foster City ASC?

Yes. The request right now is for some pilot experiments with the SOLiD™ system. Our early plans would be to get some expression profiles using deep sequencing of the micro RNAs in differentiating human stem cells. As we go forward and learn more, I'm sure there will be more specific and specialized interactions.

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