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Magnetic Cell Therapy

A new technique uses a magnetic field to guide potential therapies to stents in clogged blood vessels.

Stents are expandable stainless-steel scaffolds commonly used to prop open clogged arteries. But inserting a stent can damage an artery's inner lining, and stented arteries may reclose after several months, causing blood clots and possibly heart attacks. Now researchers at the Children's Hospital of Philadelphia have devised a way to use tiny iron-bearing nanoparticles and a magnetic field to direct cells with therapeutic properties to the sites of steel stents. The cells could help repair arterial damage and prevent clotting, among other things.

"Stents have been known to induce severe trauma," says Robert Levy, chair of pediatric cardiology at the Children's Hospital of Philadelphia. "Repairing blood vessels with cell therapy is a very important concept that can be realized with magnetic targeting."

Levy and his colleagues engineered nanoparticles, or tiny spheres, of polylactic acid, a biodegradable polymer used in sutures and other medical applications. The team then loaded each nanoparticle with a small dose of magnetically responsive iron oxide and inserted it into a bovine endothelial cell--a cell found in a blood vessel's inner lining. The bovine cells were genetically altered to express a fluorescent marker, making them easily detectable.

Next, the researchers surgically implanted small metal stents in the carotid arteries of live rats. They injected the rats with a solution of treated endothelial cells and created a steady magnetic field around each rat using two large, external electromagnetic coils. Levy says that the magnetic field he and his colleagues applied was less than a tenth of the strength of the fields generated by conventional MRI machines. After 48 hours, the team created images of the rat using bioluminescence imaging.

The researchers found that the magnetic field caused the cells to migrate to the metal stents under two scenarios: when cells were injected directly into the carotid artery, near the stent location, and when they were injected farther away, in the aortic arch, whence they could have branched out to all areas of the body. In tests that didn't use a magnetic field, the cells migrated throughout the body with little direction.

Magnetically directing cells, particularly endothelial cells, to the sites of metal stents may have a significant therapeutic effect, says Levy. During surgical implantation, stents tend to scrape off endothelial cells, whose normal functions include helping prevent blood clotting. Endothelial cells are also barriers to inflammatory cells. While inflammatory cells normally flock to an injury to help repair it, in the absence of endothelial cells, they build up excessively, creating arterial blockage. In recent years, stents have been engineered to release anticlotting drugs to prevent arteries from reclosing. But such drug-releasing stents have problems of their own, including preventing endothelial cells from regenerating.

"Two years ago, clinicians noticed that patients in significant numbers were having problems with these stents, probably because the endothelium wasn't properly healed," says Levy. "Clotting, myocardial infarctions, and sudden deaths occurred, and this has caused a big uproar over stent usage."

Levy hopes that magnetically directing new endothelial cells to blood vessels may solve many of the problems that stents currently face. His team plans to continue experimenting on rats, using endothelial cells derived from rats instead of cows, to minimize risk of rejection. Now that he has found a way to direct cells to metal stents, Levy is also looking at other potential therapies, including nitric oxide, which is known to relax and dilate blood vessels. He is currently engineering cells to genetically express enzymes that produce nitric oxide, and he will eventually load them with iron-oxide nanoparticles that will drive them to the sites of stents, further opening arteries.

Levy adds that the magnetic-based technique has applications outside of cardiovascular therapy. For example, in treating lung cancer, clinicians often use metal stents to keep airways open. However, a patient's tumor may continue to grow, eventually obstructing the passage despite the stenting. Magnetically targeted therapies could help deliver specific drugs to stent sites to treat tumors, in addition to keeping airways open.

"Metallic implants are also widely used in other areas, like orthopedics, for complex fractures, and correcting spinal curvature, where cell therapies could also be helpful," says Levy. "Steel implants are widely used in medicine, and there are all sorts of situations where applications could be used."

What's more, Levy envisions that such therapies can be applied using conventional MRI machines. The magnetic field generated by MRI cores is an order of magnitude more powerful than the ones Levy used in his experiments, so fewer iron-oxide nanoparticles could produce the same effect.

Robert Langer, Institute Professor at MIT, believes that Levy's technique is a promising step toward directed cell therapies. "They were able to localize more drugs into the targeted areas," he says. "I think it's a neat idea that has a lot of potential."

http://www.technologyreview.com/Biotech/20022/

Next Steps for Stem Cells

Searching the brain of an Alzheimer's patient for clues into the origin of the disease is like trying to find the cause of a plane crash in the wrecked aftermath. However, a recent breakthrough in stem-cell research could generate new cellular models that allow scientists to study disease with unprecedented accuracy, from its earliest inception to a cell's final biochemical demise.

Last November, two groups of scientists announced that they had independently achieved one of the stem-cell field's biggest goals: the ability to reprogram adult cells into embryonic-like stem cells without the need for human embryos. (See "Stem Cells without the Embryos.") The findings garnered extensive media attention, largely because the new method obviated the need for human embryos, a major ethical minefield that has stymied research.

But scientists at stem-cell labs around the world are excited for another reason. The technique creates cells that are genetically matched to an individual, meaning that it's now possible to create novel cell models that capture all the genetic quirks of complex diseases. "Being able to have human cells with human disease in a dish accessible for testing is a real boon to technology and to science," says Evan Snyder, director of the Stem Cells and Regeneration Program at the Burnham Institute, in La Jolla, CA.

While animal models exist for many human diseases, they typically only incorporate certain aspects of the disease and can't capture the complexity of human biology. In addition, some disorders known to have a significant genetic component, such as autism, have proved difficult to model in animals.

To reprogram cells, scientists from Wisconsin and Japan independently engineered skin cells to express four different genes known to be expressed in the developing embryo. For reasons not yet clear to scientists, this treatment turns back the developmental clock. The resulting cells are pluripotent, meaning that they can develop into any type of cell in the body, and they can apparently divide indefinitely in their undifferentiated state. The first two published studies on the new technique reprogrammed cells from a skin-cell line, while a third study, published last month, generated stem cells from the skin biopsy of a healthy volunteer.

No one has yet generated cell lines from a patient, although scientists have been talking about doing so for years. Previously, the only way to make such models for complex genetic diseases was through human therapeutic cloning, also known as nuclear transfer, which is fraught with technical and ethical issues and has not yet been achieved. (See "Stem Cells Reborn" and "The Real Stem Cell Hope.") "Assuming that these procedures are as easy to do as it seems, it's definitely more tractable than nuclear transfer," says Snyder. His own lab is trying to generate such models, as is "probably everyone else you could call on your rolodex," he says.

To generate a disease-specific cell model, scientists would take some cells from a patient with a particular disease and revert them to an embryonic state. The cells would then be prodded to develop into the tissue type damaged in that disease, such as dopamine neurons in Parkinson's disease or blood cells in sickle-cell anemia. By comparing the differentiation process in cells derived from healthy and diseased people, scientists could observe how that disease unfolds at a cellular level. They could also use the cells to test drugs that might correct those biochemical abnormalities. "We want to use these cells to ask and answer questions that can't be asked and answered any other way," says M. William Lensch, a research scientist at the Harvard Stem Cell Institute and Children's Hospital Boston.

The relative simplicity of the approach--and the fact that it can be supported by federal funding--means that many more scientists are likely to attempt reprogramming than cloning. (In 2001, President Bush limited federal funding for embryonic stem-cell research to embryonic stem-cell lines already in existence.) According to Story Landis, chair of the Stem Cell Task Force at the National Institutes of Health, in Bethesda, MD, the funding agency has already announced two programs to fund reprogramming research and would welcome applications to derive cell lines from patients.

While no one has yet announced that he or she has derived a disease-specific cell model, George Daley's lab at Harvard may be in the lead. Last month, he and his team published a paper in Nature showing that they can reprogram cells from a skin biopsy from a healthy person, and they are already trying to repeat the feat with tissue from patients. Ultimately, they are interested in developing models of sickle-cell anemia and Fanconi anemia, a hereditary disease in which the bone marrow doesn't produce enough new cells to replenish the blood.

For example, patients with Fanconi anemia often suffer from skeletal problems, and their cells show an impaired ability to repair DNA. "We don't have any idea why kids with DNA repair defect would get a blood disease, and why they sometimes get these bone abnormalities," says Lensch, who works with Daley. But with stem-cell lines developed from a patient, "we could push the cells to develop into bone and blood, and try to learn about the links between the two."

Such models could also help resolve long-held debates about specific diseases, such as Alzheimer's. By differentiating reprogrammed cells from Alzheimer's patients into neurons and comparing them with neurons derived from healthy embryonic stem cells or with cells with mutations that mimic a rare, hereditary form of the disease, scientists will be able to determine how much of Alzheimer's is due to the environment versus genes, as well as how similar the sporadic form of the disease is to the hereditary form. (Most drugs on the market for Alzheimer's were developed using models that mimic the hereditary form of the disease and have shown limited efficacy in patients.) "This is a whole new world of investigation," says Lawrence Goldstein, a neuroscientist at the University of California, San Diego, whose lab is about to begin collecting skin cells from Alzheimer's patients.

Despite the excitement, Lensch and others caution against abandoning other embryonic stem-cell research, especially therapeutic cloning. "We're in the early stages of this research, where we're excited about the possibilities but still need to show it's both useful and representative of the disease," says Snyder. In addition, he says, embryonic stem cells and perhaps cloned stem cells will be needed as controls for future studies.

Scientists also say that it's too soon to tell how easy it will be to generate stem-cell lines from patients: the genetic variations that lead to the disease could also impact the reprogramming process. "With some genetic disease, I think it will be really difficult," says Lensch.

http://www.technologyreview.com/Biotech/20007/page2/

The Year in Biotech

Genomics Gets Really PersonalThis year may be remembered as the turning point for personal genomics, when broad gene testing for individuals finally came within reach. Two genomic pioneers--James Watson, codiscover of the structure of DNA, and Craig Venter, leader of the private effort to sequence the genome--published the sequence of their own genomes, revealing personal disease risks. (See "The $2 Million Genome" and "Craig Venter's Genome.")

Taking advantage of the explosion in human genomics data, several companies launched direct-to-consumer gene-testing services that analyze an individual's genetic risk of contacting a range of diseases, including Alzheimer's, diabetes, and cancer. (See "Your Future, on a Chip" and "Your Personal Genome.") Critics say it's not yet clear how useful such tests will be in preventing disease. So with price tags ranging from about $1,000, for a microarray analysis that analyzes a million genetic variations, to $350,000, for a full genome sequence, it might be worth waiting.
The Microbial Menagerie In 2007, scientists documented the microbial world more closely than ever before, thanks to sequencing technologies that allow analysis of entire microscopic communities, an approach known as metagenomics. In the process, they have uncovered a wealth of genomic diversity that could be applied to everything from renewable energy to medicine. For example, enzymes found in microbes dwelling in the termite gut might inspire more-efficient ways of making cellulosic ethanol. (See "Termite Guts Could Boost Ethanol Efficiency.")

Our own microbial inhabitants are getting special attention, as part of the newly announced Human Microbiome Project--a massive plan sponsored by the National Institutes of Health to document the microbes that live within us and play a vital role in immune function and nutrition. (See "Our Microbial Menagerie" and "The Next Human Genome Project: Our Microbes.")
Stem Cells without Embryos Embryonic stem cell research, particularly therapeutic cloning, floundered this year, thanks to continued funding restrictions, technical issues, and ethical concerns. For example, Harvard scientist and champion cloner Kevin Eggan was given permission to start human cloning experiments almost two years ago but has not yet started due to lack of human eggs. (See "Human Therapeutic Cloning at a Standstill.")

Last month, however, scientists in Wisconsin and Japan announced an exciting potential alternative-- a relatively easy method to reprogram adult cells to behave like embryonic stem cells without the need for embryos or eggs. (See "Stem Cells without the Embryos.") Further work needs to be done to determine the exact properties of these new cells and to determine if they would be safe to use in humans.

Jump-Starting the Damaged Brain Injured soldiers returning from the war in Iraq have brought new focus to the terrible toll of brain injury. Thanks to better emergency medicine, patients with severe head trauma are more likely to survive. But they may be left with severe cognitive impairments that inhibit the ability to survey or respond to the outside world.
Nicholas Schiff, a neurologist at Weill Cornell Medical College in New York City, has provided new hope for the families of these largely forgotten patients. He found that deep brain stimulation, a technique used to treat Parkinson's disease in which electricity is delivered to specific parts of the brain, can help these patients better respond to their environment. (See "Raising Consciousness" and "Jump-Starting the Damaged Brain.")

Human Genetic VariationHumans may be more genetically varied than previous thought. Scientists have discovered that large chunks of DNA are frequently copied, deleted, or transposed and may play a role in disease. (See "Deciphering Human Differences.") In addition, a number of studies using gene arrays that scan the entire genome for SNPs or single nucleotide polymorphisms have identified specific variations linked to common complex genetic diseases, such as diabetes, Crohn's disease, and heart disease. (See "Genes for Several Common Diseases Found.")

http://www.technologyreview.com/Biotech/19982/

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