August 15, 2005
First Gene Therapy Success Against Muscular Dystrophy In Mice

A rare gene therapy success is reported in mice with muscular dystrophy.

PITTSBURGH, Aug. 15 Researchers from the University of Pittsburgh report the first study to achieve success with gene therapy for the treatment of congenital muscular dystrophy (CMD) in mice, demonstrating that the formidable scientific challenges that have cast doubt on gene therapy ever being feasible for children with muscular dystrophy can be overcome. Moreover, their results, published in this week's online edition of the Proceedings of the National Academy of Sciences (PNAS), indicate that a single treatment can have expansive reach to muscles throughout the body and significantly increase survival.

CMD is a group of some 20 inherited muscular dystrophies characterized by progressive and severe muscle wasting and weakness first noticed soon after birth. No effective treatments exist and children usually die quite young.

Despite gene therapy being among the most vigorously studied approaches for muscular dystrophy, it has been beset with uniquely difficult hurdles. The genes to replace those that are defective in CMD are larger than most, so it has not been possible to apply the same methods successfully used for delivering other types of genes. And because CMD affects all muscles, an organ that accounts for 40 percent of body weight, gene therapy can only have real therapeutic benefit if it is able to reverse genetic defects in every cell of the body's 600 muscle groups.

Aside: If a virus can get into all the muscle groups of the body it is no wonder that some kinds of viral infections make your muscles ache all over.

Think about what the passage above says about the state of gene therapy in the year 2005. Delivering a single large gene is too difficult. Well, gene therapies for rejuvenation will need to fix dozens and perhaps hundreds of genes. Some of those genes are large. We need much better gene therapy delivery vehicles.

The researchers use a couple of approaches to make the gene therapy work for CMD.

By using a miniature gene, similar in function to the one defective in CMD, and applying a newly developed method for "systemic" gene delivery, the Pitt researchers have shown that gene therapy for muscular dystrophy is both feasible and effective in a mouse model of especially profound disease. Using this approach, the team, led by Xiao Xiao, Ph.D., associate professor of orthopaedic surgery and molecular genetics and biochemistry at the University of Pittsburgh School of Medicine, report that treated mice had physiological improvements in the muscles of the heart, diaphragm, abdomen and legs; and they grew faster, were physically more active and lived four times as long as untreated animals.

The miniature gene they speak of here might be just the portion of the gene that gets translated into a protein. Many mammalian genes have what are called "introns" and "exons". The introns do not get translated into proteins but rather are spliced out in the process of going from gene to protein. The "exons" therefore can be combined into a gene that is smaller than the naturally occurring gene but which probably (barring some regulatory role for the "introns") can be made to work just as well.

"While we have much farther to go until we can say gene therapy will work in children, we have shown here a glimmer of hope by presenting the first evidence of a successful gene therapy approach that improved both the general health and longevity in mice with congenital muscular dystrophy," said Dr. Xiao.

The most common form of CMD, and also one of the most severe, is due to a genetic mutation of laminin alpha-2, a protein that is essential for maintaining the structures that surround muscle cells and is an integral link in the chain of proteins that regulate the cell's normal contraction and relaxation. If the protein is defective, or is lacking, this outside scaffold, called the extra-cellular matrix, disintegrates, and the muscle cells become vulnerable to damage.

One limit to their approach is the use of a virus to deliver genes. The adeno-associated virus (AAV) which they mention below is only 4675 DNA letters long. While the virus gets transported well into muscle cells (and muscle cells age and need gene therapy for rejuvenation) the Adeno-Associated Virus (AAV) just can not carry much DNA into a cell.

Simply replacing the defective gene with a good laminin alpha-2 gene is not possible because its size makes it impossible for researchers to get it to squeeze inside viral vectors disarmed viruses that are used to shuttle genes into cells. But the team found a good stand-in in a similar protein called agrin that when miniaturized could be inserted inside an adeno-associated virus (AAV) vector. Dr. Xiao's laboratory is known for its work developing this vector, which they have previously shown is the most efficient means for delivering genes to muscle cells.

In the current study, the authors show that two strains of AAV, AAV-1 and AAV-2, were effective in transferring the mini-agrin gene to cells in two mouse models. The AAV-1 vector was given by systemic delivery a single infusion into the abdominal cavity a method the authors only recently described and which they used for the first time in this study to transfer a therapeutic gene. The AAV-2 vector was delivered locally, given by intramuscular injection to different muscles of the leg. With both approaches, muscle cells were able to assimilate and copy the genetic instructions for making mini-agrin. Once produced, the mini-agrin protein functionally took the place of the laminin alpha-2 protein by binding to the key proteins on either end, thus restoring the cell's outside scaffolding and reestablishing the missing link to key structures inside the cell.

I'm less excited than the authors because I want vectors (delivery vehicles or carriers) for gene therapy that can delivery much more DNA into each cell for more extensive reprogramming.

Clearly, the authors are most excited about the impressive results achieved in their experiments using systemic gene delivery, which proved there could be significant therapeutic improvements and even be life-saving. Yet they say their results are far from ideal and more work lies ahead.

"It's probably not realistic to expect that we can achieve complete success using the mini-agrin gene, which while somewhat similar, is structurally unrelated to laminin alpha-2. Unless we address the underlying cause of congenital muscular dystrophy we're not likely to be able to completely arrest or cure CMD," added Chungping Qiao, M.D., Ph.D., the study's first author and a research associate fellow in Dr. Xiao's lab.

Future directions for research include finding a way to engineer the laminin alpha-2 gene. For this study, the authors chose to use the mini-agrin gene because researchers from the University of Basel, Switzerland, had already demonstrated it could improve the symptoms of muscular dystrophy in a transgenic mouse model, which has little clinical relevance. The Pitt researchers might also explore approaches that combine genes that promote both muscle and nerve growth, as well as focus on improving the AAV vectors.

In addition to Drs. Xiao and Qiao, other authors are Jianbin Li; Tong Zhu, M.D., Ph.D.; Xiaojung Ye, M.D., Ph.D.; Chunlian Chen; and Juan Li, M.D., all from the department of orthopaedic surgery; and from the department of cell biology and physiology, Romesh Draviam and Simon Watkins, Ph.D.

Gene therapy does not get as much press as cell therapy. Yet gene therapy is crucial for rejuvenation. We will be able to replace many cells and organs using stem cell therapies and also tissue engineering techniques combined with stem cells to grow replacement organs. But the toughest problem for rejuvenation is the brain. We need to rejuvenate all the cells which are already in the brain. We will do part of that job by sending in immunotherapies that attack extracellular junk. Also, stem cells will help some. But to fix aged neurons, glial cells, and blood vessels in the brain we are going to need gene therapies that can deliver lots of genetic instructions to carry out repair and to replace damaged genes.

Another problem with viral gene therapy delivery mechanisms is the immune system's tendency to quite correctly recognize viruses as invaders. One way around that problem might to be to develop methods to very briefly suppress the immune system. If the viruses move out of the bloodstream pretty quickly then immune system suppression drugs might not need to suppress the immune system so long as to put one at risk of a major infection. Or perhaps the immune system could be trained to recognize a particular and not naturally occurring variation of AAV as self.

AAV still can play a role in rejuvenation therapies. One way to get around its size limitation might be to send in lots of small genes in successive AAV packages. Other way might be to genetically engineer AAV packaging proteins to form larger cases to carry larger genes into cells. Therefore the report above is a good step in the right direction and the researchers should be applauded. We just need many more such steps to produce successively better gene therapy techniques.

Thanks to Andy Price for pointing out this report. Says Andy "The cool thing for everyone (SENS) here that this treatment gets to ALL muscle groups.". His reference is to Strategies for Engineered Negligible Senescence or SENS.

Share |      Randall Parker, 2005 August 15 08:47 PM  Biotech Gene Therapy

sr said at August 16, 2005 9:13 AM:

Micecular dystrophy!

Engineer-Poet said at August 16, 2005 10:07 AM:

The mitochondrial DNA strand consists of 16,569 base pairs divided into 37 genes.  This averages under 450 base pairs per gene; it may well be possible to substitute functional genes for many or most damaged mitochondrial genes even using a virus as small as AAV.

If we're looking for ways to fix brains, the obvious place to look for vectors is viruses which cause encephalitis.

Robert Bradbury said at August 16, 2005 1:30 PM:

First of all this blog software *sucks*. If I have
to enter information every time it is useless.

Also the "wrap" function on the entry form still doesn't work...

To the point... It was clear a decade or more ago that
one could deliver small genomic pieces into cells. The question
then becomes *what* pieces and what do you do with them?

There are people working on more than the simple pieces but they
probably cannot be discussed here.

AA2 said at August 17, 2005 2:15 AM:

This is from The Speculist:

Viral vectors have been useful for experimentation, but have shown little promise in the treatment of patients.

There has to be a better way. And scientists at the University at Buffalo may have found it. Using a form of nanoengineered silicon they're calling ORMOSIL (amino-functionalized organically modified silica) these scientists delivered genes into the brains of living mice.

A key advantage of the UB team's nanoparticle is its surface functionality, which allows it to be targeted to specific cells, explained Dhruba J. Bharali, Ph.D., a co-author on the paper...

In their first experiment the UB scientists surgically injected an ORMOSIL/DNA complex that targeted the dopamine neurons within the brain.

AA2 said at August 17, 2005 2:21 AM:

The Speculist referenced this source-

Using nanoparticles, in vivo gene therapy activates brain stem cells
26 Jul 2005

Technique may allow scientists to repair brain cells damaged by disease, trauma or stroke -....

Randall Parker said at August 17, 2005 9:23 AM:


I have been meaning to post on the U Buffalo report because I think it is quite important. Here is the U Buffalo press release on the nanoparticle delivery mechanism for gene therapy and here is the abstract from PNAS and here is the full text of the research paper.

I've had all that sitting in my browser for days (I hibernate my computer at night) and just keep forgetting to post it.

Any advance in gene therapy for the brain is incredibly important because the brain is the organ most in need of gene therapy for aging diseases and rejuvenation.

Randall Parker said at August 17, 2005 9:28 AM:


Regarding mitochondria and gene therapy: Mitochondria add an extra level of difficulty since the mitochondrial genes have to get into the mitochondria. If we follow Aubrey de Grey's proposal to make variations of mitochondrial genes that can get expressed in the nucleus then we could probably use the same gene therapy delivery mechanisms that we use to deliver other genes to the nucleus.

But of course genetically engineering mitochondrial DNA to work from the nucleus will not be easy either. The proteins whose genes are still in the mitochondrion have electrical properties that'll make it hard to get them transported into the mitochondria. But once the engineering task for each protein is solved we'd get much better results since genes in the nucleus are less likely to get damaged than genes in the mitochondrion.

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