Gene therapy sounds good, in principle, as a means of treating diseases that result from genetic defects. But there have been at least two major practical problems in making use of gene therapy in the clinic. First, it’s very important for the safety of the procedure to make changes only to the defective gene (or even just the critical part of the gene) and no other portion of the DNA. Second, there needs to be an effective way to deliver the therapy, in whatever form it takes, to exactly the right tissues in the body that are affected by the defective gene.
Solving these two problems simultaneously is especially difficult. There are a variety of techniques for modifying DNA in specific ways, using a number of specialized enzymes. But these techniques are actually usable only in a test tube. Simply setting the enzymes loose in a patient’s body doesn’t work. The usual way to get around this is by preparing the appropriate modified DNA segments and incorporating them into “vectors”, such as some sort of virus, and introducing the vector into a patient’s body, in the hope that it will reach the right tissues to deliver the DNA, without causing any other problems. That hasn’t worked very well, so far, despite a large number of attempts.
What about taking some cells from the patient, modifying their DNA in vitro and then putting them back in the body? The problem there is being able to produce enough cells with good DNA, either before or after reintroduction to the patient’s body, to make a significant difference. Most human cells just don’t reproduce very prolifically outside the body, or even inside for that matter.
At this point you should be thinking, “stem cells!” They are specialized to reproduce quickly, when needed. Up until now, there have been practical problems here, too. Adult stem cells capable of differentiating into the specific type of cell needed in a given tissue may be difficult or impossible to find in useful quantities. And embryonic stem cells, well, even any other problems aside, there’s the problem of avoiding rejection by the patient’s immune system.
The potential solution: induced pluripotent stem cells (iPSCs), made from any convenient cell type of the actual patient. It’s only been five years since iPSCs were first produced. Various practical problems have arisen along the way since then. Some have been mostly overcome; some haven’t. But progress seems to be occurring steadily – including very recently. The application to gene therapy involves making appropriate corrections to the DNA, producing a sufficient number of differentiated cells of the required type from the “fixed” iPSCs, and finally reintroducing them into the patient.
Alpha 1-antitrypsin deficiency is a genetic disorder caused by a point mutation that results in inadequate production of the alpha 1-antitrypsin (A1AT) enzyme in liver cells. The disease affects functioning of the lungs, as well as the liver. Research just published has shown that gene therapy to correct the mutation, applied to iPSCs, is successful at treating the condition in a mouse model.
Researchers from the University of Cambridge, directed by Ludovic Vallier and David Lomas, and from the Sanger Institute, coordinated by Allan Bradley, began by sampling patients’ skin cells, which were then cultured in vitro for “differentiation” before applying the properties of the pluripotent stem cells: this is the “iPS cells” stage. Through genetic engineering, scientists were then able to correct the mutation responsible for the disease. They then engaged the now “healthy” stem cells in the maturation process, leading them to differentiate to liver cells.
Scientists from the Institut Pasteur and Inserm, led by H-l-ne Strick-Marchand in the mixed Institut Pasteur/Inserm Innate Immunity unit (directed by James Di Santo), then tested new human hepatic cells thus produced on an animal model afflicted with liver failure. Their research showed that the cells were entirely functional and suited to integration in existing tissue and that they may contribute to liver regeneration in the mice treated.