![\"muscle](\"https:\/\/www.salk.edu\/wp-content\/uploads\/2011\/01\/531-BelmonteSMA.jpg\")
<\/p>\nSalk researchers reprogrammed skins cells taken from a sickle cell disease patient into induced pluripotent stem cells (iPSCs), immature cells capable of developing into any type of bodily tissue.\n<\/p>\n
\nTo assure that the skin cells were in fact reprogrammed into stem cells, the researchers coaxed them into becoming muscle cells, indicated by the presence of muscle-specific protiens (red) in this image. The cell nuclei\u2014the central compartments that contain the DNA\u2014are seen in blue.<\/p>\n
Image: Courtesy of Salk Institute for Biological Studies<\/p>\n<\/div>\n
\nSickle cell disease is a group of inherited blood disorders caused by genetic mutations in the HBB gene, resulting in abnormal hemoglobin, the iron-containing protein that normally allows blood cells to carry oxygen. This causes red blood cells to become hard and sticky and resemble a curved farm tool called a “sickle.” In the two leading disorders caused by HBB mutations, sickle cell anemia and beta thalassemia, red blood cells can’t effectively carry oxygen.\n<\/p>\n
\nSymptoms of sickle cell disease include swelling of the hands and feet, pain due to clogging of blood vessels, anemia and stroke.\n<\/p>\n
\nThe disorders are most common among people of African, Mediterranean and Middle Eastern decent. One in every 500 African Americans and one in every 30,000 Hispanic Americans are born with sickle cell disease, according to the Centers for Disease Control and Prevention.\n<\/p>\n
\nThe disease can be cured with stem cell or bone marrow transplants, but there is a high risk that recipients of transplants will reject the donated marrow or cells, which can result in serious side effects and even death.\n<\/p>\n
\nThe Salk researchers, which include co-first authors Mo Li and Keiichiro Suzuki, both research associates in Belmonte’s laboratory, set out to devise a safe method to use iPSCs to correct the HBB gene in patients who have defective copies of the gene.\n<\/p>\n
\nBecause the iPSCs come from a patient’s own body, they should carry less risk for transplant rejection. Also, about 500 other disease-causing mutations have been identified in the HBB gene, so correcting the gene could potentially cure a multitude of HBB-related diseases worldwide.\n<\/p>\n
\nHowever, traditional iPSC generation and gene therapy techniques have proven to be potentially unsafe, according to the researchers.\n<\/p>\n
\nMany have used viruses to convert adult cells to stem cells and to carry a normal HBB gene to infect and repair hematopoietic stem cells\u2014stem cells that give rise to all blood cells.\n<\/p>\n
\nBut when these repaired stem cells are given back to patients, they can include transgenes\u2014unwanted genes that have become inserted into the host genome and disrupt the normal function of DNA. The technique is also inefficient, correcting only a small percentage of gene mutations, and transplantation success has proven rare in clinical trials testing gene therapy to treat beta thalassemia.\n<\/p>\n
\n“We wanted to fix the mutation in such a way that it does not leave any unwanted traces in a patient’s genome,” Suzuki says.\n<\/p>\n
\nTo do that, the researchers used a two-step approach. First, they took adult skin cells from a patient with an HBB mutation that causes sickle cell disease. They used six genes to coax these cells to revert to iPSCs, which could then be developed into blood cells. The genes were introduced into the cells using a technique that avoids the use of viruses and insertion of transgenes into the cells’ genome.\n<\/p>\n
\nTheir next step was to repair the HBB gene mutation in the stem cells. To swap the defective gene with a normal copy in the iPSCs, the investigators used a modified adenovirus (common cold virus) that, unlike viruses used in other methods, does not replicate itself in the body and does not alter the host cells’ DNA. The viral genes were deleted and replaced with a DNA sequence that contained a normal HBB gene.\n<\/p>\n
\nThe modified virus then delivered the new genetic material inside the iPSCs, where the DNA region containing the broken gene was replaced with the sequence containing the normal gene. “It happens naturally, working like a zipper,” Li says. “The good gene just zips in perfectly, pushing the bad one out.”\n<\/p>\n
\nBy replacing a relatively large region of DNA, the technique allows the scientists to fix many gene mutations at once, which suggests the method might provide a way to treat hundreds of types of HBB-related diseases. The correction of the mutant HBB gene was also highly efficient and the research team conducted multiple tests to ensure no errant genes were integrated into the genome.\n<\/p>\n
\nThe Salk scientists now plan to make blood cells from the repaired stem cells and test their effectiveness in animals. If successful, this may lead to therapies for humans in which a patient’s stem cells will be reverted into iPSCs, then genetically repaired and transplanted back into the bone marrow of the same patient. If successful, the bone marrow will then produce all new blood cells, including normal hemoglobin.\n<\/p>\n
\nIf the technique proves effective, the researchers say, it might be used for treating other types of diseases caused by single gene mutations.\n<\/p>\n
\nThe study was funded by grants from the G. Harold and Leila Y. Mathers Charitable Foundations, Sanofi-Aventis, Ellison Medical Foundation<\/a>, Die Leona M. und Harry B. Helmsley Charitable Trust<\/a>, MICINN and Fundacion Cellex (JCIB). The study appears in the December 2011 issue of Cell Research<\/em>.<\/p>\n \u00dcber das Salk Institute for Biological Studies:<\/strong> \nDie Leistungen der Fakult\u00e4t wurden mit zahlreichen Auszeichnungen gew\u00fcrdigt, darunter Nobelpreise und Mitgliedschaften in der National Academy of Sciences. Das 1960 vom Polio-Impfstoff-Pionier Dr. Jonas Salk gegr\u00fcndete Institut ist eine unabh\u00e4ngige gemeinn\u00fctzige Organisation und ein architektonisches Wahrzeichen. <\/p>\n
\nDas Salk Institute for Biological Studies ist eine der weltweit f\u00fchrenden Institutionen f\u00fcr Grundlagenforschung, an der international renommierte Fakult\u00e4tsmitglieder grundlegende Fragen der Biowissenschaften in einem einzigartigen, kollaborativen und kreativen Umfeld untersuchen. Mit dem Fokus auf Entdeckungen und die Ausbildung zuk\u00fcnftiger Forschergenerationen leisten Salk-Wissenschaftler bahnbrechende Beitr\u00e4ge zu unserem Verst\u00e4ndnis von Krebs, Alterung, Alzheimer, Diabetes und Infektionskrankheiten durch die Untersuchung von Neurowissenschaften, Genetik, Zell- und Pflanzenbiologie sowie verwandten Disziplinen.\n<\/p>\n