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dc.contributor.authorCruz, Pedro E.
dc.contributor.authorMueller, Christian
dc.contributor.authorCossette, Travis L.
dc.contributor.authorGolant, Alexandra
dc.contributor.authorTang, Qiushi
dc.contributor.authorBeattie, Stuart G.
dc.contributor.authorBrantly, Mark
dc.contributor.authorCampbell-Thompson, Martha
dc.contributor.authorBlomenkamp, Keith S.
dc.contributor.authorTeckman, Jeffrey H.
dc.contributor.authorFlotte, Terence R.
dc.date2022-08-11T08:10:14.000
dc.date.accessioned2022-08-23T17:00:26Z
dc.date.available2022-08-23T17:00:26Z
dc.date.issued2007-09-27
dc.date.submitted2012-01-11
dc.identifier.citationLab Invest. 2007 Sep;87(9):893-902. Epub 2007 Jun 25. <a href="http://dx.doi.org/10.1038/labinvest.3700629">Link to article on publisher's site</a>
dc.identifier.issn0023-6837 (Linking)
dc.identifier.doi10.1038/labinvest.3700629
dc.identifier.pmid17592477
dc.identifier.urihttp://hdl.handle.net/20.500.14038/43797
dc.description.abstractalpha-1 Antitrypsin (AAT) deficiency is one of the most common genetic diseases in North America, with a carrier frequency of approximately 4% in the US population. Homozygosity for the most common mutation (Glu342Lys, PI(*)Z) leads to the synthesis of a mutant protein, which accumulates and polymerizes within hepatocytes rather than being efficiently secreted. This lack of secretion causes severe serum deficiency predisposing to chronic lung disease. Twelve to fifteen percent of patients with PI(*)ZZ also develop liver disease, which can be severe, even in infancy. This is thought to be due to toxic effects of the accumulated mutant Z-AAT within the hepatocyte. Thus, an approach to reduce AAT-deficient liver disease will likely require some mechanism to decrease the amount of Z-AAT within hepatocytes. In this report, we describe studies of small-interfering RNAs (siRNAs) designed to downregulate endogenous AAT within hepatocytes. Three different siRNA sequences were identified and cloned into a recombinant adeno-associated virus (rAAV) backbone, either singly or as a trifunctional (3X) construct. Each had activity independently, but the levels of AAT expression in cell culture models showed the greatest decrease with the 3X construct, resulting in levels that were five-fold lower than controls. The rAAV-3X-siRNA was then packaged into AAV8 capsids and used in vivo to transduce the livers of human Z-AAT overexpressing transgenic mice. Those studies showed a decrease in total human AAT, a clearing of Z-AAT accumulation by immunohistochemistry, and a decrease in monomer Z-AAT within the liver within 3 weeks after vector injection. The rAAV8-3X-siRNA vector may hold promise as a potential therapy for patients with AAT liver disease.
dc.language.isoen_US
dc.relation<a href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=pubmed&cmd=Retrieve&list_uids=17592477&dopt=Abstract">Link to Article in PubMed</a>
dc.relation.urlhttp://dx.doi.org/10.1038/labinvest.3700629
dc.subjectAnimals
dc.subjectCell Line, Tumor
dc.subjectDependovirus
dc.subjectDisease Models, Animal
dc.subjectDown-Regulation
dc.subject*Gene Therapy
dc.subjectGenetic Vectors
dc.subjectHepatocytes
dc.subjectHumans
dc.subjectMice
dc.subjectMice, Transgenic
dc.subjectPolymorphism, Single Nucleotide
dc.subject*RNA Interference
dc.subjectRNA, Small Interfering
dc.subjectalpha 1-Antitrypsin
dc.subjectalpha 1-Antitrypsin Deficiency
dc.subjectAllergy and Immunology
dc.subjectGenetics and Genomics
dc.subjectPediatrics
dc.subjectRespiratory Tract Diseases
dc.titleIn vivo post-transcriptional gene silencing of alpha-1 antitrypsin by adeno-associated virus vectors expressing siRNA
dc.typeJournal Article
dc.source.journaltitleLaboratory investigation; a journal of technical methods and pathology
dc.source.volume87
dc.source.issue9
dc.identifier.legacycoverpagehttps://escholarship.umassmed.edu/peds_pulmonary/13
dc.identifier.contextkey2441373
html.description.abstract<p>alpha-1 Antitrypsin (AAT) deficiency is one of the most common genetic diseases in North America, with a carrier frequency of approximately 4% in the US population. Homozygosity for the most common mutation (Glu342Lys, PI(*)Z) leads to the synthesis of a mutant protein, which accumulates and polymerizes within hepatocytes rather than being efficiently secreted. This lack of secretion causes severe serum deficiency predisposing to chronic lung disease. Twelve to fifteen percent of patients with PI(*)ZZ also develop liver disease, which can be severe, even in infancy. This is thought to be due to toxic effects of the accumulated mutant Z-AAT within the hepatocyte. Thus, an approach to reduce AAT-deficient liver disease will likely require some mechanism to decrease the amount of Z-AAT within hepatocytes. In this report, we describe studies of small-interfering RNAs (siRNAs) designed to downregulate endogenous AAT within hepatocytes. Three different siRNA sequences were identified and cloned into a recombinant adeno-associated virus (rAAV) backbone, either singly or as a trifunctional (3X) construct. Each had activity independently, but the levels of AAT expression in cell culture models showed the greatest decrease with the 3X construct, resulting in levels that were five-fold lower than controls. The rAAV-3X-siRNA was then packaged into AAV8 capsids and used in vivo to transduce the livers of human Z-AAT overexpressing transgenic mice. Those studies showed a decrease in total human AAT, a clearing of Z-AAT accumulation by immunohistochemistry, and a decrease in monomer Z-AAT within the liver within 3 weeks after vector injection. The rAAV8-3X-siRNA vector may hold promise as a potential therapy for patients with AAT liver disease.</p>
dc.identifier.submissionpathpeds_pulmonary/13
dc.contributor.departmentDepartment of Pediatrics
dc.contributor.departmentGene Therapy Center
dc.source.pages893-902


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