{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,6]],"date-time":"2026-10-06T00:24:13Z","timestamp":1791246253020,"version":"4.1.0"},"reference-count":18,"publisher":"National Academy of Sciences","issue":"34","content-domain":{"domain":["www.pnas.org"],"crossmark-restriction":true},"short-container-title":["Proc. Natl. Acad. Sci. U.S.A."],"published-print":{"date-parts":[[2011,8,23]]},"abstract":"<jats:p>Molecular knockdown of disease proteins and restoration of wild-type activity represent a promising but challenging strategy for the treatment of diseases that result from the accumulation of misfolded proteins (i.e., Huntington disease, amyotrophic lateral sclerosis, and \u03b1-1 antitrypsin deficiency). In this study we used alpha-1 antitrypsin (AAT) deficiency with the piZZ mutant phenotype as a model system to evaluate the efficiency of gene-delivery approaches that both silence the piZZ transcript (e.g., shRNA) and restore circulating wild-type AAT expression from resistant codon-optimized AAT (AAT-opt) transgene cassette using adeno-associated virus (AAV) vector delivery. After systemic injection of a self-complimentary AAV serotype 8 (scAAV8) vector encoding shRNA in piZZ transgenic mice, both mutant AAT mRNA in the liver and defected serum protein level were inhibited by 95%, whereas liver pathology, as monitored by dPAS and fibrosis staining, reversed. To restore blood AAT levels in AAV8\/shRNA-treated mice, several strategies to restore functional AAT levels were tested, including using AAV AAT-opt transgene cassettes targeted to muscle and liver, or combination vectors carrying piZZ shRNA and AAT-opt transgenes separately, or a single bicistronic AAV vector. With these molecular approaches, we observed over 90% knockdown of mutant AAT with a 13- to 30-fold increase of circulating wild-type AAT protein from the shRNA-resistant AAT-opt cassette. The molecular approaches applied in this study can simultaneously prevent liver pathology and restore blood AAT concentration in AAT deficiencies. Based on these observations, similar gene-therapy strategies could be considered for any diseases caused by accumulation of misfolded proteins.<\/jats:p>","DOI":"10.1073\/pnas.1109522108","type":"journal-article","created":{"date-parts":[[2011,8,16]],"date-time":"2011-08-16T04:34:54Z","timestamp":1313469294000},"page":"14258-14263","update-policy":"https:\/\/doi.org\/10.1073\/pnas.cm10313","source":"Crossref","is-referenced-by-count":63,"title":["Combination therapy utilizing shRNA knockdown and an optimized resistant transgene for rescue of diseases caused by misfolded proteins"],"prefix":"10.1073","volume":"108","author":[{"given":"Chengwen","family":"Li","sequence":"first","affiliation":[{"name":"Gene Therapy Center,"},{"name":"Department of Pediatrics,"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Pingjie","family":"Xiao","sequence":"additional","affiliation":[{"name":"Gene Therapy Center,"},{"name":"Department of Cell and Developmental Biology, and"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Steven James","family":"Gray","sequence":"additional","affiliation":[{"name":"Gene Therapy Center,"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Marc Scott","family":"Weinberg","sequence":"additional","affiliation":[{"name":"Gene Therapy Center,"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"R. Jude","family":"Samulski","sequence":"additional","affiliation":[{"name":"Gene Therapy Center,"},{"name":"Department of Pharmacology, University of North Carolina, Chapel Hill, NC 27599"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"341","published-online":{"date-parts":[[2011,8,15]]},"reference":[{"key":"e_1_3_3_1_2","doi-asserted-by":"publisher","DOI":"10.1038\/nrn1007"},{"key":"e_1_3_3_2_2","doi-asserted-by":"publisher","DOI":"10.1038\/sj.cgt.7700876"},{"key":"e_1_3_3_3_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.0904514106"},{"key":"e_1_3_3_4_2","doi-asserted-by":"publisher","DOI":"10.1089\/hum.2006.17.1177"},{"key":"e_1_3_3_5_2","doi-asserted-by":"publisher","DOI":"10.1038\/sj.gt.3301422"},{"key":"e_1_3_3_6_2","doi-asserted-by":"publisher","DOI":"10.1038\/sj.gt.3302134"},{"key":"e_1_3_3_7_2","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.78.12.6381-6388.2004"},{"key":"e_1_3_3_8_2","doi-asserted-by":"publisher","DOI":"10.1002\/jgm.835"},{"key":"e_1_3_3_9_2","doi-asserted-by":"publisher","DOI":"10.1038\/sj.mt.6300355"},{"key":"e_1_3_3_10_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0140-6736(09)61836-5"},{"key":"e_1_3_3_11_2","doi-asserted-by":"publisher","DOI":"10.1053\/j.gastro.2004.07.058"},{"key":"e_1_3_3_12_2","doi-asserted-by":"publisher","DOI":"10.1038\/labinvest.3700629"},{"key":"e_1_3_3_13_2","doi-asserted-by":"publisher","DOI":"10.1038\/nm1358"},{"key":"e_1_3_3_14_2","doi-asserted-by":"publisher","DOI":"10.1038\/sj.mt.6300090"},{"key":"e_1_3_3_15_2","doi-asserted-by":"publisher","DOI":"10.1038\/mt.2010.40"},{"key":"e_1_3_3_16_2","doi-asserted-by":"publisher","DOI":"10.4161\/auto.2882"},{"key":"e_1_3_3_17_2","doi-asserted-by":"publisher","DOI":"10.1083\/jcb.201002021"},{"key":"e_1_3_3_18_2","doi-asserted-by":"publisher","DOI":"10.1038\/nature04791"}],"container-title":["Proceedings of the National Academy of Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/pnas.org\/doi\/pdf\/10.1073\/pnas.1109522108","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2022,4,12]],"date-time":"2022-04-12T21:14:30Z","timestamp":1649798070000},"score":1,"resource":{"primary":{"URL":"https:\/\/pnas.org\/doi\/full\/10.1073\/pnas.1109522108"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2011,8,15]]},"references-count":18,"journal-issue":{"issue":"34","published-print":{"date-parts":[[2011,8,23]]}},"alternative-id":["10.1073\/pnas.1109522108"],"URL":"https:\/\/doi.org\/10.1073\/pnas.1109522108","relation":{},"ISSN":["0027-8424","1091-6490"],"issn-type":[{"value":"0027-8424","type":"print"},{"value":"1091-6490","type":"electronic"}],"subject":[],"published":{"date-parts":[[2011,8,15]]},"assertion":[{"value":"2011-08-15","order":2,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}]}}