{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,2,21]],"date-time":"2026-02-21T05:58:36Z","timestamp":1771653516947,"version":"3.50.1"},"reference-count":41,"publisher":"Wiley","issue":"3","license":[{"start":{"date-parts":[[2007,12,13]],"date-time":"2007-12-13T00:00:00Z","timestamp":1197504000000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["The Journal of Gene Medicine"],"published-print":{"date-parts":[[2008,3]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:sec><jats:title>Background<\/jats:title><jats:p>Systemic <jats:italic>in vivo<\/jats:italic> gene therapy has resulted in widespread correction in animal models when treated at birth. However, limited improvement was observed in postnatally treated animals with mainly targeting to the liver and bone marrow. It has been shown that an O<jats:sup>6<\/jats:sup>\u2010methylguanine\u2010DNA\u2010methyltransferase variant (MGMT<jats:sup>P140K<\/jats:sup>) mediated <jats:italic>in vivo<\/jats:italic> selection of transduced hematopoietic stem cells (HSC) in animals.<\/jats:p><\/jats:sec><jats:sec><jats:title>Methods<\/jats:title><jats:p>We investigated the feasibility of MGMT<jats:sup>P140K<\/jats:sup>\u2010mediated selection in primary hepatocytes from a mouse model of mucopolysaccharidosis type I (MPS I) <jats:italic>in vitro<\/jats:italic> using lentiviral vectors.<\/jats:p><\/jats:sec><jats:sec><jats:title>Results<\/jats:title><jats:p>We found that multiple cycles of O<jats:sup>6<\/jats:sup>\u2010benzylguanine (BG)\/1,3\u2010bis(2\u2010chloroethyl)\u20101\u2010nitrosourea (BCNU) treatment at a dosage effective for <jats:italic>ex vivo<\/jats:italic> HSC selection led to a two\u2010fold increase of MGMT\u2010expressing primary hepatocytes under culture conditions with minimum cell expansion. This enrichment level was comparable to that obtained after selection at a hepatic maximal tolerated dose of BCNU. Similar levels of increase were observed regardless of initial transduction frequency, or the position of MGMT (upstream or downstream of internal ribosome entry site) in the vector constructs. In addition, we found that elongation factor 1\u03b1 promoter was superior to the long\u2010terminal repeat promoter from spleen focus\u2010forming virus with regard to transgene expression in primary hepatocytes. Moreover, the levels of therapeutic transgene expression in transduced, enzyme\u2010deficient hepatocytes directly correlated with the doses of BCNU, leading to metabolic correction in transduced hepatocytes and metabolic cross\u2010correction in neighbouring non\u2010transduced MPS I cells.<\/jats:p><\/jats:sec><jats:sec><jats:title>Conclusions<\/jats:title><jats:p>These results demonstrate that MGMT<jats:sup>P140K<\/jats:sup> expression confers successful protection\/selection in primary hepatocytes, and provide \u2018proof of concept\u2019 to the prospect of MGMT<jats:sup>P140K<\/jats:sup>\u2010mediated co\u2010selection for hepatocytes and HSC using BG\/BCNU treatment. Copyright \u00a9 2007 John Wiley &amp; Sons, Ltd.<\/jats:p><\/jats:sec>","DOI":"10.1002\/jgm.1141","type":"journal-article","created":{"date-parts":[[2007,12,13]],"date-time":"2007-12-13T12:46:30Z","timestamp":1197549990000},"page":"249-259","source":"Crossref","is-referenced-by-count":7,"title":["Co\u2010expression of MGMT<sup>P140K<\/sup> and \u03b1\u2010<scp>L<\/scp>\u2010iduronidase in primary hepatocytes from mucopolysaccharidosis type I mice enables efficient selection with metabolic correction"],"prefix":"10.1002","volume":"10","author":[{"given":"Daren","family":"Wang","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"D. 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