{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,28]],"date-time":"2026-09-28T17:09:51Z","timestamp":1790615391645,"version":"4.1.0"},"reference-count":49,"publisher":"SAGE Publications","issue":"5","license":[{"start":{"date-parts":[[2023,7,12]],"date-time":"2023-07-12T00:00:00Z","timestamp":1689120000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by-nc\/4.0\/"},{"start":{"date-parts":[[2023,7,12]],"date-time":"2023-07-12T00:00:00Z","timestamp":1689120000000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/journals.sagepub.com\/page\/policies\/text-and-data-mining-license"}],"content-domain":{"domain":["journals.sagepub.com"],"crossmark-restriction":true},"short-container-title":["Journal of Neuromuscular Diseases"],"published-print":{"date-parts":[[2023,9,8]]},"abstract":"<jats:sec specific-use=\"heading-level-1\">\n                    <jats:title>Background:<\/jats:title>\n                    <jats:p>\n                      GNE myopathy (GNEM) is a severe muscle disease caused by mutations in the UDP-GlcNAc-2-epimerase\/ManNAc-6-kinase (\n                      <jats:italic toggle=\"yes\">GNE<\/jats:italic>\n                      ) gene, which encodes a bifunctional enzyme required for sialic acid (Sia) biosynthesis.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec specific-use=\"heading-level-1\">\n                    <jats:title>Objective:<\/jats:title>\n                    <jats:p>\n                      To develop assays to demonstrate the potency of AAV gene therapy vectors in making Sia and to define the dose required for replacement of endogenous mouse\n                      <jats:italic toggle=\"yes\">Gne<\/jats:italic>\n                      gene expression with human\n                      <jats:italic toggle=\"yes\">GNE<\/jats:italic>\n                      in skeletal muscles.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec specific-use=\"heading-level-1\">\n                    <jats:title>Methods:<\/jats:title>\n                    <jats:p>\n                      A\n                      <jats:italic toggle=\"yes\">MyoD<\/jats:italic>\n                      -inducible\n                      <jats:italic toggle=\"yes\">Gne<\/jats:italic>\n                      -deficient cell line, Lec3MyoD\n                      <jats:sup>I<\/jats:sup>\n                      , and a\n                      <jats:italic toggle=\"yes\">GNE-<\/jats:italic>\n                      deficient human muscle cell line, were made and tested to define the potency of various AAV vectors to increase binding of Sia-specific lectins, including MAA and SNA. qPCR and qRT-PCR methods were used to quantify AAV biodistribution and\n                      <jats:italic toggle=\"yes\">GNE<\/jats:italic>\n                      gene expression after intravenous delivery of AAV vectors designed with different promoters in wild-type mice.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec specific-use=\"heading-level-1\">\n                    <jats:title>Results:<\/jats:title>\n                    <jats:p>\n                      Lec3 cells showed a strong deficit in MAA binding, while\n                      <jats:italic toggle=\"yes\">GNE<\/jats:italic>\n                      <jats:sup>\u2013\/\u2013<\/jats:sup>\n                      MB135 cells did not. Overexpressing\n                      <jats:italic toggle=\"yes\">GNE<\/jats:italic>\n                      in Lec3 and Lec3MyoD\n                      <jats:sup>I<\/jats:sup>\n                      cells by AAV infection stimulated MAA binding in a dose-dependent manner. Use of a constitutive promoter, CMV, showed higher induction of MAA binding than use of muscle-specific promoters (MCK, MHCK7). rAAVrh74.CMV.\n                      <jats:italic toggle=\"yes\">GNE<\/jats:italic>\n                      stimulated human\n                      <jats:italic toggle=\"yes\">GNE<\/jats:italic>\n                      expression in muscles at levels equivalent to endogenous mouse\n                      <jats:italic toggle=\"yes\">Gne<\/jats:italic>\n                      at a dose of 1\u00d710\n                      <jats:sup>13<\/jats:sup>\n                      vg\/kg, while AAVs with muscle-specific promoters required higher doses. AAV biodistribution in skeletal muscles trended higher when CMV was used as the promoter, and this correlated with increased sialylation of its viral capsid.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec specific-use=\"heading-level-1\">\n                    <jats:title>Conclusions:<\/jats:title>\n                    <jats:p>\n                      Lec3 and Lec3MyoD\n                      <jats:sup>I<\/jats:sup>\n                      cells work well to assay the potency of AAV vectors in making Sia. Systemic delivery of rAAVrh74.CMV.\n                      <jats:italic toggle=\"yes\">GNE<\/jats:italic>\n                      can deliver\n                      <jats:italic toggle=\"yes\">GNE<\/jats:italic>\n                      gene replacement to skeletal muscles at doses that do not overwhelm non-muscle tissues, suggesting that AAV vectors that drive constitutive organ expression could be used to treat GNEM.\n                    <\/jats:p>\n                  <\/jats:sec>","DOI":"10.3233\/jnd-221596","type":"journal-article","created":{"date-parts":[[2023,7,14]],"date-time":"2023-07-14T11:42:52Z","timestamp":1689334972000},"page":"797-812","update-policy":"https:\/\/doi.org\/10.1177\/sage-journals-update-policy","source":"Crossref","is-referenced-by-count":7,"title":["Development of Assays to Measure\n                    <i>GNE<\/i>\n                    Gene Potency and Gene Replacement in Skeletal Muscle"],"prefix":"10.1177","volume":"10","author":[{"given":"Deborah A.","family":"Zygmunt","sequence":"first","affiliation":[{"name":"Abigail Wexner Research Institute","place":["USA"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Patricia","family":"Lam","sequence":"additional","affiliation":[{"name":"Abigail Wexner Research Institute","place":["USA"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Anna","family":"Ashbrook","sequence":"additional","affiliation":[{"name":"Abigail Wexner Research Institute","place":["USA"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Katherine","family":"Koczwara","sequence":"additional","affiliation":[{"name":"Yale University School of Medicine","place":["USA"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Angela","family":"Lek","sequence":"additional","affiliation":[{"name":"Yale University School of Medicine","place":["USA"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Monkol","family":"Lek","sequence":"additional","affiliation":[{"name":"Yale University School of Medicine","place":["USA"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Paul T.","family":"Martin","sequence":"additional","affiliation":[{"name":"Abigail Wexner Research Institute","place":["USA"]},{"name":"The Ohio State University College of Medicine","place":["USA"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"179","published-online":{"date-parts":[[2023,7,12]]},"reference":[{"key":"e_1_3_3_2_2","doi-asserted-by":"publisher","DOI":"10.1136\/jnnp-2013-307051"},{"key":"e_1_3_3_3_2","doi-asserted-by":"publisher","DOI":"10.1038\/ng718"},{"key":"e_1_3_3_4_2","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.272.39.24319"},{"key":"e_1_3_3_5_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.nmd.2014.03.004"},{"key":"e_1_3_3_6_2","doi-asserted-by":"publisher","DOI":"10.1212\/NXG.0000000000000308"},{"key":"e_1_3_3_7_2","doi-asserted-by":"publisher","DOI":"10.1002\/humu.22583"},{"key":"e_1_3_3_8_2","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0173261"},{"key":"e_1_3_3_9_2","doi-asserted-by":"crossref","unstructured":"CroweKE ZygmuntDA HellerK Rodino-KlapacL NoguchiS NishinoI et al .Visualizing muscle sialic acid expression in the GNED207VTgGne-\/- Cmah-\/- model of GNE myopathy: A comparison of dietary and gene therapy approaches. 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