{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,28]],"date-time":"2026-09-28T15:28:22Z","timestamp":1790609302868,"version":"4.1.0"},"reference-count":32,"publisher":"MDPI AG","issue":"21","license":[{"start":{"date-parts":[[2023,10,25]],"date-time":"2023-10-25T00:00:00Z","timestamp":1698192000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"California Institute of Regenerative Medicine","award":["13072"],"award-info":[{"award-number":["13072"]}]},{"name":"Swim Across America Foundation","award":["13072"],"award-info":[{"award-number":["13072"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Sphingosine-1-phosphate lyase insufficiency syndrome (SPLIS) is an inborn error of metabolism caused by inactivating mutations in SGPL1, the gene encoding sphingosine-1-phosphate lyase (SPL), an essential enzyme needed to degrade sphingolipids. SPLIS features include glomerulosclerosis, adrenal insufficiency, neurological defects, ichthyosis, and immune deficiency. Currently, there is no cure for SPLIS, and severely affected patients often die in the first years of life. We reported that adeno-associated virus (AAV) 9-mediated SGPL1 gene therapy (AAV-SPL) given to newborn Sgpl1 knockout mice that model SPLIS and die in the first few weeks of life prolonged their survival to 4.5 months and prevented or delayed the onset of SPLIS phenotypes. In this study, we tested the efficacy of a modified AAV-SPL, which we call AAV-SPL 2.0, in which the original cytomegalovirus (CMV) promoter driving the transgene is replaced with the synthetic \u201cCAG\u201d promoter used in several clinically approved gene therapy agents. AAV-SPL 2.0 infection of human embryonic kidney (HEK) cells led to 30% higher SPL expression and enzyme activity compared to AAV-SPL. Newborn Sgpl1 knockout mice receiving AAV-SPL 2.0 survived \u2265 5 months and showed normal neurodevelopment, 85% of normal weight gain over the first four months, and delayed onset of proteinuria. Over time, treated mice developed nephrosis and glomerulosclerosis, which likely resulted in their demise. Our overall findings show that AAV-SPL 2.0 performs equal to or better than AAV-SPL. However, improved kidney targeting may be necessary to achieve maximally optimized gene therapy as a potentially lifesaving SPLIS treatment.<\/jats:p>","DOI":"10.3390\/ijms242115560","type":"journal-article","created":{"date-parts":[[2023,10,25]],"date-time":"2023-10-25T04:14:47Z","timestamp":1698207287000},"page":"15560","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":9,"title":["AAV-SPL 2.0, a Modified Adeno-Associated Virus Gene Therapy Agent for the Treatment of Sphingosine Phosphate Lyase Insufficiency Syndrome"],"prefix":"10.3390","volume":"24","author":[{"given":"Ranjha","family":"Khan","sequence":"first","affiliation":[{"name":"Department of Pediatrics, Division of Hematology\/Oncology, University of California, San Francisco, CA 94143, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Babak","family":"Oskouian","sequence":"additional","affiliation":[{"name":"Department of Pediatrics, Division of Hematology\/Oncology, University of California, San Francisco, CA 94143, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Joanna Y.","family":"Lee","sequence":"additional","affiliation":[{"name":"Department of Pediatrics, Division of Hematology\/Oncology, University of California, San Francisco, CA 94143, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jeffrey B.","family":"Hodgin","sequence":"additional","affiliation":[{"name":"Department of Pathology, University of Michigan School of Medicine, Ann Arbor, MI 48109, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yingbao","family":"Yang","sequence":"additional","affiliation":[{"name":"Department of Pathology, University of Michigan School of Medicine, Ann Arbor, MI 48109, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Gizachew","family":"Tassew","sequence":"additional","affiliation":[{"name":"Department of Pediatrics, Division of Hematology\/Oncology, University of California, San Francisco, CA 94143, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4894-919X","authenticated-orcid":false,"given":"Julie D.","family":"Saba","sequence":"additional","affiliation":[{"name":"Department of Pediatrics, Division of Hematology\/Oncology, University of California, San Francisco, CA 94143, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2023,10,25]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"912","DOI":"10.1172\/JCI89626","article-title":"Mutations in sphingosine-1-phosphate lyase cause nephrosis with ichthyosis and adrenal insufficiency","volume":"127","author":"Lovric","year":"2017","journal-title":"J. Clin. Investig."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"942","DOI":"10.1172\/JCI90171","article-title":"Sphingosine-1-phosphate lyase mutations cause primary adrenal insufficiency and steroid-resistant nephrotic syndrome","volume":"127","author":"Prasad","year":"2017","journal-title":"J. Clin. Investig."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"173","DOI":"10.1111\/imr.12760","article-title":"Secrets and lyase: Control of sphingosine 1-phosphate distribution","volume":"289","author":"Dixit","year":"2019","journal-title":"Immunol. Rev."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"244","DOI":"10.1016\/S0076-6879(00)11087-0","article-title":"Sphingosine-1-phosphate lyase","volume":"311","year":"2000","journal-title":"Methods Enzymol."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"425","DOI":"10.1007\/s12519-022-00615-4","article-title":"Sphingosine phosphate lyase insufficiency syndrome: A systematic review","volume":"19","author":"Pournasiri","year":"2023","journal-title":"World J. Pediatr."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"711","DOI":"10.1007\/s00467-022-05656-5","article-title":"A rare cause of nephrotic syndrome-sphingosine-1-phosphate lyase (SGPL1) deficiency: 6 cases and a review of the literature","volume":"38","author":"Canpolat","year":"2023","journal-title":"Pediatr. Nephrol."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"347","DOI":"10.1111\/imm.12272","article-title":"Sphingosine-1-phosphate receptor 1 signalling in T cells: Trafficking and beyond","volume":"142","author":"Garris","year":"2014","journal-title":"Immunology"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"988","DOI":"10.1016\/j.cell.2012.01.038","article-title":"Sphingolipid metabolism cooperates with BAK and BAX to promote the mitochondrial pathway of apoptosis","volume":"148","author":"Chipuk","year":"2012","journal-title":"Cell"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"601","DOI":"10.1146\/annurev-virology-092818-015530","article-title":"Recombinant adeno-associated virus gene therapy in light of Luxturna (and Zolgensma and Glybera): Where are we, and how did we get here?","volume":"6","author":"Keeler","year":"2019","journal-title":"Annu. Rev. Virol."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"358","DOI":"10.1038\/s41573-019-0012-9","article-title":"Adeno-associated virus vector as a platform for gene therapy delivery","volume":"18","author":"Wang","year":"2019","journal-title":"Nat. Rev. Drug Discov."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"583","DOI":"10.1128\/CMR.00008-08","article-title":"Gene therapy using adeno-associated virus vectors","volume":"21","author":"Daya","year":"2008","journal-title":"Clin. Microbiol. Rev."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"107453","DOI":"10.1016\/j.pharmthera.2019.107453","article-title":"Adeno-associated viral vector-mediated immune responses: Understanding barriers to gene delivery","volume":"207","author":"Nidetz","year":"2020","journal-title":"Pharmacol. Ther."},{"key":"ref_13","unstructured":"Ungar, L. (U.S. News & World Report, 2022). Deaths in US gene therapy study sparks search for answers, U.S. News & World Report."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"e145936","DOI":"10.1172\/jci.insight.145936","article-title":"Efficacy of AAV9-mediated SGPL1 gene transfer in a mouse model of S1P lyase insufficiency syndrome","volume":"6","author":"Zhao","year":"2021","journal-title":"JCI Insight"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"1008","DOI":"10.1089\/hum.2021.29182.bfs","article-title":"Fourth Boy Dies in Clinical Trial of Astellas\u2019 AT132","volume":"32","author":"Philippidis","year":"2021","journal-title":"Hum. Gene Ther."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"908","DOI":"10.1089\/hum.2020.29133.bfs","article-title":"After Third Death, Audentes\u2019 AT132 Remains on Clinical Hold","volume":"31","author":"Philippidis","year":"2020","journal-title":"Hum. Gene Ther."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"20","DOI":"10.1016\/j.gene.2008.09.033","article-title":"Epigenetic regulation of cytomegalovirus major immediate-early promoter activity in transgenic mice","volume":"428","author":"Mehta","year":"2009","journal-title":"Gene"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"605380","DOI":"10.3389\/fonc.2020.605380","article-title":"Strategies for Targeting Gene Therapy in Cancer Cells With Tumor-Specific Promoters","volume":"10","year":"2020","journal-title":"Front. Oncol."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"269","DOI":"10.1016\/0378-1119(89)90209-6","article-title":"Expression vector system based on the chicken beta-actin promoter directs efficient production of interleukin-5","volume":"79","author":"Miyazaki","year":"1989","journal-title":"Gene"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"193","DOI":"10.1016\/0378-1119(91)90434-D","article-title":"Efficient selection for high-expression transfectants with a novel eukaryotic vector","volume":"108","author":"Niwa","year":"1991","journal-title":"Gene"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"52","DOI":"10.1038\/ng1922","article-title":"PDGF signaling specificity is mediated through multiple immediate early genes","volume":"39","author":"Schmahl","year":"2007","journal-title":"Nat. Genet."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"127","DOI":"10.3389\/fendo.2014.00127","article-title":"Podocyte pathology and nephropathy\u2014Sphingolipids in glomerular diseases","volume":"5","author":"Merscher","year":"2014","journal-title":"Front. Endocrinol."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"76","DOI":"10.1038\/nrneph.2014.216","article-title":"Causes and pathogenesis of focal segmental glomerulosclerosis","volume":"11","author":"Fogo","year":"2015","journal-title":"Nat. Rev. Nephrol."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"1421","DOI":"10.1038\/nm.2250","article-title":"STAT3-induced S1PR1 expression is crucial for persistent STAT3 activation in tumors","volume":"16","author":"Lee","year":"2010","journal-title":"Nat. Med."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"88","DOI":"10.1097\/MNH.0000000000000079","article-title":"JAK inhibition and progressive kidney disease","volume":"24","author":"Brosius","year":"2015","journal-title":"Curr. Opin. Nephrol. Hypertens."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"973","DOI":"10.2215\/CJN.11010919","article-title":"JAK-STAT activity in peripheral blood cells and kidney tissue in IgA nephropathy","volume":"15","author":"Tao","year":"2020","journal-title":"Clin. J. Am. Soc. Nephrol."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"F1151","DOI":"10.1152\/ajprenal.00034.2019","article-title":"Targeting STAT3 signaling in kidney disease","volume":"316","author":"Pace","year":"2019","journal-title":"Am. J. Physiol. Renal Physiol."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"1661","DOI":"10.1007\/s00467-009-1163-4","article-title":"Expression of JAKs\/STATs pathway molecules in rat model of rapid focal segmental glomerulosclerosis","volume":"24","author":"Liang","year":"2009","journal-title":"Pediatr. Nephrol."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"384","DOI":"10.1016\/j.trsl.2015.03.002","article-title":"Janus kinase 2\/signal transducer and activator of transcription 3 inhibitors attenuate the effect of cardiotrophin-like cytokine factor 1 and human focal segmental glomerulosclerosis serum on glomerular filtration barrier","volume":"166","author":"Sharma","year":"2015","journal-title":"Transl. Res."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"15055","DOI":"10.1038\/mtm.2015.55","article-title":"AAV9-mediated central nervous system-targeted gene delivery via cisterna magna route in mice","volume":"3","author":"Lukashchuk","year":"2016","journal-title":"Mol. Ther. Methods Clin. Dev."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"101","DOI":"10.1016\/j.chemphyslip.2015.09.006","article-title":"A facile stable-isotope dilution method for determination of sphingosine phosphate lyase activity","volume":"194","author":"Suh","year":"2016","journal-title":"Chem. Phys. Lipids"},{"key":"ref_32","first-page":"469","article-title":"Sphingosine-1-phosphate in inflammatory bowel disease and colitis-associated colon cancer: The fat\u2019s in the fire","volume":"4","author":"Suh","year":"2015","journal-title":"Transl. Cancer Res."}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/24\/21\/15560\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T21:11:24Z","timestamp":1760130684000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/24\/21\/15560"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2023,10,25]]},"references-count":32,"journal-issue":{"issue":"21","published-online":{"date-parts":[[2023,11]]}},"alternative-id":["ijms242115560"],"URL":"https:\/\/doi.org\/10.3390\/ijms242115560","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2023,10,25]]}}}