{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,8,3]],"date-time":"2025-08-03T04:24:02Z","timestamp":1754195042559,"version":"3.37.3"},"reference-count":46,"publisher":"Public Library of Science (PLoS)","issue":"12","license":[{"start":{"date-parts":[[2022,12,30]],"date-time":"2022-12-30T00:00:00Z","timestamp":1672358400000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/100000057","name":"National Institute of General Medical Sciences","doi-asserted-by":"publisher","award":["GM115866"],"award-info":[{"award-number":["GM115866"]}],"id":[{"id":"10.13039\/100000057","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100001790","name":"Ara Parseghian Medical Research Foundation","doi-asserted-by":"publisher","id":[{"id":"10.13039\/100001790","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000097","name":"National Center for Research Resources","doi-asserted-by":"publisher","award":["S10DO20029"],"award-info":[{"award-number":["S10DO20029"]}],"id":[{"id":"10.13039\/100000097","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["www.plosone.org"],"crossmark-restriction":false},"short-container-title":["PLoS ONE"],"abstract":"<jats:p>Niemann-Pick Type C is a rare metabolic disorder characterized by the cellular accumulation of cholesterol within endosomal and lysosomal compartments. 2-Hydroxypropyl-\u03b2-cyclodextrin (HP-\u03b2-CD) containing polyrotaxanes represent an attractive approach for treating this disease due to their ability to circulate in the blood stream for longer periods of time as a prodrug form of HP-\u03b2-CD. Once inside the cell, the macromolecular structure is thought to break down into the Pluronic precursor and the active cyclodextrin agent that promotes cholesterol mobilization from the aberrant accumulations within NPC-deficient cells. We now report that both cholesterol and decaarginine (R<jats:sub>10<\/jats:sub>) endcapped polyrotaxanes are able to remove cholesterol from NPC1 patient fibroblasts. R<jats:sub>10<\/jats:sub> endcapped materials enter these cells and are localized within endosomes after 16 h. The cholesterol mobilization from endo-lysosomal compartments of NPC1 cells by the polyrotaxanes was directly related to their extent of endcapping and their threading efficiency. Incorporation of 4-sulfobutylether-\u03b2-cyclodextrin (SBE-\u03b2-CD) significantly improved cholesterol mobilization due to the improved solubility of the compounds. Additionally, in our efforts to scale-up the synthesis for preclinical studies, we prepared a library of polyrotaxanes using a solid phase synthesis method. These compounds also led to significant cholesterol mobilization from the cells, however, cytotoxicity studies showed that they were substantially more toxic than those prepared by the solvent-assisted method, thus limiting the therapeutic utility of agents prepared by this expedited method. Our findings demonstrate that complete endcapping of the polyrotaxanes and improved solubility are important design features for delivering high copy numbers of therapeutic \u03b2-CD to promote enhanced sterol clearance in human NPC1-deficient cells.<\/jats:p>","DOI":"10.1371\/journal.pone.0268613","type":"journal-article","created":{"date-parts":[[2022,12,30]],"date-time":"2022-12-30T18:59:54Z","timestamp":1672426794000},"page":"e0268613","update-policy":"https:\/\/doi.org\/10.1371\/journal.pone.corrections_policy","source":"Crossref","is-referenced-by-count":1,"title":["Structure-function relationships of cholesterol mobilization from the endo-lysosome compartment of NPC1-deficient human cells by \u03b2-CD polyrotaxanes"],"prefix":"10.1371","volume":"17","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-4406-8790","authenticated-orcid":true,"given":"Shayak","family":"Samaddar","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Debosreeta","family":"Bose","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Bradley P.","family":"Loren","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Joseph L.","family":"Skulsky","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0807-9158","authenticated-orcid":true,"given":"Olga","family":"Ilnytska","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Zachary J.","family":"Struzik","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Judith","family":"Storch","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0746-1526","authenticated-orcid":true,"given":"David H.","family":"Thompson","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"340","published-online":{"date-parts":[[2022,12,30]]},"reference":[{"issue":"22","key":"pone.0268613.ref001","doi-asserted-by":"crossref","first-page":"20917","DOI":"10.1074\/jbc.R400040200","article-title":"Niemann-Pick Type C Disease and Intracellular Cholesterol Trafficking","volume":"280","author":"T-Y Chang","year":"2005","journal-title":"Journal of Biological Chemistry"},{"issue":"3","key":"pone.0268613.ref002","doi-asserted-by":"crossref","first-page":"307","DOI":"10.1016\/S1096-7192(03)00136-7","article-title":"Fabry disease: D313Y is an \u03b1-galactosidase A sequence variant that causes pseudodeficient activity in plasma","volume":"80","author":"R Froissart","year":"2003","journal-title":"Molecular Genetics and Metabolism"},{"issue":"1","key":"pone.0268613.ref003","doi-asserted-by":"crossref","first-page":"16","DOI":"10.1186\/1750-1172-5-16","article-title":"Niemann-Pick disease type C","volume":"5","author":"MT Vanier","year":"2010","journal-title":"Orphanet Journal of Rare Diseases"},{"issue":"5323","key":"pone.0268613.ref004","doi-asserted-by":"crossref","first-page":"228","DOI":"10.1126\/science.277.5323.228","article-title":"Niemann-Pick C1 Disease Gene: Homology to Mediators of Cholesterol Homeostasis","volume":"277","author":"ED Carstea","year":"1997","journal-title":"Science"},{"issue":"40","key":"pone.0268613.ref005","doi-asserted-by":"crossref","first-page":"15287","DOI":"10.1073\/pnas.0807328105","article-title":"NPC2 facilitates bidirectional transfer of cholesterol between NPC1 and lipid bilayers, a step in cholesterol egress from lysosomes","volume":"105","author":"RE Infante","year":"2008","journal-title":"Proceedings of the National Academy of Sciences"},{"issue":"4","key":"pone.0268613.ref006","doi-asserted-by":"crossref","first-page":"309","DOI":"10.1203\/PDR.0b013e3181ee4dd2","article-title":"Weekly cyclodextrin administration normalizes cholesterol metabolism in nearly every organ of the Niemann-Pick type C1 mouse and markedly prolongs life","volume":"68","author":"CM Ramirez","year":"2010","journal-title":"Pediatric research"},{"issue":"1","key":"pone.0268613.ref007","doi-asserted-by":"crossref","first-page":"52","DOI":"10.1203\/PDR.0b013e3181df4623","article-title":"2-hydroxypropyl-beta-cyclodextrin raises hearing threshold in normal cats and in cats with Niemann-Pick type C disease","volume":"68","author":"Ward S","year":"2010","journal-title":"Pediatric research"},{"key":"pone.0268613.ref008","doi-asserted-by":"crossref","first-page":"5477","DOI":"10.1073\/pnas.0914309107","article-title":"Endocytosis of beta-cyclodextrins is responsible for cholesterol reduction in Niemann-Pick type C mutant cells","volume":"107","author":"AI Rosenbaum","year":"2010","journal-title":"Proc. Natl. Acad. Sci"},{"issue":"10104","key":"pone.0268613.ref009","doi-asserted-by":"crossref","first-page":"1758","DOI":"10.1016\/S0140-6736(17)31465-4","article-title":"Intrathecal 2-hydroxypropyl-\u03b2-cyclodextrin decreases neurological disease progression in Niemann-Pick disease, type C1: a non-randomised, open-label, phase 1\u20132 trial","volume":"390","author":"DS Ory","year":"2017","journal-title":"Lancet (London, England)."},{"key":"pone.0268613.ref010","doi-asserted-by":"crossref","first-page":"355","DOI":"10.3389\/fncel.2017.00355","article-title":"Cyclodextrins and Iatrogenic Hearing Loss: New Drugs with Significant Risk.","volume":"11","author":"MA Crumling","year":"2017","journal-title":"Front Cell Neurosci"},{"issue":"2","key":"pone.0268613.ref011","doi-asserted-by":"crossref","first-page":"231","DOI":"10.1016\/j.ymgme.2013.04.003","article-title":"Lung toxicity of hydroxypropyl-\u03b2-cyclodextrin infusion","volume":"109","author":"Y-H Chien","year":"2013","journal-title":"Molecular Genetics and Metabolism"},{"issue":"12","key":"pone.0268613.ref012","doi-asserted-by":"crossref","first-page":"4189","DOI":"10.1021\/bm400922a","article-title":"Synthesis of 2-Hydroxypropyl-\u03b2-cyclodextrin\/Pluronic-Based Polyrotaxanes via Heterogeneous Reaction as Potential Niemann-Pick Type C Therapeutics.","volume":"14","author":"YA Mondjinou","year":"2013","journal-title":"Biomacromolecules"},{"issue":"19","key":"pone.0268613.ref013","doi-asserted-by":"crossref","first-page":"3242","DOI":"10.1021\/bi3010889","article-title":"Synthesis, characterization, and evaluation of pluronic-based beta-cyclodextrin polyrotaxanes for mobilization of accumulated cholesterol from Niemann-Pick type C fibroblasts","volume":"52","author":"CJ Collins","year":"2013","journal-title":"Biochemistry"},{"issue":"9","key":"pone.0268613.ref014","doi-asserted-by":"crossref","first-page":"2777","DOI":"10.1021\/acs.biomac.6b00508","article-title":"Influence of Molecular Structure on the In Vivo Performance of Flexible Rod Polyrotaxanes.","volume":"17","author":"CJ Collins","year":"2016","journal-title":"Biomacromolecules"},{"key":"pone.0268613.ref015","doi-asserted-by":"crossref","first-page":"46737","DOI":"10.1038\/srep46737","article-title":"Pluronic based \u03b2-cyclodextrin polyrotaxanes for treatment of Niemann-Pick Type C disease.","volume":"7","author":"CJ Collins","year":"2017","journal-title":"Scientific Reports."},{"key":"pone.0268613.ref016","doi-asserted-by":"crossref","first-page":"148","DOI":"10.1016\/j.jconrel.2017.11.016","article-title":"Polyrotaxane-based systemic delivery of \u03b2-cyclodextrins for potentiating therapeutic efficacy in a mouse model of Niemann-Pick type C disease","volume":"269","author":"A Tamura","year":"2018","journal-title":"Journal of Controlled Release"},{"issue":"4","key":"pone.0268613.ref017","doi-asserted-by":"crossref","first-page":"528","DOI":"10.1039\/C8TB02950D","article-title":"Synthesis of the anionic hydroxypropyl-\u03b2-cyclodextrin:poly(decamethylenephosphate) polyrotaxane and evaluation of its cholesterol efflux potential in Niemann\u2013Pick C1 cells.","volume":"7","author":"K Egele","year":"2019","journal-title":"Journal of Materials Chemistry B"},{"issue":"3","key":"pone.0268613.ref018","doi-asserted-by":"crossref","first-page":"1025","DOI":"10.1021\/cr940089p","article-title":"Solvent-Free Organic Synthesis.","volume":"100","author":"K Tanaka","year":"2000","journal-title":"Chemical Reviews"},{"issue":"43","key":"pone.0268613.ref019","doi-asserted-by":"crossref","first-page":"23831","DOI":"10.1021\/acsami.5b01016","article-title":"Impact of Mixed \u03b2-Cyclodextrin Ratios on Pluronic Rotaxanation Efficiency and Product Solubility.","volume":"7","author":"YA Mondjinou","year":"2015","journal-title":"ACS Applied Materials & Interfaces."},{"issue":"8","key":"pone.0268613.ref020","doi-asserted-by":"crossref","first-page":"1710","DOI":"10.1194\/jlr.M700125-JLR200","article-title":"Receptor-mediated and bulk-phase endocytosis cause macrophage and cholesterol accumulation in Niemann-Pick C disease","volume":"48","author":"B Liu","year":"2007","journal-title":"Journal of Lipid Research"},{"issue":"25","key":"pone.0268613.ref021","doi-asserted-by":"crossref","first-page":"16010","DOI":"10.1074\/jbc.272.25.16010","article-title":"A truncated HIV-1 Tat protein basic domain rapidly translocates through the plasma membrane and accumulates in the cell nucleus","volume":"272","author":"E Vives","year":"1997","journal-title":"The Journal of biological chemistry"},{"issue":"9","key":"pone.0268613.ref022","doi-asserted-by":"crossref","first-page":"1806","DOI":"10.1016\/j.febslet.2009.11.046","article-title":"Arginine-rich cell-penetrating peptides","volume":"584","author":"N Schmidt","year":"2010","journal-title":"FEBS Letters"},{"key":"pone.0268613.ref023","first-page":"2011","article-title":"Mechanisms of Cellular Uptake of Cell-Penetrating Peptides.","author":"F Madani","year":"2011","journal-title":"J Biophys"},{"issue":"10","key":"pone.0268613.ref024","doi-asserted-by":"crossref","first-page":"1029","DOI":"10.1039\/c3bm00189j","article-title":"Microfluidic assembly of cationic-\u03b2-cyclodextrin:hyaluronic acid-adamantane host:guest pDNA nanoparticles","volume":"1","author":"A Kulkarni","year":"2013","journal-title":"Biomaterials Science"},{"issue":"10","key":"pone.0268613.ref025","doi-asserted-by":"crossref","first-page":"e110632","DOI":"10.1371\/journal.pone.0110632","article-title":"Selective DNA Delivery to Tumor Cells Using an Oligoarginine-LTVSPWY Peptide.","volume":"9","author":"C Gong","year":"2014","journal-title":"PLOS ONE"},{"issue":"6","key":"pone.0268613.ref026","doi-asserted-by":"crossref","first-page":"772","DOI":"10.1002\/bip.21681","article-title":"Enhanced cellular uptake and metabolic stability of lipo-oligoarginine peptides","volume":"96","author":"JS Lee","year":"2011","journal-title":"Peptide Science"},{"issue":"9","key":"pone.0268613.ref027","doi-asserted-by":"crossref","first-page":"2438","DOI":"10.1021\/bi035933x","article-title":"Pathway for polyarginine entry into mammalian cells","volume":"43","author":"SM Fuchs","year":"2004","journal-title":"Biochemistry"},{"issue":"7","key":"pone.0268613.ref028","doi-asserted-by":"crossref","first-page":"2692","DOI":"10.1021\/bi048046i","article-title":"Binding of Oligoarginine to Membrane Lipids and Heparan Sulfate:\u2009 Structural and Thermodynamic Characterization of a Cell-Penetrating Peptide","volume":"44","author":"E Gon\u00e7alves","year":"2005","journal-title":"Biochemistry"},{"issue":"5","key":"pone.0268613.ref029","doi-asserted-by":"crossref","first-page":"855","DOI":"10.1039\/c2mb25467k","article-title":"Cell-penetrating peptides (CPPs) as a vector for the delivery of siRNAs into cells.","volume":"9","author":"I Nakase","year":"2013","journal-title":"Molecular BioSystems."},{"issue":"4","key":"pone.0268613.ref030","doi-asserted-by":"crossref","first-page":"2437","DOI":"10.1074\/jbc.M110017200","article-title":"Possible Existence of Common Internalization Mechanisms among Arginine-rich Peptides","volume":"277","author":"T Suzuki","year":"2002","journal-title":"Journal of Biological Chemistry"},{"issue":"37","key":"pone.0268613.ref031","doi-asserted-by":"crossref","first-page":"35109","DOI":"10.1074\/jbc.M301726200","article-title":"Antennapedia and HIV Transactivator of Transcription (TAT) \u201cProtein Transduction Domains\u201d Promote Endocytosis of High Molecular Weight Cargo upon Binding to Cell Surface Glycosaminoglycans.","volume":"278","author":"S Console","year":"2003","journal-title":"Journal of Biological Chemistry"},{"key":"pone.0268613.ref032","first-page":"357","volume-title":"Methods in Cell Biology","author":"MT Vanier","year":"2015"},{"key":"pone.0268613.ref033","doi-asserted-by":"crossref","first-page":"5620","DOI":"10.1073\/pnas.1014890108","article-title":"Histone deacetylase inhibitor treatment dramatically reduces cholesterol accumulation in Niemann-Pick type C1 mutant human fibroblasts","volume":"108","author":"NH Pipalia","year":"2011","journal-title":"Proc. Natl. Acad. Sci"},{"key":"pone.0268613.ref034","doi-asserted-by":"crossref","first-page":"e48561","DOI":"10.1371\/journal.pone.0048561","article-title":"Quantitative comparison of the efficacy of various compounds in lowering intracellular cholesterol levels in Niemann-Pick type C fibroblasts.","volume":"7","author":"ZT Wehrmann","year":"2012","journal-title":"PLOS ONE."},{"key":"pone.0268613.ref035","doi-asserted-by":"crossref","first-page":"284","DOI":"10.1194\/jlr.M500388-JLR200","article-title":"Automated microscopy screening for compounds that partially revert cholesterol accumulation in Niemann-Pick C cells","volume":"47","author":"NH Pipalia","year":"2006","journal-title":"Journal of Lipid Research"},{"key":"pone.0268613.ref036","doi-asserted-by":"crossref","first-page":"877","DOI":"10.2147\/TCRM.S5777","article-title":"New therapies in the management of Niemann-Pick type C disease: clinical utility of miglustat.","volume":"5","author":"JE Wrait","year":"2009","journal-title":"Therapeutics and Clinical Risk Management"},{"issue":"2","key":"pone.0268613.ref037","doi-asserted-by":"crossref","first-page":"422","DOI":"10.1016\/j.bbrc.2014.02.019","article-title":"Distribution of the LDL receptor within clathrin-coated pits and caveolae in rat and human liver","volume":"445","author":"S Ivaturi","year":"2014","journal-title":"Biochemical and Biophysical Research Communications"},{"issue":"4","key":"pone.0268613.ref038","doi-asserted-by":"crossref","first-page":"389","DOI":"10.1111\/j.1600-0854.2007.00540.x","article-title":"Internalization and trafficking of cell surface proteoglycans and proteoglycan-binding ligands","volume":"8","author":"CK Payne","year":"2007","journal-title":"Traffic (Copenhagen, Denmark)."},{"key":"pone.0268613.ref039","doi-asserted-by":"crossref","first-page":"3550","DOI":"10.1021\/acs.bioconjchem.8b00525","article-title":"Gd3+:DOTA-modified 2-hydroxypropyl-\u03b2-cyclodextrin\/4-sulfobutyl ether-\u03b2-cyclodextrin-based polyrotaxanes as long circulating high relaxivity MRI contrast agents.","volume":"29","author":"YA Mondjinou","year":"2018","journal-title":"Bioconjugate Chemistry"},{"issue":"1","key":"pone.0268613.ref040","doi-asserted-by":"crossref","first-page":"139","DOI":"10.1016\/0021-9673(94)87083-7","article-title":"Effect of the degree of substitution of (2-hydroxy)propyl-\u03b2-cyclodextrin on the enantioseparation of organic acids by capillary electrophoresis.","volume":"678","author":"IE Valk\u00f3","year":"1994","journal-title":"Journal of Chromatography A"},{"issue":"5","key":"pone.0268613.ref041","doi-asserted-by":"crossref","first-page":"1092","DOI":"10.1007\/s11095-007-9506-y","article-title":"Structural behaviour of 2-hydroxypropyl-beta-cyclodextrin in water: molecular dynamics simulation studies","volume":"25","author":"CW Yong","year":"2008","journal-title":"Pharmaceutical research"},{"key":"pone.0268613.ref042","doi-asserted-by":"crossref","first-page":"24567","DOI":"10.1038\/srep24567","article-title":"Size Dependent Cellular Uptake of Rod-like Bionanoparticles with Different Aspect Ratios.","volume":"6","author":"X Liu","year":"2016","journal-title":"Scientific Reports."},{"issue":"12","key":"pone.0268613.ref043","doi-asserted-by":"crossref","first-page":"1251","DOI":"10.1093\/glycob\/cwl045","article-title":"Constitutive and vitamin C-induced, NO-catalyzed release of heparan sulfate from recycling glypican-1 in late endosomes","volume":"16","author":"F Cheng","year":"2006","journal-title":"Glycobiology"},{"issue":"5","key":"pone.0268613.ref044","doi-asserted-by":"crossref","first-page":"1644","DOI":"10.1523\/JNEUROSCI.5491-10.2011","article-title":"Heparan sulphate proteoglycan and the low-density lipoprotein receptor-related protein 1 constitute major pathways for neuronal amyloid-beta uptake.","volume":"31","author":"T Kanekiyo","year":"2011","journal-title":"The Journal of neuroscience: the official journal of the Society for Neuroscience."},{"issue":"33","key":"pone.0268613.ref045","first-page":"E3138","article-title":"Heparan sulfate proteoglycans mediate internalization and propagation of specific proteopathic seeds","volume":"110","author":"BB Holmes","year":"2013","journal-title":"Proceedings of the National Academy of Sciences of the United States of America"},{"key":"pone.0268613.ref046","doi-asserted-by":"crossref","first-page":"637","DOI":"10.1038\/sj.cdd.4401647","article-title":"Heparan sulfate proteoglycan binding promotes APRIL-induced tumor cell proliferation","volume":"12","author":"J Hendriks","year":"2005","journal-title":"Cell Death And Differentiation"}],"container-title":["PLOS ONE"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/dx.plos.org\/10.1371\/journal.pone.0268613","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2022,12,30]],"date-time":"2022-12-30T19:00:24Z","timestamp":1672426824000},"score":1,"resource":{"primary":{"URL":"https:\/\/dx.plos.org\/10.1371\/journal.pone.0268613"}},"subtitle":[],"editor":[{"given":"Andrea","family":"Dardis","sequence":"first","affiliation":[],"role":[{"role":"editor","vocabulary":"crossref"}]}],"short-title":[],"issued":{"date-parts":[[2022,12,30]]},"references-count":46,"journal-issue":{"issue":"12","published-online":{"date-parts":[[2022,12,30]]}},"URL":"https:\/\/doi.org\/10.1371\/journal.pone.0268613","relation":{},"ISSN":["1932-6203"],"issn-type":[{"type":"electronic","value":"1932-6203"}],"subject":[],"published":{"date-parts":[[2022,12,30]]}}}