title: Niemann-Pick C1 affects the gene delivery efficacy of degradable polymeric nanoparticles.
abstract: Despite intensive research effort, the rational design of improved nanoparticulate drug carriers remains challenging, in part due to a limited understanding of the determinants of nanoparticle entry and transport in target cells. Recent studies have shown that Niemann-Pick C1 (NPC1), the lysosome membrane protein that mediates trafficking of cholesterol in cells, is involved in the endosomal escape and subsequent infection caused by filoviruses, and that its absence promotes the retention and efficacy of lipid nanoparticles encapsulating siRNA. Here, we report that NPC1 deficiency results in dramatic reduction in internalization and transfection efficiency mediated by degradable cationic gene delivery polymers, poly(β-amino ester)s (PBAEs). PBAEs utilized cholesterol and dynamin-dependent endocytosis pathways, and these were found to be heavily compromised in NPC1-deficient cells. In contrast, the absence of NPC1 had minor effects on DNA uptake mediated by polyethylenimine or Lipofectamine 2000. Strikingly, stable overexpression of human NPC1 in chinese hamster ovary cells was associated with enhanced gene uptake (3-fold) and transfection (10-fold) by PBAEs. These findings reveal a role of NPC1 in the regulation of endocytic mechanisms affecting nanoparticle trafficking. We hypothesize that in-depth understanding sites of entry and endosomal escape may lead to highly efficient nanotechnologies for drug delivery.
authors.affiliation: Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology , Cambridge, Massachusetts 02139, United States.
authors.forename: Ahmed A
authors.initials: AA
authors.lastname: Eltoukhy
authors.forename: Gaurav
authors.initials: G
authors.lastname: Sahay
authors.forename: James M
authors.initials: JM
authors.lastname: Cunningham
authors.forename: Daniel G
authors.initials: DG
authors.lastname: Anderson
mesh_terms: D000736:Androstenes / Q000494:pharmacology; D000818:Animals; D016466:CHO Cells; D002352:Carrier Proteins / Q000378:metabolism*; D002784:Cholesterol / Q000378:metabolism; D006224:Cricetinae; D003412:Cricetulus; D004247:DNA / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D004337:Drug Carriers / Q000737:chemistry* / Q000378:metabolism*; D004705:Endocytosis / Q000187:drug effects; D005347:Fibroblasts / Q000378:metabolism; D005786:Gene Expression Regulation; D006801:Humans; D047908:Intracellular Signaling Peptides and Proteins; D008562:Membrane Glycoproteins / Q000172:deficiency / Q000378:metabolism*; D051379:Mice; D053758:Nanoparticles; D000077296:Niemann-Pick C1 Protein; D011108:Polymers / Q000737:chemistry* / Q000378:metabolism*; D014162:Transfection
chemical_list: D000736:Androstenes; D002352:Carrier Proteins; D004337:Drug Carriers; D047908:Intracellular Signaling Peptides and Proteins; D008562:Membrane Glycoproteins; C106881:NPC1 protein, human; D000077296:Niemann-Pick C1 Protein; D011108:Polymers; C507253:poly(beta-amino ester); C006261:3-beta-(2-(diethylamino)ethoxy)androst-5-en-17-one; D004247:DNA; D002784:Cholesterol