title: A combinatorial approach for targeted delivery using small molecules and reversible masking to bypass nonspecific uptake in vivo.
abstract: We have developed a multi-disciplinary approach combining molecular biology, delivery technology, combinatorial chemistry and reversible masking to create improved systemic, targeted delivery of plasmid DNA while avoiding nonspecific uptake in vivo. We initially used a well-characterized model targeting the asialolglycoprotein receptor in the liver. Using our bilamellar invaginated vesicle (BIV) liposomal delivery system with reversible masking, we increased expression in the liver by 76-fold, nearly equaling expression in first-pass organs using non-targeted complexes, with no expression in other organs. The same technology was then applied to efficiently target delivery to a human tumor microenvironment model. We achieved efficient, targeted delivery by attachment of specific targeting ligands to the surface of our BIV complexes in conjunction with reversible masking to bypass nonspecific tissues and organs. We identified ligands that target a human tumor microenvironment created in vitro by co-culturing primary human endothelial cells with human lung or pancreatic cancer cells. The model was confirmed by increased expression of tumor endothelial phenotypes including CD31 and vascular endothelial growth factor-A, and prolonged survival of endothelial capillary-like structures. The co-cultures were used for high-throughput screening of a specialized small molecule library to identify ligands specific for human tumor-associated endothelial cells in vitro. We identified small molecules that enhanced the transfection efficiency of tumor-associated endothelial cells, but not normal human endothelial cells or cancer cells. Intravenous (i.v.) injection of our targeted, reversibly masked complexes into mice, bearing human pancreatic tumor and endothelial cells, specifically increased transfection to this tumor microenvironment approximately 200-fold. Efficacy studies using our optimized targeted delivery of a plasmid encoding thrombospondin-1 eliminated tumors completely after five i.v. injections administered once every week.
authors.affiliation: Department of Molecular Physiology and Biophysics, Baylor College of Medicine, Houston, TX, USA.
authors.forename: Q
authors.initials: Q
authors.lastname: Shi
authors.forename: A T
authors.initials: AT
authors.lastname: Nguyen
authors.forename: Y
authors.initials: Y
authors.lastname: Angell
authors.forename: D
authors.initials: D
authors.lastname: Deng
authors.forename: C-R
authors.initials: CR
authors.lastname: Na
authors.forename: K
authors.initials: K
authors.lastname: Burgess
authors.forename: D D
authors.initials: DD
authors.lastname: Roberts
authors.forename: F C
authors.initials: FC
authors.lastname: Brunicardi
authors.forename: N S
authors.initials: NS
authors.lastname: Templeton
mesh_terms: D000818:Animals; D037263:Asialoglycoprotein Receptor / Q000378:metabolism; D045744:Cell Line, Tumor; D002478:Cells, Cultured; D018920:Coculture Techniques; D042783:Endothelial Cells / Q000378:metabolism; D015316:Genetic Therapy / Q000379:methods*; D006801:Humans; D008081:Liposomes / Q000737:chemistry / Q000378:metabolism; D051379:Mice; D009369:Neoplasms / Q000628:therapy*; D010957:Plasmids / Q000235:genetics / Q000378:metabolism; D019408:Platelet Endothelial Cell Adhesion Molecule-1 / Q000378:metabolism; D014162:Transfection; D042461:Vascular Endothelial Growth Factor A / Q000378:metabolism
chemical_list: D037263:Asialoglycoprotein Receptor; D008081:Liposomes; D019408:Platelet Endothelial Cell Adhesion Molecule-1; D042461:Vascular Endothelial Growth Factor A
references.citation: Am J Pathol. 2004 Aug;165(2):601-8
references.pmid: 15277233
references.citation: Science. 2000 Aug 18;289(5482):1197-202
references.pmid: 10947988
references.citation: Thromb Haemost. 2004 Dec;92(6):1438-45
references.pmid: 15583754
references.citation: J Virol. 2004 Aug;78(15):8271-80
references.pmid: 15254199
references.citation: Nat Rev Cancer. 2009 Mar;9(3):182-94
references.pmid: 19194382
references.citation: Mol Pharmacol. 2000 Feb;57(2):385-91
references.pmid: 10648649
references.citation: J Transl Med. 2004 Jun 25;2(1):22
references.pmid: 15219236
references.citation: J Gene Med. 2003 Apr;5(4):311-23
references.pmid: 12692865
references.citation: Nat Rev Drug Discov. 2004 May;3(5):391-400
references.pmid: 15136787
references.citation: J Virol. 2007 Mar;81(6):2688-99
references.pmid: 17192304
references.citation: Nat Rev Drug Discov. 2007 Sep;6(9):734-45
references.pmid: 17690708
references.citation: Cancer Sci. 2008 Mar;99(3):459-66
references.pmid: 18167133
references.citation: FASEB J. 1999 Jan;13(1):9-22
references.pmid: 9872925
references.citation: Nat Biotechnol. 1997 Jul;15(7):647-52
references.pmid: 9219267
references.citation: Ann N Y Acad Sci. 2003 May;995:151-61
references.pmid: 12814947
references.citation: Cancer Res. 2004 Nov 15;64(22):8249-55
references.pmid: 15548691
references.citation: N Engl J Med. 2004 Jun 3;350(23):2335-42
references.pmid: 15175435
references.citation: Clin Cancer Res. 2004 Jan 15;10(2):415-27
references.pmid: 14760060
references.citation: Diabetes. 2003 Dec;52(12):2959-68
references.pmid: 14633857
references.citation: Cancer Res. 2001 Sep 15;61(18):6649-55
references.pmid: 11559528
references.citation: Mol Genet Metab. 1998 Feb;63(2):103-9
references.pmid: 9562963
references.citation: Nat Methods. 2008 May;5(5):439-45
references.pmid: 18391960
references.citation: Mol Ther. 2004 Apr;9(4):489-95
references.pmid: 15093179
references.citation: J Virol. 2003 Apr;77(7):4449-56
references.pmid: 12634405
references.citation: Virology. 2008 Sep 15;379(1):125-34
references.pmid: 18672260
references.citation: Cancer Gene Ther. 2010 Apr;17(4):244-55
references.pmid: 19798121
references.citation: J Mol Biol. 2009 Mar 13;386(5):1368-81
references.pmid: 19063898
references.citation: APMIS. 2004 Jul-Aug;112(7-8):508-25
references.pmid: 15563313
references.citation: Proc Natl Acad Sci U S A. 2009 Jan 6;106(1):44-9
references.pmid: 19118196
references.citation: Science. 1998 Jan 16;279(5349):377-80
references.pmid: 9430587
references.citation: Nature. 2004 Jun 10;429(6992):629-35
references.pmid: 15190345
references.citation: Eur J Cancer. 2006 Dec;42(18):3127-39
references.pmid: 17098419
references.citation: Cancer Gene Ther. 2005 Feb;12(2):175-84
references.pmid: 15375381
references.citation: Nature. 2004 Feb 26;427(6977):787
references.pmid: 14985739
references.citation: Acc Chem Res. 2001 Oct;34(10):826-35
references.pmid: 11601967
references.citation: Clin Cancer Res. 2007 Jul 1;13(13):3968-76
references.pmid: 17606731
references.citation: Semin Cancer Biol. 2002 Apr;12(2):105-12
references.pmid: 12027582
references.citation: Virology. 1997 Jun 23;233(1):105-17
references.pmid: 9201220
references.citation: Genes Dev. 2007 Jun 15;21(12):1546-58
references.pmid: 17575055
references.citation: Nat Rev Cancer. 2007 Jun;7(6):475-85
references.pmid: 17522716
references.citation: Mol Genet Metab. 1998 Jul;64(3):193-7
references.pmid: 9719628
references.citation: J Clin Invest. 1998 Apr 1;101(7):1401-13
references.pmid: 9525983
references.citation: J Am Chem Soc. 2008 Jan 16;130(2):556-65
references.pmid: 18088119
references.citation: Cancer Res. 2000 Feb 1;60(3):722-7
references.pmid: 10676659
references.citation: J Neurosci. 2004 Sep 15;24(37):8009-18
references.pmid: 15371501
references.citation: Cancer. 2004 Jun 15;100(12):2491-9
references.pmid: 15197790
references.citation: Cancer Res. 2005 Feb 1;65(3):972-81
references.pmid: 15705898
references.citation: Cancer Res. 1994 Dec 15;54(24):6504-11
references.pmid: 7527299
references.citation: Ann Surg. 2001 May;233(5):603-11
references.pmid: 11323498
references.citation: Nat Rev Cancer. 2005 Jun;5(6):436-46
references.pmid: 15928674
references.citation: Neoplasia. 2008 Aug;10(8):886-96
references.pmid: 18670646
references.citation: J Gen Virol. 1995 Aug;76 ( Pt 8):1901-7
references.pmid: 7636471
references.citation: FEBS Lett. 2009 Jun 18;583(12):2100-7
references.pmid: 19481546
references.citation: Mol Ther. 2001 Mar;3(3):337-50
references.pmid: 11273776
references.citation: N Engl J Med. 2008 May 8;358(19):2039-49
references.pmid: 18463380
references.citation: Invest Ophthalmol Vis Sci. 2004 Sep;45(9):2915-21
references.pmid: 15326102
references.citation: Curr Opin Oncol. 2004 Jan;16(1):44-9
references.pmid: 14685092
references.citation: Mol Ther. 2002 Jun;5(6):755-61
references.pmid: 12027560
references.citation: Mol Ther. 2002 Mar;5(3):233-41
references.pmid: 11863412
references.citation: Mol Ther. 2008 Mar;16(3):516-24
references.pmid: 18195720
references.citation: J Virol. 1997 Feb;71(2):1453-65
references.pmid: 8995671
references.citation: Neoplasia. 2003 May-Jun;5(3):205-17
references.pmid: 12869304
references.citation: J Comb Chem. 2001 May-Jun;3(3):257-66
references.pmid: 11350249
references.citation: Science. 2002 Jun 28;296(5577):2404-7
references.pmid: 12089446
references.citation: Leuk Res. 2003 Aug;27(8):701-8
references.pmid: 12801528
references.citation: Pancreas. 2008 Aug;37(2):210-20
references.pmid: 18665085