A combinatorial approach for targeted delivery using small molecules and reversible masking to bypass nonspecific uptake in vivo.
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.
Shi
Q
Q
Department of Molecular Physiology and Biophysics, Baylor College of Medicine, Houston, TX, USA.
Nguyen
A T
AT
Angell
Y
Y
Deng
D
D
Na
C-R
CR
Burgess
K
K
Roberts
D D
DD
Brunicardi
F C
FC
Templeton
N S
NS
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
D037263:Asialoglycoprotein Receptor; D008081:Liposomes; D019408:Platelet Endothelial Cell Adhesion Molecule-1; D042461:Vascular Endothelial Growth Factor A
Am J Pathol. 2004 Aug;165(2):601-8
15277233
Science. 2000 Aug 18;289(5482):1197-202
10947988
Thromb Haemost. 2004 Dec;92(6):1438-45
15583754
J Virol. 2004 Aug;78(15):8271-80
15254199
Nat Rev Cancer. 2009 Mar;9(3):182-94
19194382
Mol Pharmacol. 2000 Feb;57(2):385-91
10648649
J Transl Med. 2004 Jun 25;2(1):22
15219236
J Gene Med. 2003 Apr;5(4):311-23
12692865
Nat Rev Drug Discov. 2004 May;3(5):391-400
15136787
J Virol. 2007 Mar;81(6):2688-99
17192304
Nat Rev Drug Discov. 2007 Sep;6(9):734-45
17690708
Cancer Sci. 2008 Mar;99(3):459-66
18167133
FASEB J. 1999 Jan;13(1):9-22
9872925
Nat Biotechnol. 1997 Jul;15(7):647-52
9219267
Ann N Y Acad Sci. 2003 May;995:151-61
12814947
Cancer Res. 2004 Nov 15;64(22):8249-55
15548691
N Engl J Med. 2004 Jun 3;350(23):2335-42
15175435
Clin Cancer Res. 2004 Jan 15;10(2):415-27
14760060
Diabetes. 2003 Dec;52(12):2959-68
14633857
Cancer Res. 2001 Sep 15;61(18):6649-55
11559528
Mol Genet Metab. 1998 Feb;63(2):103-9
9562963
Nat Methods. 2008 May;5(5):439-45
18391960
Mol Ther. 2004 Apr;9(4):489-95
15093179
J Virol. 2003 Apr;77(7):4449-56
12634405
Virology. 2008 Sep 15;379(1):125-34
18672260
Cancer Gene Ther. 2010 Apr;17(4):244-55
19798121
J Mol Biol. 2009 Mar 13;386(5):1368-81
19063898
APMIS. 2004 Jul-Aug;112(7-8):508-25
15563313
Proc Natl Acad Sci U S A. 2009 Jan 6;106(1):44-9
19118196
Science. 1998 Jan 16;279(5349):377-80
9430587
Nature. 2004 Jun 10;429(6992):629-35
15190345
Eur J Cancer. 2006 Dec;42(18):3127-39
17098419
Cancer Gene Ther. 2005 Feb;12(2):175-84
15375381
Nature. 2004 Feb 26;427(6977):787
14985739
Acc Chem Res. 2001 Oct;34(10):826-35
11601967
Clin Cancer Res. 2007 Jul 1;13(13):3968-76
17606731
Semin Cancer Biol. 2002 Apr;12(2):105-12
12027582
Virology. 1997 Jun 23;233(1):105-17
9201220
Genes Dev. 2007 Jun 15;21(12):1546-58
17575055
Nat Rev Cancer. 2007 Jun;7(6):475-85
17522716
Mol Genet Metab. 1998 Jul;64(3):193-7
9719628
J Clin Invest. 1998 Apr 1;101(7):1401-13
9525983
J Am Chem Soc. 2008 Jan 16;130(2):556-65
18088119
Cancer Res. 2000 Feb 1;60(3):722-7
10676659
J Neurosci. 2004 Sep 15;24(37):8009-18
15371501
Cancer. 2004 Jun 15;100(12):2491-9
15197790
Cancer Res. 2005 Feb 1;65(3):972-81
15705898
Cancer Res. 1994 Dec 15;54(24):6504-11
7527299
Ann Surg. 2001 May;233(5):603-11
11323498
Nat Rev Cancer. 2005 Jun;5(6):436-46
15928674
Neoplasia. 2008 Aug;10(8):886-96
18670646
J Gen Virol. 1995 Aug;76 ( Pt 8):1901-7
7636471
FEBS Lett. 2009 Jun 18;583(12):2100-7
19481546
Mol Ther. 2001 Mar;3(3):337-50
11273776
N Engl J Med. 2008 May 8;358(19):2039-49
18463380
Invest Ophthalmol Vis Sci. 2004 Sep;45(9):2915-21
15326102
Curr Opin Oncol. 2004 Jan;16(1):44-9
14685092
Mol Ther. 2002 Jun;5(6):755-61
12027560
Mol Ther. 2002 Mar;5(3):233-41
11863412
Mol Ther. 2008 Mar;16(3):516-24
18195720
J Virol. 1997 Feb;71(2):1453-65
8995671
Neoplasia. 2003 May-Jun;5(3):205-17
12869304
J Comb Chem. 2001 May-Jun;3(3):257-66
11350249
Science. 2002 Jun 28;296(5577):2404-7
12089446
Leuk Res. 2003 Aug;27(8):701-8
12801528
Pancreas. 2008 Aug;37(2):210-20
18665085