The endothelium forms the critical barrier restricting the movement of drugs and endogenous molecules from the circulating blood to the underlying tissue cells. Little molecular information is available about the luminal endothelial cell surface. The applicant s laboratory has: i) developed a new technique for purifying this cell surface directly from tissue; ii) elucidated basic selective transport mechanisms that move proteins into and across endothelium; and iii) purified specialized vesicles in endothelium called caveolae that contain the molecular machinery necessary for this transport. With this knowledge, site-directed delivery systems will be created in order to confer accessibility to the tissue cells by overcoming the endothelial barrier in an organ-specific manner. Endothelial membranes will be purified from many tissues to create a protein map that identifies tissue- and caveolae-specific targets. Monoclonal antibodies achieved in vivo tissue-specific targeting by intravenous injection. By design, the antibodies target the endothelial caveolae in one rat tissue and thereby rapidly accumulate in the intended target organ. The applicant proposes to study the molecular topography of the endothelial cell surface in vivo as a means of creating tissue-specific immunotargeting agents for the endothelium and its caveolae. He will test the in vivo targeting of our antibodies and determine how the endothelium transports them. The applicant also aims to purify and sequence the key targeting proteins. It appears that the quest to acquire basic knowledge at the molecular and cellular level about the endothelium and its caveolae in transport, the applicant has discovered a direct application towards the creation of tissue-specific targeting agents potentially useful in drug and gene therapy of many diseases.

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Research Project (R01)
Project #
5R01HL058216-04
Application #
6330127
Study Section
Pathology A Study Section (PTHA)
Program Officer
Applebaum-Bowden, Deborah
Project Start
1997-12-01
Project End
2002-11-30
Budget Start
2000-12-01
Budget End
2002-11-30
Support Year
4
Fiscal Year
2001
Total Cost
$435,195
Indirect Cost
Name
Sidney Kimmel Cancer Center
Department
Type
DUNS #
City
San Diego
State
CA
Country
United States
Zip Code
92121
Oh, Phil; Horner, Thierry; Witkiewicz, Halina et al. (2012) Endothelin induces rapid, dynamin-mediated budding of endothelial caveolae rich in ET-B. J Biol Chem 287:17353-62
Chrastina, Adrian; Massey, Kerri A; Schnitzer, Jan E (2011) Overcoming in vivo barriers to targeted nanodelivery. Wiley Interdiscip Rev Nanomed Nanobiotechnol 3:421-37
Valadon, Philippe; Darsow, Bryan; Buss, Tim N et al. (2010) Designed auto-assembly of nanostreptabodies for rapid tissue-specific targeting in vivo. J Biol Chem 285:713-22
Massey, Kerri A; Schnitzer, Jan E (2009) Targeting and imaging signature caveolar molecules in lungs. Proc Am Thorac Soc 6:419-30
Testa, Jacqueline E; Chrastina, Adrian; Li, Yan et al. (2009) Ubiquitous yet distinct expression of podocalyxin on vascular surfaces in normal and tumor tissues in the rat. J Vasc Res 46:311-24
Li, Yan; Yu, Jingyi; Wang, Yipeng et al. (2009) Enhancing identifications of lipid-embedded proteins by mass spectrometry for improved mapping of endothelial plasma membranes in vivo. Mol Cell Proteomics 8:1219-35
Testa, Jacqueline E; Chrastina, Adrian; Oh, Phil et al. (2009) Immunotargeting and cloning of two CD34 variants exhibiting restricted expression in adult rat endothelia in vivo. Am J Physiol Lung Cell Mol Physiol 297:L251-62
Oh, Phil; Borgstrom, Per; Witkiewicz, Halina et al. (2007) Live dynamic imaging of caveolae pumping targeted antibody rapidly and specifically across endothelium in the lung. Nat Biotechnol 25:327-37
Valadon, Philippe; Garnett, Jeff D; Testa, Jacqueline E et al. (2006) Screening phage display libraries for organ-specific vascular immunotargeting in vivo. Proc Natl Acad Sci U S A 103:407-12
Durr, Eberhard; Yu, Jingyi; Krasinska, Karolina M et al. (2004) Direct proteomic mapping of the lung microvascular endothelial cell surface in vivo and in cell culture. Nat Biotechnol 22:985-92

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