Subtractive proteomic mapping of the endothelial surface in lung and solid tumours for tissue-specific therapy

Abstract

The molecular complexity of tissues and the inaccessibility of most cells within a tissue limit the discovery of key targets for tissue-specific delivery of therapeutic and imaging agents in vivo. Here, we describe a hypothesis-driven, systems biology approach to identifying a small subset of proteins induced at the tissue–blood interface that are inherently accessible to antibodies injected intravenously. We use subcellular fractionation, subtractive proteomics and bioinformatics to identify endothelial cell surface proteins exhibiting restricted tissue distribution and apparent tissue modulation. Expression profiling and γ-scintigraphic imaging with antibodies establishes two of these proteins, aminopeptidase-P and annexin A1, as selective in vivo targets for antibodies in lungs and solid tumours, respectively. Radio-immunotherapy to annexin A1 destroys tumours and increases animal survival. This analytical strategy can map tissue- and disease-specific expression of endothelial cell surface proteins to uncover novel accessible targets useful for imaging and therapy.

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Figure 1: Molecular heterogeneity of endothelial cell surface in multiple rat organs in vivo.
Figure 2: In vivo γ-scintigraphic imaging of APP antibody targeting in normal and tumour-bearing rats.
Figure 3: Mapping rat lung tumours and organs for differential endothelial cell surface protein expression in vivo.
Figure 4: Tumour-specific targeting of 125I-AnnA1 antibodies after intravenous injection into tumour-bearing rats.
Figure 5: AnnA1 targeted radio-immunotherapy increases survival in rats.
Figure 6: Expression of AnnA1 on vascular endothelium of multiple primary human solid tumours.

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Acknowledgements

We thank A. Wempren, K. Hearn, M. Bourne, L. Randall and T. Smith for technical assistance. This research was supported by the National Institute of Health (Heart, Lung and Blood), National Cancer Institute, Sidney Kimmel, Schutz Foundation, California Tobacco-related Disease Research Program and California Breast Cancer Research Program.

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Correspondence to Jan E. Schnitzer.

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Supplementary Methods

Includes information on: materials; expression profiling in vivo of candidate proteins; rat tumor models; gamma scintigraphic imaging and biodistribution analysis. (DOC 28 kb)

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Oh, P., Li, Y., Yu, J. et al. Subtractive proteomic mapping of the endothelial surface in lung and solid tumours for tissue-specific therapy. Nature 429, 629–635 (2004). https://doi.org/10.1038/nature02580

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