The goal of this renewal proposal is to continue our approach that combines phage antibody display with laser capture micro dissection (LCM) to identify novel internalizing human single chain antibodies (scFvs) that target metastatic tumor cells in situ residing in their natural tissue microenvironment. The work is built on our previous work where we developed and used this novel approach to identify antibodies targeting primary prostate tumor. This proposal aims to advance our study to prostate tumor bone metastases, a major challenge for treatment and management of the disease. This proposal aims to use our novel LCM-based human antibody library selection strategy to identify novel scFvs that target tumor metastases-associated cell surface antigens expressed by tumor cells in situ for imaging and therapy development. In addition, based on novel findings of the previous R01 study where we found that tumor antigens such as ALCAM are actively cleaved in metastatic prostate cancer cells, we will select antibodies that target cell surface neoepitopes on the post-cleavage transmembrane remnants of ALCAM. Specifically, we aim (1) to develop novel human antibodies that target ALCAM post-cleavage transmembrane remnants. We will use these antibodies to study the prevalence of ALCAM transmembrane remnants on prostate tumor bone metastases by immunohistochemistry, and evaluate utility of anti-remnant scFvs in tumor targeting in vivo using SPECT/CT imaging techniques. (2) To identify rapidly internalizing antibodies that are capable of high-level accumulation in target metastatic prostate cancer cells residing in their tissue microenvironment. We propose to use our previously developed and validated novel LCM-based antibody library selection strategies to identify novel antibodies targeting prostate tumor bone metastases with the following properties: (a) binding to tumor cells in situ. (b) Binding to live tumor cells and thus recognizing tumor antigens in their native conformations. (c) rapidly internalizing and accumulating to high levels inside tumor cells. (d) Exhibiting high-level tumor uptake in vivo and minimal uptake in off target tissues in both localized (subcutaneous xenograft) and disseminated prostate tumor models. (3) To identify tumor antigens bound by our novel antibodies. We will screen a large yeast surface displayed cDNA library developed in the lab to identify tumor antigens bound by our novel phage antibodies. In parallel, we will identify tumor antigens by immunoprecipitation and mass spectrometry analysis that allow identification of post- translational modified antigens.

Public Health Relevance

The goal of this renewal proposal is to continue our novel approach that combines phage antibody display with laser capture microdissection to identify rapidly internalizing human single chain antibodies that target metastatic tumor cells in situ residing in their natural tissue microenvironment. The work is built on our previous work where we developed and used this novel approach to identify antibodies targeting primary prostate tumor. This proposal aims to advance our study to prostate tumor bone metastases, a major challenge for treatment and management of the disease.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Project (R01)
Project #
5R01CA118919-07
Application #
8277976
Study Section
Special Emphasis Panel (ZRG1-DTCS-A (81))
Program Officer
Mccarthy, Susan A
Project Start
2006-03-07
Project End
2014-03-31
Budget Start
2012-04-01
Budget End
2013-03-31
Support Year
7
Fiscal Year
2012
Total Cost
$307,484
Indirect Cost
$108,465
Name
University of California San Francisco
Department
Anesthesiology
Type
Schools of Medicine
DUNS #
094878337
City
San Francisco
State
CA
Country
United States
Zip Code
94143
Ha, Kevin D; Bidlingmaier, Scott M; Liu, Bin (2016) Macropinocytosis Exploitation by Cancers and Cancer Therapeutics. Front Physiol 7:381
Bidlingmaier, Scott; Ha, Kevin; Lee, Nam-Kyung et al. (2016) Proteome-wide Identification of Novel Ceramide-binding Proteins by Yeast Surface cDNA Display and Deep Sequencing. Mol Cell Proteomics 15:1232-45
Sherbenou, Daniel W; Aftab, Blake T; Su, Yang et al. (2016) Antibody-drug conjugate targeting CD46 eliminates multiple myeloma cells. J Clin Invest 126:4640-4653
Bidlingmaier, Scott; Liu, Bin (2015) Utilizing Yeast Surface Human Proteome Display Libraries to Identify Small Molecule-Protein Interactions. Methods Mol Biol 1319:203-14
Bidlingmaier, Scott; Su, Yang; Liu, Bin (2015) Combining Phage and Yeast Cell Surface Antibody Display to Identify Novel Cell Type-Selective Internalizing Human Monoclonal Antibodies. Methods Mol Biol 1319:51-63
Sherbenou, Daniel W; Behrens, Christopher R; Su, Yang et al. (2015) The development of potential antibody-based therapies for myeloma. Blood Rev 29:81-91
Bidlingmaier, Scott; Liu, Bin (2015) Identification of Novel Protein-Ligand Interactions by Exon Microarray Analysis of Yeast Surface Displayed cDNA Library Selection Outputs. Methods Mol Biol 1319:179-92
Bidlingmaier, Scott; Liu, Bin (2015) Identification of Posttranslational Modification-Dependent Protein Interactions Using Yeast Surface Displayed Human Proteome Libraries. Methods Mol Biol 1319:193-202
Ha, Kevin D; Bidlingmaier, Scott M; Zhang, Yafeng et al. (2014) High-content analysis of antibody phage-display library selection outputs identifies tumor selective macropinocytosis-dependent rapidly internalizing antibodies. Mol Cell Proteomics 13:3320-31
Behrens, Christopher R; Liu, Bin (2014) Methods for site-specific drug conjugation to antibodies. MAbs 6:46-53

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