The goal of this project is to develop a new technology to systematically delineate disease mechanisms of genetic mutations at the protein level. Identification of the large number of disease-associated mutations has shifted the research challenge of defining underlying disease mechanisms from genomics to functional proteomics. One of the examples is tubby-like protein 1 (Tulp1) that has 23 known mutations associated with retinal degeneration. Although various technologies of functional proteomics have been used to identify a few Tulp1-binding proteins, three challenges are: (a) how to sensitively identify less abundant protein-protein interactions (PPIs);(b) how to quantify binding activity of the entire PPI dataset;and (c) how to systematically distinguish disease-relevant PPIs. Consequently, disease relevance of Tulp1 PPIs is yet to be defined, and its pathological mechanisms remain elusive. We developed open reading frame phage display (OPD) as a new technology of functional proteomics to complement the existing technologies. Owing to its capacity to enrich clones displaying binding proteins through multi-round selection and amplification, OPD should be more sensitive to identify less abundant PPIs than other technologies that lack protein amplification capacity. Other advantages of OPD include its unique capacity to globally map mutation-specific PPIs by next generation sequencing (NGS) and to quantitatively compare the binding activity of entire PPI datasets to wild-type proteins and their pathogenic mutants for reliable delineation of mutation-specific PPIs. The hypothesis is that OPD is a new technology that can be used for global mapping of functionally-relevant and disease-relevant binding proteins of Tulp1.
The specific aims are: (1) To test the working hypothesis that OPD systematically identifies mutation-specific PPIs;(2) to test the working hypothesis that OPD-NGS hybrid technology globally maps total PPIs and mutation-specific PPIs. (3) To test the working hypothesis that OPD reliably delineates mutation- specific and biologically-relevant PPIs in a biochemical pathway. This project will facilitate delineation of Tulp1 disease mechanisms of retinal degeneration. Given its broad applicability, OPD-NGS as the only technology for global mapping of mutation-specific PPIs will advance our understanding of disease mechanisms for many other proteins and their genetic mutations.

Public Health Relevance

The goal of this project is to develop a new technology to systematically delineate disease mechanisms of genetic mutations at the protein level. The new technology will be developed to identify and characterize disease-relevant binding proteins and pathological mechanisms for an eye protein that has 23 known blindness-causing mutations. This technology is applicable to other proteins and their pathogenic mutations. Thus, this study has broad impact on our understanding of disease mechanisms associated with genetic mutations.

National Institute of Health (NIH)
National Institute of General Medical Sciences (NIGMS)
Research Project (R01)
Project #
Application #
Study Section
Enabling Bioanalytical and Imaging Technologies Study Section (EBIT)
Program Officer
Edmonds, Charles G
Project Start
Project End
Budget Start
Budget End
Support Year
Fiscal Year
Total Cost
Indirect Cost
University of Miami School of Medicine
Schools of Medicine
Coral Gables
United States
Zip Code
Wang, Weiwen; LeBlanc, Michelle E; Chen, Xiuping et al. (2017) Pathogenic role and therapeutic potential of pleiotrophin in mouse models of ocular vascular disease. Angiogenesis 20:479-492
LeBlanc, Michelle E; Wang, Weiwen; Chen, Xiuping et al. (2017) Secretogranin III as a disease-associated ligand for antiangiogenic therapy of diabetic retinopathy. J Exp Med 214:1029-1047
LeBlanc, Michelle E; Wang, Weiwen; Chen, Xiuping et al. (2016) The regulatory role of hepatoma-derived growth factor as an angiogenic factor in the eye. Mol Vis 22:374-86
Chen, Xiuping; Guo, Feiye; LeBlanc, Michelle E et al. (2016) Mesd extrinsically promotes phagocytosis by retinal pigment epithelial cells. Cell Biol Toxicol 32:347-58
Ding, Ying; Caberoy, Nora B; Guo, Feiye et al. (2015) Reticulocalbin-1 facilitates microglial phagocytosis. PLoS One 10:e0126993
Guo, Feiye; Ding, Ying; Caberoy, Nora et al. (2015) ABCF1 extrinsically regulates retinal pigment epithelial cell phagocytosis. Mol Biol Cell 26:2311-20
Guo, Feiye; Ding, Ying; Caberoy, Nora B et al. (2015) Lyar Is a New Ligand for Retinal Pigment Epithelial Phagocytosis. J Cell Biochem 116:2177-87
LeBlanc, Michelle E; Wang, Weiwen; Caberoy, Nora B et al. (2015) Hepatoma-derived growth factor-related protein-3 is a novel angiogenic factor. PLoS One 10:e0127904
Li, Wei (2013) Phagocyte dysfunction, tissue aging and degeneration. Ageing Res Rev 12:1005-12
Caberoy, Nora B; Li, Wei (2012) Unraveling the molecular mystery of retinal pigment epithelium phagocytosis. Adv Exp Med Biol 723:693-9

Showing the most recent 10 out of 12 publications