This project aims to develop a novel platform that would help accelerate the development of novel antibody- based therapeutics. Numerous antibody therapies undergoing clinical trials are concerned with immunological and oncological targets. The proposed project is innovative and differs from previous studies by using deep sequencing to yeast display to systematically screen antibodies binding different epitopes on therapeutically relevant target proteins. Our technology generates unprecedented sequence-function maps that cover the entire sequence of protein binders [1]. We hypothesize that this same method can be used to develop rapid fine epitope maps of antibody-antigen interactions. Thus, the overall goal of the project is to develop the deep scanning methodology on proteins target proteins whose structures or active domains are currently unknown. To demonstrate that our novel platform can identify functional antibodies that target such proteins, we will evaluate their ability to inhibit the function of TROP2 (a.k.a tumor-associated calcium signal transducer 2, TACSTD2), our model protein, from promoting growth and metastasis. The project involves two aims.
The first aim i s to develop a novel epitope mapping strategy that couples yeast display with deep sequencing to generate unprecedented sequence-function maps of protein binders.
The second aim will develop and screen potential novel antibodies to TROP2 for their ability to inhibit cell proliferation and metastasis through in vitro and in vivo experiments. Completion of this project will provide a platform that can identify functional antibodies that inhibit the activty of target proteins for which structures are not available. The proposed methodology will aid in the discovery and development of new therapeutics not only for cancer but other diseases, including autoimmune diseases that are amendable to antibody therapeutics.

Public Health Relevance

This project aims to develop a novel platform that could accelerate the discovery and development of new antibody based therapeutics. Antibodies are widely recognized as therapeutic molecules for many diseases, such as autoimmune diseases (e.g. Crohn's disease, multiple sclerosis, rheumatoid arthritis and psoriasis, etc.) and cancers. The proposed project differs from previous studies in this field by using deep sequencing to rapidly map epitopes of novel antibodies.

National Institute of Health (NIH)
National Cancer Institute (NCI)
Exploratory/Developmental Grants (R21)
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Special Emphasis Panel ()
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Muszynski, Karen
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Michigan State University
Engineering (All Types)
Schools of Engineering
East Lansing
United States
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Klesmith, Justin R; Whitehead, Timothy A (2016) High-throughput evaluation of synthetic metabolic pathways. Technology (Singap World Sci) 4:9-14
Stapleton, James A; Kim, Jeongwoon; Hamilton, John P et al. (2016) Haplotype-Phased Synthetic Long Reads from Short-Read Sequencing. PLoS One 11:e0147229
Thompson, Ryan; Chan, Christina (2016) Signal transduction of the physical environment in the neural differentiation of stem cells. Technology (Singap World Sci) 4:1-8
Vocelle, Daniel; Chesniak, Olivia M; Malefyt, Amanda P et al. (2016) Dextran functionalization enhances nanoparticle-mediated siRNA delivery and silencing. Technology (Singap World Sci) 4:
Nath, Aritro; Chan, Christina (2016) Genetic alterations in fatty acid transport and metabolism genes are associated with metastatic progression and poor prognosis of human cancers. Sci Rep 6:18669
Vocelle, Daniel; Chan, Christina; Walton, S Patrick (2015) How can novel microscopic approaches shed light on the function of nucleic acid-based drugs? Future Med Chem 7:1623-5
Nath, Aritro; Li, Irene; Roberts, Lewis R et al. (2015) Elevated free fatty acid uptake via CD36 promotes epithelial-mesenchymal transition in hepatocellular carcinoma. Sci Rep 5:14752
Kowalsky, Caitlin A; Faber, Matthew S; Nath, Aritro et al. (2015) Rapid fine conformational epitope mapping using comprehensive mutagenesis and deep sequencing. J Biol Chem 290:26457-70
Zhang, Linxia; Liu, Li; Thompson, Ryan et al. (2014) CREB modulates calcium signaling in cAMP-induced bone marrow stromal cells (BMSCs). Cell Calcium 56:257-68
Liu, Li; Chan, Christina (2014) The role of inflammasome in Alzheimer's disease. Ageing Res Rev 15:6-15