Publication Date

2018-05-14

Document Type

Honors Project

Degree Name

Bachelor of Arts

Department

Biochemistry

Advisors

Sarah J. Moore

Keywords

Mesothelin, Cancer, Targeted cancer therapy, Fibronectin, Protein engineering, Cytotoxicity, Cancer-Treatment, Drug targeting, Antineoplastic agents, Mesothelium

Abstract

Traditional cancer therapies involve the indiscriminate killing of all rapidly dividing cells, subsequently targeting healthy cells that are naturally dividing at a fast rate. To minimize the egregious side effects that result from such therapies, there is sustained effort to develop a class of more targeted cancer therapeutics. A tumor biomarker currently under investigation for targeted cancer therapy is a 40 kDa cell surface glycoprotein called mesothelin (MSLN). MSLN is overexpressed on the surfaces of some breast, ovarian, pancreatic, liver and lung cancers. The interaction between MSLN and MUC16, an established tumor biomarker, has been implicated in the progression, aggressiveness, and metastasis of cancer. While there are ongoing clinical trials for antibodies that recognize MSLN, no MSLN-targeting therapies have received FDA approval for routine clinical use. There remains an unmet need for the development of therapeutics targeting MSLN. We report novel non-antibody proteins, based on the fibronectin type III (Fn3) scaffold, that have been engineered using directed evolution to bind to MSLN for development as diagnostics and therapeutics. Equilibrium tumor cell binding studies using flow cytometry reveal a moderate binding affinity with dissociation constants in the nanomolar range. We are currently using a variety of assays to measure the therapeutic potential of the engineered proteins. We have observed that the engineered proteins are internalized upon binding to MSLN and colocalize with early endosomes. The internalization of our proteins makes them promising targeting molecules as part of a drug delivery system. We are developing assays to measure the cytotoxic potential of the engineered proteins and their ability to trigger apoptosis. Further, we intend to measure if the engineered proteins are capable of inhibiting cell migration, with implications for blocking metastasis. Our current data indicate the promise of the engineered Fn3 proteins for further development as targeted cancer therapeutics.

Rights

2018 Daniela Amoakohene Deny. Access limited to the Smith College community and other researchers while on campus. Smith College community members also may access from off-campus using a Smith College log-in. Other off-campus researchers may request a copy through Interlibrary Loan for personal use.

Language

English

Comments

viii, 78 pages : color illustrations. Includes bibliographical references (pages 73-78)

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