Overexpression of thymidylate synthase (TS) is observed in a wide range of human cancers where it is associated with advanced disease and resistance to commercially available TS inhibitors. My Lab demonstrated that activated expression of TS, an essential enzyme for DNA synthesis and repair, plays a direct role in promoting tumorigenesis. This was an unexpected observation as elevated TS levels were previously believed to reflect a passive biomarker for tumor cell growth. Our observation changed the paradigm of TS as a biomarker and has great significance for cancer treatment as it refocuses attention on the importance of TS as a tumor-promoting signal. While our published data proposed that TS plays an active role in promoting tumorigenesis, questions remain whether elevated TS cooperates with other common genetic lesions to enhance the neoplastic process. To address this question we developed a new genetically engineered mouse model (GEMM) with Ink4a/Arf-/- mutation and overexpression of human TS (designated hTS/Ink4a/Arf-/-). We showed for the first time that TS activation in Ink4a/Arf null mice is associated with accelerated tumor growth, larger tumor size and more advanced stage of the disease. We also showed that activated TS promote the growth and metastasis of previously established tumors rather than inducing development of a new tumor spectrum. Ink4a/Arf is one of the most commonly mutated loci in human cancer and its products encode tumor suppressor proteins required for G1 arrest. The objectives of this proposal are: i) to determine how TS overexpression promotes tumorigenicity in hTS/Ink4a/Arf-/-mice. We hypothesize that TS overexpression drives DNA replication stress by deregulating replication factors and nucleotide pool ratios that result in double strands break and genomic instability in the absence of INK4a/ARF checkpoints. ii) to develop new potent TS allosteric inhibitor analogues. We have discovered a new strategy that takes advantage of high TS subunit cooperativity to identify small molecule inhibitors of TS. Targeting TS dimer interface with allosteric inhibitors overstabilize the dimer structure resulting in loss of subunit cooperativity and inhibition of TS enzymatic function. We will identify potent derivatives to prevent or reverse drug resistance that limits the effectiveness of current therapy. Our new TS allosteric inhibitors do not cause TS overexpression thus these new small molecules may overcome the problem of TS overexpression associated resistance and iii) to test anti-tumor activity of allosteric inhibitors alone and in combination with conventional TS inhibitors in our new GEMM with activated hTS. In summary, this research project proposes a new approach to control TS mediated tumorigenesis with the goal to reduce drug resistance and optimize TS inhibition in cancer treatment. Determination of how TS overexpression promotes tumorigenicity and the development of novel TS inhibitors will provide a new focus toward targeting TS for therapeutic intervention and for cancer treatment.

Public Health Relevance

Thymidylate synthase (TS) is a cell cycle regulated DNA replication enzyme that is aberrantly over-expressed in a broad range of common adult cancers such as lung, gastrointestinal, lymphoma and sarcomas. This research project proposes a new paradigm to define the role of TS activation not as a biomarker but as a driving force in tumor development and to develop more effective cancer treatment by targeting TS inhibition.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Project (R01)
Project #
5R01CA188132-02
Application #
9199072
Study Section
Special Emphasis Panel (ZRG1)
Program Officer
Alley, Michael C
Project Start
2016-01-01
Project End
2020-12-31
Budget Start
2017-01-01
Budget End
2017-12-31
Support Year
2
Fiscal Year
2017
Total Cost
Indirect Cost
Name
University of Florida
Department
Anatomy/Cell Biology
Type
Schools of Medicine
DUNS #
969663814
City
Gainesville
State
FL
Country
United States
Zip Code
32611
Chen, Min; Maeng, Kyungah; Nawab, Akbar et al. (2017) Efficient Gene Delivery and Expression in Pancreas and Pancreatic Tumors by Capsid-Optimized AAV8 Vectors. Hum Gene Ther Methods 28:49-59