The goal of this proposal is to identify citrullinated biomarkers of cancer, rheumatoid arthritis and colitis. This goal will be achieved by developing a highly practical and versatile technology to detect and characterize citrullinated proteins in complex proteomes. Protein citrullination is increased in virtually all inflammatory diseases, as well as cancer, and the evidence indicates that this post-translational modification is a major contributor to disease pathogenesis. In fact, inhibition of protein citrullination decreases diseas severity in animal models of rheumatoid arthritis, ulcerative colitis, nerve damage, and cancer. Thus, citrullinated biomarkers most likely exist and exploiting these citrullination events for disease diagnostics presents a single provocative strategy that is applicable to multiple diseases. The approach proposed herein is innovative because a new paradigm is being tested - that citrullinated proteins are disease biomarkers. Additionally, an enabling technology will be developed that can be used to identify citrullinated biomarkers in numerous disease states. The platform being developed is faster and more versatile than existing technologies, providing a unique strategy to interrogate the role of citrullination in these diseases. Furthermore, this technology can be extended to the identification and quantification of citrullinated proteins by MS, which is not possible with existing technologies. This new platform is therefore not only applicable to screening patient samples to identify biomarkers, but can be adapted to enrich and identify specific citrullinated proteins, thereby providing deeper insight into the functional roleof citrullination. This technology, coupled with the disease models and collaborator expertise in this grant, enables the exploration of a potential paradigm-shifting outcome, whereby the detection of citrullination events provides a common diagnostic for a multitude of divergent diseases. The impact of this proposal is multifold. First, the identification of citrullinated biomarkers will alow for better, more tailored treatments, because they can be used to diagnose and monitor the efficacy of treatments for these diseases. Second, these efforts will define the 'citrullinome', thereby facilitating a greater understanding of how this modification contributes to normal and pathological processes. Third, these findings will determine if there is a common link between these disparate diseases. In summary, the funding of this proposal will ultimately provide powerful chemical probes for scientists interested in the in vivo role of protein citrullination an will have a strong translational impact on the development of diagnostics and therapeutics for numerous diseases.

Public Health Relevance

The identification of disease biomarkers has been identified by the NIH as a high priority research area. Such biomarkers can be used to diagnose and monitor the efficacy of therapeutic interventions. The citrullinated protein biomarkers that we plan to identify will be useful for diagnosing and monitoring the progression of a variety of inflammatory diseases and cancers.

National Institute of Health (NIH)
National Institute of General Medical Sciences (NIGMS)
Research Project (R01)
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Synthetic and Biological Chemistry A Study Section (SBCA)
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Fabian, Miles
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University of Massachusetts Medical School Worcester
Schools of Medicine
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Clancy, Kathleen W; Weerapana, Eranthie; Thompson, Paul R (2016) Detection and identification of protein citrullination in complex biological systems. Curr Opin Chem Biol 30:1-6
Fuhrmann, Jakob; Thompson, Paul R (2016) Protein Arginine Methylation and Citrullination in Epigenetic Regulation. ACS Chem Biol 11:654-68
Kawalkowska, Joanna; Quirke, Anne-Marie; Ghari, Fatemeh et al. (2016) Abrogation of collagen-induced arthritis by a peptidyl arginine deiminase inhibitor is associated with modulation of T cell-mediated immune responses. Sci Rep 6:26430
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Fuhrmann, Jakob; Clancy, Kathleen W; Thompson, Paul R (2015) Chemical biology of protein arginine modifications in epigenetic regulation. Chem Rev 115:5413-61
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Slade, Daniel J; Horibata, Sachi; Coonrod, Scott A et al. (2014) A novel role for protein arginine deiminase 4 in pluripotency: the emerging role of citrullinated histone H1 in cellular programming. Bioessays 36:736-40

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