PROTEOMIC DISSECTION OF LPS-INDUCIBLE PHF8-DEPENDENT SECRETOME REVEALS NOVEL ROLES OF PHF8 IN TLR4-INDUCED ACUTE INFLAMMATION AND ACTIVATION OF ADAPTIVE IMMUNITY Public Deposited

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  • March 20, 2019
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  • Erdogan, Ozgun
    • Affiliation: School of Medicine, Department of Biochemistry and Biophysics
Abstract
  • This dissertation examines the role of PHF8 in the regulation of LPS-induced inflammatory response in macrophages and activation of adaptive immunity. I extend prior work on PP2Ac-dependent activation of endotoxin tolerance to reveal that PHF8 is negatively regulated by chronic-active PP2Ac in endotoxin tolerance, leading to our observation that PHF8 broadly regulates LPS-induced acute inflammation response in macrophages. This research offers four conclusions: (1) PHF8 enzymatic activity is important for LPS-induced cytokine expression in macrophages, (2) PHF8 targets H3K9me1/2 for demethylation to promote LPS-induced gene-specific transcription, (3) PHF8 binds to, and regulates NFκB transcription factor activity, and (4) PHF8 broadly regulates the secretion of molecules involved in activation of adaptive immunity. In addition to the biological and biochemical experiments that provide evidence for PHF8-dependent H3K9me1/2 demethylation, cytokine expression, and NFκB activation, we conducted proteomic dissection of LPS-induced macrophage secretome revealing important roles of PHF8 in activation of adaptive immunity. Moreover, the secretome analysis illustrates that PHF8 is a major regulator of LPS-induced secretion of molecules involved in activation of adaptive immunity. Further, through T cell proliferation and activation assays, I provide evidence that the activation of T cells by macrophages depends on the LPS-induced PHF8 activity. Our findings advance our understanding of regulation of LPS response by PHF8, and reveal that PHF8 bridges the innate immunity, and adaptive immunity responses. These results also offer insights to differential regulation of acute- vs chronic-inflammation by the antagonistic activities of chromatin writers and erasers, namely H3K9me1/2 methyltransferase G9a and H3K9me1/2 demethylase PHF8, respectively.
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  • In Copyright
Advisor
  • Errede, Beverly
  • Chen, Xian
  • Parise, Leslie
  • Wan, Yisong
  • Marzluff, William
Degree
  • Doctor of Philosophy
Degree granting institution
  • University of North Carolina at Chapel Hill Graduate School
Graduation year
  • 2016
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