Kevin Lee

Publications (9)163.97 Total impact

• Article: Suppression of the antiviral response by an influenza histone mimic.

  Ivan Marazzi, Jessica S Y Ho, Jaehoon Kim, Balaji Manicassamy, Scott Dewell, Randy A Albrecht, Chris W Seibert, Uwe Schaefer, Kate L Jeffrey, Rab K Prinjha, Kevin Lee, Adolfo García-Sastre, Robert G Roeder, Alexander Tarakhovsky
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  ABSTRACT: Viral infection is commonly associated with virus-driven hijacking of host proteins. Here we describe a novel mechanism by which influenza virus affects host cells through the interaction of influenza non-structural protein 1 (NS1) with the infected cell epigenome. We show that the NS1 protein of influenza A H3N2 subtype possesses a histone-like sequence (histone mimic) that is used by the virus to target the human PAF1 transcription elongation complex (hPAF1C). We demonstrate that binding of NS1 to hPAF1C depends on the NS1 histone mimic and results in suppression of hPAF1C-mediated transcriptional elongation. Furthermore, human PAF1 has a crucial role in the antiviral response. Loss of hPAF1C binding by NS1 attenuates influenza infection, whereas hPAF1C deficiency reduces antiviral gene expression and renders cells more susceptible to viruses. We propose that the histone mimic in NS1 enables the influenza virus to affect inducible gene expression selectively, thus contributing to suppression of the antiviral response.
  Nature 03/2012; 483(7390):428-33. · 36.28 Impact Factor
• Source

  Available from: Terry Fang

  Article: Histone H3 lysine 9 di-methylation as an epigenetic signature of the interferon response.

  Terry C Fang, Uwe Schaefer, Ingrid Mecklenbrauker, Astrid Stienen, Scott Dewell, Marie S Chen, Inmaculada Rioja, Valentino Parravicini, Rab K Prinjha, Rohit Chandwani, Margaret R MacDonald, Kevin Lee, Charles M Rice, Alexander Tarakhovsky
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  ABSTRACT: Effective antiviral immunity depends on the ability of infected cells or cells triggered with virus-derived nucleic acids to produce type I interferon (IFN), which activates transcription of numerous antiviral genes. However, disproportionately strong or chronic IFN expression is a common cause of inflammatory and autoimmune diseases. We describe an epigenetic mechanism that determines cell type-specific differences in IFN and IFN-stimulated gene (ISG) expression in response to exogenous signals. We identify di-methylation of histone H3 at lysine 9 (H3K9me2) as a suppressor of IFN and IFN-inducible antiviral gene expression. We show that levels of H3K9me2 at IFN and ISG correlate inversely with the scope and amplitude of IFN and ISG expression in fibroblasts and dendritic cells. Accordingly, genetic ablation or pharmacological inactivation of lysine methyltransferase G9a, which is essential for the generation of H3K9me2, resulted in phenotypic conversion of fibroblasts into highly potent IFN-producing cells and rendered these cells resistant to pathogenic RNA viruses. In summary, our studies implicate H3K9me2 and enzymes controlling its abundance as key regulators of innate antiviral immunity.
  Journal of Experimental Medicine 03/2012; 209(4):661-9. · 13.85 Impact Factor
• Article: Identification of a novel series of BET family bromodomain inhibitors: binding mode and profile of I-BET151 (GSK1210151A).

  Jonathan Seal, Yann Lamotte, Frédéric Donche, Anne Bouillot, Olivier Mirguet, Françoise Gellibert, Edwige Nicodeme, Gael Krysa, Jorge Kirilovsky, Soren Beinke, [......], Chun-Wa Chung, Laurie Gordon, Toni Lewis, Ann L Walker, Leanne Cutler, David Lugo, David M Wilson, Jason Witherington, Kevin Lee, Rab K Prinjha
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  ABSTRACT: A novel series of quinoline isoxazole BET family bromodomain inhibitors are discussed. Crystallography is used to illustrate binding modes and rationalize their SAR. One member, I-BET151 (GSK1210151A), shows good oral bioavailability in both the rat and minipig as well as demonstrating efficient suppression of bacterial induced inflammation and sepsis in a murine in vivo endotoxaemia model.
  Bioorganic & medicinal chemistry letters 02/2012; 22(8):2968-72. · 2.65 Impact Factor
• Article: Bone morphogenetic protein 7 (BMP7) reverses obesity and regulates appetite through a central mTOR pathway.

  Kristy L Townsend, Ryo Suzuki, Tian Lian Huang, Enxuan Jing, Tim J Schulz, Kevin Lee, Cullen M Taniguchi, Daniel O Espinoza, Lindsay E McDougall, Hongbin Zhang, Tong-Chuan He, Efi Kokkotou, Yu-Hua Tseng
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  ABSTRACT: Body weight is regulated by coordinating energy intake and energy expenditure. Transforming growth factor β (TGFβ)/bone morphogenetic protein (BMP) signaling has been shown to regulate energy balance in lower organisms, but whether a similar pathway exists in mammals is unknown. We have previously demonstrated that BMP7 can regulate brown adipogenesis and energy expenditure. In the current study, we have uncovered a novel role for BMP7 in appetite regulation. Systemic treatment of diet-induced obese mice with BMP7 resulted in increased energy expenditure and decreased food intake, leading to a significant reduction in body weight and improvement of metabolic syndrome. Similar degrees of weight loss with reduced appetite were also observed in BMP7-treated ob/ob mice, suggesting a leptin-independent mechanism utilized by BMP7. Intracerebroventricular administration of BMP7 to mice led to an acute decrease in food intake, which was mediated, at least in part, by a central rapamycin-sensitive mTOR-p70S6 kinase pathway. Together, these results underscore the importance of BMP7 in regulating both food intake and energy expenditure, and suggest new therapeutic approaches for obesity and its comorbidities.
  The FASEB Journal 02/2012; 26(5):2187-96. · 5.71 Impact Factor
• Article: Selective analysis of cell-free DNA in maternal blood for evaluation of fetal trisomy.

  Andrew B Sparks, Eric T Wang, Craig A Struble, Wade Barrett, Renee Stokowski, Celeste McBride, Jacob Zahn, Kevin Lee, Naiping Shen, Jigna Doshi, Michel Sun, Jill Garrison, Jay Sandler, Desiree Hollemon, Patrick Pattee, Aoy Tomita-Mitchell, Michael Mitchell, John Stuelpnagel, Ken Song, Arnold Oliphant
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  ABSTRACT: To develop a novel prenatal assay based on selective analysis of cell-free DNA in maternal blood for evaluation of fetal Trisomy 21 (T21) and Trisomy 18 (T18). Two hundred ninety-eight pregnancies, including 39 T21 and seven T18 confirmed fetal aneuploidies, were analyzed using a novel, highly multiplexed assay, termed digital analysis of selected regions (DANSR™). Cell-free DNA from maternal blood samples was analyzed using DANSR assays for loci on chromosomes 21 and 18. Products from 96 separate patients were pooled and sequenced together. A standard Z-test of chromosomal proportions was used to distinguish aneuploid samples from average-risk pregnancy samples. DANSR aneuploidy discrimination was evaluated at various sequence depths. At the lowest sequencing depth, corresponding to 204,000 sequencing counts per sample, average-risk cases where distinguished from T21 and T18 cases, with Z statistics for all cases exceeding 3.6. Increasing the sequencing depth to 410,000 counts per sample substantially improved separation of aneuploid and average-risk cases. A further increase to 620,000 counts per sample resulted in only marginal improvement. This depth of sequencing represents less than 5% of that required by massively parallel shotgun sequencing approaches. Digital analysis of selected regions enables highly accurate, cost efficient, and scalable noninvasive fetal aneuploidy assessment.
  Prenatal Diagnosis 01/2012; 32(1):3-9. · 2.11 Impact Factor
• Article: Inhibition of BET recruitment to chromatin as an effective treatment for MLL-fusion leukaemia.

  Mark A Dawson, Rab K Prinjha, Antje Dittmann, George Giotopoulos, Marcus Bantscheff, Wai-In Chan, Samuel C Robson, Chun-wa Chung, Carsten Hopf, Mikhail M Savitski, [......], Kurt R Auger, Olivier Mirguet, Konstanze Doehner, Ruud Delwel, Alan K Burnett, Phillip Jeffrey, Gerard Drewes, Kevin Lee, Brian J P Huntly, Tony Kouzarides
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  ABSTRACT: Recurrent chromosomal translocations involving the mixed lineage leukaemia (MLL) gene initiate aggressive forms of leukaemia, which are often refractory to conventional therapies. Many MLL-fusion partners are members of the super elongation complex (SEC), a critical regulator of transcriptional elongation, suggesting that aberrant control of this process has an important role in leukaemia induction. Here we use a global proteomic strategy to demonstrate that MLL fusions, as part of SEC and the polymerase-associated factor complex (PAFc), are associated with the BET family of acetyl-lysine recognizing, chromatin 'adaptor' proteins. These data provided the basis for therapeutic intervention in MLL-fusion leukaemia, via the displacement of the BET family of proteins from chromatin. We show that a novel small molecule inhibitor of the BET family, GSK1210151A (I-BET151), has profound efficacy against human and murine MLL-fusion leukaemic cell lines, through the induction of early cell cycle arrest and apoptosis. I-BET151 treatment in two human leukaemia cell lines with different MLL fusions alters the expression of a common set of genes whose function may account for these phenotypic changes. The mode of action of I-BET151 is, at least in part, due to the inhibition of transcription at key genes (BCL2, C-MYC and CDK6) through the displacement of BRD3/4, PAFc and SEC components from chromatin. In vivo studies indicate that I-BET151 has significant therapeutic value, providing survival benefit in two distinct mouse models of murine MLL-AF9 and human MLL-AF4 leukaemia. Finally, the efficacy of I-BET151 against human leukaemia stem cells is demonstrated, providing further evidence of its potent therapeutic potential. These findings establish the displacement of BET proteins from chromatin as a promising epigenetic therapy for these aggressive leukaemias.
  Nature 10/2011; 478(7370):529-33. · 36.28 Impact Factor
• Article: Discovery and characterization of small molecule inhibitors of the BET family bromodomains.

  Chun-Wa Chung, Herve Coste, Julia H White, Olivier Mirguet, Jonathan Wilde, Romain L Gosmini, Chris Delves, Sylvie M Magny, Robert Woodward, Stephen A Hughes, [......], Iain J Uings, Jerome Toum, Catherine A Clement, Anne-Benedicte Boullay, Rachel L Grimley, Florence M Blandel, Rab K Prinjha, Kevin Lee, Jorge Kirilovsky, Edwige Nicodeme
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  ABSTRACT: Epigenetic mechanisms of gene regulation have a profound role in normal development and disease processes. An integral part of this mechanism occurs through lysine acetylation of histone tails which are recognized by bromodomains. While the biological and structural characterization of many bromodomain containing proteins has advanced considerably, the therapeutic tractability of this protein family is only now becoming understood. This paper describes the discovery and molecular characterization of potent (nM) small molecule inhibitors that disrupt the function of the BET family of bromodomains (Brd2, Brd3, and Brd4). By using a combination of phenotypic screening, chemoproteomics, and biophysical studies, we have discovered that the protein-protein interactions between bromodomains and acetylated histones can be antagonized by selective small molecules that bind at the acetylated lysine recognition pocket. X-ray crystal structures of compounds bound into bromodomains of Brd2 and Brd4 elucidate the molecular interactions of binding and explain the precisely defined stereochemistry required for activity.
  Journal of Medicinal Chemistry 06/2011; 54(11):3827-38. · 4.80 Impact Factor
• Source

  Available from: stanford.edu

  Article: Lysine methylation of the NF-κB subunit RelA by SETD6 couples activity of the histone methyltransferase GLP at chromatin to tonic repression of NF-κB signaling.

  Dan Levy, Alex J Kuo, Yanqi Chang, Uwe Schaefer, Christopher Kitson, Peggie Cheung, Alexsandra Espejo, Barry M Zee, Chih Long Liu, Stephanie Tangsombatvisit, [......], Katrin F Chua, Paul J Utz, Xiaobing Shi, Rab K Prinjha, Kevin Lee, Benjamin A Garcia, Mark T Bedford, Alexander Tarakhovsky, Xiaodong Cheng, Or Gozani
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  ABSTRACT: Signaling via the methylation of lysine residues in proteins has been linked to diverse biological and disease processes, yet the catalytic activity and substrate specificity of many human protein lysine methyltransferases (PKMTs) are unknown. We screened over 40 candidate PKMTs and identified SETD6 as a methyltransferase that monomethylated chromatin-associated transcription factor NF-κB subunit RelA at Lys310 (RelAK310me1). SETD6-mediated methylation rendered RelA inert and attenuated RelA-driven transcriptional programs, including inflammatory responses in primary immune cells. RelAK310me1 was recognized by the ankryin repeat of the histone methyltransferase GLP, which under basal conditions promoted a repressed chromatin state at RelA target genes through GLP-mediated methylation of histone H3 Lys9 (H3K9). NF-κB-activation-linked phosphorylation of RelA at Ser311 by protein kinase C-ζ (PKC-ζ) blocked the binding of GLP to RelAK310me1 and relieved repression of the target gene. Our findings establish a previously uncharacterized mechanism by which chromatin signaling regulates inflammation programs.
  Nature Immunology 01/2011; 12(1):29-36. · 26.01 Impact Factor
• Source

  Available from: reactionbiology.com

  Article: Suppression of inflammation by a synthetic histone mimic.

  Edwige Nicodeme, Kate L Jeffrey, Uwe Schaefer, Soren Beinke, Scott Dewell, Chun-Wa Chung, Rohit Chandwani, Ivan Marazzi, Paul Wilson, Hervé Coste, Julia White, Jorge Kirilovsky, Charles M Rice, Jose M Lora, Rab K Prinjha, Kevin Lee, Alexander Tarakhovsky
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  ABSTRACT: Interaction of pathogens with cells of the immune system results in activation of inflammatory gene expression. This response, although vital for immune defence, is frequently deleterious to the host due to the exaggerated production of inflammatory proteins. The scope of inflammatory responses reflects the activation state of signalling proteins upstream of inflammatory genes as well as signal-induced assembly of nuclear chromatin complexes that support mRNA expression. Recognition of post-translationally modified histones by nuclear proteins that initiate mRNA transcription and support mRNA elongation is a critical step in the regulation of gene expression. Here we present a novel pharmacological approach that targets inflammatory gene expression by interfering with the recognition of acetylated histones by the bromodomain and extra terminal domain (BET) family of proteins. We describe a synthetic compound (I-BET) that by 'mimicking' acetylated histones disrupts chromatin complexes responsible for the expression of key inflammatory genes in activated macrophages, and confers protection against lipopolysaccharide-induced endotoxic shock and bacteria-induced sepsis. Our findings suggest that synthetic compounds specifically targeting proteins that recognize post-translationally modified histones can serve as a new generation of immunomodulatory drugs.
  Nature 11/2010; 468(7327):1119-23. · 36.28 Impact Factor