MECHANISMS OF ACTIVATION OF ICE-LIKE CYSTEINE PROTEASES

Summary

Principal Investigator: EMAD ALNEMRI
Affiliation: Thomas Jefferson University
Country: USA
Abstract: DESCRIPTION (provided by applicant): Caspase-1 is an important component of the innate immune response against pathogenic infection and cellular stresses. Activation of caspase-1 can lead to processing of the inactive pro-IL-12 and pro-IL-18 to produce the active cytokines IL-12 and IL-18, respectively, which are potent mediators of inflammation that stimulate fever, recruitment and activation of immune cells, production of secondary cytokines and cellular proliferation. Caspase-1 activation can also lead to an inflammatory form of cell death called pyroptosis. The applicant's laboratory demonstrated recently that this form of cell death is mediated by the caspase-1 adaptor protein ASC, which assembles a supramolecular structure termed the pyroptosome that activates caspase-1 during pyroptosis in response to diverse pro-inflammatory and stress signals. They have also elucidated the role of pyrin, a protein mutated in familial Mediterranean fever, in caspase-1 activation and inflammation in the human auto-inflammatory PAPA syndrome, and demonstrated that pyrin is a homotrimeric cytosolic receptor for the cytoskeleton- organizing protein PSTPIP1. Ligation by PSTPIP1 activates pyrin by unmasking of its PYD domain, which then interacts with ASC and facilitates ASC oligomerization into the active pyroptosome. In this application specific aims are proposed to extend these ongoing studies to further investigate the regulation and molecular determinants of the interaction between PSTPIP1 and pyrin. In particular, the aims will focus on characterizing the exact role of the protein-tyrosine phosphatase PTP-PEST and tyrosine kinase c-Abl, both of which have been shown to interact with PSTPIP1, in the pyrin-mediated caspase-1 activation pathway. It is also proposed to characterize novel upstream activators of the ASC pyroptosome and further investigate the role TLR2, TLR4, cryopyrin and caspase-1-mediated proteolysis in macrophage pyroptosis. Finally, studies are proposed to investigate the potential use of the in vitro ASC pyroptosome assembly as a high throughput screen to identify small molecule therapeutics to inhibit and treat inflammation. PUBLIC HEALTH RELEVANCE: Inflammation is a key process in the innate immune response to infections and cellular stress. This research focuses on understanding the role of key cellular proteins in the complex molecular and cellular inflammatory mechanisms. The results of this research will help in the design and discovery of effective therapeutics to treat inflammation associated with aging, auto-inflammatory disease and chronic inflammatory diseases.
Funding Period: ----------------1998 - ---------------2013-
more information: NIH RePORT

Top Publications

  1. ncbi Cryopyrin and pyrin activate caspase-1, but not NF-kappaB, via ASC oligomerization
    J W Yu
    Department of Microbiology and Immunology, Center for Apoptosis Research, Kimmel Cancer Institute, Thomas Jefferson University, Philadelphia, PA 19107, USA
    Cell Death Differ 13:236-49. 2006
  2. pmc Cutting edge: TLR signaling licenses IRAK1 for rapid activation of the NLRP3 inflammasome
    Teresa Fernandes-Alnemri
    Department of Biochemistry and Molecular Biology, Kimmel Cancer Center, Thomas Jefferson University, Philadelphia, PA 19107
    J Immunol 191:3995-9. 2013
  3. pmc Ribotoxic stress through p38 mitogen-activated protein kinase activates in vitro the human pyrin inflammasome
    Je Wook Yu
    Department of Biochemistry and Molecular Biology and the Kimmel Cancer Center, Thomas Jefferson University, Philadelphia, Pennsylvania 19107, USA
    J Biol Chem 288:11378-83. 2013
  4. pmc Restoration of ASC expression sensitizes colorectal cancer cells to genotoxic stress-induced caspase-independent cell death
    Sujeong Hong
    Department of Microbiology, Yonsei University College of Medicine, Seoul, Republic of Korea
    Cancer Lett 331:183-91. 2013
  5. doi Translation termination factor eRF3 is targeted for caspase-mediated proteolytic cleavage and degradation during DNA damage-induced apoptosis
    Yoshifumi Hashimoto
    Department of Biological Chemistry, Graduate School of Pharmaceutical Sciences, Nagoya City University, Nagoya 467 8603, Japan
    Apoptosis 17:1287-99. 2012
  6. pmc Loss of HtrA2/Omi activity in non-neuronal tissues of adult mice causes premature aging
    S Kang
    Department of Biochemistry and Molecular Biology, The Kimmel Cancer Center, Thomas Jefferson University, Philadelphia, PA 19107, USA
    Cell Death Differ 20:259-69. 2013
  7. pmc Non-transcriptional priming and deubiquitination regulate NLRP3 inflammasome activation
    Christine Juliana
    Department of Biochemistry and Molecular Biology and the Kimmel Cancer Center, Thomas Jefferson University, Philadelphia, Pennsylvania 19107, USA
    J Biol Chem 287:36617-22. 2012
  8. pmc The mitochondrial antiviral protein MAVS associates with NLRP3 and regulates its inflammasome activity
    Sangjun Park
    Department of Microbiology, Brain Korea 21 Project for Medical Science, Institute for Immunology and Immunological Diseases, Yonsei University College of Medicine, Seoul 120 752, Korea
    J Immunol 191:4358-66. 2013
  9. pmc The pyroptosome: a supramolecular assembly of ASC dimers mediating inflammatory cell death via caspase-1 activation
    T Fernandes-Alnemri
    Department of Biochemistry and Molecular Biology, Center for Apoptosis Research, Kimmel Cancer Institute, Thomas Jefferson University, Philadelphia, PA 19107, USA
    Cell Death Differ 14:1590-604. 2007
  10. pmc Pyrin activates the ASC pyroptosome in response to engagement by autoinflammatory PSTPIP1 mutants
    Je Wook Yu
    Department of Biochemistry and Molecular Biology, Center for Apoptosis Research, Kimmel Cancer Institute, Thomas Jefferson University, Philadelphia, PA 19107, USA
    Mol Cell 28:214-27. 2007

Scientific Experts

  • Emad S Alnemri
  • Teresa Fernandes-Alnemri
  • Je Wook Yu
  • Christine Juliana
  • Jianghong Wu
  • Pinaki Datta
  • T Fernandes-Alnemri
  • Inhwa Hwang
  • Sangjun Park
  • Sujeong Hong
  • J W Yu
  • Seokwon Kang
  • Leobaldo Solorzano
  • P Datta
  • S Kang
  • Andrew Farias
  • Yoshifumi Hashimoto
  • Katherine A Fitzgerald
  • Rendong Fang
  • Tyler K Ulland
  • Eicke Latz
  • Franz G Bauernfeind
  • Margaret McCormick
  • I Jeru
  • J Wu
  • Yun Sun Lee
  • D Martinez
  • A S Zervos
  • D S Strayer
  • Won Keun Lee
  • J P Louboutin
  • Junji Sagara
  • Connor Anderson
  • You Sun Kim
  • C P Landel
  • Nao Hosoda
  • Fengsong Qin
  • Shin Ichi Hoshino
  • Yanting Xu
  • Eduardo Hernandez-Cuellar
  • Takeshi Yamamoto
  • Sita R Dewamitta
  • Masao Mitsuyama
  • Shunsuke Sakai
  • Kohsuke Tsuchiya
  • Yanna Shen
  • Huixin Qu
  • RuiLi Yang
  • Ikuo Kawamura
  • Hideki Hara
  • Andrew A Quong
  • Rong Meng
  • Michael A Apicella
  • Bradley D Jones
  • Charles P Scott
  • Carlisle P Landel
  • David K Meyerholz
  • Fayyaz S Sutterwala
  • Margaret R Ketterer
  • Blake W Buchan
  • William M Nauseef
  • Erin McDermott
  • Laurence Eisenlohr
  • Lan Huang
  • Brian G Monks
  • Veit Hornung
  • David Speert
  • Andrea Stutz
  • Kelly MacDonald
  • Gabor Horvath
  • K McElreavey
  • V Hentgen
  • S Amselem
  • S Marlin
  • E Grimprel
  • G Grateau
  • J Landman-Parker
  • E Cochet
  • P Duquesnoy
  • T Sarkisian
  • M Lackmy-Port-Lis
  • ZhiJia Zhang
  • W Jankowski
  • J Zhang
  • B Miller
  • S Rosenberg
  • Z Zhang
  • I Ibrahimi
  • J Sagara
  • S Taniguchi

Detail Information

Publications20

  1. ncbi Cryopyrin and pyrin activate caspase-1, but not NF-kappaB, via ASC oligomerization
    J W Yu
    Department of Microbiology and Immunology, Center for Apoptosis Research, Kimmel Cancer Institute, Thomas Jefferson University, Philadelphia, PA 19107, USA
    Cell Death Differ 13:236-49. 2006
    ..Thus, we propose that pyrin could function as a proinflammatory molecule...
  2. pmc Cutting edge: TLR signaling licenses IRAK1 for rapid activation of the NLRP3 inflammasome
    Teresa Fernandes-Alnemri
    Department of Biochemistry and Molecular Biology, Kimmel Cancer Center, Thomas Jefferson University, Philadelphia, PA 19107
    J Immunol 191:3995-9. 2013
    ..These findings define the pathways that lead to rapid NLRP3 activation and identify IRAK1 as a critical mediator of a transcription-independent,inflammasome-dependent early warning response to pathogenic infection. ..
  3. pmc Ribotoxic stress through p38 mitogen-activated protein kinase activates in vitro the human pyrin inflammasome
    Je Wook Yu
    Department of Biochemistry and Molecular Biology and the Kimmel Cancer Center, Thomas Jefferson University, Philadelphia, Pennsylvania 19107, USA
    J Biol Chem 288:11378-83. 2013
    ..Together, our results indicate that ribotoxic stress activates the human pyrin inflammasome through a mechanism that requires p38 MAPK signaling and microtubule stability...
  4. pmc Restoration of ASC expression sensitizes colorectal cancer cells to genotoxic stress-induced caspase-independent cell death
    Sujeong Hong
    Department of Microbiology, Yonsei University College of Medicine, Seoul, Republic of Korea
    Cancer Lett 331:183-91. 2013
    ..Thus, our data suggest that ASC expression in cancer cells is an important factor in determining their susceptibility to chemotherapy...
  5. doi Translation termination factor eRF3 is targeted for caspase-mediated proteolytic cleavage and degradation during DNA damage-induced apoptosis
    Yoshifumi Hashimoto
    Department of Biological Chemistry, Graduate School of Pharmaceutical Sciences, Nagoya City University, Nagoya 467 8603, Japan
    Apoptosis 17:1287-99. 2012
    ..This is the first report showing that eRF3 could serve as a target in the regulation of gene expression...
  6. pmc Loss of HtrA2/Omi activity in non-neuronal tissues of adult mice causes premature aging
    S Kang
    Department of Biochemistry and Molecular Biology, The Kimmel Cancer Center, Thomas Jefferson University, Philadelphia, PA 19107, USA
    Cell Death Differ 20:259-69. 2013
    ..Our results provide direct genetic evidence linking mitochondrial protein quality control to mtDNA deletions and aging in mammals...
  7. pmc Non-transcriptional priming and deubiquitination regulate NLRP3 inflammasome activation
    Christine Juliana
    Department of Biochemistry and Molecular Biology and the Kimmel Cancer Center, Thomas Jefferson University, Philadelphia, Pennsylvania 19107, USA
    J Biol Chem 287:36617-22. 2012
    ....
  8. pmc The mitochondrial antiviral protein MAVS associates with NLRP3 and regulates its inflammasome activity
    Sangjun Park
    Department of Microbiology, Brain Korea 21 Project for Medical Science, Institute for Immunology and Immunological Diseases, Yonsei University College of Medicine, Seoul 120 752, Korea
    J Immunol 191:4358-66. 2013
    ..Taken together, our results suggest that MAVS facilitates the recruitment of NLRP3 to the mitochondria and may enhance its oligomerization and activation by bringing it in close proximity to mitochondrial reactive oxygen species. ..
  9. pmc The pyroptosome: a supramolecular assembly of ASC dimers mediating inflammatory cell death via caspase-1 activation
    T Fernandes-Alnemri
    Department of Biochemistry and Molecular Biology, Center for Apoptosis Research, Kimmel Cancer Institute, Thomas Jefferson University, Philadelphia, PA 19107, USA
    Cell Death Differ 14:1590-604. 2007
    ..Our results establish that macrophage pyroptosis is mediated by a unique pyroptosome, distinct from the inflammasome...
  10. pmc Pyrin activates the ASC pyroptosome in response to engagement by autoinflammatory PSTPIP1 mutants
    Je Wook Yu
    Department of Biochemistry and Molecular Biology, Center for Apoptosis Research, Kimmel Cancer Institute, Thomas Jefferson University, Philadelphia, PA 19107, USA
    Mol Cell 28:214-27. 2007
    ..Therefore, constitutive ligation and activation of pyrin by mutant PSTPIP1 proteins explain the autoinflammatory phenotype seen in PAPA syndrome...
  11. pmc Mutations in NALP12 cause hereditary periodic fever syndromes
    I Jeru
    Institut National de la Santé et de la Recherche Médicale U 654, Universite Paris 6 Pierre et Marie Curie, 75012 Paris, France
    Proc Natl Acad Sci U S A 105:1614-9. 2008
    ....
  12. ncbi Assembly, purification, and assay of the activity of the ASC pyroptosome
    Teresa Fernandes-Alnemri
    Department of Biochemistry and Molecular Biology, Center for Apoptosis Research, Kimmel Cancer Institute, Thomas Jefferson University, Philadelphia, Pennsylvania, USA
    Methods Enzymol 442:251-70. 2008
    ..Finally, it describes methods for the in vitro reconstitution of a functional ASC pyroptosome from the recombinant ASC protein produced in Escherichia coli...
  13. pmc The AIM2 inflammasome is critical for innate immunity to Francisella tularensis
    Teresa Fernandes-Alnemri
    Department of Biochemistry and Molecular Biology, Thomas Jefferson University, Philadelphia, Pennsylvania, USA
    Nat Immunol 11:385-93. 2010
    ..tularensis infection or the presence of cytoplasmic DNA. Our study identifies AIM2 as a crucial sensor of F. tularensis infection and provides genetic proof of its critical role in host innate immunity to intracellular pathogens...
  14. pmc AIM2 activates the inflammasome and cell death in response to cytoplasmic DNA
    Teresa Fernandes-Alnemri
    Department of Biochemistry and Molecular Biology, Center for Apoptosis Research, Kimmel Cancer Institute, Thomas Jefferson University, Philadelphia, Pennsylvania 19107, USA
    Nature 458:509-13. 2009
    ..This study identifies AIM2 as an important inflammasome component that senses potentially dangerous cytoplasmic DNA, leading to activation of the ASC pyroptosome and caspase-1...
  15. pmc Cutting edge: NF-kappaB activating pattern recognition and cytokine receptors license NLRP3 inflammasome activation by regulating NLRP3 expression
    Franz G Bauernfeind
    Department of Infectious Diseases and Immunology, University of Massachusetts Medical School, Worcester, MA 01605, USA
    J Immunol 183:787-91. 2009
    ..Signals provided by NF-kappaB activators are necessary but not sufficient for NLRP3 activation, and a second stimulus such as ATP or crystal-induced damage is required for NLRP3 activation...
  16. pmc Critical roles of ASC inflammasomes in caspase-1 activation and host innate resistance to Streptococcus pneumoniae infection
    Rendong Fang
    Department of Microbiology, Kyoto University Graduate School of Medicine, Kyoto 606 8501, Japan
    J Immunol 187:4890-9. 2011
    ..pneumoniae, with impaired secretion of IL-1β and IL-18 into the bronchoalveolar lavage after intranasal infection, suggesting that ASC inflammasomes contribute to the protection of host from infection with PLY-producing S. pneumoniae...
  17. pmc Cutting edge: mutation of Francisella tularensis mviN leads to increased macrophage absent in melanoma 2 inflammasome activation and a loss of virulence
    Tyler K Ulland
    Graduate Program in Immunology, University of Iowa, Iowa City, IA 52242, USA
    J Immunol 185:2670-4. 2010
    ..This study demonstrates that mviN limits F. tularensis-induced absent in melanoma 2 inflammasome activation, which is critical for its virulence in vivo...
  18. pmc Anti-inflammatory compounds parthenolide and Bay 11-7082 are direct inhibitors of the inflammasome
    Christine Juliana
    Department of Biochemistry and Molecular Biology, Kimmel Cancer Center, Thomas Jefferson University, Philadelphia, Pennsylvania 19107, USA
    J Biol Chem 285:9792-802. 2010
    ..Our results thus elucidate the molecular mechanism for the therapeutic anti-inflammatory activity of parthenolide and identify vinyl sulfones as a new class of potential therapeutics that target the NLRP3 inflammasome...
  19. pmc Sensing cytoplasmic danger signals by the inflammasome
    Emad S Alnemri
    Department of Biochemistry and Molecular Biology, Thomas Jefferson University, Philadelphia, PA, 19107, USA
    J Clin Immunol 30:512-9. 2010
    ..Absent in melanoma 2 (AIM2) is a newly discovered PRR involved in the sensing of dangerous cytosolic DNA produced by infection with DNA viruses...
  20. pmc Involvement of the AIM2, NLRC4, and NLRP3 inflammasomes in caspase-1 activation by Listeria monocytogenes
    Jianghong Wu
    Department of Biochemistry and Molecular Biology, Thomas Jefferson University, Philadelphia, PA 19107, USA
    J Clin Immunol 30:693-702. 2010
    ..Taken together, our results indicate that Listeria infection is sensed by multiple inflammasomes that collectively orchestrate a robust caspase-1 activation and proinflammatory response...