DOCSLIB.ORG

Tumor Necrosis Factor‑Α‑Induced Protein‑8 Like 2 Regulates

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Tumor Necrosis Factor‑Α‑Induced Protein‑8 Like 2 Regulates 6346 MOLECULAR MEDICINE REPORTS 16: 6346-6353, 2017 Tumor necrosis factor‑α‑induced protein‑8 like 2 regulates lipopolysaccharide‑induced rat rheumatoid arthritis immune responses and is associated with Rac activation and interferon regulatory factor 3 phosphorylation CHUNYAN SHI1,2*, YUE WANG1*, GUOHONG ZHUANG1*, ZHONGQUAN QI1, YANPING LI1 and PING YIN3 1Organ Transplantation Institute, Anti‑Cancer Research Center, Medical College, Xiamen University, Xiamen, Fujian 361100; 2The Department of Oncology, Jiujiang People's Hospital, Jiujiang, Jiangxi 332000; 3Department of Pathology, Zhongshan Hospital Affiliated to Xiamen University, Xiamen, Fujian 361100, P.R. China Received October 28, 2016; Accepted June 19, 2017 DOI: 10.3892/mmr.2017.7311 Abstract. The endogenously activated rheumatoid arthritis Introduction (R A) synovial fibroblasts (RSFs) a re li kely to be the key to cur ing the disease. RSFs express Toll‑like receptors (TLRs) rendering Rheumatoid arthritis (RA) is a chronic inflammatory disease them prone to activation by exogenous and endogenous TLR characterized by articular inflammation and leads to joint ligands, resulting in the production of chemokines and cyto- destruction (1,2). RA pathogenesis involves a complex kines Germline deletion of tumor necrosis factor‑α-induced humoral and cellular immune response, including the infiltra- protein‑8 like 2 (TIPE2, also known as TNFAIP8L2) results tion of lymphocytes and monocytes into the synovium. These in fatal inflammation and hypersensitivity to TLR and T cell infiltrating cells and synoviocytes release numerous proin- receptor stimulation. The present study demonstrates an flammatory cytokine and chemokines, including interleukin inverse association between TIPE2 and cytokine gene expres- IL‑6, IL‑1 and tumor necrosis factor‑α (TNF‑α), which may sion in RSFs following lipopolysaccharide (LPS) stimulation. serve a significant role in RA pathogenesis (3,4), and may Enhanced TIPE2 expression decreased Ras‑related C3 botu- determine the activation and proliferation of the synovial linum toxin substrate (Rac) activation and interferon regulatory lining and the recruitment of inflammatory cells that induce factor 3 phosphorylation, and phosphoinositide 3‑kinase and inflammation and promote destruction. Elevated levels of Rac inhibition significantly diminished LPS‑induced cytokine proinflammatory cytokines have been observed in the sera gene expression in RSFs. In conclusion, the findings of the and synovial fluids of patients with RA, such as IL‑6, IL‑8, present study demonstrate that TIPE2 serves a negative role in TNF‑α, IL-20, IL-17 and IL-33 (5-9). Neutralizing these activating the Rac signaling pathway and in the initiation of the cytokines with monoclonal antibodies or soluble receptors has immune response by decreasing the activity of proinflamma- previously been developed as a novel treatment for RA (10). tory cytokines. These results may be useful in designing novel The TNF‑α‑inducible protein 8 (TNFAIP8) family consists strategies for the prevention and treatment of RA. of TNFAIP8, TNF‑α‑induced protein 8‑like 1 (TIPE1, also termed TNFAIP8L1), TIPE2 (also termed TNFAIP8L2) and TIPE3 (also termed TNFAIP8L3) (11). These are recently identified proteins that share considerable sequence homology for the regulation of cellular and immune homeostasis (12). Correspondence to: Dr Guohong Zhuang, Organ Transplantation TIPE2 is preferentially expressed in lymphoid tissues and Institute, Anti‑Cancer Research Center, Medical College, Xiamen hematopoietic cells, and negatively regulates immunity (12,13). University, Xiangan South Road, Xiamen, Fujian 361100, P.R. China E‑mail: [email protected] TIPE2 deletion in mice leads to multi‑organ inflammation, splenomegaly and premature death (14-16). TIPE2‑deficient Dr Ping Yin, Department of Pathology, Zhongshan Hospital cells hyperactivate toll‑like receptors (TLRs) and T cell recep- Affiliated to Xiamen University, Hubin South Road, Xiamen, tors (11,12). TIPE2 is significantly downregulated in during Fujian 361100, P.R. China infection or autoimmune disease (17,18). TIPE2‑deficient cells E‑mail: [email protected] lead to the production of various kinds of proinflammatory *Contributed equally cytokines and activation of the phosphoinositide 3‑kinase (PI3K)‑Ras‑related C3 botulinum toxin substrate (Rac) Key words: rheumatoid arthritis, toll‑like receptors, tumor necrosis signaling pathway, which enhances protein kinase B (AKT), factor‑α‑induced protein‑8 like 2, Ras‑related C3 botulinum toxin Rac and interferon (IFN) regulatory factor 3 activities (19-21). substrate 1 In the present study, TIPE2 regulated lipopolysaccharide (LPS)‑induced RA immune responses by targeting Rac SHI et al: TIPE2 REGULATES RHEUMATOID ARTHRITIS IMMUNE RESPONSES IN RATS 6347 GTPases in a PI3K‑dependent manner. Tipe2 protein has the were as follows: 5'‑GTT GAC AGC CAC TGC CTT C C ‑ 3 ' potential to be used to treated RA (19,21-25). (forward) and 5'‑CTG ACA GTG C A T C ATC GCT G‑3' (reverse) for IL‑6; 5 ' ‑ C C T CTC CAT CGA CTA CAA GC‑3' (forward) and Materials and methods 5 ' ‑ C T C TTC TCC ATC TGT GAC GG‑3' (reverse) for IFNG; 5 ' ‑ C C A GGA GAA AGT CAG CCT CC‑3' (forward) and 5'‑GTT Cell lines and plasmids. The induction of adjuvant arthritis GAC CTC AGC GCT GAG C‑3' (reverse) for TNF; 5'‑CTG TGA in rats, the preparation of rheumatoid arthritis (RA) syno- TAG GCC TCT CAA GG‑3' (forward) and 5'‑CAC CAT GGC vial fibroblasts (RSF) and identification techniques were CAA TGT AGG TG‑3' (reverse) for TLR2; 5 ' ‑ C C A GCT GTT performed according to previous publications (10,26). AGC AAC CAG C‑3' (forward) and 5'‑CGA GGG ACA GAT Normal synovial fibroblasts (NSFs) and RSFs were grown ACT TCA GG‑3' (reverse) for TLR3; 5 ' ‑ G T A GCC GCT CTG in Dulbecco's modified Eagle's medium (Gibco; Thermo GCA TCA TC‑3' (forward) and 5'‑CTC CCC AGA GCA TTG Fisher Scientific, Inc., Waltham, MA, USA) supplemented TCC TC‑3' (reverse) for TLR4; 5 ' ‑ T G C GTG ACA TCA AAG with 10% fetal bovine serum (Hangzhou Sijiqing Biological AGA AG‑3' (forward) and 5'‑TCC ATA CCC AAG AAG GAA Engineering Materials Co., Ltd., Hangzhou, China), penicillin GG‑3' (reverse) for β‑actin 5'‑TGA TTC TAC CCA CGG CAA and streptomycin. To generate stable cell lines, 5x105 adju- GTT‑3' (forward) and 5'‑TGA TGG GTT TCC CAT TGA T G A ‑ 3 ' vant arthritis fibroblast‑like synoviocytes were transduced (reverse) for GAPDH; 5'‑CCT TGT GCA AGT GTC TGA with MIGR1/TIPE2‑overexpression recombinant lentiviral AGC‑3' (forward) and 5'‑CCC AAG TCA AGG GCT TGG A A ‑ 3 ' vectors and a MIGR1 lentiviral vector was used as the (reverse) for IL‑1; and, 5'‑GGG AAC ATC CAA GGC A A G ‑ 3 ' negative control (gift from Dr Chen Youhai, University of (forward) and 5'‑AGC TCA TCT AGC ACC TCA CT‑3' (reverse) Pennsylvania School of Medicine, Philadelphia, PA, USA). for TNFAIP8L2. The lentiviral vectors contained human and rat inserts for the gene encoding TIPE2 and the sequences are at: http://www. LPS stimulation assay. Control RFSs and TIPE2‑ addgene.org/27490/sequences/#addgene‑partial. The cells over exp r ess e d RSFs wer e t r e at e d by L PS (10 0 ng /ml; 9001‑62‑1; were cultured at 37˚C and the culture medium was replaced Sigma‑Aldrich; Merck KGaA, Darmstadt, Germany) at 24 h after transduction. After an additional 48 h culture at 3 time‑points (0 min, 2, 8 h). After LPS treatment the cells 37˚C, transduced cells were observed visually under a fluo- were collected and Total RNA was isolated using TRIzol rescent microscope, 12 fields were counted and infected cells reagent (Invitrogen; Thermo Fisher Scientific, Inc.). Analysis constituted >90% of the total cell count. The infected cells of relative gene expression data was performed using the 2-ΔΔCq were selected by flow cytometry and identified by reverse tran- method (27). scription‑quantitative polymerase chain reaction (RT‑qPCR) and western blot analysis (11,26). PI3K and Rac inhibition assay. Control RSFs were treated with or without the indicated concentrations of LY294002 RT‑qPCR. Total RNA was isolated using TRIzol reagent or NSC23766 inhibitors for 20 min prior to stimulation with (Invitrogen; Thermo Fisher Scientific, Inc.) from NSFs LPS (100 ng/ml) for 2 h. Cytokine expression was determined and RSFs and purified with RNeasy Mini kits (Qiagen, using RT‑qPCR (19,26). Inc., Valencia, CA, USA) according to the manufacturer's protocol (11,26). Western blot analysis. The protein was isolated from RSFs After processing with RNase‑free DNase I (Invitrogen; and NSFs using the ProteoPrep Total Extraction Sample Thermo Fisher Scientific, Inc.), RNA samples were reverse kit (PROTTOT‑1KT; Sigma‑Aldrich; Merck KGaA), and transcribed with oligo (dT) and SuperScript II transcrip- the protein concentration was determined by BCA assay tase (Invitrogen; Thermo Fisher Scientific, Inc.). PCR was (p0012 BCA; Beyotime Institute of Biotechnology; Shanghai, performed using an Applied Biosystems 7500 system and China). Then, 0.6 µg aliquots of synovial cell lysates were Power SYBR‑Green PCR Master mix (Applied Biosystems, loaded and separated by 12% SDS‑PAGE, transferred onto Thermo Fisher Scientific, Inc.). Relative gene expression levels a polyvinylidene difluoride membrane, blocked by 5% were determined using GAPDH as the control. The PCR FBS (Hangzhou Sijiqing Biological Engineering Materials products were run in an agarose gel and were in all cases Co., Ltd.) for 1 h at room temperature and probed with the confined to a single band of the expected size. A melting‑curve following primary antibodies for 75 min at room tempera- analysis was also performed to ensure specificity of the prod- ture: rabbit anti‑β‑actin (m1210‑5; 1:1,000; Hangzhou ucts (11,24). Analysis of relative gene expression data was Huaan Biotechnology Co., Ltd., Hangzhou, China), rabbit performed using the 2-ΔΔCq method (27).
Load more

Recommended publications
  • Protein Interactions in the Cancer Proteome† Cite This: Mol
    Molecular BioSystems View Article Online PAPER View Journal | View Issue Small-molecule binding sites to explore protein– protein interactions in the cancer proteome† Cite this: Mol. BioSyst., 2016, 12,3067 David Xu,ab Shadia I. Jalal,c George W. Sledge Jr.d and Samy O. Meroueh*aef The Cancer Genome Atlas (TCGA) offers an unprecedented opportunity to identify small-molecule binding sites on proteins with overexpressed mRNA levels that correlate with poor survival. Here, we analyze RNA-seq and clinical data for 10 tumor types to identify genes that are both overexpressed and correlate with patient survival. Protein products of these genes were scanned for binding sites that possess shape and physicochemical properties that can accommodate small-molecule probes or therapeutic agents (druggable). These binding sites were classified as enzyme active sites (ENZ), protein–protein interaction sites (PPI), or other sites whose function is unknown (OTH). Interestingly, the overwhelming majority of binding sites were classified as OTH. We find that ENZ, PPI, and OTH binding sites often occurred on the same structure suggesting that many of these OTH cavities can be used for allosteric modulation of Creative Commons Attribution 3.0 Unported Licence. enzyme activity or protein–protein interactions with small molecules. We discovered several ENZ (PYCR1, QPRT,andHSPA6)andPPI(CASC5, ZBTB32,andCSAD) binding sites on proteins that have been seldom explored in cancer. We also found proteins that have been extensively studied in cancer that have not been previously explored with small molecules that harbor ENZ (PKMYT1, STEAP3,andNNMT) and PPI (HNF4A, MEF2B,andCBX2) binding sites. All binding sites were classified by the signaling pathways to Received 29th March 2016, which the protein that harbors them belongs using KEGG.
    [Show full text]
  • Tumor Necrosis Factor-Α-Induced Protein 8-Like-2 Is Involved in the Activation of Macrophages by Astragalus Polysaccharides in Vitro
    7428 MOLECULAR MEDICINE REPORTS 17: 7428-7434, 2018 Tumor necrosis factor-α-induced protein 8-like-2 is involved in the activation of macrophages by Astragalus polysaccharides in vitro JIE ZHU1, YUANYUAN ZHANG2, FANGHUA FAN1, GUOYOU WU1, ZHEN XIAO1 and HUANQIN ZHOU1 1Department of Clinical Laboratory, Zhejiang Hospital, Hangzhou, Zhejiang 310013; 2Department of Pulmonology, The Children's Hospital of Zhejiang University School of Medicine, Hangzhou, Zhejiang 310052, P.R. China Received April 6, 2017; Accepted September 11, 2017 DOI: 10.3892/mmr.2018.8730 Abstract. In previous years, studies have shown that Introduction Astragalus polysaccharides (APS) can improve cellular immu- nity and humoral immune function, which has become a focus Macrophages are important immune cells, which are involved of investigations. Tumor necrosis factor-α-induced protein in the stimulation of the innate immune system, including the 8-like 2 (TIPE2) is a negative regulator of immune reactions. release of inflammatory cytokines and inflammatory molecules, However, the effect and underlying mechanisms of TIPE2 on including tumor necrosis-factor (TNF)-α, interleukin (IL)-1β, the APS-induced immune response remains to be fully eluci- IL-6 and nitric oxide (NO) (1,2). These cytokines and inflam- dated. The present study aimed to examine the role of TIPE2 matory molecules are often regulated by the mitogen-activated and its underlying mechanisms in the APS-induced immune protein kinase (MAPK) signaling pathway (3,4). response. The production of nitric oxide (NO) was detected TNF-α-induced protein-8 like 2 (TNFAIP8L2, also known in macrophages in vitro following APS stimulation. In addi- as TIPE2), is a novel member of the TNFAIP8 (TIPE) family.
    [Show full text]
  • Supplementary Materials
    Supplementary materials Supplementary Table S1: MGNC compound library Ingredien Molecule Caco- Mol ID MW AlogP OB (%) BBB DL FASA- HL t Name Name 2 shengdi MOL012254 campesterol 400.8 7.63 37.58 1.34 0.98 0.7 0.21 20.2 shengdi MOL000519 coniferin 314.4 3.16 31.11 0.42 -0.2 0.3 0.27 74.6 beta- shengdi MOL000359 414.8 8.08 36.91 1.32 0.99 0.8 0.23 20.2 sitosterol pachymic shengdi MOL000289 528.9 6.54 33.63 0.1 -0.6 0.8 0 9.27 acid Poricoic acid shengdi MOL000291 484.7 5.64 30.52 -0.08 -0.9 0.8 0 8.67 B Chrysanthem shengdi MOL004492 585 8.24 38.72 0.51 -1 0.6 0.3 17.5 axanthin 20- shengdi MOL011455 Hexadecano 418.6 1.91 32.7 -0.24 -0.4 0.7 0.29 104 ylingenol huanglian MOL001454 berberine 336.4 3.45 36.86 1.24 0.57 0.8 0.19 6.57 huanglian MOL013352 Obacunone 454.6 2.68 43.29 0.01 -0.4 0.8 0.31 -13 huanglian MOL002894 berberrubine 322.4 3.2 35.74 1.07 0.17 0.7 0.24 6.46 huanglian MOL002897 epiberberine 336.4 3.45 43.09 1.17 0.4 0.8 0.19 6.1 huanglian MOL002903 (R)-Canadine 339.4 3.4 55.37 1.04 0.57 0.8 0.2 6.41 huanglian MOL002904 Berlambine 351.4 2.49 36.68 0.97 0.17 0.8 0.28 7.33 Corchorosid huanglian MOL002907 404.6 1.34 105 -0.91 -1.3 0.8 0.29 6.68 e A_qt Magnogrand huanglian MOL000622 266.4 1.18 63.71 0.02 -0.2 0.2 0.3 3.17 iolide huanglian MOL000762 Palmidin A 510.5 4.52 35.36 -0.38 -1.5 0.7 0.39 33.2 huanglian MOL000785 palmatine 352.4 3.65 64.6 1.33 0.37 0.7 0.13 2.25 huanglian MOL000098 quercetin 302.3 1.5 46.43 0.05 -0.8 0.3 0.38 14.4 huanglian MOL001458 coptisine 320.3 3.25 30.67 1.21 0.32 0.9 0.26 9.33 huanglian MOL002668 Worenine
    [Show full text]
  • Lineage-Specific Programming Target Genes Defines Potential for Th1 Temporal Induction Pattern of STAT4
    Downloaded from http://www.jimmunol.org/ by guest on October 1, 2021 is online at: average * The Journal of Immunology published online 26 August 2009 from submission to initial decision 4 weeks from acceptance to publication J Immunol http://www.jimmunol.org/content/early/2009/08/26/jimmuno l.0901411 Temporal Induction Pattern of STAT4 Target Genes Defines Potential for Th1 Lineage-Specific Programming Seth R. Good, Vivian T. Thieu, Anubhav N. Mathur, Qing Yu, Gretta L. Stritesky, Norman Yeh, John T. O'Malley, Narayanan B. Perumal and Mark H. Kaplan Submit online. Every submission reviewed by practicing scientists ? is published twice each month by http://jimmunol.org/subscription Submit copyright permission requests at: http://www.aai.org/About/Publications/JI/copyright.html Receive free email-alerts when new articles cite this article. Sign up at: http://jimmunol.org/alerts http://www.jimmunol.org/content/suppl/2009/08/26/jimmunol.090141 1.DC1 Information about subscribing to The JI No Triage! Fast Publication! Rapid Reviews! 30 days* • Why • • Material Permissions Email Alerts Subscription Supplementary The Journal of Immunology The American Association of Immunologists, Inc., 1451 Rockville Pike, Suite 650, Rockville, MD 20852 Copyright © 2009 by The American Association of Immunologists, Inc. All rights reserved. Print ISSN: 0022-1767 Online ISSN: 1550-6606. This information is current as of October 1, 2021. Published August 26, 2009, doi:10.4049/jimmunol.0901411 The Journal of Immunology Temporal Induction Pattern of STAT4 Target Genes Defines Potential for Th1 Lineage-Specific Programming1 Seth R. Good,2* Vivian T. Thieu,2† Anubhav N. Mathur,† Qing Yu,† Gretta L.
    [Show full text]
  • TNFAIP8L2/TIPE2 Polyclonal Antibody, FITC Conjugated Catalog No
    Product Name: TNFAIP8L2/TIPE2 Polyclonal Antibody, FITC Conjugated Catalog No. : TAP01-88965R-FITC Intended Use: For Research Use Only. Not for used in diagnostic procedures. Size 100ul Concentration 1ug/ul Gene ID ISO Type Rabbit IgG Clone N/A Immunogen Range Conjugation FITC Subcellular Locations Applications IF(IHC-P) Cross Reactive Species Human, Mouse, Rat Source KLH conjugated synthetic peptide derived from human TNFAIP8L2/TIPE2 Applications with IF(IHC-P)(1:50-200) Dilutions Purification Purified by Protein A. Background TNF Alpha-IP 8L2 (tumor necrosis factor, alpha-induced protein 8-like 2), also known as TIPE2, is a 184 amino acid protein that shares 94% identity with its mouse counterpart and belongs to the TNFAIP8 family. Expressed in spleen, thymus, small intestineand lymph node with lower levels present in colon, lung and skin, TNF Alpha-IP 8L2 plays a role in maintaining immune homeostasis, specifically by acting as a negative regulator of both innate and adaptive immunity. In addition, TNF?IP 8L2 functions as anegative regulator of T-cell receptor function and is thought to promote Fas-induced apoptosis. The gene encoding TNF?IP 8L2 maps to human chromosome 1, which spans 260 million base pairs, contains over 3,000 genes and comprises nearly 8% of the human genome. Synonyms Tumor necrosis factor alpha-induced protein 8-like protein 2; Inflammation factor protein 20; TIPE 2; TIPE-2; TNF alpha-induced protein 8-like protein 2; TNFAIP8-like protein 2; TNFAIP8L2; TP8L2_HUMAN; Tumor necrosis factor alpha-induced protein 8-like protein 2; AW610835; FLJ23467. Storage Aqueous buffered solution containing 1% BSA, 50% glycerol and 0.09% sodium azide.
    [Show full text]
  • KRAS Mutations Are Negatively Correlated with Immunity in Colon Cancer
    www.aging-us.com AGING 2021, Vol. 13, No. 1 Research Paper KRAS mutations are negatively correlated with immunity in colon cancer Xiaorui Fu1,2,*, Xinyi Wang1,2,*, Jinzhong Duanmu1, Taiyuan Li1, Qunguang Jiang1 1Department of Gastrointestinal Surgery, The First Affiliated Hospital of Nanchang University, Nanchang, Jiangxi, People's Republic of China 2Queen Mary College, Medical Department, Nanchang University, Nanchang, Jiangxi, People's Republic of China *Equal contribution Correspondence to: Qunguang Jiang; email: [email protected] Keywords: KRAS mutations, immunity, colon cancer, tumor-infiltrating immune cells, inflammation Received: March 27, 2020 Accepted: October 8, 2020 Published: November 26, 2020 Copyright: © 2020 Fu et al. This is an open access article distributed under the terms of the Creative Commons Attribution License (CC BY 3.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. ABSTRACT The heterogeneity of colon cancer tumors suggests that therapeutics targeting specific molecules may be effective in only a few patients. It is therefore necessary to explore gene mutations in colon cancer. In this study, we obtained colon cancer samples from The Cancer Genome Atlas, and the International Cancer Genome Consortium. We evaluated the landscape of somatic mutations in colon cancer and found that KRAS mutations, particularly rs121913529, were frequent and had prognostic value. Using ESTIMATE analysis, we observed that the KRAS-mutated group had higher tumor purity, lower immune score, and lower stromal score than the wild- type group. Through single-sample Gene Set Enrichment Analysis and Gene Set Enrichment Analysis, we found that KRAS mutations negatively correlated with enrichment levels of tumor infiltrating lymphocytes, inflammation, and cytolytic activities.
    [Show full text]
  • Identification of TNFAIP8L1 Binding Partners Through Co-Immunoprecipitation and Mass Spectrometry Audrey Hoyle University of Maine
    The University of Maine DigitalCommons@UMaine Honors College Spring 5-2018 Identification of TNFAIP8L1 Binding Partners Through Co-Immunoprecipitation and Mass Spectrometry Audrey Hoyle University of Maine Follow this and additional works at: https://digitalcommons.library.umaine.edu/honors Part of the Biochemistry Commons, and the Microbiology Commons Recommended Citation Hoyle, Audrey, "Identification of TNFAIP8L1 Binding Partners Through Co-Immunoprecipitation and Mass Spectrometry" (2018). Honors College. 335. https://digitalcommons.library.umaine.edu/honors/335 This Honors Thesis is brought to you for free and open access by DigitalCommons@UMaine. It has been accepted for inclusion in Honors College by an authorized administrator of DigitalCommons@UMaine. For more information, please contact [email protected]. IDENTIFICATION OF TNFAIP8L1 BINDING PARTNERS THROUGH CO- IMMUNOPRECIPITATION AND MASS SPECTROMETRY by Audrey Hoyle A Thesis Submitted in Partial Fulfillment of the Requirements for a Degree with Honors (Biochemistry and Microbiology) The Honors College University of Maine May 2018 Advisory Committee: Con Sullivan, Ph.D., Assistant Research Professor (Advisor) Benjamin L. King, Ph.D., Assistant Professor of Bioinformatics Melissa S. Maginnis, Ph.D., Assistant Professor of Microbiology Sally Molloy, Ph.D., Assistant Professor of Genomics Mary S. Tyler, PhD., Professor of Zoology ©2018 Audrey Hoyle All Rights Reserved ABSTRACT The expanded understanding of the gene families and mechanisms governing tumorigenesis pathways has enormous potential for improving current cancer therapies and patient prognoses. One such gene family that participates in the regulation of tumorigenesis is the tumor necrosis factor alpha-induced protein 8 (TNFAIP8) gene family, which is comprised of four members: TNFAIP8, TNFAIP8L1, TNFAIP8L2, and TNFAIP8L3.
    [Show full text]
  • Evolutionary Divergence of the Vertebrate TNFAIP8 Gene Family: Applying the Spotted Gar Orthology Bridge to Understand Ohnolog Loss in Teleosts
    RESEARCH ARTICLE Evolutionary divergence of the vertebrate TNFAIP8 gene family: Applying the spotted gar orthology bridge to understand ohnolog loss in teleosts Con Sullivan1,2*, Christopher R. Lage3, Jeffrey A. Yoder4, John H. Postlethwait5, Carol H. Kim1,2* a1111111111 1 Department of Molecular and Biomedical Sciences, University of Maine, Orono, Maine, United States of America, 2 Graduate School of Biomedical Sciences and Engineering, University of Maine, Orono, Maine, a1111111111 United States of America, 3 Program in Biology, University of Maine - Augusta, Augusta, Maine, United a1111111111 States of America, 4 Department of Molecular Biomedical Sciences, North Carolina State University, Raleigh, a1111111111 North Carolina, United States of America, 5 Institute of Neuroscience, University of Oregon, Eugene, Oregon, a1111111111 United States of America * [email protected] (CS); [email protected] (CHK) OPEN ACCESS Abstract Citation: Sullivan C, Lage CR, Yoder JA, Postlethwait JH, Kim CH (2017) Evolutionary Comparative functional genomic studies require the proper identification of gene orthologs divergence of the vertebrate TNFAIP8 gene family: to properly exploit animal biomedical research models. To identify gene orthologs, compre- Applying the spotted gar orthology bridge to hensive, conserved gene synteny analyses are necessary to unwind gene histories that understand ohnolog loss in teleosts. PLoS ONE 12 are convoluted by two rounds of early vertebrate genome duplication, and in the case of the (6): e0179517. https://doi.org/10.1371/journal. pone.0179517 teleosts, a third round, the teleost genome duplication (TGD). Recently, the genome of the spotted gar, a holostean outgroup to the teleosts that did not undergo this third genome Editor: Pierre Boudinot, INRA, FRANCE duplication, was sequenced and applied as an orthology bridge to facilitate the identification Received: February 3, 2017 of teleost orthologs to human genes and to enhance the power of teleosts as biomedical Accepted: May 30, 2017 models.
    [Show full text]
  • Roles of Tnfaip8 Protein in Cell Death, Listeriosis, and Colitis
    University of Pennsylvania ScholarlyCommons Publicly Accessible Penn Dissertations 2014 Roles of Tnfaip8 Protein in Cell Death, Listeriosis, and Colitis Thomas Porturas University of Pennsylvania, [email protected] Follow this and additional works at: https://repository.upenn.edu/edissertations Part of the Cell Biology Commons, and the Molecular Biology Commons Recommended Citation Porturas, Thomas, "Roles of Tnfaip8 Protein in Cell Death, Listeriosis, and Colitis" (2014). Publicly Accessible Penn Dissertations. 1409. https://repository.upenn.edu/edissertations/1409 This paper is posted at ScholarlyCommons. https://repository.upenn.edu/edissertations/1409 For more information, please contact [email protected]. Roles of Tnfaip8 Protein in Cell Death, Listeriosis, and Colitis Abstract TNF-alpha-induced protein 8 (TNFAIP8 or TIPE) is a newly described regulator of cancer and infection. However, its precise roles and mechanisms of actions are not well understood. Here I report the generation of TNFAIP8 knockout mice and describe their increased responsiveness to colonic inflammation and esistancer to lethal Listeria monocytogenes infection. TNFAIP8 knockout mice were generated by germ line gene targeting and were born without noticeable developmental abnormalities. Their major organs including those of the immune and digestive systems were macroscopically and microscopically normal. However, compared to wild type mice, TNFAIP8 knockout mice exhibited significant differences in the development of listeriosis and experimentally induced colitis. I discovered that TNFAIP8 regulates L. monocytogenes infection by controlling pathogen invasion and host cell apoptosis potentially in a RAC1 GTPase-dependent manner. TNFAIP8 knockout mice had reduced bacterial load in the liver and spleen and TNFAIP8 knockdown in murine liver HEPA1-6 cells increased apoptosis, reduced bacterial invasion into cells, and resulted in dysregulated RAC1 activation.
    [Show full text]
  • Pan-Cancer Analysis of Frequent DNA Co-Methylation Patterns Reveals
    The Author(s) BMC Genomics 2017, 18(Suppl 1):1045 DOI 10.1186/s12864-016-3259-0 RESEARCH Open Access Pan-cancer analysis of frequent DNA co- methylation patterns reveals consistent epigenetic landscape changes in multiple cancers Jie Zhang and Kun Huang* From The 27th International Conference on Genome Informatics Shanghai, China. 3-5 October 2016 Abstract Background: DNA methylation is the major form of epigenetic modifications through which the cell regulates the gene expression and silencing. There have been extensive studies on the roles of DNA methylation in cancers, and several cancer drugs were developed targeting this process. However, DNA co-methylation cluster has not been examined in depth, and co-methylation in multiple cancer types has never been studied previously. Results: In this study, we applied newly developed lmQCM algorithm to mine co-methylation clusters using methylome data from 11 cancer types in TCGA database, and found frequent co-methylated gene clusters exist in these cancer types. Among the four identified frequent clusters, two of them separate the tumor sample from normal sample in 10 out of 11 cancer types, which indicates that consistent epigenetic landscape changes exist in multiple cancer types. Conclusion: This discovery provides new insight on the epigenetic regulation in cancers and leads to potential new direction for epigenetic biomarker and cancer drug discovery. We also found that genes commonly believed to be silenced via hypermethylation in cancers may still display highly variable methylation levels among cancer cells, and should be considered while using them as epigenetic biomarkers. Keywords: DNA co-methylation, Pan-cancer methylation, Frequent network mining, Epigenetics Background methylome) is affected by cell age, tissue types, and DNA methylation is the most extensively studied form many environmental factors such as nutrients and car- of epigenetic modification in the cell.
    [Show full text]
  • Rare Disruptive Variants in the DISC1 Interactome and Regulome: Association with Cognitive Ability and Schizophrenia
    OPEN Molecular Psychiatry (2018) 23, 1270–1277 www.nature.com/mp ORIGINAL ARTICLE Rare disruptive variants in the DISC1 Interactome and Regulome: association with cognitive ability and schizophrenia S Teng1,2,10, PA Thomson3,4,10, S McCarthy1,10, M Kramer1, S Muller1, J Lihm1, S Morris3, DC Soares3, W Hennah5, S Harris3,4, LM Camargo6, V Malkov7, AM McIntosh8, JK Millar3, DH Blackwood8, KL Evans4, IJ Deary4,9, DJ Porteous3,4 and WR McCombie1 Schizophrenia (SCZ), bipolar disorder (BD) and recurrent major depressive disorder (rMDD) are common psychiatric illnesses. All have been associated with lower cognitive ability, and show evidence of genetic overlap and substantial evidence of pleiotropy with cognitive function and neuroticism. Disrupted in schizophrenia 1 (DISC1) protein directly interacts with a large set of proteins (DISC1 Interactome) that are involved in brain development and signaling. Modulation of DISC1 expression alters the expression of a circumscribed set of genes (DISC1 Regulome) that are also implicated in brain biology and disorder. Here we report targeted sequencing of 59 DISC1 Interactome genes and 154 Regulome genes in 654 psychiatric patients and 889 cognitively-phenotyped control subjects, on whom we previously reported evidence for trait association from complete sequencing of the DISC1 locus. Burden analyses of rare and singleton variants predicted to be damaging were performed for psychiatric disorders, cognitive variables and personality traits. The DISC1 Interactome and Regulome showed differential association across the phenotypes tested. After family-wise error correction across all traits (FWERacross), an increased burden of singleton disruptive variants in the Regulome was associated with SCZ (FWERacross P=0.0339).
    [Show full text]
  • Epigenetic Changes in Human Model KMT2A Leukemias Highlight Early Events During Leukemogenesis
    Epigenetic changes in human model KMT2A leukemias highlight early events during leukemogenesis Thomas Milan1, Magalie Celton1, Karine Lagacé1, Élodie Roques1, Safia Safa-Tahar-Henni1, Eva Bresson2,3,4, Anne Bergeron2,3,4, Josée Hebert5,6, Soheil Meshinchi7, Sonia Cellot8, Frédéric Barabé2,3,4, 1,6 Brian T Wilhelm Author contributions: BTW and TM designed the study, analyzed data and drafted the manuscript. TM and KL performed ChIP-seq and ATAC-seq, analysed the data and generated figures. MC performed Methyl-seq experiments, analysed data, generated figures and drafted the manuscript. SS-T-H assisted with the ATAC-seq analysis and generation of figures. KL, ER, EB, and AB helped with molecular biology work, cloning and cell culture. EB, AB and FB generated the model on mice from CD34+ cells. SM provided expression and clinical data for patient samples and JH and SC collected, characterized, and banked the local patient samples used in this study. Affiliations: 1Laboratory for High Throughput Biology, Institute for Research in Immunology and Cancer, Montréal, QC, Canada 2Centre de recherche en infectiologie du CHUL, Centre de recherche du CHU de Québec – Université Laval, Québec City, QC, Canada, 3CHU de Québec – Université Laval – Hôpital Enfant-Jésus; Québec City, QC, Canada; 4Department of Medicine, Université Laval, Quebec City, QC, Canada 5Division of Hematology-Oncology and Leukemia Cell Bank of Quebec, Maisonneuve-Rosemont Hospital, Montréal, QC, Canada, 6Department of Medicine, Université de Montréal, Montréal, QC, Canada, 7Clinical Research Division, Fred Hutchinson Cancer Research Center, Seattle, Washington, USA 8Department of pediatrics, division of Hematology, Ste-Justine Hospital, Montréal, QC, Canada Running head: Early epigenetic changes in KM3-AML *Correspondence to: Brian T Wilhelm Institute for Research in Immunology and Cancer Université de Montréal P.O.
    [Show full text]