DOCSLIB.ORG

TCL1 Mice Represent a Model for Immunotherapeutic Reversal of Chronic Lymphocytic Leukemia-Induced T-Cell Dysfunction

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
TCL1 Mice Represent a Model for Immunotherapeutic Reversal of Chronic Lymphocytic Leukemia-Induced T-Cell Dysfunction E␮-TCL1 mice represent a model for immunotherapeutic reversal of chronic lymphocytic leukemia-induced T-cell dysfunction Gullu Gorguna,1, Alan G. Ramsayb,1, Tobias A. W. Holderrieda,1, David Zahriehc, Rifca Le Dieub, Fenglong Liuc, John Quackenbushc, Carlo M. Croced, and John G. Gribbenb,2 aDepartments of Medical Oncology and cBiostatistics and Computational Biology, The Dana-Farber Cancer Institute, Harvard Medical School, Boston, MA 02115; dDepartment of Molecular Virology, Immunology, and Medical Genetics, Ohio State University, Columbus, OH 43210; and bInstitute of Cancer, Barts and London School of Medicine, University of London, London EC1M 6BQ, United Kingdom Contributed by Carlo M. Croce, February 21, 2009 (sent for review November 26, 2008) Preclinical animal models have largely ignored the immune-sup- impairment of T-cell function and alteration of gene expression pressive mechanisms that are important in human cancers. The in CD4 and CD8 T cells to that observed in human patients with identification and use of such models should allow better predic- this disease. The causal relationship between leukemia and the tions of successful human responses to immunotherapy. As a induction of T-cell changes is demonstrated in vivo by the finding model for changes induced in nonmalignant cells by cancer, we that infusion of CLL B cells into nonleukemia-bearing E␮-TCL1 examined T-cell function in the chronic lymphocytic leukemia (CLL) mice rapidly induces these changes, demonstrating a causal E␮-TCL1 transgenic mouse model. With development of leukemia, relationship. This model allows dissection of the molecular E␮-TCL1 transgenic mice developed functional T-cell defects and changes induced in CD4 and CD8 T cells by interaction with alteration of gene and protein expression closely resembling leukemia cells, and further supports the concept that cancer changes seen in CLL human patients. Furthermore, infusion of CLL results in complex abnormalities in the immune microenviron- cells into young E␮-TCL1 mice induced defects comparable to those ment. The similarities with human CLL, including reversible seen in mice with developed leukemia, demonstrating a causal immunological synapse dysfunction with an immunomodulating relationship between leukemia and the T-cell defects. Altered drug (1), validates the use of E␮-TCL1 mice as a model for pathways involved genes regulating actin remodeling, and T cells further analyses of ways to prevent and reverse cancer-induced exhibited dysfunctional immunological synapse formation and immune dysfunction. T-cell signaling, which was reversed by the immunomodulatory drug lenalidomide. These results further demonstrate the utility of Results this animal model of CLL and define a versatile model to investigate Dysfunctional T-Cell Function in E␮-TCL1 Mice That Have Developed both the molecular mechanisms of cancer-induced immune sup- Leukemia. To test the hypothesis that development of leukemia pression and immunotherapeutic repair strategies. would induce changes in the nonmalignant cells of the immune system, we examined changes in T-cell function in E␮-TCL1 ancer cells induce changes in the tumor microenvironment. transgenic mice (6, 7) as they developed leukemia from 12- CThe mechanisms whereby this occurs are poorly understood, months of age [see supporting information (SI) Fig. S1]. Trans- but include direct cancer cell-T cell interactions, production of genic expression of TCL1 in B cells has no demonstrable effect immune-suppressive factors by cancer cells, and induction of on T cells, because before development of leukemia (tested at regulatory T-cell subsets (1, 2). Understanding the elusive 3–4 months) transgenic mice had normal T-cell numbers, CD4/ mechanisms of tumor-driven immune evasion will aid the re- CD8 ratio, cytokine production on mitogen, and antigenic finement of existing cancer immunotherapy strategies and iden- stimulation (data not shown), and as described below, similar tify novel treatments. To date, preclinical animal models that gene-expression profiles to WT mice. closely model human cancer and, in particular, include exami- However, once the mice developed CLL they had decreased nation of the immune-suppressive mechanisms used by cancer in vivo antigen-specific T-cell activation, suppressed T-cell mi- cells have been under-characterized. The identification and use togenic proliferation, and impaired induction of idiotype- of such models should allow better predictions of successful specific CD8 T cells capable of killing CLL cells (Fig. 1) human responses to immunotherapy. compared to age-matched control WT mice or E␮-TCL1 trans- Chronic lymphocytic leukemia (CLL) is an ideal model cancer genic mice without CLL. Leukemic mice had dysfunctional to study immune T cells that have been exposed to circulating T-cell lymphokine production (Th2-preponderant) with in- tumor B cells and is associated with immune dysfunction. CLL creased IL1, IL4, and IL6 but decreased IL2, IFN-␥, and IL-12␤ is the most common adult leukemia in North America and production. These mice also had impaired proliferation and IL-2 Europe and is currently incurable. Immunotherapeutic strategies production following cross-linking of CD3/CD28 (data not are believed to represent a promising treatment modality for this shown). These data identify defective T-cell function with the disease if immune suppression can be controlled (3). Extensive research has been carried out using CLL human patient cells, Author contributions: G.G., A.G.R., T.A.W.H., R.L.D., C.M.C., and J.G.G. designed research; including the characterization of altered gene and protein- G.G., A.G.R., T.A.W.H., R.L.D., and J.G.G. performed research; C.M.C. contributed new expression profiles (4) and suppressed functional responses in reagents/analytic tools; D.Z., F.L., and J.Q. analyzed data; and G.G., A.G.R., T.A.W.H., R.L.D., patient T cells compared to healthy-donor T cells (1, 5). and J.G.G. wrote the paper. We sought to determine whether development of leukemia in The authors declare no conflict of interest. the well-established E␮-TCL1 transgenic murine model of CLL Data deposition: The data reported in this paper have been deposited in the Gene (6) would induce changes in nonmalignant T cells. To examine Expression Omnibus (GEO) database, www.ncbi.nlm.nih.gov/geo (accession no. GSE8836). this, we examined changes in gene-expression profile, protein 1G.G., A.G.R., and T.A.W.H. contributed equally to this work. expression, and function in T cells from E␮-TCL1 transgenic 2To whom correspondence should be addressed. E-mail: [email protected]. mice as they developed CLL. We demonstrate that development This article contains supporting information online at www.pnas.org/cgi/content/full/ of CLL in these transgenic mice is associated with similar 0901166106/DCSupplemental. 6250–6255 ͉ PNAS ͉ April 14, 2009 ͉ vol. 106 ͉ no. 15 www.pnas.org͞cgi͞doi͞10.1073͞pnas.0901166106 Downloaded by guest on September 28, 2021 A B C Wild type Eµ-Tcl1 mice without CLL 60 Wild type Eμ-Tcl1 without CLL 10000 Eµ-Tcl1 mice with CLL Eμ-Tcl1 without CLL 20 Eμ-Tcl1 with CLL 50 Eμ-Tcl1 with CLL 3 8000 40 6000 30 10 p=0.0003 % lysis CPM x10 CPM 4000 20 2000 p=0.05 10 (CPM) 0 0 [3H]-Thymidine incorporation 0 0.5 1.0 Unstimulated PMA-Ionomycin 3 10 30 Peptide ng/ml Effector:Target Ratio Fig. 1. Development of CLL in E␮-TCL1 mice induces suppressed T-cell function. (A) Decreased in vivo gp33 antigen-specific T-cell activation in mice with CLL. WT mice and mice with and without CLL were immunized with gp33 peptide. On day 8, spleen cells from immunized mice (n ϭ 3 in each group) were stimulated with indicated concentrations of KAVYNFATM peptide in triplicate for 3 days before [3H]-Thymidine incorporation was measured. (B) Decreased response to mitogen activators in mononuclear cells from E␮TCL1 mice with CLL compared to those without CLL. Mononuclear cells were stimulated with 100-ng/ml PMA and 4-␮g/ml ionomycin for 3 days. Proliferation was measured using [3H]-Thymidine. (C) Decreased idiotype-specific cytotoxicity against CLL cells by CD8 T cells. CD8 cells were stimulated in vitro with dendritic cells (DCs) pulsed with Ig heavy chain (IgVH)-derived peptide GSAINYAPSI weekly for 4 weeks. Killing of CLL cells expressing this IgVH by the resulting T-cell lines was assessed at the effector:target ratios shown from wild type and E␮-TCL1 mice with and without CLL. (A–C) Results are the mean Ϯ SD and are representative of at least 3 independent experiments. development of CLL by using this mouse model that is not a regulatory subset. Changes in expression of chemokines and result of aging or the expression of the transgene alone. chemokine receptors in CLL mice were similar to those in CLL patients (4). Molecular Basis for Defective T-Cell Function in the E␮-TCL1 Mouse Model. To understand the molecular basis for the observed Comparison of T-Cell Defects Between E␮-TCL1 Mouse Model and functional defects and to compare changes seen in mice and Human Patients. An important question was whether the changes patients with CLL (4), we performed gene-expression profiling. in gene-expression profiles observed in the mice with CLL were Unsupervised analysis of highly purified CD4 and CD8 T cells similar to those observed in human patients with this disease. To in CLL mice demonstrated altered gene-expression profiles examine this, analysis of gene-expression changes in T cells in compared to WT mice or to young E␮-TCL1 mice without CLL mice compared with those in patients (4) was performed disease (Fig. S2). These findings again support the hypothesis using RESOURCERER (8, 9), a database for annotating and that the altered T-cell gene-expression is not a result of aging or linking microarray resources within and across species.
Load more

Recommended publications
  • Focus on Cdc42 in Breast Cancer: New Insights, Target Therapy Development and Non-Coding Rnas
    Review Focus on Cdc42 in Breast Cancer: New Insights, Target Therapy Development and Non-Coding RNAs Yu Zhang †, Jun Li †, Xing-Ning Lai, Xue-Qiao Jiao, Jun-Ping Xiong and Li-Xia Xiong * Department of Pathophysiology, Jiangxi Province Key Laboratory of Tumor Pathogenesis and Molecular Pathology, Medical College, Nanchang University, 461 Bayi Road, Nanchang 330006, China; [email protected] (Y.Z.); [email protected] (J.L.); [email protected] (X.-N.L.); [email protected] (X.-Q.J.); [email protected] (J.-P.X.) * Correspondence: [email protected]; Tel.: +86-791-8636-0556 † These authors contributed equally to this work. Received: 30 December 2018; Accepted: 8 February 2019; Published: 11 February 2019 Abstract: Breast cancer is the most common malignant tumors in females. Although the conventional treatment has demonstrated a certain effect, some limitations still exist. The Rho guanosine triphosphatase (GTPase) Cdc42 (Cell division control protein 42 homolog) is often upregulated by some cell surface receptors and oncogenes in breast cancer. Cdc42 switches from inactive guanosine diphosphate (GDP)-bound to active GTP-bound though guanine-nucleotide- exchange factors (GEFs), results in activation of signaling cascades that regulate various cellular processes such as cytoskeletal changes, proliferation and polarity establishment. Targeting Cdc42 also provides a strategy for precise breast cancer therapy. In addition, Cdc42 is a potential target for several types of non-coding RNAs including microRNAs and lncRNAs. These non-coding RNAs is extensively involved in Cdc42-induced tumor processes, while many of them are aberrantly expressed. Here, we focus on the role of Cdc42 in cell morphogenesis, proliferation, motility, angiogenesis and survival, introduce the Cdc42-targeted non-coding RNAs, as well as present current development of effective Cdc42-targeted inhibitors in breast cancer.
    [Show full text]
  • Small Rho Gtpase Family Member Cdc42 and Its Role in Neuronal Survival and Apoptosis
    University of Denver Digital Commons @ DU Electronic Theses and Dissertations Graduate Studies 1-1-2017 Small Rho GTPase Family Member Cdc42 and Its Role in Neuronal Survival and Apoptosis Noelle Christine Punessen University of Denver Follow this and additional works at: https://digitalcommons.du.edu/etd Part of the Biology Commons, and the Genetics and Genomics Commons Recommended Citation Punessen, Noelle Christine, "Small Rho GTPase Family Member Cdc42 and Its Role in Neuronal Survival and Apoptosis" (2017). Electronic Theses and Dissertations. 1337. https://digitalcommons.du.edu/etd/1337 This Thesis is brought to you for free and open access by the Graduate Studies at Digital Commons @ DU. It has been accepted for inclusion in Electronic Theses and Dissertations by an authorized administrator of Digital Commons @ DU. For more information, please contact [email protected],[email protected]. Small Rho GTPase Family Member Cdc42 and its Role in Neuronal Survival and Apoptosis A Thesis Presented to the Faculty of Natural Sciences and Mathematics University of Denver In Partial Fulfillment of the Requirements for the Degree Master of Science by Noelle C. Punessen August 2017 Advisor: Dr. Daniel A. Linseman Author: Noelle C. Punessen Title: Small Rho GTPase Family Member Cdc42 and its Role in Neuronal Survival and Apoptosis Advisor: Dr. Daniel A. Linseman Degree Date: August 2017 Abstract Neurodegenerative diseases such as amyotrophic lateral sclerosis (ALS), Alzheimer’s and Parkinson’s disease are caused by a progressive and aberrant destruction of neurons in the brain and spinal cord. These disorders lack effective long term treatments, and existing options focus primarily on either delaying disease onset or alleviating symptomology.
    [Show full text]
  • Supplementary Information Method CLEAR-CLIP. Mouse Keratinocytes
    Supplementary Information Method CLEAR-CLIP. Mouse keratinocytes of the designated genotype were maintained in E-low calcium medium. Inducible cells were treated with 3 ug/ml final concentration doxycycline for 24 hours before performing CLEAR-CLIP. One 15cm dish of confluent cells was used per sample. Cells were washed once with cold PBS. 10mls of cold PBS was then added and cells were irradiated with 300mJ/cm2 UVC (254nM wavelength). Cells were then scraped from the plates in cold PBS and pelleted by centrifugation at 1,000g for 2 minutes. Pellets were frozen at -80oC until needed. Cells were then lysed on ice with occasional vortexing in 1ml of lysis buffer (50mM Tris-HCl pH 7.4, 100mM NaCl, 1mM MgCl2, 0.1 mM CaCl2, 1% NP-40, 0.5% Sodium Deoxycholate, 0.1% SDS) containing 1X protease inhibitors (Roche #88665) and RNaseOUT (Invitrogen #10777019) at 4ul/ml final concentration. Next, TurboDNase (Invitrogen #AM2238, 10U), RNase A (0.13ug) and RNase T1 (0.13U) were added and samples were incubated at 37oC for 5 minutes with occasional mixing. Samples were immediately placed on ice and then centrifuged at 16,160g at 4oC for 20 minutes to clear lysate. 25ul of Protein-G Dynabeads (Invitrogen #10004D) were used per IP. Dynabeads were pre-washed with lysis buffer and pre- incubated with 3ul of Wako Anti-Mouse-Ago2 (2D4) antibody. The dynabead/antibody mixture was added to the lysate and rocked for 2 hours at 4oC. All steps after the IP were done on bead until samples were loaded into the polyacrylamide gel.
    [Show full text]
  • G Protein Regulation of MAPK Networks
    Oncogene (2007) 26, 3122–3142 & 2007 Nature Publishing Group All rights reserved 0950-9232/07 $30.00 www.nature.com/onc REVIEW G Protein regulation of MAPK networks ZG Goldsmith and DN Dhanasekaran Fels Institute for Cancer Research and Molecular Biology, Temple University School of Medicine, Philadelphia, PA, USA G proteins provide signal-coupling mechanisms to hepta- the a-subunits has been used as a basis for the helical cell surface receptors and are criticallyinvolved classification of G proteins into Gs,Gi,Gq and G12 in the regulation of different mitogen-activated protein families in which the a-subunits that show more than kinase (MAPK) networks. The four classes of G proteins, 50% homology are grouped together (Simon et al., defined bythe G s,Gi,Gq and G12 families, regulate 1991). In G-protein-coupled receptor (GPCR)-mediated ERK1/2, JNK, p38MAPK, ERK5 and ERK6 modules by signaling pathways, ligand-activated receptors catalyse different mechanisms. The a- as well as bc-subunits are the exchange of the bound GDP to GTP in the a-subunit involved in the regulation of these MAPK modules in a following which the GTP-bound a-subunit disassociate context-specific manner. While the a- and bc-subunits from the receptor as well as the bg-subunit. The GTP- primarilyregulate the MAPK pathwaysvia their respec- bound a-subunit and the bg-subunit stimulate distinct tive effector-mediated signaling pathways, recent studies downstream effectors including enzymes, ion channels have unraveled several novel signaling intermediates and small GTPase, thus regulating multiple signaling including receptor tyrosine kinases and small GTPases pathways including those involved in the activation of through which these G-protein subunits positivelyas well mitogen-activated protein kinase (MAPK) modules as negativelyregulate specific MAPK modules.
    [Show full text]
  • DNA Modifications in Models of Alcohol Use Disorders
    Alcohol 60 (2017) 19e30 Contents lists available at ScienceDirect Alcohol journal homepage: http://www.alcoholjournal.org/ DNA modifications in models of alcohol use disorders * Christopher T. Tulisiak a, R. Adron Harris a, b, Igor Ponomarev a, b, a Waggoner Center for Alcohol and Addiction Research, The University of Texas at Austin, 2500 Speedway, A4800, Austin, TX 78712, USA b The College of Pharmacy, The University of Texas at Austin, 2409 University Avenue, A1900, Austin, TX 78712, USA article info abstract Article history: Chronic alcohol use and abuse result in widespread changes to gene expression, some of which Received 2 September 2016 contribute to the development of alcohol-use disorders (AUD). Gene expression is controlled, in part, by a Received in revised form group of regulatory systems often referred to as epigenetic factors, which includes, among other 3 November 2016 mechanisms, chemical marks made on the histone proteins around which genomic DNA is wound to Accepted 5 November 2016 form chromatin, and on nucleotides of the DNA itself. In particular, alcohol has been shown to perturb the epigenetic machinery, leading to changes in gene expression and cellular functions characteristic of Keywords: AUD and, ultimately, to altered behavior. DNA modifications in particular are seeing increasing research Alcohol fi Epigenetics in the context of alcohol use and abuse. To date, studies of DNA modi cations in AUD have primarily fi fi fi DNA methylation looked at global methylation pro les in human brain and blood, gene-speci c methylation pro les in DNMT animal models, methylation changes associated with prenatal ethanol exposure, and the potential DNA hydroxymethylation therapeutic abilities of DNA methyltransferase inhibitors.
    [Show full text]
  • Gadd45b Acts As Neuroprotective Effector in Global Ischemia- Induced Neuronal Death
    INTERNATIONAL NEUROUROLOGY JOURNAL INTERNATIONAL INJ pISSN 2093-4777 eISSN 2093-6931 Original Article Int Neurourol J 2019;23(Suppl 1):S11-21 NEU INTERNATIONAL RO UROLOGY JOURNAL https://doi.org/10.5213/inj.1938040.020 pISSN 2093-4777 · eISSN 2093-6931 Volume 19 | Number 2 June 2015 Volume pages 131-210 Official Journal of Korean Continence Society / Korean Society of Urological Research / The Korean Children’s Continence and Enuresis Society / The Korean Association of Urogenital Tract Infection and Inflammation einj.org Mobile Web Gadd45b Acts as Neuroprotective Effector in Global Ischemia- Induced Neuronal Death Chang Hoon Cho1,*, Hyae-Ran Byun1,*, Teresa Jover-Mengual1,2, Fabrizio Pontarelli1, Christopher Dejesus1, Ah-Rhang Cho3, R. Suzanne Zukin1, Jee-Yeon Hwang1,4 1Dominick P. Purpura Department of Neuroscience, Albert Einstein College of Medicine, New York, NY, USA 2 Unidad Mixta de Investigación Cerebrovascular, Instituto de Investigación Sanitaria La Fe - Universidad de Valencia; Departamento de Fisiología, Universidad de Vale---ncia, Valencia, Spain 3Department of Beauty-Art, Seo-Jeong University, Seoul, Korea 4Department of Pharmacology, Creighton University School of Medicine, Omaha, NE, USA Purpose: Transient global ischemia arising in human due to cardiac arrest causes selective, delayed neuronal death in hippo- campal CA1 and cognitive impairment. Growth arrest and DNA-damage-inducible protein 45 beta (Gadd45b) is a well- known molecule in both DNA damage-related pathogenesis and therapies. Emerging evidence suggests that Gadd45b is an anti-apoptotic factor in nonneuronal cells and is an intrinsic neuroprotective molecule in neurons. However, the mechanism of Gadd45b pathway is not fully examined in neurodegeneration associated with global ischemia.
    [Show full text]
  • Gadd45b Deficiency Promotes Premature Senescence and Skin Aging
    www.impactjournals.com/oncotarget/ Oncotarget, Vol. 7, No. 19 Gadd45b deficiency promotes premature senescence and skin aging Andrew Magimaidas1, Priyanka Madireddi1, Silvia Maifrede1, Kaushiki Mukherjee1, Barbara Hoffman1,2 and Dan A. Liebermann1,2 1 Fels Institute for Cancer Research and Molecular Biology, Temple University School of Medicine, Philadelphia, PA, USA 2 Department of Medical Genetics and Molecular Biochemistry, Temple University School of Medicine, Philadelphia, PA, USA Correspondence to: Dan A. Liebermann, email: [email protected] Keywords: Gadd45b, senescence, oxidative stress, DNA damage, cell cycle arrest, Gerotarget Received: March 17, 2016 Accepted: April 12, 2016 Published: April 20, 2016 ABSTRACT The GADD45 family of proteins functions as stress sensors in response to various physiological and environmental stressors. Here we show that primary mouse embryo fibroblasts (MEFs) from Gadd45b null mice proliferate slowly, accumulate increased levels of DNA damage, and senesce prematurely. The impaired proliferation and increased senescence in Gadd45b null MEFs is partially reversed by culturing at physiological oxygen levels, indicating that Gadd45b deficiency leads to decreased ability to cope with oxidative stress. Interestingly, Gadd45b null MEFs arrest at the G2/M phase of cell cycle, in contrast to other senescent MEFs, which arrest at G1. FACS analysis of phospho-histone H3 staining showed that Gadd45b null MEFs are arrested in G2 phase rather than M phase. H2O2 and UV irradiation, known to increase oxidative stress, also triggered increased senescence in Gadd45b null MEFs compared to wild type MEFs. In vivo evidence for increased senescence in Gadd45b null mice includes the observation that embryos from Gadd45b null mice exhibit increased senescence staining compared to wild type embryos.
    [Show full text]
  • CDC42 and Three Newly Identified Genes Including the Ras-Related
    Proc. Natl. Acad. Sci. USA Vol. 86, pp. 9976-9980, December 1989 Genetics Multicopy suppression of the cdc24 budding defect in yeast by CDC42 and three newly identified genes including the ras-related gene RSRI (cell polarity/cell cycle/morphogenesis/Saccharomyces cerevisiae/guanine nucleotide-binding protein) ALAN BENDER AND JOHN R. PRINGLE Department of Biology, The University of Michigan, Ann Arbor, MI 48109 Communicated by Leland Hartwell, July 31, 1989 ABSTRACT Genes CDC24, CDC42, and CDC43 are re- Johnson, and J.R.P., unpublished data), and overproduction quired for the establishment ofcell polarity and the localization of the CDC42 product can produce a mislocalization of of secretion in Saccharomyces cerevisiae; mutants defective in budding sites like that seen in some cdc24 mutants (D. these genes fail to form buds and display isotropic expansion of Johnson and J.R.P., unpublished data). Sequencing of the cell surface. To identify other genes that may be involved CDC42 (D. Johnson and J.R.P., unpublished data) revealed in these processes, we screened yeast genomic DNA libraries for that it is a member of the rho family (11) of ras oncogene- heterologous genes that, when overexpressed from a plasmid, related genes and encodes typical domains for GTP binding can suppress a temperature-sensitive cdc24 mutation. We and hydrolysis. Moreover, its C-terminal sequence suggests identified four such genes. One of these proved to be CDC42, that the CDC42 product, like the ras products, may be which has previously been shown to be a member of the rho modified and thence membrane-associated. The available (ras-homologous) family of genes, and a second is a newly observations suggest a tentative model in which the products identified ras-related gene that we named RSR1.
    [Show full text]
  • Genetic Deletion Ofgadd45b, a Regulator of Active DNA
    The Journal of Neuroscience, November 28, 2012 • 32(48):17059–17066 • 17059 Brief Communications Genetic Deletion of gadd45b, a Regulator of Active DNA Demethylation, Enhances Long-Term Memory and Synaptic Plasticity Faraz A. Sultan,1 Jing Wang,1 Jennifer Tront,2 Dan A. Liebermann,2 and J. David Sweatt1 1Department of Neurobiology and Evelyn F. McKnight Brain Institute, University of Alabama at Birmingham, Birmingham, Alabama 35294 and 2Fels Institute for Cancer Research and Molecular Biology, Temple University, Philadelphia, Pennsylvania 19140 Dynamic epigenetic mechanisms including histone and DNA modifications regulate animal behavior and memory. While numerous enzymes regulating these mechanisms have been linked to memory formation, the regulation of active DNA demethylation (i.e., cytosine-5 demethylation) has only recently been investigated. New discoveries aim toward the Growth arrest and DNA damage- inducible 45 (Gadd45) family, particularly Gadd45b, in activity-dependent demethylation in the adult CNS. This study found memory- associated expression of gadd45b in the hippocampus and characterized the behavioral phenotype of gadd45b Ϫ/Ϫ mice. Results indicate normal baseline behaviors and initial learning but enhanced persisting memory in mutants in tasks of motor performance, aversive conditioning and spatial navigation. Furthermore, we showed facilitation of hippocampal long-term potentiation in mutants. These results implicate Gadd45b as a learning-induced gene and a regulator of memory formation and are consistent with its potential role in active DNA demethylation in memory. Introduction along with the finding of activity-induced gadd45b in the hip- Alterations in neuronal gene expression play a necessary role pocampus led us to hypothesize that Gadd45b modulates mem- in memory consolidation (Miyashita et al., 2008).
    [Show full text]
  • Ingenuity Canonical Pathways
    Supplementary Table 4: Canonical pathways of differentially expressed genes between cases and controls in combined microarray and RT- qPCR experiments Ingenuity Canonical Pathways -log(p-value) Ratio Molecules Complement System 6.35 0.111 CD55,CD59,ITGAM,C3AR1 Granulocyte Adhesion and Diapedesis 5.02 0.0303 IL1R2,GNAI3,ITGAM,IL1RN,MMP9 LXR/RXR Activation 4.24 0.0331 IL1R2,IL1RN,SERPINA1,MMP9 IL-10 Signaling 3.64 0.0441 IL1R2,MAPK14,IL1RN Leukocyte Extravasation Signaling 3.36 0.0195 GNAI3,MAPK14,ITGAM,MMP9 p38 MAPK Signaling 2.95 0.0256 IL1R2,MAPK14,IL1RN Atherosclerosis Signaling 2.88 0.0242 IL1RN,SERPINA1,MMP9 IL-6 Signaling 2.85 0.0236 IL1R2,MAPK14,IL1RN Inhibition of Angiogenesis by TSP1 2.84 0.0625 MAPK14,MMP9 Glucocorticoid Receptor Signaling 2.83 0.0141 IL1R2,MAPK14,BAG1,IL1RN IL-17A Signaling in Fibroblasts 2.77 0.0571 MAPK14,LCN2 Notch Signaling 2.72 0.0541 NUMB,PSEN1 Acute Phase Response Signaling 2.5 0.0179 MAPK14,IL1RN,SERPINA1 Amyloid Processing 2.46 0.04 MAPK14,PSEN1 Agranulocyte Adhesion and Diapedesis 2.45 0.0171 GNAI3,IL1RN,MMP9 NF-κB Signaling 2.44 0.0169 IL1R2,LCK,IL1RN Molecular Mechanisms of Cancer 2.42 0.0109 GNAI3,MAPK14,CFLAR,PSEN1 IL-8 Signaling 2.31 0.0153 GNAI3,ITGAM,MMP9 LPS/IL-1 Mediated Inhibition of RXR Function 2.24 0.0144 IL1R2,IL1RN,ACSL1 CCR5 Signaling in Macrophages 2.21 0.0299 GNAI3,MAPK14 Chemokine Signaling 2.2 0.0294 GNAI3,MAPK14 Caveolar-mediated Endocytosis Signaling 2.16 0.0282 CD55,ITGAM Arginine Degradation I (Arginase Pathway) 2.15 0.25 ARG1 Acetate Conversion to Acetyl-CoA 2.15 0.25 ACSL1
    [Show full text]
  • The Small GTP-Binding Proteins Racl, and Cdc42 Regulate the Activity of the JNK/SAPK Signaling Pathway
    View metadata, citation and similar papers at core.ac.uk brought to you by CORE provided by Elsevier - Publisher Connector Cell, Vol. 81, 1137-1146, June 30, 1995, Copyright © 1995 by Cell Press The Small GTP-Binding Proteins Racl, and Cdc42 Regulate the Activity of the JNK/SAPK Signaling Pathway Omar A. Coso,* Mario Chiariello,* Jin-Chen Yu,t closely related to MAPKs have been identified. One class Hidemi Teramoto,* Piero Crespo,* Ningzhi Xu,* presents extended similarity to the Saccharomyces cere- Toru Miki,t and J. Silvio Gutkind* visiae HOG1 kinase (Han et al., 1994), which is involved *Molecular Signaling Unit in protecting S. cerevisiae from hyperosmotic solutions Laboratory of Cellular Development and Oncology (reviewed by Herskowitz, 1995). The role of this mamma- National Institute of Dental Research lian HOG1 homolog is largely unknown. Although it can tLaboratory of Cellular and Molecular Biology also be activated by changes in osmolarity, it appears to National Cancer Institute participate in the inflammatory response to lipopolysac- National Institutes of Health charides or to inflammatory mediators such as interleu- Bethesda, Maryland 20892 kin-1 (IL-1) (Han et al., 1994; Freshney et al., 1994). The other class of MAPKs represents a family of closely related enzymes activated by cellular stress, which have been Summary named stress-activated protein kinases (SAPKs) (Kyriakis et al., 1994). SAPKs were independently identified by vir- c-Jun amino-terminal kinases (JNKs) and mitogen- tue of their ability to phosphorylate the amino terminus of activated protein kinases (MAPKs) are closely related; the c-Jun transcription factor; hence, they have been also however, they are independently regulated by a variety termed c-Jun amino-terminal kinases (JNKs) (D~rijard et of environmental stimuli.
    [Show full text]
  • Identification of Key Genes and Pathways for Alzheimer's Disease
    Biophys Rep 2019, 5(2):98–109 https://doi.org/10.1007/s41048-019-0086-2 Biophysics Reports RESEARCH ARTICLE Identification of key genes and pathways for Alzheimer’s disease via combined analysis of genome-wide expression profiling in the hippocampus Mengsi Wu1,2, Kechi Fang1, Weixiao Wang1,2, Wei Lin1,2, Liyuan Guo1,2&, Jing Wang1,2& 1 CAS Key Laboratory of Mental Health, Institute of Psychology, Chinese Academy of Sciences, Beijing 100101, China 2 Department of Psychology, University of Chinese Academy of Sciences, Beijing 10049, China Received: 8 August 2018 / Accepted: 17 January 2019 / Published online: 20 April 2019 Abstract In this study, combined analysis of expression profiling in the hippocampus of 76 patients with Alz- heimer’s disease (AD) and 40 healthy controls was performed. The effects of covariates (including age, gender, postmortem interval, and batch effect) were controlled, and differentially expressed genes (DEGs) were identified using a linear mixed-effects model. To explore the biological processes, func- tional pathway enrichment and protein–protein interaction (PPI) network analyses were performed on the DEGs. The extended genes with PPI to the DEGs were obtained. Finally, the DEGs and the extended genes were ranked using the convergent functional genomics method. Eighty DEGs with q \ 0.1, including 67 downregulated and 13 upregulated genes, were identified. In the pathway enrichment analysis, the 80 DEGs were significantly enriched in one Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway, GABAergic synapses, and 22 Gene Ontology terms. These genes were mainly involved in neuron, synaptic signaling and transmission, and vesicle metabolism. These processes are all linked to the pathological features of AD, demonstrating that the GABAergic system, neurons, and synaptic function might be affected in AD.
    [Show full text]