DOCSLIB.ORG

Histone Acetyltransferase KAT6A Upregulates PI3K/AKT Signaling Through TRIM24 Binding Deguan Lv1,2, Feng Jia3, Yanli Hou4, Youzhou Sang2, Angel A

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Histone Acetyltransferase KAT6A Upregulates PI3K/AKT Signaling Through TRIM24 Binding Deguan Lv1,2, Feng Jia3, Yanli Hou4, Youzhou Sang2, Angel A Published OnlineFirst October 11, 2017; DOI: 10.1158/0008-5472.CAN-17-1388 Cancer Tumor and Stem Cell Biology Research Histone Acetyltransferase KAT6A Upregulates PI3K/AKT Signaling through TRIM24 Binding Deguan Lv1,2, Feng Jia3, Yanli Hou4, Youzhou Sang2, Angel A. Alvarez5, Weiwei Zhang2, Wei-Qiang Gao2,BoHu5, Shi-Yuan Cheng2,5, Jianwei Ge3, Yanxin Li6, and Haizhong Feng2 Abstract Lysine acetyltransferase KAT6A is a chromatin regulator that PI3K/AKT signaling and tumorigenesis. Overexpressing activated contributes to histone modification and cancer, but the basis of its AKT or PIK3CA rescued the growth inhibition due to KAT6A actions are not well understood. Here, we identify a KAT6A silencing. Conversely, the pan-PI3K inhibitor LY294002 abrogat- signaling pathway that facilitates glioblastoma (GBM), where it ed the growth-promoting effect of KAT6A. Overexpression is upregulated. KAT6A expression was associated with GBM of KAT6A or TRIM24, but not KAT6A acetyltransferase activity– patient survival. KAT6A silencing suppressed cell proliferation, deficient mutants or TRIM24 mutants lacking H3K23ac-bind- cell migration, colony formation, and tumor development in an ing sites, promoted PIK3CA expression, AKT phosphorylation, orthotopic mouse xenograft model system. Mechanistic investi- and cell proliferation. Taken together, our results define gations demonstrated that KAT6A acetylates lysine 23 of histone an essential role of KAT6A in glioma formation, rationalizing H3 (H3K23), which recruits the nuclear receptor binding protein its candidacy as a therapeutic target for GBM treatment. TRIM24 to activate PIK3CA transcription, thereby enhancing Cancer Res; 77(22); 6190–201. Ó2017 AACR. Introduction deregulated in cancer, including glioma, partially due to aberrant histone modifications and mutations (2–4). However, the Glioblastoma (GBM) is the most common primary malignant mechanisms by which histone modifiers regulate gliomagenesis cancer of the central nervous system with a grim median survival are still being elucidated. of less than 1.5 years upon diagnosis (1). Recent global genomic Lysine acetyltransferase 6A (KAT6A, also known as MYST3 and and transcriptome analyses reveal that the epigenetic landscape is MOZ) is a MYST-family histone acetyltransferase, which controls fundamental cellular processes, including gene transcription (5), cellular senescence (6), cardiac septum development (7), memory 1State Key Laboratory of Oncogenes and Related Genes, Renji-Med X Clinical T-cell diversity (8), and maintenance of normal hematopoietic Stem Cell Research Center, Ren Ji Hospital, School of Biomedical Engineering, stem cells (9). Dysregulation of the KAT6A histone acetyltransfer- Shanghai Jiao Tong University, Shanghai, China. 2Renji-Med X Clinical Stem Cell 3 ase activity or aberrant expression of KAT6A has been associated Research Center, Ren Ji Hospital, Shanghai, China. Department of Neurosur- – gery, Ren Ji Hospital, Shanghai, China. 4Department of Radiotherapy, Ren Ji with oncogenesis in leukemia (10 13) and breast cancer (14). fi fi Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai, China. KAT6A's oncogenic activity was rst identi ed as a recurrent 5Department of Neurology, Northwestern Brain Tumor Institute, The Robert H. fusion partner of the CREB binding protein (CBP) as a conse- Lurie Comprehensive Cancer Center, Northwestern University Feinberg School quence of gene translocation, t(8;16)(p11;p13) in the FAB 6 of Medicine, Chicago, Illinois. Key Laboratory of Pediatric Hematology and M4/M5 subtype of acute myeloid leukemia (AML; ref. 13). Then, Oncology Ministry of Health, Pediatric Translational Medicine Institute, Shanghai the KAT6A-TIF2 fusion was identified as a consequence of another Children's Medical Center, School of Medicine, Shanghai Jiao Tong University, Shanghai, China. translocation, inv(8)(p11;p13) in AML. This AML fusion event requires the KAT6A nucleosome binding motif and TIF2-medi- Note: Supplementary data for this article are available at Cancer Research ated recruitment of CBP (10). KAT6A is composed of tandem Online (http://cancerres.aacrjournals.org/). PHD fingers, a MYST domain, an acidic region, and a Ser/Met D. Lv, F. Jia, and Y. Hou contributed equally to this article. (SM)-rich domain (15). KAT6A binds and acetylates histones via Corresponding Authors: Haizhong Feng, State Key Laboratory of Oncogenes its MYST domain (15). In addition, KAT6A and the related factor and Related Genes, Renji-Med X Clinical Stem Cell Research Center, Ren Ji Lysine acetyltransferase 6B (KAT6B, also known as MORF) form a Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai 200127, tetrameric complex with ING5 (Inhibitor of growth 5), EAF6 China; Phone: 8621-6838-3921; Fax: 8621-6838-3916; E-mail: [email protected]; Yanxin Li, Pediatric Translational Medicine Institute, (Esa1-associated factor 6 ortholog), and the bromodomain-PHD fi Shanghai Children's Medical Center, School of Medicine, Shanghai Jiao Tong nger protein BRPF1 (15). Increasing evidence suggests that University, Shanghai 200127, China. Phone: 8621-3862-6295; E-mail: KAT6A is implicated in regulating tumor progression (16), but [email protected]; and Jianwei Ge, Department of Neurosurgery, Ren Ji mechanisms that facilitate its oncogenic activity independent of Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai 200127, gene fusion remain elusive. China. Phone: 8621-6838-3768; E-mail: [email protected] In this study, we firstly examined KAT6A expression in glioma doi: 10.1158/0008-5472.CAN-17-1388 cells and clinical GBM specimens. We then assessed the role of Ó2017 American Association for Cancer Research. KAT6A in cell proliferation, cell migration, and tumor growth in 6190 Cancer Res; 77(22) November 15, 2017 Downloaded from cancerres.aacrjournals.org on September 30, 2021. © 2017 American Association for Cancer Research. Published OnlineFirst October 11, 2017; DOI: 10.1158/0008-5472.CAN-17-1388 KAT6A Promotes Glioma Tumorigenesis gliomas using cell culture and orthotopic xenograft models. Colony formation assay Finally, we determined the mechanisms by which KAT6A regu- Soft agar colony formation assays were performed as we lates glioma tumorigenesis. described previously (21). Briefly, 1 Â 104 cells were seeded in a 0.4% noble agar top layer with a bottom layer of 0.8% noble agar Materials and Methods in each of the triplicate wells of a 12-well plate. Cell culture media Cell lines was changed every 3 days thereafter. Colonies were scored after – LN428, LN340, U87, LN229, D54, T98G, U251, and LN444 2 3 weeks and data were analyzed. GBM cells were from ATCC, and were cultured in 10% FBS/ DMEM. All cell lines in this study were authenticated using STR shRNA knockdown and transfection DNA fingerprinting, most recently in March 2017 by Shanghai Lentiviruses were produced by cotransfecting various Biowing Applied Biotechnology Co., Ltd (Shanghai, China), and cDNA and packaging plasmids into HEK293T cells using mycoplasma infection was detected using LookOut Mycoplasma Lipofectamine 2000 reagent according to manufacturer's PCR Detection Kit (Sigma-Aldrich). Only lower passage cell lines instruction (#52758, Invitrogen). Forty-eight hours after trans- were used for the study. fection, the viral-containing supernatants were filtered and added into the culture media supplemented with 8 mg/mL polybrene. Transduced human GBM cells were harvested and Plasmids expression of exogenous proteins was validated by Western Flag-TRIM24 was a gift from Michelle Barton (University of blot analysis. Texas M.D. Anderson Cancer Center, Houston, TX; Addgene plasmid # 28138; ref. 17), pCIG-PIK3CA was a gift from Joseph RNA isolation and qRT-PCR Gleeson (Rockefeller University, New York, NY; Addgene plasmid Total RNA was isolated from cells with TRIzol Plus RNA # 73056; ref. 18), and pLNCX-Myr-AKT was a gift from Joan Purification Kit (Thermo Fisher Scientific) for reverse transcrip- Brugge (Harvard Medical School, Boston, MA; Addgene plasmid # tion with Reverse Transcription Kit (Takara) according to the 17245; ref. 19). Wild-type KAT6A cDNA was amplified from U87 manufacturer's instructions. qRT-PCR was performed with the cells, and then inserted into a pcDNA3 vector (Invitrogen). Power SYBR Green Master Mix (Life Technologies) on the Applied KAT6AG657E, KAT6AC543G/G657E, and TRIM24F979A/N980A point Biosystems StepOne Plus Real-Time Thermal Cycling Block. mutations were generated using a site-directed mutagenesis kit ÀDDC Results were analyzed using the 2 t method. Primers are listed (Invitrogen) following the manufacturer's protocol. KAT6A in Supplementary Table S1. shRNAs were purchased from Genechem, Inc. Chromatin immunoprecipitation–qPCR Immunoprecipitation and Western blotting assays Chromatin immunoprecipitation (ChIP) was performed using Cells were lysed in an immunoprecipitation (IP) buffer the Chromatin Immunoprecipitation Kit (Millipore-Upstate) (20 mmol/L Tris-HCl, pH 7.5, 150 mmol/L NaCl, 1 mmol/L according to the manufacturer's instructions. Immunoprecipi- EDTA, 2 mmol/L Na3VO4, 5 mmol/L NaF, 1%Triton X-100, and fi tated DNA was puri ed after phenol extraction and was used for protease inhibitor cocktail) at 4 C for 30 minutes. The lysates were qRT-PCR. Primers are listed in Supplementary Table S1. centrifuged, and then protein concentrations were determined. Equal amounts of cell lysates were immunoprecipitated with Tumorigenesis studies specific antibodies and protein G-agarose beads (Invitrogen). All
Load more

Recommended publications
  • Functional Roles of Bromodomain Proteins in Cancer
    cancers Review Functional Roles of Bromodomain Proteins in Cancer Samuel P. Boyson 1,2, Cong Gao 3, Kathleen Quinn 2,3, Joseph Boyd 3, Hana Paculova 3 , Seth Frietze 3,4,* and Karen C. Glass 1,2,4,* 1 Department of Pharmaceutical Sciences, Albany College of Pharmacy and Health Sciences, Colchester, VT 05446, USA; [email protected] 2 Department of Pharmacology, Larner College of Medicine, University of Vermont, Burlington, VT 05405, USA; [email protected] 3 Department of Biomedical and Health Sciences, University of Vermont, Burlington, VT 05405, USA; [email protected] (C.G.); [email protected] (J.B.); [email protected] (H.P.) 4 University of Vermont Cancer Center, Burlington, VT 05405, USA * Correspondence: [email protected] (S.F.); [email protected] (K.C.G.) Simple Summary: This review provides an in depth analysis of the role of bromodomain-containing proteins in cancer development. As readers of acetylated lysine on nucleosomal histones, bromod- omain proteins are poised to activate gene expression, and often promote cancer progression. We examined changes in gene expression patterns that are observed in bromodomain-containing proteins and associated with specific cancer types. We also mapped the protein–protein interaction network for the human bromodomain-containing proteins, discuss the cellular roles of these epigenetic regu- lators as part of nine different functional groups, and identify bromodomain-specific mechanisms in cancer development. Lastly, we summarize emerging strategies to target bromodomain proteins in cancer therapy, including those that may be essential for overcoming resistance. Overall, this review provides a timely discussion of the different mechanisms of bromodomain-containing pro- Citation: Boyson, S.P.; Gao, C.; teins in cancer, and an updated assessment of their utility as a therapeutic target for a variety of Quinn, K.; Boyd, J.; Paculova, H.; cancer subtypes.
    [Show full text]
  • KAT6A Amplifications Are Associated with Shorter Progression-Free Survival and Overall Survival in Patients with Endometrial Serous Carcinoma
    PLOS ONE RESEARCH ARTICLE KAT6A amplifications are associated with shorter progression-free survival and overall survival in patients with endometrial serous carcinoma 1 2 2 2 2 Ozlen Saglam , Zhenya Tang , Guilin Tang , L. Jeffrey Medeiros , Gokce A. TorunerID * 1 Department of Surgical Pathology, Moffitt Cancer Center, Tampa, Florida, United States of America, 2 Department of Hematopathology, Section of Clinical Cytogenetics, The University of Texas MD Anderson Cancer Center, Houston, Texas, United States of America * [email protected] a1111111111 a1111111111 Abstract a1111111111 a1111111111 Somatic copy number alterations (CNA) are common in endometrial serous carcinoma a1111111111 (ESC). We used the Tumor Cancer Genome Atlas Pan Cancer dataset (TCGA Pan Can) to explore the impact of somatic CNA and gene expression levels (mRNA) of cancer-related genes in ESC. Results were correlated with clinico-pathologic parameters such as age of onset, disease stage, progression-free survival (PFS) and overall survival (OS) (n = 108). OPEN ACCESS 1,449 genes with recurrent somatic CNA were identified, observed in 10% or more tumor Citation: Saglam O, Tang Z, Tang G, Medeiros LJ, samples. Somatic CNA and mRNA expression levels were highly correlated (r> = 0.6) for Toruner GA (2020) KAT6A amplifications are 383 genes. Among these, 45 genes were classified in the Tier 1 category of Cancer associated with shorter progression-free survival and overall survival in patients with endometrial Genome Census-Catalogue of Somatic Mutations in Cancer. Eighteen of 45 Tier 1 genes serous carcinoma. PLoS ONE 15(9): e0238477. had highly correlated somatic CNA and mRNA expression levels including ARNT, PIK3CA, https://doi.org/10.1371/journal.pone.0238477 TBLXR1, ASXL1, EIF4A2, HOOK3, IKBKB, KAT6A, TCEA1, KAT6B, ERBB2, BRD4, Editor: JesuÂs Maria Paramio, CIEMAT, SPAIN KEAP1, PRKACA, DNM2, SMARCA4, AKT2, SS18L1.
    [Show full text]
  • Recognition of Cancer Mutations in Histone H3K36 by Epigenetic Writers and Readers Brianna J
    EPIGENETICS https://doi.org/10.1080/15592294.2018.1503491 REVIEW Recognition of cancer mutations in histone H3K36 by epigenetic writers and readers Brianna J. Kleina, Krzysztof Krajewski b, Susana Restrepoa, Peter W. Lewis c, Brian D. Strahlb, and Tatiana G. Kutateladzea aDepartment of Pharmacology, University of Colorado School of Medicine, Aurora, CO, USA; bDepartment of Biochemistry & Biophysics, The University of North Carolina School of Medicine, Chapel Hill, NC, USA; cWisconsin Institute for Discovery, University of Wisconsin, Madison, WI, USA ABSTRACT ARTICLE HISTORY Histone posttranslational modifications control the organization and function of chromatin. In Received 30 May 2018 particular, methylation of lysine 36 in histone H3 (H3K36me) has been shown to mediate gene Revised 1 July 2018 transcription, DNA repair, cell cycle regulation, and pre-mRNA splicing. Notably, mutations at or Accepted 12 July 2018 near this residue have been causally linked to the development of several human cancers. These KEYWORDS observations have helped to illuminate the role of histones themselves in disease and to clarify Histone; H3K36M; cancer; the mechanisms by which they acquire oncogenic properties. This perspective focuses on recent PTM; methylation advances in discovery and characterization of histone H3 mutations that impact H3K36 methyla- tion. We also highlight findings that the common cancer-related substitution of H3K36 to methionine (H3K36M) disturbs functions of not only H3K36me-writing enzymes but also H3K36me-specific readers. The latter case suggests that the oncogenic effects could also be linked to the inability of readers to engage H3K36M. Introduction from yeast to humans and has been shown to have a variety of functions that range from the control Histone proteins are main components of the of gene transcription and DNA repair, to cell cycle nucleosome, the fundamental building block of regulation and nutrient stress response [8].
    [Show full text]
  • Further Delineation of the Clinical Spectrum of KAT6B Disorders and Allelic Series of Pathogenic Variants
    ARTICLE © American College of Medical Genetics and Genomics Further delineation of the clinical spectrum of KAT6B disorders and allelic series of pathogenic variants Li Xin Zhang1, Gabrielle Lemire, MD2, Claudia Gonzaga-Jauregui, PhD3, Sirinart Molidperee, Gr. Cert1, Carolina Galaz-Montoya, MD3, David S. Liu, MD3, Alain Verloes, MD PhD4, Amelle G. Shillington, DO5, Kosuke Izumi, MD PhD6, Alyssa L. Ritter, MS LCGC7, Beth Keena, MS LCGC6, Elaine Zackai, MD7,8, Dong Li, PhD9, Elizabeth Bhoj, MD PhD7, Jennifer M. Tarpinian, MS CGC10, Emma Bedoukian, MS LCGC10, Mary K. Kukolich, MD11, A. Micheil Innes, MD12, Grace U. Ediae, BSc13, Sarah L. Sawyer, MD PhD14, Karippoth Mohandas Nair, MD15, Para Chottil Soumya, MD15, Kinattinkara R. Subbaraman, MD15, Frank J. Probst, MD PhD3,16, Jennifer A. Bassetti, MD3,16, Reid V. Sutton, MD3,16, Richard A. Gibbs, PhD3, Chester Brown, MD PhD17, Philip M. Boone, MD PhD18, Ingrid A. Holm, MD MPH18, Marco Tartaglia, PhD19, Giovanni Battista Ferrero, MD PhD20, Marcello Niceta, MD PhD19, Maria Lisa Dentici, MD19, Francesca Clementina Radio, MD PhD19, Boris Keren, MD21, Constance F. Wells, MD22, Christine Coubes, MD22, Annie Laquerrière, MD PhD23, Jacqueline Aziza, MD24, Charlotte Dubucs, MD24, Sheela Nampoothiri, MD25, David Mowat, MD26, Millan S. Patel, MD27, Ana Bracho, MD28, Francisco Cammarata-Scalisi, MD29, Alper Gezdirici, MD30, Alberto Fernandez-Jaen, MD31, Natalie Hauser, MD32, Yuri A. Zarate, MD33, Katherine A. Bosanko, MS CGC33, Klaus Dieterich, MD PhD34, John C. Carey, MD MPH35, Jessica X. Chong, PhD36,37, Deborah A. Nickerson, PhD37,38, Michael J. Bamshad, MD36,37, Brendan H. Lee, MD PhD3, Xiang-Jiao Yang, PhD39, James R.
    [Show full text]
  • A Set of Regulatory Genes Co-Expressed in Embryonic Human Brain Is Implicated in Disrupted Speech Development
    Molecular Psychiatry https://doi.org/10.1038/s41380-018-0020-x ARTICLE A set of regulatory genes co-expressed in embryonic human brain is implicated in disrupted speech development 1 1 1 2 3 Else Eising ● Amaia Carrion-Castillo ● Arianna Vino ● Edythe A. Strand ● Kathy J. Jakielski ● 4,5 6 7 8 9 Thomas S. Scerri ● Michael S. Hildebrand ● Richard Webster ● Alan Ma ● Bernard Mazoyer ● 1,10 4,5 6,11 6,12 13 Clyde Francks ● Melanie Bahlo ● Ingrid E. Scheffer ● Angela T. Morgan ● Lawrence D. Shriberg ● Simon E. Fisher 1,10 Received: 22 September 2017 / Revised: 3 December 2017 / Accepted: 2 January 2018 © The Author(s) 2018. This article is published with open access Abstract Genetic investigations of people with impaired development of spoken language provide windows into key aspects of human biology. Over 15 years after FOXP2 was identified, most speech and language impairments remain unexplained at the molecular level. We sequenced whole genomes of nineteen unrelated individuals diagnosed with childhood apraxia of speech, a rare disorder enriched for causative mutations of large effect. Where DNA was available from unaffected parents, CHD3 SETD1A WDR5 fi 1234567890();,: we discovered de novo mutations, implicating genes, including , and . In other probands, we identi ed novel loss-of-function variants affecting KAT6A, SETBP1, ZFHX4, TNRC6B and MKL2, regulatory genes with links to neurodevelopment. Several of the new candidates interact with each other or with known speech-related genes. Moreover, they show significant clustering within a single co-expression module of genes highly expressed during early human brain development. This study highlights gene regulatory pathways in the developing brain that may contribute to acquisition of proficient speech.
    [Show full text]
  • Recombinant BRPF1 (627-746) Protein Catalog No: 31375, 31875 Quantity: 100, 1000 Μg Lot No: 32717002 Concentration: 1.3 Μg/Μl Expressed In: E
    Recombinant BRPF1 (627-746) protein Catalog No: 31375, 31875 Quantity: 100, 1000 µg Lot No: 32717002 Concentration: 1.3 µg/µl Expressed In: E. coli Source: Human Buffer Contents: Recombinant BRPF1 (627-746) protein expressed in E. coli and provided in 25 mM Tris pH 8, 300 mM NaCl and 10% glycerol. Background: Bromodomain and PHD finger containing (BRPF1) protein is a component of the MOZ/MORF complex which has histone H3 acetyltransferase activity. The bromodomain of BRPF1 recognizes acetylated histone lysine residues and function as a ‘reader’ of these epigenetic histone marks to regulate chromatin structure and gene expression by linking associated proteins to the recognized acetylated nucleosomal targets. BRPF1 is known to positively regulate the transcription of RUNX1 and RUNX2. Protein Details: The peptide corresponding to amino acids 627 - 746 that contains the bromodomain sequences of BRPF1 (accession number NM_001003694.1) was expressed in E. coli and contains an N-terminal His-Tag and C- terminal FLAG-Tag with an observed molecular weight of 20.3 kDa. The recombinant protein is >90% pure by SDS- PAGE. Application Notes: Recombinant BRPF1 (627-746) is suitable for use in binding assays, inhibitor screening, and selectivity profiling. Storage and Guarantee: Recombinant proteins in solution are temperature sensitive and must be stored at -80°C to prevent degradation. Avoid repeated freeze/thaw cycles and keep on ice when not in storage. This product is guaranteed for 6 months from date of receipt. This product is for research use only and is not for use in diagnostic procedures. North America 877 222 9543 • Europe +32 (0)2 653 0001 • Japan +81 (0)3 5225 3638 • www.activemotif.com Recombinant BRPF1 (627-746) protein gel.
    [Show full text]
  • De Novo Nonsense Mutations in KAT6A, a Lysine
    Please cite this article in press as: Arboleda et al., De Novo Nonsense Mutations in KAT6A, a Lysine Acetyl-Transferase Gene, Cause a Syn- drome Including Microcephal..., The American Journal of Human Genetics (2015), http://dx.doi.org/10.1016/j.ajhg.2015.01.017 REPORT De Novo Nonsense Mutations in KAT6A, a Lysine Acetyl-Transferase Gene, Cause a Syndrome Including Microcephaly and Global Developmental Delay Valerie A. Arboleda,1 Hane Lee,1 Naghmeh Dorrani,2 Neda Zadeh,3,4 Mary Willis,5 Colleen Forsyth Macmurdo,6 Melanie A. Manning,6,7 Andrea Kwan,6,8 Louanne Hudgins,6 Florian Barthelemy,9 M. Carrie Miceli,9 Fabiola Quintero-Rivera,1 Sibel Kantarci,1 Samuel P. Strom,1 Joshua L. Deignan,1 UCLA Clinical Genomics Center,1 Wayne W. Grody,1,2,10 Eric Vilain,2,10 and Stanley F. Nelson1,10,* Chromatin remodeling through histone acetyltransferase (HAT) and histone deactylase (HDAC) enzymes affects fundamental cellular processes including the cell-cycle, cell differentiation, metabolism, and apoptosis. Nonsense mutations in genes that are involved in his- tone acetylation and deacetylation result in multiple congenital anomalies with most individuals displaying significant developmental delay, microcephaly and dysmorphism. Here, we report a syndrome caused by de novo heterozygous nonsense mutations in KAT6A (a.k.a., MOZ, MYST3) identified by clinical exome sequencing (CES) in four independent families. The same de novo nonsense mutation (c.3385C>T [p.Arg1129*]) was observed in three individuals, and the fourth individual had a nearby de novo nonsense mutation (c.3070C>T [p.Arg1024*]). Neither of these variants was present in 1,815 in-house exomes or in public databases.
    [Show full text]
  • Weighted Burden Analysis of Exome-Sequenced Case-Control Sample Implicates Synaptic Genes in Schizophrenia Aetiology
    Behavior Genetics (2018) 48:198–208 https://doi.org/10.1007/s10519-018-9893-3 ORIGINAL RESEARCH Weighted Burden Analysis of Exome-Sequenced Case-Control Sample Implicates Synaptic Genes in Schizophrenia Aetiology David Curtis1,2 · Leda Coelewij1 · Shou‑Hwa Liu1 · Jack Humphrey1,3 · Richard Mott1 Received: 22 November 2017 / Accepted: 13 March 2018 / Published online: 21 March 2018 © The Author(s) 2018 Abstract A previous study of exome-sequenced schizophrenia cases and controls reported an excess of singleton, gene-disruptive vari- ants among cases, concentrated in particular gene sets. The dataset included a number of subjects with a substantial Finnish contribution to ancestry. We have reanalysed the same dataset after removal of these subjects and we have also included non- singleton variants of all types using a weighted burden test which assigns higher weights to variants predicted to have a greater effect on protein function. We investigated the same 31 gene sets as previously and also 1454 GO gene sets. The reduced dataset consisted of 4225 cases and 5834 controls. No individual variants or genes were significantly enriched in cases but 13 out of the 31 gene sets were significant after Bonferroni correction and the “FMRP targets” set produced a signed log p value (SLP) of 7.1. The gene within this set with the highest SLP, equal to 3.4, was FYN, which codes for a tyrosine kinase which phosphorylates glutamate metabotropic receptors and ionotropic NMDA receptors, thus modulating their trafficking, subcellular distribution and function. In the most recent GWAS of schizophrenia it was identified as a “prioritized candidate gene”.
    [Show full text]
  • Mouse Kat6a Conditional Knockout Project (CRISPR/Cas9)
    https://www.alphaknockout.com Mouse Kat6a Conditional Knockout Project (CRISPR/Cas9) Objective: To create a Kat6a conditional knockout Mouse model (C57BL/6J) by CRISPR/Cas-mediated genome engineering. Strategy summary: The Kat6a gene (NCBI Reference Sequence: NM_001081149 ; Ensembl: ENSMUSG00000031540 ) is located on Mouse chromosome 8. 18 exons are identified, with the ATG start codon in exon 3 and the TGA stop codon in exon 18 (Transcript: ENSMUST00000044331). Exon 4 will be selected as conditional knockout region (cKO region). Deletion of this region should result in the loss of function of the Mouse Kat6a gene. To engineer the targeting vector, homologous arms and cKO region will be generated by PCR using BAC clone RP23-327N5 as template. Cas9, gRNA and targeting vector will be co-injected into fertilized eggs for cKO Mouse production. The pups will be genotyped by PCR followed by sequencing analysis. Note: Homozygous null mice display perinatal lethality, cyanosis, decreased hematopoietic progenitor cell numbers, and severely impaired spleen and thymus development, but are not anemic. Heterozygotes display strain background dependent reductions in fertility. Exon 4 starts from about 10.03% of the coding region. The knockout of Exon 4 will result in frameshift of the gene. The size of intron 3 for 5'-loxP site insertion: 40306 bp, and the size of intron 4 for 3'-loxP site insertion: 4429 bp. The size of effective cKO region: ~873 bp. The cKO region does not have any other known gene. Page 1 of 7 https://www.alphaknockout.com Overview of the Targeting Strategy Wildtype allele gRNA region 5' gRNA region 3' 1 4 18 Targeting vector Targeted allele Constitutive KO allele (After Cre recombination) Legends Exon of mouse Kat6a Homology arm cKO region loxP site Page 2 of 7 https://www.alphaknockout.com Overview of the Dot Plot Window size: 10 bp Forward Reverse Complement Sequence 12 Note: The sequence of homologous arms and cKO region is aligned with itself to determine if there are tandem repeats.
    [Show full text]
  • Deficiency of Intellectual Disability-Related Gene Brpf1 Reduced Inhibitory
    bioRxiv preprint doi: https://doi.org/10.1101/2021.01.11.426233; this version posted January 12, 2021. The copyright holder for this preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under aCC-BY-NC-ND 4.0 International license. 1 Deficiency of intellectual disability-related gene Brpf1 reduced inhibitory 2 neurotransmission and Map2k7 expression in GABAergic interneurons 3 Jingli Cao1, Weiwei Xian1, Maierdan Palihati1, Yu Zhu1, Guoxiang Wang2, Yunli Xie2, 4 Guomin Zhou1,3#, Linya You1# 5 6 1Department of Human Anatomy & Histoembryology, School of Basic Medical 7 Sciences, Fudan University, Shanghai 200032, China 8 2Institutes of Brain Sciences, Fudan University, Shanghai 200032, China 9 3Key Laboratory of Medical Imaging Computing and Computer Assisted Intervention 10 of Shanghai, Shanghai 200032, China 11 12 #Correspondences to: [email protected]; [email protected]. 13 14 Conflict of interest: The authors declare no conflicts of interest. 15 16 Running Title: Brpf1 loss reduced neurotransmission 17 18 Keywords: intellectual disability; Brpf1; MGE-derived GABAergic interneurons; 19 differentiation; dendritic morphology, electrophysiology; in vivo migration; 20 mRNA-Seq 21 22 bioRxiv preprint doi: https://doi.org/10.1101/2021.01.11.426233; this version posted January 12, 2021. The copyright holder for this preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under aCC-BY-NC-ND 4.0 International license. 23 Abstract 24 Intellectual disability is closely related to impaired GABA neurotransmission.
    [Show full text]
  • Transcriptional Regulators Are Upregulated in the Substantia Nigra
    Journal of Emerging Investigators Transcriptional Regulators are Upregulated in the Substantia Nigra of Parkinson’s Disease Patients Marianne Cowherd1 and Inhan Lee2 1Community High School, Ann Arbor, MI 2miRcore, Ann Arbor, MI Summary neurological conditions is an established practice (3). Parkinson’s disease (PD) affects approximately 10 Significant gene expression dysregulation in the SN and million people worldwide with tremors, bradykinesia, in the striatum has been described, particularly decreased apathy, memory loss, and language issues. Though such expression in PD synapses. Protein degradation has symptoms are due to the loss of the substantia nigra (SN) been found to be upregulated (4). Mutations in SNCA brain region, the ultimate causes and complete pathology are unknown. To understand the global gene expression (5), LRRK2 (6), and GBA (6) have also been identified changes in SN, microarray expression data from the SN as familial markers of PD. SNCA encodes alpha- tissue of 9 controls and 16 PD patients were compared, synuclein, a protein found in presynaptic terminals that and significantly upregulated and downregulated may regulate vesicle presence and dopamine release. genes were identified. Among the upregulated genes, Eighteen SNCA mutations have been associated with a network of 33 interacting genes centered around the PD and, although the exact pathogenic mechanism is cAMP-response element binding protein (CREBBP) was not confirmed, mutated alpha-synuclein is the major found. The downstream effects of increased CREBBP- component of protein aggregates, called Lewy bodies, related transcription and the resulting protein levels that are often found in PD brains and may contribute may result in PD symptoms, making CREBBP a potential therapeutic target due to its central role in the interactive to cell death.
    [Show full text]
  • FOXP2 and Language Alterations in Psychiatric Pathology Salud Mental, Vol
    Salud mental ISSN: 0185-3325 Instituto Nacional de Psiquiatría Ramón de la Fuente Muñiz Castro Martínez, Xochitl Helga; Moltó Ruiz, María Dolores; Morales Marin, Mirna Edith; Flores Lázaro, Julio César; González Fernández, Javier; Gutiérrez Najera, Nora Andrea; Alvarez Amado, Daniel Eduardo; Nicolini Sánchez, José Humberto FOXP2 and language alterations in psychiatric pathology Salud mental, vol. 42, no. 6, 2019, pp. 297-308 Instituto Nacional de Psiquiatría Ramón de la Fuente Muñiz DOI: 10.17711/SM.0185-3325.2019.039 Available in: http://www.redalyc.org/articulo.oa?id=58262364007 How to cite Complete issue Scientific Information System Redalyc More information about this article Network of Scientific Journals from Latin America and the Caribbean, Spain and Journal's webpage in redalyc.org Portugal Project academic non-profit, developed under the open access initiative REVIEW ARTICLE Volume 42, Issue 6, November-December 2019 doi: 10.17711/SM.0185-3325.2019.039 FOXP2 and language alterations in psychiatric pathology Xochitl Helga Castro Martínez,1 María Dolores Moltó Ruiz,2,3 Mirna Edith Morales Marin,1 Julio César Flores Lázaro,4 Javier González Fernández,2 Nora Andrea Gutiérrez Najera,1 Daniel Eduardo Alvarez Amado,5 José Humberto Nicolini Sánchez1 1 Laboratorio de Genómica de Enfer- ABSTRACT medades Psiquiátricas y Neurode- generativas, Instituto Nacional de From the first reports of the linguist Noam Chomsky it has become clear that the development Medicina Genómica, Ciudad de Background. México, México. of language has an important genetic component. Several reports in families have shown the relationship 2 Departamento de Genética. Univer- between language disorders and genetic polymorphisms. The FOXP2 gene has been a fundamental piece sitat de València.
    [Show full text]