Nonaminoglycoside compounds induce readthrough of nonsense mutations

被引：111

作者：

Du, Liutao ^{[1
]}

Damoiseaux, Robert ^{[5
]}

Nahas, Shareef ^{[1
]}

Gao, Kun ^{[2
]}

Hu, Hailiang ^{[1
]}

Pollard, Julianne M. ^{[1
]}

Goldstine, Jimena ^{[3
]}

Jung, Michael E. ^{[6
]}

Henning, Susanne M. ^{[2
]}

Bertoni, Carmen ^{[4
]}

Gatti, Richard A. ^{[1
]}

机构：

[1] Univ Calif Los Angeles, Dept Pathol & Lab Med, Los Angeles, CA 90095 USA

[2] Univ Calif Los Angeles, Ctr Human Nutr, Los Angeles, CA 90095 USA

[3] Univ Calif Los Angeles, Dept Human Genet, Los Angeles, CA 90095 USA

[4] Univ Calif Los Angeles, Dept Neurol, David Geffen Sch Med, Los Angeles, CA 90095 USA

[5] Univ Calif Los Angeles, Calif NanoSyst Inst, Los Angeles, CA 90095 USA

[6] Univ Calif Los Angeles, Dept Chem & Biochem, Los Angeles, CA 90095 USA

来源：

JOURNAL OF EXPERIMENTAL MEDICINE | 2009年 / 206卷 / 10期

基金：

美国国家卫生研究院;

关键词：

PREMATURE STOP MUTATIONS; DUCHENNE MUSCULAR-DYSTROPHY; CYSTIC-FIBROSIS PATIENTS; PROTEIN TRUNCATION TEST; HUMAN GENETIC-DISEASES; ATAXIA-TELANGIECTASIA; IN-VITRO; READ-THROUGH; GENTAMICIN; SUPPRESSION;

D O I：

10.1084/jem.20081940

中图分类号：

R392 [医学免疫学]; Q939.91 [免疫学];

学科分类号：

071005 [微生物学]; 100108 [医学免疫学];

摘要：

Large numbers of genetic disorders are caused by nonsense mutations for which compound-induced readthrough of premature termination codons (PTCs) might be exploited as a potential treatment strategy. We have successfully developed a sensitive and quantitative high-throughput screening (HTS) assay, protein transcription/translation (PTT)-enzymelinked immunosorbent assay (ELISA), for identifying novel PTC-readthrough compounds using ataxia-telangiectasia (A-T) as a genetic disease model. This HTS PTT-ELISA assay is based on a coupled PTT that uses plasmid templates containing prototypic A-T mutated (ATM) mutations for HTS. The assay is luciferase independent. We screened. 34,000 compounds and identified 12 low-molecular-mass nonaminoglycosides with potential PTC-readthrough activity. From these, two leading compounds consistently induced functional ATM protein in ATM-deficient cells containing disease-causing nonsense mutations, as demonstrated by direct measurement of ATM protein, restored ATM kinase activity, and colony survival assays for cellular radiosensitivity. The two compounds also demonstrated readthrough activity in mdx mouse myotube cells carrying a nonsense mutation and induced significant amounts of dystrophin protein.

引用

页码：2285 / 2297

页数：13

共 54 条

[1]

Mechanism of PTC124 activity in cell-based luciferase assays of nonsense codon suppression [J].

Auld, Douglas S. ;

Thorne, Natasha ;

Maguire, William F. ;

Inglese, James .

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2009, 106 (09) :3585-3590

[2]

Aminoglycoside antibiotics restore dystrophin function to skeletal muscles of mdx mice [J].

Barton-Davis, ER ;

Cordier, L ;

Shoturma, DI ;

Leland, SE ;

Sweeney, HL .

JOURNAL OF CLINICAL INVESTIGATION, 1999, 104 (04) :375-381

[3]

Suppression of a CFTR premature stop mutation in a bronchial epithelial cell line [J].

Bedwell, DM ;

Kaenjak, A ;

Benos, DJ ;

Bebok, Z ;

Bubien, JK ;

Hong, J ;

Tousson, A ;

Clancy, JP ;

Sorscher, EJ .

NATURE MEDICINE, 1997, 3 (11) :1280-1284

[4]

Dystrophin gene repair in mdx muscle precursor cells in vitro and in vivo mediated by RNA-DNA chimeric oligonucleotides [J].

Bertoni, C ;

Rando, TA .

HUMAN GENE THERAPY, 2002, 13 (06) :707-718

[5]

Immunoassay to measure ataxia-telangiectasia mutated protein in cellular lysates [J].

Butch, AW ;

Chun, HH ;

Nahas, SA ;

Gatti, RA .

CLINICAL CHEMISTRY, 2004, 50 (12) :2302-2308

[6]

Ataxia-telangiectasia, an evolving phenotype [J].

Chun, HH ;

Gatti, RA .

DNA REPAIR, 2004, 3 (8-9) :1187-1196

[7]

ATM protein purified from vaccinia virus expression system: DNA binding requirements for kinase activation [J].

Chun, HH ;

Cary, RB ;

Lansigan, F ;

Whitelegge, J ;

Rawlings, DJ ;

Gatti, RA .

BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 2004, 322 (01) :74-81

[8]

Improved diagnostic testing for ataxia-telangiectasia by immunoblotting of nuclear lysates for ATM protein expression [J].

Chun, HH ;

Sun, X ;

Nahas, SA ;

Teraoka, S ;

Lai, CH ;

Concannon, P ;

Gatti, RA .

MOLECULAR GENETICS AND METABOLISM, 2003, 80 (04) :437-443

[9]

Evidence that systemic gentamicin suppresses premature stop mutations in patients with cystic fibrosis [J].

Clancy, JP ;

Bobök, Z ;

Ruiz, F ;

King, C ;

Jones, J ;

Walker, L ;

Greer, H ;

Hong, J ;

Wing, L ;

Macaluso, M ;

Lyrene, R ;

Sorscher, EJ ;

Bedwell, DM .

AMERICAN JOURNAL OF RESPIRATORY AND CRITICAL CARE MEDICINE, 2001, 163 (07) :1683-1692

[10]

Rapid screen for truncating ATM mutations by PTT-ELISA [J].

Du, Liutao ;

Lai, Chih-Hung ;

Concannon, Patrick ;

Gatti, Richard A. .

MUTATION RESEARCH-FUNDAMENTAL AND MOLECULAR MECHANISMS OF MUTAGENESIS, 2008, 640 (1-2) :139-144

← 1 2 3 4 5 6 →