title: Detection of human survival motor neuron (SMN) protein in mice containing the SMN2 transgene: applicability to preclinical therapy development for spinal muscular atrophy.
abstract: Spinal muscular atrophy (SMA), the leading genetic cause of infant death results from loss of spinal motor neurons causing atrophy of skeletal muscle. SMA is caused by loss of the Survival Motor Neuron 1 (SMN1) gene, however, an identically coding gene called SMN2 is retained, but is alternatively spliced to produce approximately 90% truncated protein. Most SMA translational and preclinical drug development has relied on the use of SMA mice to determine changes in SMN protein levels. However, the SMA mouse models are relatively severe and analysis of SMN-inducing compounds is confounded by the early mortality of these animals. An antibody that could detect SMN protein on a Smn background could circumvent this limitation and allow unaffected, heterozygous animals to be examined. Here we describe the generation and characterization of a monoclonal anti-SMN antibody, 4F11, which specifically recognizes human SMN protein. 4F11 detects SMN (human) but not native Smn (mouse) protein in SMN2 transgenic mice and in SMA cell lines. We demonstrate the feasibility of using 4F11 to detect changes in SMN2-derived SMN protein in SMA patient fibroblasts and in healthy SMN2 transgenic mice. This antibody is, therefore, an excellent tool for examining SMN2-inducing therapeutics in patient cells as well as in transgenic mice.
authors.affiliation: Department of Veterinary Pathobiology, University of Missouri, Columbia, MO 65211, USA. Vbm7aa@mizzou.edu
authors.forename: Virginia B
authors.initials: VB
authors.lastname: Mattis
authors.forename: Matthew E R
authors.initials: ME
authors.lastname: Butchbach
authors.forename: Christian L
authors.initials: CL
authors.lastname: Lorson
mesh_terms: D017398:Alternative Splicing / Q000235:genetics; D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000378:metabolism; D002478:Cells, Cultured; D004195:Disease Models, Animal; D004791:Enzyme Inhibitors; D018604:Epitope Mapping / Q000379:methods; D005347:Fibroblasts / Q000187:drug effects / Q000378:metabolism; D005786:Gene Expression Regulation / Q000187:drug effects; D006801:Humans; D006918:Hydroxyurea / Q000627:therapeutic use; D007211:Indoles; D051379:Mice; D008822:Mice, Transgenic / Q000378:metabolism*; D009134:Muscular Atrophy, Spinal / Q000188:drug therapy / Q000378:metabolism* / Q000473:pathology; D055532:SMN Complex Proteins / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D013116:Spinal Cord / Q000187:drug effects / Q000378:metabolism; D055540:Survival of Motor Neuron 2 Protein; D014635:Valproic Acid / Q000627:therapeutic use
chemical_list: D000911:Antibodies, Monoclonal; D004791:Enzyme Inhibitors; D007211:Indoles; D055532:SMN Complex Proteins; C526477:SMN2 protein, human; D055540:Survival of Motor Neuron 2 Protein; C007293:DAPI; D014635:Valproic Acid; D006918:Hydroxyurea
references.citation: Hum Mol Genet. 2003 Oct 1;12(19):2481-9
references.pmid: 12915451
references.citation: J Clin Invest. 2007 Mar;117(3):659-71
references.pmid: 17318264
references.citation: Ann Neurol. 2003 Nov;54(5):647-54
references.pmid: 14595654
references.citation: Hum Genet. 2006 Aug;120(1):101-10
references.pmid: 16724231
references.citation: Hum Mol Genet. 2001 Nov 15;10(24):2841-9
references.pmid: 11734549
references.citation: Am J Hum Genet. 1997 Jun;60(6):1411-22
references.pmid: 9199562
references.citation: Hum Mol Genet. 1999 Jul;8(7):1177-83
references.pmid: 10369862
references.citation: J Cell Biol. 2003 Jan 6;160(1):41-52
references.pmid: 12515823
references.citation: Hum Mol Genet. 2005 May 1;14(9):1199-210
references.pmid: 15790598
references.citation: Am J Hum Genet. 2002 Feb;70(2):358-68
references.pmid: 11791208
references.citation: Neuron. 2005 Dec 22;48(6):885-96
references.pmid: 16364894
references.citation: J Med Chem. 2008 Feb 14;51(3):449-69
references.pmid: 18205293
references.citation: Eur J Hum Genet. 2004 Jan;12(1):59-65
references.pmid: 14560316
references.citation: Proc Natl Acad Sci U S A. 1999 May 25;96(11):6307-11
references.pmid: 10339583
references.citation: Chem Biol. 2004 Nov;11(11):1489-93
references.pmid: 15555999
references.citation: Proc Natl Acad Sci U S A. 2001 Aug 14;98(17):9808-13
references.pmid: 11504946
references.citation: Nat Genet. 1997 Jul;16(3):265-9
references.pmid: 9207792
references.citation: Hum Mol Genet. 2008 Jan 1;17(1):52-70
references.pmid: 17913700
references.citation: J Neurochem. 2006 Jul;98(1):193-202
references.pmid: 16805808
references.citation: Hum Mol Genet. 2000 Oct;9(16):2451-7
references.pmid: 11005801
references.citation: Proc Natl Acad Sci U S A. 1997 Sep 2;94(18):9920-5
references.pmid: 9275227
references.citation: Neuromuscul Disord. 1992;2(5-6):423-8
references.pmid: 1300191
references.citation: Hum Genet. 2008 Jan;122(6):635-43
references.pmid: 17962980
references.citation: Cell. 1995 Jan 13;80(1):155-65
references.pmid: 7813012
references.citation: Neurobiol Dis. 2007 Aug;27(2):207-19
references.pmid: 17561409
references.citation: Hum Mol Genet. 2000 Feb 12;9(3):333-9
references.pmid: 10655541
references.citation: Ann Neurol. 2005 Aug;58(2):194-202
references.pmid: 16049920
references.citation: J Biol Chem. 2004 Apr 9;279(15):14803-11
references.pmid: 14742439
references.citation: Neurobiol Dis. 1996 Apr;3(2):97-110
references.pmid: 9173917
references.citation: Mol Med. 2000 Nov;6(11):957-68
references.pmid: 11147573
references.citation: Gene Ther. 2001 Oct;8(20):1532-8
references.pmid: 11704813
references.citation: J Child Neurol. 2007 Aug;22(8):1004-12
references.pmid: 17761656
references.citation: Hum Mol Genet. 2005 Jul 15;14(14):2003-18
references.pmid: 15944201
references.citation: Hum Mol Genet. 2005 Mar 15;14(6):845-57
references.pmid: 15703193
references.citation: Hum Mol Genet. 1997 Aug;6(8):1205-14
references.pmid: 9259265
references.citation: J Neurosci Methods. 2007 Apr 15;161(2):285-90
references.pmid: 17161463
references.citation: Hum Genet. 2006 Nov;120(4):589-601
references.pmid: 16951947