Study Links Motor Neuron Disease to New Receptor Tyrosine Kinase

Known genetic variants explain only 75 percent of familial amyotrophic lateral sclerosis cases, leaving more ALS genes to find. However, since the largest ALS families already have been scoured by gene-hunters, researchers now resort to studying smaller kindreds. In the October 10 American Journal of Human Genetics online, researchers from Japan reported that one such family led them to a new ALS gene candidate. Members of the family harbor mutations in a gene that goes by the mouthful v-erb-a erythroblastic leukemia viral oncogene homolog 4 (ErbB4). The small size of the family means the genetic statistics are less than a slam-dunk, but finding two unrelated cases of ALS with mutations in the same gene strengthens the connection, said senior author Shoji Tsuji of the University of Tokyo.

First author Yuji Takahashi, who has since moved to the National Center of Neurology and Psychiatry in Tokyo, and colleagues first heard about the kindred from the family's primary-care physicians. Three of the eight children in the family had ALS, and the father likely had it, too, although he was never diagnosed before he died at the age of 74. Tsuji and colleagues checked one son with ALS for known genetic causes, but finding none, they decided to look for novel mutations.

First, Tsuji and colleagues genotyped single-nucleotide polymorphisms in the mother, whom they deemed free of the disease gene, and all but one of the siblings (the one they did not test had never been diagnosed with ALS, but he had died so DNA was unavailable). Linkage analysis was difficult because the group was so small, and because any of the young, healthy siblings might be a carrier yet to develop symptoms. To determine linkage, the researchers had to assume 80 percent penetrance for any novel genetic variant—that is, they assumed at least one of the healthy siblings was actually a carrier. This analysis led them to seven genetic regions that associated with the disease with a logarithm of odds (LOD) score of 0.7 or higher. This is not a very high LOD score, noted John Landers of the University of Massachusetts Medical School in Worcester, who was not part of the study group. The National Human Genome Research Institute says a LOD score of three or higher typically indicates linkage. Tsuji conceded the statistics were weak, but said it was the best the researchers could expect when working with a small family.

Next, the researchers used whole-genome sequencing to look for novel variants in seven regions. This led them to three candidates that were predicted to change amino acid sequences. Of the three, only the ErbB4 variant, an arginine-for-glutamine swap at amino acid 927 (R927Q), was absent in 477 control samples collected in Japan, North America, and Australia.

While it is certainly plausible that the ErbB4 variant causesALS in this family, it remains plausible that something else is to blame, Landers said. He noted that the National Heart, Lung, and Blood Institute’s Exome Variant Server lists dozens of variants in ErbB4, and it is possible Tsuji and colleagues had discovered one that segregated with disease by chance, while missing out on the true ALS mutation.

Landers did agree that subsequent analyses boost the chance that ErbB4 is an ALS gene. Tsuji and colleagues checked ErbB4 in 364 people with familial ALS, and 818 with sporadic ALS, from the same geographic locales as the controls. The same R927Q variant appeared in a Canadian familial ALS case, but no DNA from that person’s family was available to check if the variant segregated with disease. In addition, a Japanese person with sporadic ALS carried an arginine-1275-tryptophan variant; this must have been a new mutation because it did not turn up in either parent. Though Tsuji conceded the linkage analysis could be stronger, he felt these additional cases confirmed the association between ErbB4 and ALS. The affected amino acids are conserved across species, indicating their importance for ErbB4 function.

Philip Wong of Johns Hopkins University in Baltimore, Maryland, who was not involved in the study, agreed that the small size of the family weakened the human genetics. What would really clinch the ALS connection, he suggested, would be a mouse model in which the ErbB4 mutations led to motor neuron disease. No such model exists, though a complete knockout of ErbB4 kills mouse embryos before birth, causing both cardiac and neural defects (Gassmann et al., 1995). Researchers also have made a conditional knockout mouse for ErbB4 in the nervous system, which exhibited weak motor defects (Golub et al., 2004 ).

The receptor is expressed in nervous system gray matter and spinal motor neurons. Neuregulins bind ErbB4, initiating autophosphorylation downstream signaling. By adding neuregulin to cultured monkey kidney cells, Tsuji and colleagues observed that the ALS-linked ErbB4 mutants exhibited less autophosphorylation than the wild-type receptor on a Western blot. However, this single experiment alone offers fairly minimal data, and the researchers did not quantitate the amount of phosphorylation, Landers noted. Neuregulin-ErbB4 signaling promotes myelination, which Wong speculated might explain the link between the receptor and ALS (see Alzforum News story on Roy et al., 2007 and Alzforum News story on Michailov et al., 2004).

Neuregulins are a substrate for BACE1, which cleaves the protein to release the functional signaling fragment. Might BACE inhibitors compromise motor neurons? Probably not, Wong said. For one, only neuregulin 1 type III is a BACE1 substrate, and other neuregulins might still activate ErbB4. In addition, BACE1 knockout animals do not exhibit motor neuron disease, he noted. “I personally do not think that BACE1 inhibitor will cause ALS-like effect,” Riqiang Yan of the Cleveland Clinic in Ohio wrote in an email to Alzforum.

ErbB4 variants also have been linked to schizophrenia ( Agim et al., 2013, Stefansson et al., 2002), and the receptor kinase is expressed at high levels in apoptotic neurons of people with Alzheimer’s (Woo et al., 2010). The cases in this study did not have schizophrenia or Alzheimer’s, displaying fairly typical ALS, Tsuji said.

Tsuji suggested that upregulating the neuregulin-ErbB4 pathway could be a therapeutic approach for ALS. Such a treatment might be applied broadly, even for people without ErbB4 mutations, he suggested, because people with sporadic ALS exhibit unusually low neuregulin 1 type III levels in their spinal motor neurons (Song et al., 2012).—Amber Dance

Comments

  1. This new discovery is exciting. In addition to the welcome identification of another critical player in the pathogenesis of ALS, ErbB receptor tyrosine kinases and neuregulins are much-studied groups of molecules, so there should be a lot of resources (mice, cell lines, receptor agonists, and antagonists) already available to launch the basic and translational studies needed to follow up the gene identification. Further, the apparent lack of dementia is important considering the current focus on the emergence of FTD in a large segment of both familial and sporadic ALS. Is the loss of non-motor frontal neurons just very subtle in these patients, or in fact are there two quite distinct pathways for ALS—one involving additional prefrontal loss, and one not?

    View all comments by Denise A. Figlewicz

  2. A new study by Takahashi et al. in the American Journal of Human Genetics identifies a role for loss-of-function mutations in the gene ERBB4 in familial amyotrophic lateral sclerosis (FALS). The mutation was discovered by whole-genome sequencing (WGS) in a Japanese family and confirmed in a Canadian patient. ErbB4 is a receptor tyrosine kinase that is activated by the neuregulin family of signaling factors, notably neuregulin-1 (NRG1) (Mei and Xiong, 2008). In the brain, NRG1- ErbB4 signaling is crucial to the development and maintenance of inhibitory circuitry, particularly by regulating the migration and synaptic plasticity of fast-spiking interneurons, in which ErbB4 is specifically expressed (Fazzari et al., 2010). Mutations in ERBB4, including SNPs (Hatzimanolis et al., 2013) and structural variants (Walsh et al., 2008), have been linked to schizophrenia (SZ) in multiple populations. Recent studies in mouse models show that cell type-specific ablation of ErbB4 in cortical interneurons results in SZ-like behavioral phenotypes (Del Pino et al., 2013). It is therefore intriguing that disrupted NRG1-ErbB4 signaling in a different location and cellular population (i.e., spinal motor neurons) underlies FALS—could this finding indicate that the cellular pathophysiologies of SZ and FALS are more closely related than previously thought?

    Frontotemporal dementia (FTD) may represent an important domain that could possibly link SZ to FALS. In recent years, ALS and FTD have been associated with mutations in several genes, such as C90RF72 (Dejesus-Hernandez et al., 2011). SZ and FTD exhibit some degree of phenotypic similarity, which can cause diagnostic confusion, particularly in early stages of FTD (Cooper and Ovsiew, 2013). More than one report has indicated that family members of patients with SZ or schizoaffective disorder are more likely to suffer from FTD, indicating that susceptibility for both diseases may be heritable (Chow et al., 1999). These observations have led investigators to suspect a common genetic basis for SZ and FALS; however, a recent study did not find C9ORF72 mutations in an SZ population (Huey et al., 2013). The results reported by Takahashi et al. therefore might represent an interesting first step toward identifying common genetic factors for these devastating disorders.

    References:

    Mei L, Xiong WC. Neuregulin 1 in neural development, synaptic plasticity and schizophrenia. Nat Rev Neurosci. 2008 Jun;9(6):437-52. PubMed.

    Fazzari P, Paternain AV, Valiente M, Pla R, Luján R, Lloyd K, Lerma J, Marín O, Rico B. Control of cortical GABA circuitry development by Nrg1 and ErbB4 signalling. Nature. 2010 Apr 29;464(7293):1376-80. Epub 2010 Apr 14 PubMed.

    Hatzimanolis A, McGrath JA, Wang R, Li T, Wong PC, Nestadt G, Wolyniec PS, Valle D, Pulver AE, Avramopoulos D. Multiple variants aggregate in the neuregulin signaling pathway in a subset of schizophrenia patients. Transl Psychiatry. 2013;3:e264. PubMed.

    Walsh T, McClellan JM, McCarthy SE, Addington AM, Pierce SB, Cooper GM, Nord AS, Kusenda M, Malhotra D, Bhandari A, Stray SM, Rippey CF, Roccanova P, Makarov V, Lakshmi B, Findling RL, Sikich L, Stromberg T, Merriman B, Gogtay N, Butler P, Eckstrand K, Noory L, Gochman P, Long R, Chen Z, Davis S, Baker C, Eichler EE, Meltzer PS, Nelson SF, Singleton AB, Lee MK, Rapoport JL, King MC, Sebat J. Rare structural variants disrupt multiple genes in neurodevelopmental pathways in schizophrenia. Science. 2008 Apr 25;320(5875):539-43. PubMed.

    Del Pino I, García-Frigola C, Dehorter N, Brotons-Mas JR, Alvarez-Salvado E, Martínez de Lagrán M, Ciceri G, Gabaldón MV, Moratal D, Dierssen M, Canals S, Marín O, Rico B. Erbb4 deletion from fast-spiking interneurons causes schizophrenia-like phenotypes. Neuron. 2013 Sep 18;79(6):1152-68. PubMed.

    DeJesus-Hernandez M, Mackenzie IR, Boeve BF, Boxer AL, Baker M, Rutherford NJ, Nicholson AM, Finch NA, Flynn H, Adamson J, Kouri N, Wojtas A, Sengdy P, Hsiung GY, Karydas A, Seeley WW, Josephs KA, Coppola G, Geschwind DH, Wszolek ZK, Feldman H, Knopman DS, Petersen RC, Miller BL, Dickson DW, Boylan KB, Graff-Radford NR, Rademakers R. Expanded GGGGCC hexanucleotide repeat in noncoding region of C9ORF72 causes chromosome 9p-linked FTD and ALS. Neuron. 2011 Oct 20;72(2):245-56. Epub 2011 Sep 21 PubMed.

    Cooper JJ, Ovsiew F. The relationship between schizophrenia and frontotemporal dementia. J Geriatr Psychiatry Neurol. 2013 Sep;26(3):131-7. PubMed.

    Chow TW, Miller BL, Hayashi VN, Geschwind DH. Inheritance of frontotemporal dementia. Arch Neurol. 1999 Jul;56(7):817-22. PubMed.

    Huey ED, Nagy PL, Rodriguez-Murillo L, Manoochehri M, Goldman J, Lieberman J, Karayiorgou M, Mayeux R. C9ORF72 repeat expansions not detected in a group of patients with schizophrenia. Neurobiol Aging. 2013 Apr;34(4):1309.e9-10. PubMed.

    View all comments by Saurav Seshadri

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References

News Citations

  1. Neuregulin/ErbB4 Mutant Mice Have Myelin, Synaptic Deficits
  2. Wrap it Up! Neuregulin Directs Axonal Myelination

Paper Citations

  1. Gassmann M, Casagranda F, Orioli D, Simon H, Lai C, Klein R, Lemke G. Aberrant neural and cardiac development in mice lacking the ErbB4 neuregulin receptor. Nature. 1995 Nov 23;378(6555):390-4. PubMed.
  2. Golub MS, Germann SL, Lloyd KC. Behavioral characteristics of a nervous system-specific erbB4 knock-out mouse. Behav Brain Res. 2004 Aug 12;153(1):159-70. PubMed.
  3. Roy K, Murtie JC, El-Khodor BF, Edgar N, Sardi SP, Hooks BM, Benoit-Marand M, Chen C, Moore H, O'Donnell P, Brunner D, Corfas G. Loss of erbB signaling in oligodendrocytes alters myelin and dopaminergic function, a potential mechanism for neuropsychiatric disorders. Proc Natl Acad Sci U S A. 2007 May 8;104(19):8131-6. PubMed.
  4. Michailov GV, Sereda MW, Brinkmann BG, Fischer TM, Haug B, Birchmeier C, Role L, Lai C, Schwab MH, Nave KA. Axonal neuregulin-1 regulates myelin sheath thickness. Science. 2004 Apr 30;304(5671):700-3. PubMed.
  6. Stefansson H, Sigurdsson E, Steinthorsdottir V, Bjornsdottir S, Sigmundsson T, Ghosh S, Brynjolfsson J, Gunnarsdottir S, Ivarsson O, Chou TT, Hjaltason O, Birgisdottir B, Jonsson H, Gudnadottir VG, Gudmundsdottir E, Bjornsson A, Ingvarsson B, Ingason A, Sigfusson S, Hardardottir H, Harvey RP, Lai D, Zhou M, Brunner D, Mutel V, Gonzalo A, Lemke G, Sainz J, Johannesson G, Andresson T, Gudbjartsson D, Manolescu A, Frigge ML, Gurney ME, Kong A, Gulcher JR, Petursson H, Stefansson K. Neuregulin 1 and susceptibility to schizophrenia. Am J Hum Genet. 2002 Oct;71(4):877-92. PubMed.
  7. Woo RS, Lee JH, Yu HN, Song DY, Baik TK. Expression of ErbB4 in the apoptotic neurons of Alzheimer's disease brain. Anat Cell Biol. 2010 Dec;43(4):332-9. PubMed.
  8. Song F, Chiang P, Wang J, Ravits J, Loeb JA. Aberrant neuregulin 1 signaling in amyotrophic lateral sclerosis. J Neuropathol Exp Neurol. 2012 Feb;71(2):104-15. PubMed.

External Citations

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Further Reading

Papers

  1. Ryu J, Yu HN, Cho H, Kim HS, Baik TK, Lee SJ, Woo RS. Neuregulin-1 exerts protective effects against neurotoxicities induced by C-terminal fragments of APP via ErbB4 receptor. J Pharmacol Sci. 2012;119(1):73-81. PubMed.
  2. Fleck D, Garratt AN, Haass C, Willem M. BACE1 dependent neuregulin processing: review. Curr Alzheimer Res. 2012 Feb;9(2):178-83. PubMed.
  3. Woo RS, Lee JH, Kim HS, Baek CH, Song DY, Suh YH, Baik TK. Neuregulin-1 protects against neurotoxicities induced by Swedish amyloid precursor protein via the ErbB4 receptor. Neuroscience. 2012 Jan 27;202:413-23. PubMed.
  4. Woo RS, Lee JH, Yu HN, Song DY, Baik TK. Expression of ErbB4 in the neurons of Alzheimer's disease brain and APP/PS1 mice, a model of Alzheimer's disease. Anat Cell Biol. 2011 Jun;44(2):116-27. PubMed.
  5. Min SS, An J, Lee JH, Seol GH, Im JH, Kim HS, Baik TK, Woo RS. Neuregulin-1 prevents amyloid β-induced impairment of long-term potentiation in hippocampal slices via ErbB4. Neurosci Lett. 2011 Nov 7;505(1):6-9. PubMed.

Primary Papers

  1. Takahashi Y, Fukuda Y, Yoshimura J, Toyoda A, Kurppa K, Moritoyo H, Belzil VV, Dion PA, Higasa K, Doi K, Ishiura H, Mitsui J, Date H, Ahsan B, Matsukawa T, Ichikawa Y, Moritoyo T, Ikoma M, Hashimoto T, Kimura F, Murayama S, Onodera O, Nishizawa M, Yoshida M, Atsuta N, Sobue G, , Fifita JA, Williams KL, Blair IP, Nicholson GA, Gonzalez-Perez P, Brown RH, Nomoto M, Elenius K, Rouleau GA, Fujiyama A, Morishita S, Goto J, Tsuji S. ERBB4 Mutations that Disrupt the Neuregulin-ErbB4 Pathway Cause Amyotrophic Lateral Sclerosis Type 19. Am J Hum Genet. 2013 Oct 8; PubMed.

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