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Identification of ALK as a major familial neuroblastoma predisposition gene

Nature volume 455, pages 930–935 (2008)Cite this article

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Abstract

Neuroblastoma is a childhood cancer that can be inherited, but the genetic aetiology is largely unknown. Here we show that germline mutations in the anaplastic lymphoma kinase (ALK) gene explain most hereditary neuroblastomas, and that activating mutations can also be somatically acquired. We first identified a significant linkage signal at chromosome bands 2p23–24 using a whole-genome scan in neuroblastoma pedigrees. Resequencing of regional candidate genes identified three separate germline missense mutations in the tyrosine kinase domain of ALK that segregated with the disease in eight separate families. Resequencing in 194 high-risk neuroblastoma samples showed somatically acquired mutations in the tyrosine kinase domain in 12.4% of samples. Nine of the ten mutations map to critical regions of the kinase domain and were predicted, with high probability, to be oncogenic drivers. Mutations resulted in constitutive phosphorylation, and targeted knockdown of ALK messenger RNA resulted in profound inhibition of growth in all cell lines harbouring mutant or amplified ALK, as well as in two out of six wild-type cell lines for ALK. Our results demonstrate that heritable mutations of ALK are the main cause of familial neuroblastoma, and that germline or acquired activation of this cell-surface kinase is a tractable therapeutic target for this lethal paediatric malignancy.

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Figure 1: Eight neuroblastoma pedigrees with ALK mutations.
Figure 2: Germline and somatic ALK mutations.
Figure 3: Representative ALK copy number alterations in five neuroblastoma primary tumours.
Figure 4: ALK is highly expressed and the kinase is phosphorylated in neuroblastoma cell lines harbouring activating mutations.
Figure 5: ALK knockdown results in growth inhibition of ALK mutated or amplified neuroblastoma cell lines.

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Primary accessions

GenBank/EMBL/DDBJ

Data deposits

All sequence variations have been deposited to GenBank under accession numbers EU660517, EU660518, EU660519, EU660520, EU660521, EU660522, EU660523, EU660524, EU660525, EU660526 and EU660527.

Change history

  • 16 October 2008

    The AOP version of this paper contained an affiliation error and an erroneous sentence in the Discussion. These were corrected for print on 16 October 2008.

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Acknowledgements

We acknowledge the families and children that participated in this research study, and the Children’s Oncology Group for providing specimens. We thank W. London for providing statistical analyses related to the Children’s Oncology Group tumour set, H. Rydbeck for his assistance with the linkage analysis, M. Wasik for technical assistance, and J. Felgenhauer, N. Van Roy and C. McConville for providing neuroblastoma pedigrees. This work was supported in part by National Institutes of Health grants K08-111733 (Y.P.M.), R01-CA78454 (J.M.M.), R01-CA87847 (J.M.M.), an American Society of Clinical Oncology Career Development Award (Y.P.M.), the Foerderer-Murray Fund (Y.P.M.), the Carly Hillman Fund (Y.P.M.), the Alex’s Lemonade Stand Foundation (J.M.M.), the Andrew’s Army Foundation (J.M.M.), the Giulio D’Angio Endowed Chair (J.M.M.), the Italian Neuroblastoma Foundation (L.L.), the Center for Applied Genomics at the Joseph Stokes Research Institute (H.H.), Scripps Genomic Medicine (A.T., N.J.S.), the Scripps Dickinson Scholarship (A.T.), and the Abramson Family Cancer Research Institute (J.M.M.).

Author Contributions Y.P.M. and J.M.M. designed the experiments and wrote the manuscript. Y.P.M., M.L., J.M.M., G.L., F.S., P.P. and G.P.T. collected the families for the linkage analysis. Y.P.M., M.L., L.L., C.K., C.H., E.R., H.H. and M.D. performed the genome-wide genotyping and linkage analysis. M.L. performed the DNA sequencing and analyses. K.A.C., A.W. and M.J.L. performed the siRNA experiments. E.F.A., H.H. and Y.P.M. performed the tumour SNP genotyping/copy number analyses. J.E.L., K.A.C. and A.W. performed the expression analyses. K.A.C., R.S. and M.L. performed the protein work. G.M.B. initiated the collection of neuroblastoma pedigrees. A.T. and N.J.S. performed the structural analysis of ALK coding mutations.

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Authors and Affiliations

  1. Division of Oncology and Center for Childhood Cancer Research, Department of Pediatrics, Children’s Hospital of Philadelphia, University of Pennsylvania School of Medicine, Philadelphia, Pennsylvania 19104, USA,

    Yaël P. Mossé, Marci Laudenslager, Kristina A. Cole, Andrew Wood, Edward F. Attiyeh, Michael J. Laquaglia, Rachel Sennett, Jill E. Lynch, Cuiping Hou, Garrett M. Brodeur, Eric Rappaport & John M. Maris

  2. Translational Pediatric Oncology, National Institute for Cancer Research and Italian Neuroblastoma Foundation, National Institute for Cancer Research, Genoa 16132, Italy

    Luca Longo, Patrizia Perri & Gian P. Tonini

  3. Advanced Biotechnology Center, Genoa 16132, Italy

    Patrizia Perri

  4. Center for Medical Genetics, Ghent University Hospital, Gent B-9000, Belgium

    Geneviève Laureys & Frank Speleman

  5. The Center for Applied Genomics, Children’s Hospital of Philadelphia, Philadelphia, Pennsylvania 19104, USA ,

    Cecilia Kim, Cuiping Hou & Hakon Hakonarson

  6. Scripps Genomic Medicine and the Scripps Research Institute, La Jolla, California 92037, USA ,

    Ali Torkamani & Nicholas J. Schork

  7. Division of Genetics, Department of Pediatrics and CCEB, Department of Biostatistics and Epidemiology, The Children’s Hospital of Philadelphia, University of Pennsylvania School of Medicine, Philadelphia, Pennsylvania 19104, USA,

    Hakon Hakonarson & Marcella Devoto

  8. Department of Experimental Medicine, University of Rome “La Sapienza”, Rome 00161, Italy

    Marcella Devoto

  9. Abramson Family Cancer Research Institute, University of Pennsylvania School of Medicine, Philadelphia, Pennsylvania 19104, USA ,

    John M. Maris

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  1. Yaël P. Mossé
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Correspondence to John M. Maris.

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Mossé, Y., Laudenslager, M., Longo, L. et al. Identification of ALK as a major familial neuroblastoma predisposition gene. Nature 455, 930–935 (2008). https://doi.org/10.1038/nature07261

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