Neuroblastoma in advanced stages is one of the most intractable paediatric
cancers, even with recent therapeutic advances.
Neuroblastoma harbours a variety of genetic changes, including a high frequency of MYCN amplification, loss of heterozygosity at 1p36 and 11q, and gain of genetic material from 17q, all of which have been implicated in the pathogenesis of
neuroblastoma. However, the scarcity of reliable molecular targets has hampered the development of effective therapeutic agents targeting
neuroblastoma. Here we show that the
anaplastic lymphoma kinase (ALK), originally identified as a fusion
kinase in a subtype of
non-Hodgkin's lymphoma (
NPM-ALK) and more recently in
adenocarcinoma of lung (EML4-ALK), is also a frequent target of genetic alteration in advanced
neuroblastoma. According to our genome-wide scans of genetic lesions in 215 primary
neuroblastoma samples using high-density single-nucleotide polymorphism genotyping microarrays, the ALK locus, centromeric to the MYCN locus, was identified as a recurrent target of copy number gain and gene amplification. Furthermore,
DNA sequencing of ALK revealed eight novel missense mutations in 13 out of 215 (6.1%) fresh tumours and 8 out of 24 (33%)
neuroblastoma-derived cell lines. All but one mutation in the primary samples (12 out of 13) were found in stages 3-4 of the disease and were harboured in the
kinase domain. The mutated
kinases were autophosphorylated and displayed increased
kinase activity compared with the wild-type
kinase. They were able to transform NIH3T3 fibroblasts as shown by their colony formation ability in soft
agar and their capacity to form tumours in nude mice. Furthermore, we demonstrate that downregulation of ALK through RNA interference suppresses proliferation of
neuroblastoma cells harbouring mutated ALK. We anticipate that our findings will provide new insights into the pathogenesis of advanced
neuroblastoma and that ALK-specific
kinase inhibitors might improve its clinical outcome.