Amplification and overexpression of CACNA1E correlates with relapse in favorable histology Wilms' tumors.
OBJECTIVE
The most well established molecular markers of poor outcome in Wilms' tumor are loss of heterozygosity at chromosomes 1p and/or 16q, although to date no specific genes at these loci have been identified. We have previously shown a link between genomic gain of chromosome 1q and tumor relapse and sought to further elucidate the role of genes on 1q in treatment failure.


METHODS
Microarray-based comparative genomic hybridization identified a microamplification harboring a single gene (CACNA1E) at 1q25.3 in 6 of 76 (7.9%) Wilms' tumors, correlating with a shorter relapse-free survival (P = 0.0044, log-rank test). Further characterization of this gene was carried out by measuring mRNA and protein expression as well as stable transfection of HEK293 cells.


RESULTS
Overexpression of the CACNA1E transcript was associated with DNA copy number (P = 0.0204, ANOVA) and tumor relapse (P = 0.0851, log-rank test). Immunohistochemistry against the protein product Ca(V)2.3 revealed expression localized to the apical membrane in the distal tubules of normal kidney but not to the metanephric blastemal cells of fetal kidney from which Wilms' tumors arise. Nuclear localization in 99 of 160 (61.9%) Wilms' tumor cases correlated with a reduced relapse-free survival, particularly in cases treated with preoperative chemotherapy (P = 0.009, log-rank test). Expression profiling of stably transfected HEK293 cells revealed specific up-regulation of the immediate early response genes EGR1/EGR2/EGR3 and FOS/FOSB, mediated by activation of the MEK/ERK5/Nur77 pathway.


CONCLUSIONS
These data identify a unique genetic aberration with direct clinical relevance in Wilms' tumor relapse and provide evidence for a potential novel mechanism of treatment resistance in these tumors.
Natrajan
Rachael
R
Paediatric Oncology, Institute of Cancer Research/Royal Marsden NHS Trust, Sutton, Surrey, United Kingdom.
Little
Suzanne E
SE
Reis-Filho
Jorge S
JS
Hing
Lara
L
Messahel
Boo
B
Grundy
Paul E
PE
Dome
Jeffrey S
JS
Schneider
Toni
T
Vujanic
Gordan M
GM
Pritchard-Jones
Kathy
K
Jones
Chris
C
D020908:Calcium Channels, R-Type / Q000235:genetics* / Q000378:metabolism*; D027682:Cation Transport Proteins / Q000235:genetics* / Q000378:metabolism*; D002478:Cells, Cultured; D002878:Chromosomes, Human, Pair 1; D018572:Disease-Free Survival; D051766:Early Growth Response Protein 1 / Q000378:metabolism; D051767:Early Growth Response Protein 2 / Q000378:metabolism; D051776:Early Growth Response Protein 3 / Q000378:metabolism; D005784:Gene Amplification; D015972:Gene Expression Regulation, Neoplastic; D016162:Genes, Wilms Tumor; D006801:Humans; D007668:Kidney / Q000166:cytology / Q000196:embryology / Q000378:metabolism; D007680:Kidney Neoplasms / Q000235:genetics / Q000378:metabolism* / Q000473:pathology* / Q000628:therapy; D048671:MAP Kinase Kinase 5 / Q000378:metabolism; D009367:Neoplasm Staging; D020411:Oligonucleotide Array Sequence Analysis; D016760:Proto-Oncogene Proteins c-fos / Q000378:metabolism; D012008:Recurrence; D009396:Wilms Tumor / Q000235:genetics / Q000378:metabolism* / Q000473:pathology* / Q000628:therapy
C423009:CACNA1E protein, human; D020908:Calcium Channels, R-Type; D027682:Cation Transport Proteins; C497325:EGR1 protein, human; C497326:EGR2 protein, human; C079945:EGR3 protein, human; D051766:Early Growth Response Protein 1; D051767:Early Growth Response Protein 2; D016760:Proto-Oncogene Proteins c-fos; D051776:Early Growth Response Protein 3; D048671:MAP Kinase Kinase 5; C482563:MAP2K5 protein, human