Somatic mutations of theERBB4kinase domain in human cancers

Abstract

The EGFR family consists of 4 receptor tyrosine kinases, EGFR (ERBB1), ERBB2 (HER2), ERBB3 (HER3) and ERBB4 (HER4). Recent reports revealed that the kinase domains of both EGFR (ERBB1) and ERBB2 gene were somatically mutated in human cancers, raising the possibility that the other ERBB members possess somatic mutations in human cancers. Here, we performed mutational analysis of the ERBB4 kinase domain by polymerase chain reaction-single-strand conformation polymorphism assay in 595 cancer tissues from stomach, lung, colon and breast. We detected the ERBB4 somatic mutations in 3 of 180 gastric carcinomas (1.7%), 3 of 104 colorectal carcinomas (2.9%), 5 of 217 nonsmall cell lung cancers (2.3%) and 1 of 94 breast carcinomas (1.1%). The 12 ERBB4 mutations consisted of 1 in-frame duplication mutation and 8 missense mutations in the exons, and 3 mutations in the introns. We simultaneously analyzed the somatic mutations of EGFR, ERBB2, K-RAS, PIK3CA and BRAF genes in the 12 samples with the ERBB4 mutations and found that 1 gastric carcinoma with ERBB4 mutation also harbored K-RAS gene mutation. Our study demonstrated that in addition to EGFR and ERBB2, somatic mutation of the kinase domain of ERBB4 occurs in the common human cancers, and suggested that alterations of ERBB4-mediated signaling pathway by ERBB4 mutations may contribute to the development of human cancers. ' 2005 Wiley-Liss, Inc.

Somatic Mutations of ErbB4

“…MDA-MB-468 cells were maintained in Dulbecco's modified Eagle's medium (Invitrogen) supplemented with 10% fetal calf serum (Promocell), 50 g/ml streptomycin (Sigma), and 100 IU/ml penicillin (Sigma). Mutations targeting the tyrosine kinase or cytoplasmic domains of ErbB4 reported by Soung et al (8) and Parsons et al (11) were introduced into pcDNA3.1ErbB4JM-aCYT-1, pcDNA3.1ErbB4JM-aCYT-2, or pBABE-puroErbB4JM-aCYT-2 using the QuikChange site-directed mutagenesis kit (Stratagene). Similarly, a conserved lysine residue (Lys-751) within the ATP-binding site of ErbB4 was mutated to arginine in pcDNA3.1ErbB4JM-aCYT-2-HA to generate kinase-dead pcDNA3.1ErbB4JM-aCYT-2K751R-HA.…”

“…Lack of knowledge of tumor-associated genetic changes in ErbB4 has precluded addressing their potential lossof-function or gain-of-function phenotypes. Recently, nine different somatic mutations targeting the ErbB4 kinase domain were reported in patients with either breast, gastric, colorectal, or non-small cell lung cancer (8). In EGFR, similar mutations targeting the kinase domain sensitize patients to treatment with tyrosine kinase inhibitors (9,10).…”

“…Here, we exploited the recently reported crystal structure of ErbB4 kinase domain (4) to predict the functionality of 9 somatic ErbB4 kinase domain mutants (8). In addition, we analyzed the basal and ligand-induced phosphotyrosine content of the 9-kinase domain and 1-cytoplasmic domain mutations (8,11) in different cell backgrounds.…”

“…In addition, we analyzed the basal and ligand-induced phosphotyrosine content of the 9-kinase domain and 1-cytoplasmic domain mutations (8,11) in different cell backgrounds. Our data demonstrate that 2 of the 10 mutations disrupt the catalytic activity of the ErbB4 tyrosine kinase, and there are clear structural reasons for the observed effects.…”

Formation of Neu/ErbB2-induced mammary tumors is unaffected by loss of ErbB4

“…ErbB4 overexpression is associated with poor prognosis for thyroid cancer and childhood medulloblastoma (Haugen et al, 1996;Gilbertson et al, 1997). Somatic mutations in the ErbB4 kinase domain were detected at a low frequency in gastric carcinomas, colorectal carcinomas, non-small lung cancers and breast carcinomas (Soung et al, 2006). One early survey revealed that ErbB4 expression is lower in breast and prostate cancer specimens than in normal tissue, and, in contrast to ErbB2, tended to be associated with differentiating tissue, rather than proliferating tissue (Srinivasan et al, 1998).…”