High-throughput sequence analysis of the tyrosine kinome in acute myeloid leukemia.
To determine whether aberrantly activated tyrosine kinases other than FLT3 and c-KIT contribute to acute myeloid leukemia (AML) pathogenesis, we used high-throughput (HT) DNA sequence ana-lysis to screen exons encoding the activation loop and juxtamembrane domains of 85 tyrosine kinase genes in 188 AML patients without FLT3 or c-KIT mutations. The screen identified 30 nonsynonymous sequence variations in 22 different kinases not previously reported in single-nucleotide polymorphism (SNP) databases. These included a novel FLT3 activating allele and a previously described activating mutation in MET (METT1010I). The majority of novel sequence variants were stably expressed in factor-dependent Ba/F3 cells. Apart from one FLT3 allele, none of the novel variants showed constitutive phosphorylation by immunoblot analysis and none transformed Ba/F3 cells to factor-independent growth. These findings indicate the majority of these alleles are not potent tyrosine kinase activators in this cellular context and that a significant proportion of nonsynonymous sequence variants identified in HT DNA sequencing screens may not have functional significance. Although some sequence variants may represent SNPs, these data are consistent with recent reports that a significant fraction of such sequence variants are "passenger" rather than "driver" alleles and underscore the importance of functional assessment of candidate disease alleles.
Loriaux
Marc M
MM
Division of Hematology and Medical Oncology, Oregon Health & Science University Cancer Institute, Portland, OR 97239, USA.
Levine
Ross L
RL
Tyner
Jeffrey W
JW
Fröhling
Stefan
S
Scholl
Claudia
C
Stoffregen
Eric P
EP
Wernig
Gerlinde
G
Erickson
Heidi
H
Eide
Christopher A
CA
Berger
Roland
R
Bernard
Olivier A
OA
Griffin
James D
JD
Stone
Richard M
RM
Lee
Benjamin
B
Meyerson
Matthew
M
Heinrich
Michael C
MC
Deininger
Michael W
MW
Gilliland
D Gary
DG
Druker
Brian J
BJ
D001483:Base Sequence; D004252:DNA Mutational Analysis; D006801:Humans; D015470:Leukemia, Myeloid, Acute / Q000201:enzymology* / Q000209:etiology; D020641:Polymorphism, Single Nucleotide; D011505:Protein-Tyrosine Kinases / Q000235:genetics*; D051941:fms-Like Tyrosine Kinase 3
D011505:Protein-Tyrosine Kinases; D051941:fms-Like Tyrosine Kinase 3