Somatic mutation as a mechanism of Wnt/β-catenin pathway activation in CLL.
One major goal of cancer genome sequencing is to identify key genes and pathways that drive tumor pathogenesis. Although many studies have identified candidate driver genes based on recurrence of mutations in individual genes, subsets of genes with nonrecurrent mutations may also be defined as putative drivers if they affect a single biological pathway. In this fashion, we previously identified Wnt signaling as significantly mutated through large-scale massively parallel DNA sequencing of chronic lymphocytic leukemia (CLL). Here, we use a novel method of biomolecule delivery, vertical silicon nanowires, to efficiently introduce small interfering RNAs into CLL cells, and interrogate the effects of 8 of 15 mutated Wnt pathway members identified across 91 CLLs. In HEK293T cells, mutations in 2 genes did not generate functional changes, 3 led to dysregulated pathway activation, and 3 led to further activation or loss of repression of pathway activation. Silencing 4 of 8 mutated genes in CLL samples harboring the mutated alleles resulted in reduced viability compared with leukemia samples with wild-type alleles. We demonstrate that somatic mutations in CLL can generate dependence on this pathway for survival. These findings support the notion that nonrecurrent mutations at different nodes of the Wnt pathway can contribute to leukemogenesis.
Cancer Vaccine Center, and Department of Medical Oncology, Dana-Farber Cancer Institute, Boston, MA;
Lili
L
Wang
Department of Chemistry and Chemical Biology, Harvard University, Cambridge, MA;
Alex K
AK
Shalek
Broad Institute, Cambridge, MA;
Mike
M
Lawrence
Department of Chemistry and Chemical Biology, Harvard University, Cambridge, MA;
Ruihua
R
Ding
Department of Chemistry and Chemical Biology, Harvard University, Cambridge, MA;
Jellert T
JT
Gaublomme
Broad Institute, Cambridge, MA;
Nathalie
N
Pochet
Broad Institute, Cambridge, MA;
Petar
P
Stojanov
Broad Institute, Cambridge, MA;
Carrie
C
Sougnez
Cancer Vaccine Center, and Department of Medical Oncology, Dana-Farber Cancer Institute, Boston, MA;
Sachet A
SA
Shukla
Biostatistics and Computational Biology, Dana-Farber Cancer Institute, Boston, MA;
Kristen E
KE
Stevenson
Cancer Vaccine Center, and Department of Medical Oncology, Dana-Farber Cancer Institute, Boston, MA;
Wandi
W
Zhang
Cancer Vaccine Center, and Department of Medical Oncology, Dana-Farber Cancer Institute, Boston, MA;
Jessica
J
Wong
Cancer Vaccine Center, and Department of Medical Oncology, Dana-Farber Cancer Institute, Boston, MA;
Quinlan L
QL
Sievers
F. M. Kirby Neurobiology Center, Boston Children's Hospital, Boston, MA; Department of Neurology, Harvard Medical School, Boston, MA;
Bryan T
BT
MacDonald
Department of Medical Oncology, Dana-Farber Cancer Institute, Boston, MA;
Alexander R
AR
Vartanov
Department of Medical Oncology, Dana-Farber Cancer Institute, Boston, MA;
Natalie R
NR
Goldstein
Biostatistics and Computational Biology, Dana-Farber Cancer Institute, Boston, MA;
Donna
D
Neuberg
F. M. Kirby Neurobiology Center, Boston Children's Hospital, Boston, MA; Department of Neurology, Harvard Medical School, Boston, MA;
Xi
X
He
Broad Institute, Cambridge, MA;
Eric
E
Lander
Broad Institute, Cambridge, MA; Division of Allergy, Immunology and Rheumatology, Department of Medicine, Massachusetts General Hospital, Boston MA; and.
Nir
N
Hacohen
Broad Institute, Cambridge, MA;
Aviv
A
Regev
Broad Institute, Cambridge, MA;
Gad
G
Getz
Department of Medical Oncology, Dana-Farber Cancer Institute, Boston, MA; Department of Medicine, Brigham and Women's Hospital, Harvard Medical School, Boston, MA.
Jennifer R
JR
Brown
Department of Chemistry and Chemical Biology, Harvard University, Cambridge, MA;
Hongkun
H
Park
Cancer Vaccine Center, and Department of Medical Oncology, Dana-Farber Cancer Institute, Boston, MA; Department of Medicine, Brigham and Women's Hospital, Harvard Medical School, Boston, MA.
Catherine J
CJ
Wu
D000328:Adult; D045744:Cell Line, Tumor; D002470:Cell Survival / Q000235:genetics; D002478:Cells, Cultured; D020869:Gene Expression Profiling; D015973:Gene Expression Regulation, Leukemic; D057809:HEK293 Cells; D006801:Humans; D015451:Leukemia, Lymphocytic, Chronic, B-Cell / Q000235:genetics* / Q000378:metabolism / Q000473:pathology; D009154:Mutation; D020411:Oligonucleotide Array Sequence Analysis; D034622:RNA Interference; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015398:Signal Transduction / Q000235:genetics*; D060449:Wnt Signaling Pathway / Q000235:genetics*; D051176:beta Catenin / Q000378:metabolism*
D051176:beta Catenin