PMID- 21289283
OWN - NLM
STAT- MEDLINE
DCOM- 20110506
LR  - 20211020
IS  - 1091-6490 (Electronic)
IS  - 0027-8424 (Print)
IS  - 0027-8424 (Linking)
VI  - 108
IP  - 8
DP  - 2011 Feb 22
TI  - RNAi screen of the protein kinome identifies checkpoint kinase 1 (CHK1) as a 
      therapeutic target in neuroblastoma.
PG  - 3336-41
LID - 10.1073/pnas.1012351108 [doi]
AB  - Neuroblastoma is a childhood cancer that is often fatal despite intense 
      multimodality therapy. In an effort to identify therapeutic targets for this 
      disease, we performed a comprehensive loss-of-function screen of the protein 
      kinome. Thirty kinases showed significant cellular cytotoxicity when depleted, 
      with loss of the cell cycle checkpoint kinase 1 (CHK1/CHEK1) being the most 
      potent. CHK1 mRNA expression was higher in MYC-Neuroblastoma-related 
      (MYCN)-amplified (P < 0.0001) and high-risk (P = 0.03) tumors. Western blotting 
      revealed that CHK1 was constitutively phosphorylated at the ataxia telangiectasia 
      response kinase target site Ser345 and the autophosphorylation site Ser296 in 
      neuroblastoma cell lines. This pattern was also seen in six of eight high-risk 
      primary tumors but not in control nonneuroblastoma cell lines or in seven of 
      eight low-risk primary tumors. Neuroblastoma cells were sensitive to the two CHK1 
      inhibitors SB21807 and TCS2312, with median IC(50) values of 564 nM and 548 nM, 
      respectively. In contrast, the control lines had high micromolar IC(50) values, 
      indicating a strong correlation between CHK1 phosphorylation and CHK1 inhibitor 
      sensitivity (P = 0.0004). Furthermore, cell cycle analysis revealed that CHK1 
      inhibition in neuroblastoma cells caused apoptosis during S-phase, consistent 
      with its role in replication fork progression. CHK1 inhibitor sensitivity 
      correlated with total MYC(N) protein levels, and inducing MYCN in retinal 
      pigmented epithelial cells resulted in CHK1 phosphorylation, which caused growth 
      inhibition when inhibited. These data show the power of a functional RNAi screen 
      to identify tractable therapeutical targets in neuroblastoma and support CHK1 
      inhibition strategies in this disease.
FAU - Cole, Kristina A
AU  - Cole KA
AD  - Division of Oncology, Children's Hospital of Philadelphia, Philadelphia, PA 
      19104-4318, USA.
FAU - Huggins, Jonathan
AU  - Huggins J
FAU - Laquaglia, Michael
AU  - Laquaglia M
FAU - Hulderman, Chase E
AU  - Hulderman CE
FAU - Russell, Mike R
AU  - Russell MR
FAU - Bosse, Kristopher
AU  - Bosse K
FAU - Diskin, Sharon J
AU  - Diskin SJ
FAU - Attiyeh, Edward F
AU  - Attiyeh EF
FAU - Sennett, Rachel
AU  - Sennett R
FAU - Norris, Geoffrey
AU  - Norris G
FAU - Laudenslager, Marci
AU  - Laudenslager M
FAU - Wood, Andrew C
AU  - Wood AC
FAU - Mayes, Patrick A
AU  - Mayes PA
FAU - Jagannathan, Jayanti
AU  - Jagannathan J
FAU - Winter, Cynthia
AU  - Winter C
FAU - Mosse, Yael P
AU  - Mosse YP
FAU - Maris, John M
AU  - Maris JM
LA  - eng
SI  - GEO/GSE19274
GR  - U10-CA98543/CA/NCI NIH HHS/United States
GR  - K08 CA136979-01/CA/NCI NIH HHS/United States
GR  - R01-CA87847/CA/NCI NIH HHS/United States
GR  - U10 CA098543/CA/NCI NIH HHS/United States
GR  - K08 CA136979/CA/NCI NIH HHS/United States
GR  - R01 CA087847/CA/NCI NIH HHS/United States
PT  - Journal Article
PT  - Research Support, N.I.H., Extramural
PT  - Research Support, Non-U.S. Gov't
DEP - 20110202
PL  - United States
TA  - Proc Natl Acad Sci U S A
JT  - Proceedings of the National Academy of Sciences of the United States of America
JID - 7505876
RN  - 0 (MYCN protein, human)
RN  - 0 (N-Myc Proto-Oncogene Protein)
RN  - 0 (Nuclear Proteins)
RN  - 0 (Oncogene Proteins)
RN  - 0 (Protein Kinase Inhibitors)
RN  - 0 (RNA, Messenger)
RN  - 0 (RNA, Small Interfering)
RN  - EC 2.7.- (Protein Kinases)
RN  - EC 2.7.11.1 (CHEK1 protein, human)
RN  - EC 2.7.11.1 (Checkpoint Kinase 1)
SB  - IM
MH  - Apoptosis/drug effects
MH  - Checkpoint Kinase 1
MH  - Drug Delivery Systems
MH  - Drug Evaluation, Preclinical
MH  - Humans
MH  - N-Myc Proto-Oncogene Protein
MH  - Neuroblastoma/*drug therapy/pathology
MH  - Nuclear Proteins/analysis
MH  - Oncogene Proteins/analysis
MH  - Phosphorylation
MH  - Protein Kinase Inhibitors/pharmacology
MH  - Protein Kinases/*genetics/*metabolism
MH  - RNA, Messenger
MH  - RNA, Small Interfering/*pharmacology
MH  - S Phase/drug effects
PMC - PMC3044382
COIS- The authors declare no conflict of interest.
EDAT- 2011/02/04 06:00
MHDA- 2011/05/07 06:00
PMCR- 2011/08/22
CRDT- 2011/02/04 06:00
PHST- 2011/02/04 06:00 [entrez]
PHST- 2011/02/04 06:00 [pubmed]
PHST- 2011/05/07 06:00 [medline]
PHST- 2011/08/22 00:00 [pmc-release]
AID - 1012351108 [pii]
AID - 201012351 [pii]
AID - 10.1073/pnas.1012351108 [doi]
PST - ppublish
SO  - Proc Natl Acad Sci U S A. 2011 Feb 22;108(8):3336-41. doi: 
      10.1073/pnas.1012351108. Epub 2011 Feb 2.