TY - JOUR
T1 - A genome-wide CRISPR screen identifies genes critical for resistance to FLT3 inhibitor AC220
AU - Hou, Panpan
AU - Wu, Chao
AU - Wang, Yuchen
AU - Qi, Rui
AU - Bhavanasi, Dheeraj
AU - Zuo, Zhixiang
AU - Dos Santos, Cedric
AU - Chen, Shuliang
AU - Chen, Yu
AU - Zheng, Hong
AU - Wang, Hong
AU - Perl, Alexander
AU - Guo, Deyin
AU - Huang, Jian
N1 - Funding Information:
J. Huang has been awarded a grant from the NHLBI (R00 HL107747-04) and seed grant from Temple University Lewis Katz School of Medicine.
Funding Information:
We thank Dr. Martin Carroll, Peter Klein at University of Pennsylvania, and Drs. Yuri Persidsky, Jean-Pierre Issa, and Yi Zhang at Temple University School of Medicine for their insightful comments and discussion. We specially thank Dr. Hong Tian for technical support to our study. We thank all the members of Klein lab for their help and discussions. We thank the Stem cell and Xenograft core facility of UPenn for providing us with the AML samples, especially Dr. Gwenn Danet-Desnoyers. We greatly thank Dr. Yuesheng Li from Genomic facility of Fox Chase Cancer Center for help with the deep sequencing. We specially thank Xiang Yu at University of Pennsylvania and Tian Tian at New Jersey Institute of Technology for assistance of bioinformatic analysis.
Publisher Copyright:
©2017 AACR.
PY - 2017/8/15
Y1 - 2017/8/15
N2 - Acute myeloid leukemia (AML) is a malignant hematopoietic disease and the most common type of acute leukemia in adults. The mechanisms underlying drug resistance in AML are poorly understood. Activating mutations in FMS-like tyrosine kinase 3 (FLT3) are the most common molecular abnormality in AML. Quizartinib (AC220) is a potent and selective second-generation inhibitor of FLT3. It is in clinical trials for the treatment of relapsed or refractory FLT3-ITD–positive and –negative AML patients and as maintenance therapy. To understand the mechanisms of drug resistance to AC220, we undertook an unbiased approach with a novel CRISPR-pooled library to screen new genes whose loss of function confers resistance to AC220. We identified SPRY3, an intracellular inhibitor of FGF signaling and GSK3 a canonical Wnt signaling antagonist, and demonstrated reactivation of downstream FGF/Ras/ERK and Wnt signaling as major mechanisms of resistance to AC220. We confirmed these findings in primary AML patient samples. Expression of SPRY3 and GSK3A was dramatically reduced in AC220-resistant AML samples, and SPRY3-deleted primary AML cells were resistant to AC220. Intriguingly, expression of SPRY3 was greatly reduced in GSK3 knockout AML cells, which positioned SPRY3 downstream of GSK3 in the resistance pathway. Taken together, our study identified novel genes whose loss of function conferred resistance to a selective FLT3 inhibitor, providing new insight into signaling pathways that contribute to acquired resistance in AML.
AB - Acute myeloid leukemia (AML) is a malignant hematopoietic disease and the most common type of acute leukemia in adults. The mechanisms underlying drug resistance in AML are poorly understood. Activating mutations in FMS-like tyrosine kinase 3 (FLT3) are the most common molecular abnormality in AML. Quizartinib (AC220) is a potent and selective second-generation inhibitor of FLT3. It is in clinical trials for the treatment of relapsed or refractory FLT3-ITD–positive and –negative AML patients and as maintenance therapy. To understand the mechanisms of drug resistance to AC220, we undertook an unbiased approach with a novel CRISPR-pooled library to screen new genes whose loss of function confers resistance to AC220. We identified SPRY3, an intracellular inhibitor of FGF signaling and GSK3 a canonical Wnt signaling antagonist, and demonstrated reactivation of downstream FGF/Ras/ERK and Wnt signaling as major mechanisms of resistance to AC220. We confirmed these findings in primary AML patient samples. Expression of SPRY3 and GSK3A was dramatically reduced in AC220-resistant AML samples, and SPRY3-deleted primary AML cells were resistant to AC220. Intriguingly, expression of SPRY3 was greatly reduced in GSK3 knockout AML cells, which positioned SPRY3 downstream of GSK3 in the resistance pathway. Taken together, our study identified novel genes whose loss of function conferred resistance to a selective FLT3 inhibitor, providing new insight into signaling pathways that contribute to acquired resistance in AML.
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U2 - 10.1158/0008-5472.CAN-16-1627
DO - 10.1158/0008-5472.CAN-16-1627
M3 - Article
C2 - 28625976
AN - SCOPUS:85028321894
VL - 77
SP - 4402
EP - 4413
JO - Cancer Research
JF - Cancer Research
SN - 0008-5472
IS - 16
ER -