Epigenetic Input Dictates the Threshold of Targeting of the Integrin-Dependent Pathway in Non-small Cell Lung Cancer

We investigated the therapeutic potential of targeting integrin/FAK-dependent signaling, an adhesion receptor-mediated pathway that has been increasingly linked to non-small cell lung cancer (NSCLC) malignancy. Our analysis of the TCGA cohort showed that a subset of pro-tumorigenic integrins, includ...

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Main Authors: Yang Zhang, Kai Cheng, Bingwei Xu, Junfeng Shi, Jun Qiang, Shujin Shi, Yuanqin Yi, Hongxia Li, Tengchuan Jin, Ruihua Guo, Yadi Wu, Zeyi Liu, Xiaowei Wei, Jian-An Huang, Xiuwei H. Yang
Format: Article
Language:English
Published: Frontiers Media S.A. 2020-07-01
Series:Frontiers in Cell and Developmental Biology
Subjects:
FAK
Online Access:https://www.frontiersin.org/article/10.3389/fcell.2020.00652/full
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author Yang Zhang
Yang Zhang
Yang Zhang
Kai Cheng
Bingwei Xu
Bingwei Xu
Junfeng Shi
Jun Qiang
Jun Qiang
Shujin Shi
Yuanqin Yi
Yuanqin Yi
Yuanqin Yi
Hongxia Li
Hongxia Li
Hongxia Li
Hongxia Li
Tengchuan Jin
Ruihua Guo
Yadi Wu
Yadi Wu
Zeyi Liu
Xiaowei Wei
Jian-An Huang
Xiuwei H. Yang
Xiuwei H. Yang
spellingShingle Yang Zhang
Yang Zhang
Yang Zhang
Kai Cheng
Bingwei Xu
Bingwei Xu
Junfeng Shi
Jun Qiang
Jun Qiang
Shujin Shi
Yuanqin Yi
Yuanqin Yi
Yuanqin Yi
Hongxia Li
Hongxia Li
Hongxia Li
Hongxia Li
Tengchuan Jin
Ruihua Guo
Yadi Wu
Yadi Wu
Zeyi Liu
Xiaowei Wei
Jian-An Huang
Xiuwei H. Yang
Xiuwei H. Yang
Epigenetic Input Dictates the Threshold of Targeting of the Integrin-Dependent Pathway in Non-small Cell Lung Cancer
Frontiers in Cell and Developmental Biology
lung cancer
FAK
BRD4
integrins
KRAS targeted therapy
author_facet Yang Zhang
Yang Zhang
Yang Zhang
Kai Cheng
Bingwei Xu
Bingwei Xu
Junfeng Shi
Jun Qiang
Jun Qiang
Shujin Shi
Yuanqin Yi
Yuanqin Yi
Yuanqin Yi
Hongxia Li
Hongxia Li
Hongxia Li
Hongxia Li
Tengchuan Jin
Ruihua Guo
Yadi Wu
Yadi Wu
Zeyi Liu
Xiaowei Wei
Jian-An Huang
Xiuwei H. Yang
Xiuwei H. Yang
author_sort Yang Zhang
title Epigenetic Input Dictates the Threshold of Targeting of the Integrin-Dependent Pathway in Non-small Cell Lung Cancer
title_short Epigenetic Input Dictates the Threshold of Targeting of the Integrin-Dependent Pathway in Non-small Cell Lung Cancer
title_full Epigenetic Input Dictates the Threshold of Targeting of the Integrin-Dependent Pathway in Non-small Cell Lung Cancer
title_fullStr Epigenetic Input Dictates the Threshold of Targeting of the Integrin-Dependent Pathway in Non-small Cell Lung Cancer
title_full_unstemmed Epigenetic Input Dictates the Threshold of Targeting of the Integrin-Dependent Pathway in Non-small Cell Lung Cancer
title_sort epigenetic input dictates the threshold of targeting of the integrin-dependent pathway in non-small cell lung cancer
publisher Frontiers Media S.A.
series Frontiers in Cell and Developmental Biology
issn 2296-634X
publishDate 2020-07-01
description We investigated the therapeutic potential of targeting integrin/FAK-dependent signaling, an adhesion receptor-mediated pathway that has been increasingly linked to non-small cell lung cancer (NSCLC) malignancy. Our analysis of the TCGA cohort showed that a subset of pro-tumorigenic integrins, including α1β1, α2β1, α3β1, α5β1, and α6β4, were frequently amplified or upregulated at the genomic or mRNA level in KRAS or EGFR mutation/overexpression-enriched adenocarcinomas. These alterations appeared complementary, correlated with poor patient survival (p < 0.0072), and were collaborative with KRAS mutation-coupled αv integrins (p < 0.00159). Since integrin/FAK-dependent signaling is tightly coupled with normal human physiology, we sought to use a synthetic lethal-type targeting comprising of VS-6063, a chemical inhibitor of integrin-mediated FAK activity, and A549 cells, which carry a KRAS mutation and EGFR overexpression. Our screening analysis revealed that JQ1 and IBET-762, inhibitors of epigenetic reader BRD4, and LBH589, a pan inhibitor of histone deacetylases (HDACs), exhibited synergy with VS-6063 in mitigating tumor cell viability. This epigenetic link was corroborated by strong effects of additional inhibitors and RNAi-mediated knockdown of FAK and BRD4 or its downstream effector, c-Myc. Low doses of JQ1 (≤0.5 μM) markedly escalated efficacy of VS-6063 across a panel of 10 NSCLC cell lines. This catalyst-like effect is in line with the oncogenic landscape in the TCGA cohort since c-Myc falls downstream of the KRAS and EGFR oncogenes. Mechanistically, co-inhibiting the integrin-FAK and BRD4/c-Myc axes synergistically induced apoptotic cell death and DNA damage response, and impaired stemness-associated tumorsphere formation. These effects were accompanied by a marked inhibition of Akt- and p130Cas/Src-dependent signaling, but not Erk1/2 activity. Meanwhile, JQ1 alone or in combination with VS-6063 attenuated cell-cell adhesion and extracellular matrix (ECM)-dependent cell spreading, which is reminiscent of phenotype induced by malfunctional E-cadherin or integrins. Paradoxically, this phenotypic impact coincided with downregulation of epithelial-mesenchymal transition (EMT)-inducting transcription factor ZEB1 or Snail. Finally, we showed that the effect of the VS-6063/JQ1 combination was nearly equivalent to that of VS-6063 plus Carboplatin or Osimertinib. Overall, our study indicates that the integrin/FAK and BRD4/c-Myc axes cooperatively drive NSCLC virulence, and a co-targeting may provide a line of therapy capable of overcoming EGFR/KRAS-driven malignancy.
topic lung cancer
FAK
BRD4
integrins
KRAS targeted therapy
url https://www.frontiersin.org/article/10.3389/fcell.2020.00652/full
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spelling doaj-a81bd69401dc44c9830497613399ff302020-11-25T03:42:45ZengFrontiers Media S.A.Frontiers in Cell and Developmental Biology2296-634X2020-07-01810.3389/fcell.2020.00652560485Epigenetic Input Dictates the Threshold of Targeting of the Integrin-Dependent Pathway in Non-small Cell Lung CancerYang Zhang0Yang Zhang1Yang Zhang2Kai Cheng3Bingwei Xu4Bingwei Xu5Junfeng Shi6Jun Qiang7Jun Qiang8Shujin Shi9Yuanqin Yi10Yuanqin Yi11Yuanqin Yi12Hongxia Li13Hongxia Li14Hongxia Li15Hongxia Li16Tengchuan Jin17Ruihua Guo18Yadi Wu19Yadi Wu20Zeyi Liu21Xiaowei Wei22Jian-An Huang23Xiuwei H. Yang24Xiuwei H. Yang25Department of Respiratory Medicine, First Affiliated Hospital of Soochow University, Suzhou, ChinaDepartment of Pharmacology and Nutritional Sciences, University of Kentucky, Lexington, KY, United StatesMarkey Cancer Center, University of Kentucky, Lexington, KY, United StatesDepartment of Pathology, Nanjing Jinling Hospital, Nanjing University School of Medicine, Nanjing, ChinaDepartment of Pharmacology and Nutritional Sciences, University of Kentucky, Lexington, KY, United StatesMarkey Cancer Center, University of Kentucky, Lexington, KY, United StatesDepartment of Oncology, Nanjing Medical University, Nanjing, ChinaMinistry of Agriculture, Freshwater Fisheries Research Center, Chinese Academy of Fishery Sciences, Wuxi, ChinaFisheries College, Nanjing Agricultural University, Wuxi, ChinaDepartment of Oncology, Nanjing Medical University, Nanjing, ChinaDepartment of Pharmacology and Nutritional Sciences, University of Kentucky, Lexington, KY, United StatesMarkey Cancer Center, University of Kentucky, Lexington, KY, United StatesCancer Institute, The First Affiliated Hospital of China Medical University, Shenyang, ChinaDepartment of Pharmacology and Nutritional Sciences, University of Kentucky, Lexington, KY, United StatesMarkey Cancer Center, University of Kentucky, Lexington, KY, United StatesMinistry of Agriculture, Freshwater Fisheries Research Center, Chinese Academy of Fishery Sciences, Wuxi, ChinaFisheries College, Nanjing Agricultural University, Wuxi, ChinaLaboratory of Structural Immunology, Division of Life Sciences and Medicine, University of Science and Technology of China, Hefei, China0College of Food Science and Technology, Shanghai Ocean University, Shanghai, ChinaDepartment of Pharmacology and Nutritional Sciences, University of Kentucky, Lexington, KY, United StatesMarkey Cancer Center, University of Kentucky, Lexington, KY, United StatesDepartment of Respiratory Medicine, First Affiliated Hospital of Soochow University, Suzhou, ChinaDepartment of Oncology, Nanjing Medical University, Nanjing, ChinaDepartment of Respiratory Medicine, First Affiliated Hospital of Soochow University, Suzhou, ChinaDepartment of Pharmacology and Nutritional Sciences, University of Kentucky, Lexington, KY, United StatesMarkey Cancer Center, University of Kentucky, Lexington, KY, United StatesWe investigated the therapeutic potential of targeting integrin/FAK-dependent signaling, an adhesion receptor-mediated pathway that has been increasingly linked to non-small cell lung cancer (NSCLC) malignancy. Our analysis of the TCGA cohort showed that a subset of pro-tumorigenic integrins, including α1β1, α2β1, α3β1, α5β1, and α6β4, were frequently amplified or upregulated at the genomic or mRNA level in KRAS or EGFR mutation/overexpression-enriched adenocarcinomas. These alterations appeared complementary, correlated with poor patient survival (p < 0.0072), and were collaborative with KRAS mutation-coupled αv integrins (p < 0.00159). Since integrin/FAK-dependent signaling is tightly coupled with normal human physiology, we sought to use a synthetic lethal-type targeting comprising of VS-6063, a chemical inhibitor of integrin-mediated FAK activity, and A549 cells, which carry a KRAS mutation and EGFR overexpression. Our screening analysis revealed that JQ1 and IBET-762, inhibitors of epigenetic reader BRD4, and LBH589, a pan inhibitor of histone deacetylases (HDACs), exhibited synergy with VS-6063 in mitigating tumor cell viability. This epigenetic link was corroborated by strong effects of additional inhibitors and RNAi-mediated knockdown of FAK and BRD4 or its downstream effector, c-Myc. Low doses of JQ1 (≤0.5 μM) markedly escalated efficacy of VS-6063 across a panel of 10 NSCLC cell lines. This catalyst-like effect is in line with the oncogenic landscape in the TCGA cohort since c-Myc falls downstream of the KRAS and EGFR oncogenes. Mechanistically, co-inhibiting the integrin-FAK and BRD4/c-Myc axes synergistically induced apoptotic cell death and DNA damage response, and impaired stemness-associated tumorsphere formation. These effects were accompanied by a marked inhibition of Akt- and p130Cas/Src-dependent signaling, but not Erk1/2 activity. Meanwhile, JQ1 alone or in combination with VS-6063 attenuated cell-cell adhesion and extracellular matrix (ECM)-dependent cell spreading, which is reminiscent of phenotype induced by malfunctional E-cadherin or integrins. Paradoxically, this phenotypic impact coincided with downregulation of epithelial-mesenchymal transition (EMT)-inducting transcription factor ZEB1 or Snail. Finally, we showed that the effect of the VS-6063/JQ1 combination was nearly equivalent to that of VS-6063 plus Carboplatin or Osimertinib. Overall, our study indicates that the integrin/FAK and BRD4/c-Myc axes cooperatively drive NSCLC virulence, and a co-targeting may provide a line of therapy capable of overcoming EGFR/KRAS-driven malignancy.https://www.frontiersin.org/article/10.3389/fcell.2020.00652/fulllung cancerFAKBRD4integrinsKRAS targeted therapy