CRISPR Screen of Druggable Targets in Small Cell Lung Cancer Identified ATM Inhibitor (AZD1390) as a Radiosensitizer

放射增敏剂 辐射敏感性 克隆形成试验 癌症研究 抗辐射性 活力测定 DNA损伤 生物 细胞培养 医学 放射治疗 内科学 遗传学 DNA
作者
Xiaozhuo Ran,Bell Xi Wu,Mary Shi,Lifang Song,Kevin C. Nixon,Vivek M. Philip,Housheng Hansen He,Ming‐Sound Tsao,Benjamin H. Lok
出处
期刊:International Journal of Radiation Oncology Biology Physics [Elsevier]
卷期号:118 (5): 1308-1314 被引量:1
标识
DOI:10.1016/j.ijrobp.2023.12.011
摘要

Small cell lung cancer (SCLC) is an aggressive and lethal form of lung cancer and the overall 5-year survival (OS) for patients is a dismal 7%. Radiation therapy (RT) provides some benefit for selected patients with SCLC but could be improved with radiosensitizing agents. In this study, we identified novel radiosensitizers for SCLC by a CRISPR-Cas9 screen and evaluated the efficacy of ATM inhibitor AZD1390 as a radiosensitizer of SCLC.We transduced the SCLC cell line SBC5 with a custom CRISPR sgRNA library focused on druggable gene targets and treated cells with RT. Cells collected at multiple timepoints were subjected to next-generation sequencing. We determined radiosensitization both in vitro with cell lines assessed by short-term viability and clonogenic assays, and in vivo mouse models by tumor growth delay. Pharmacodynamic effects of AZD1390 were quantified by ATM-Ser1981 phosphorylation, and RT-induced DNA damage by comet assay.Using a CRISPR dropout screen, we identified multiple radiosensitizing genes for SCLC at various timepoints with ATM as a top determinant gene for radiosensitivity. Validation by ATM knockout (KO) demonstrated increased radiosensitivity by short-term viability assay (dose modification factor [DMF]50 = 3.25-3.73 in SBC5 ATM-KO) and clonogenic assays (DMF37 1.25-1.65 in SBC5 ATM-KO). ATM inhibition by AZD1390 effectively abrogated ATM Ser1981 phosphorylation in SCLC cell lines and increased RT-induced DNA damage. AZD1390 synergistically increased the radiosensitivity of SCLC cell lines (cell viability assay: SBC5 DMF37 = 2.19, SHP77 DMF37 = 1.56, H446 DMF37 = 3.27, KP1 DMF37 = 1.65 at 100nM; clonogenic assay: SBC5 DMF37 = 4.23, H1048 DMF37 = 1.91), and in vivo murine syngeneic, KP1, and patient-derived xenograft (PDX) models, JHU-LX108 and JHU-LX33.In this study, we demonstrated that genetically and pharmacologically (AZD1390) inhibiting ATM markedly enhanced RT against SCLC, providing a novel pharmacologically tractable radiosensitizing strategy for patients with SCLC.
最长约 10秒,即可获得该文献文件

科研通智能强力驱动
Strongly Powered by AbleSci AI
更新
大幅提高文件上传限制,最高150M (2024-4-1)

科研通是完全免费的文献互助平台,具备全网最快的应助速度,最高的求助完成率。 对每一个文献求助,科研通都将尽心尽力,给求助人一个满意的交代。
实时播报
1秒前
1秒前
2秒前
CWYY完成签到,获得积分10
2秒前
2秒前
JYQ发布了新的文献求助10
3秒前
共享精神应助无限的雨梅采纳,获得10
3秒前
典雅的静发布了新的文献求助10
4秒前
4秒前
cm发布了新的文献求助10
4秒前
薰硝壤应助yunduan采纳,获得50
5秒前
5秒前
5秒前
思源应助YUMI采纳,获得10
6秒前
7秒前
xiaogui完成签到,获得积分10
7秒前
大锤应助宝贝充电站采纳,获得10
7秒前
8秒前
CipherSage应助66采纳,获得10
8秒前
EShan发布了新的文献求助10
8秒前
jin发布了新的文献求助10
8秒前
EmmaEmma发布了新的文献求助10
9秒前
SciGPT应助zhouyupeng采纳,获得10
10秒前
科研小白发布了新的文献求助10
11秒前
11秒前
小欢完成签到,获得积分10
11秒前
Jeneration发布了新的文献求助10
11秒前
上官若男应助贵贵采纳,获得10
12秒前
13秒前
anti1988完成签到,获得积分10
13秒前
13秒前
wang完成签到,获得积分10
14秒前
123完成签到,获得积分10
14秒前
cm发布了新的文献求助10
15秒前
15秒前
禹无极完成签到,获得积分10
15秒前
学术混子完成签到,获得积分10
16秒前
所所应助可靠听荷采纳,获得10
16秒前
17秒前
17秒前
高分求助中
Sustainability in Tides Chemistry 2800
The Young builders of New china : the visit of the delegation of the WFDY to the Chinese People's Republic 1000
Rechtsphilosophie 1000
Bayesian Models of Cognition:Reverse Engineering the Mind 888
Le dégorgement réflexe des Acridiens 800
Defense against predation 800
Very-high-order BVD Schemes Using β-variable THINC Method 568
热门求助领域 (近24小时)
化学 医学 生物 材料科学 工程类 有机化学 生物化学 物理 内科学 纳米技术 计算机科学 化学工程 复合材料 基因 遗传学 催化作用 物理化学 免疫学 量子力学 细胞生物学
热门帖子
关注 科研通微信公众号,转发送积分 3136000
求助须知:如何正确求助?哪些是违规求助? 2786769
关于积分的说明 7779614
捐赠科研通 2443019
什么是DOI,文献DOI怎么找? 1298798
科研通“疑难数据库(出版商)”最低求助积分说明 625232
版权声明 600870