The role of NFIB-MAST1 signaling in mediating adaptive cisplatin resistance in SCLC

NFIB-MAST1信号在介导SCLC适应性顺铂耐药中的作用

• 批准号: 10553970
• 负责人: Lingtao Jin
• 金额: $ 36.77万
• 依托单位: UNIVERSITY OF TEXAS HLTH SCIENCE CENTER
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-02-01 至 2025-01-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10553970
• 关键词: ASCL1 gene; Affect; Antineoplastic Agents; Automobile Driving; Binding; Biological Assay; Biological Markers; Cancer Patient; Cancer cell line; Cell Cycle Progression; Chemoresistance; Cisplatin; Clinical; Clinical Trials; Data; Databases; Disease Progression; Drug Targeting; Drug resistance; Failure; Genetic Transcription; In Vitro; Lead; Mass Spectrum Analysis; Mediating; Microtubules; Mining; Mitotic; Modeling; Molecular; NFIB gene; Neoplasm Metastasis; PLK1 gene; Patients; Pharmaceutical Preparations; Phosphorylation; Phosphotransferases; Platinum; Primary Neoplasm; Promoter Regions; Protein-Serine-Threonine Kinases; Proteins; Proteomics; Recovery; Recurrence; Reporter; Resistance; Resistance development; Role; Signal Pathway; Signal Transduction; Site; TP53 gene; Testing; Therapeutic; Toxic effect; Tumor-Derived; base; chromatin immunoprecipitation; data mining; drug repurposing; efficacy evaluation; improved; in vivo; inhibitor; kinase inhibitor; knock-down; lung cancer cell; lung small cell carcinoma; mRNA Expression; mimetics; novel; overexpression; patient derived xenograft model; patient stratification; prevent; promoter; recruit; response; screening; small hairpin RNA; small molecule; standard care; therapeutic target; transcription factor; tumor xenograft; ubiquitin-protein ligase

PROJECT SUMMARY Cisplatin is one of the most effective and widely used anti-cancer drugs. For small cell lung cancer (SCLC), the current platinum-based standard treatment has not changed for more than three decades. Robust initial clinical response is usually observed in SCLC patients but the majority of patients succumb to chemoresistant recurrence. Despite tremendous efforts have been made to understand how SCLC cells develop cisplatin resistance, the precise mechanism remains elusive. As a part of our larger effort to decipher the mechanism of chemo-resistance, we employed a kinome wide shRNA screening and identified microtubule-associated serine/threonine kinase 1 (MAST1) as a “synthetic lethal” partner of cisplatin in SCLC. Using both SCLC cell lines and patient-derived tumors (PDX), we have demonstrated that MAST1 knockdown sensitizes ASCL1-high SCLC cells to cisplatin treatment in vitro and in vivo. Mining of CCLE database further shows that MAST1 expression is elevated in ASCL1-high subtype SCLC and positively correlates with cisplatin resistance in ASCL1-high SCLC cell lines. Through transcription factor profiling and unbiased datamining, we discovered that cisplatin physically binds and stabilize NFIB protein, a SCLC driver associated with disease progression, to promote MAST1 expression. Indeed, knockdown of NFIB blocked MAST1 induction by cisplatin, suggesting that MAST1 may be a downstream target of NFIB in SCLC. In addition, to identify the potential downstream effectors of MAST1 that contributes to cisplatin resistance in SCLC, we performed mass spectrometry-based proteomic studies and identified PLK1 as a potential binding partner of MAST1. In vitro kinase assay demonstrated that MAST1 directly phosphorylates PLK1 activation site T210. These findings identified MAST1 as a promising target to overcome cisplatin resistance in SCLC. However, no small molecule drug targeting MAST1 is currently available. Thus, we employed a “drug repurposing” strategy and identified clinical trial-staged kinase inhibitor lestaurtinib as a novel MAST1 inhibitor. Lestaurtinib significantly restored cisplatin sensitivity in SCLC cell lines and PDX models with minimal toxicity. Our central hypothesis is that cisplatin stabilizes NFIB protein to promote MAST1-PLK1 signaling, leading to cisplatin resistance in SCLC. Thus, targeting MAST1-PLK1 signaling represents a promising anti-SCLC target in combination with cisplatin, particularly for ASCL1-high subtype SCLC. We will test our hypothesis through the following aims: (1) To determine whether and how cisplatin stabilizes NFIB to promote MAST1 expression and consequently lead to cisplatin resistance in SCLC. (2) To determine whether MAST1 confers cisplatin resistance to SCLC by activating PLK1 and promoting cell cycle progression in the presence of cisplatin. (3) To validate NFIB-MAST1-PLK1 axis as a therapeutic target for cisplatin-resistant SCLC.

项目摘要 顺铂是最有效，最广泛使用的抗癌药物之一。对于小细胞肺癌（SCLC）， 目前基于铂金的标准治疗已经超过三十年没有改变。强大的初始临床 通常在SCLC患者中观察到反应，但大多数患者屈服于化学抗性 复发。尽管已经做出了巨大的努力来了解SCLC细胞如何发展顺铂 阻力，确切的机制仍然难以捉摸。 作为我们为破译化学耐药机制的更大努力的一部分，我们工作了宽度 shRNA筛选并鉴定出微管相关的丝氨酸/苏氨酸激酶1（MAST1）作为“合成 SCLC中顺铂的致命合作伙伴。使用SCLC细胞系和患者衍生的肿瘤（PDX），我们有 证明MAST1敲低的体外和在 体内。 CCLE数据库的挖掘进一步表明，在ASCL1高亚型SCLC中，MAST1表达升高 并与ASCL1高SCLC细胞系中的顺铂抗性正相关。通过转录因子 分析和无偏的数据材料，我们发现顺铂具有物理结合并稳定NFIB蛋白，A 与疾病进展相关的SCLC驱动器，以促进MAST1表达。确实，NFIB的敲除 顺铂阻断了MAST1诱导，这表明MAST1可能是SCLC中NFIB的下游靶标。 此外，要确定MAST1的潜在下游效应，这有助于顺铂抗性 SCLC，我们进行了基于质谱的蛋白质组学研究，并将PLK1鉴定为潜在的结合 Mast1的合作伙伴。体外激酶测定表明MAST1直接磷酸化PLK1激活位点 T210。这些发现将MAST1确定为克服SCLC中顺铂耐药性的承诺靶标。 但是，目前没有小分子药物靶向MAST1。那我们工作了一个毒品 重新利用”策略，并确定临床试验阶段的激酶抑制剂lestaurtinib是一种新型的MAST1抑制剂。 Lestaurtinib在SCLC细胞系和具有最小毒性的PDX模型中显着恢复了顺铂敏感性。 我们的中心假设是，顺铂稳定NFIB蛋白以促进MAST1-PLK1信号传导，导致 SCLC中的顺铂抗性。靶向MAST1-PLK1信号传导代表一个有希望的抗SCLC目标 与顺铂结合，特别是对于ASCL1高亚型SCLC。我们将通过 以下目的：（1）确定顺铂是否以及如何稳定NFIB以促进MAST1表达和 因此导致SCLC中的顺铂抗性。 （2）确定MAST1是否赋予顺铂抗性 通过激活PLK1并在顺铂存在下促进细胞周期进程来促进SCLC。 （3）验证 NFIB-MAST1-PLK1轴是抗顺铂的SCLC的治疗靶标。