PO.ET01.04 · 实验与分子治疗
MARK2/MARK3/MARK4激酶是与YAP-TEAD通路存在共依赖性的人头颈部鳞状细胞癌的治疗靶点
MARK2/MARK3/MARK4 kinases are therapeutic targets for human head and neck squamous cell carcinoma with co-dependencies of YAP-TEAD pathway
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摘要 Abstract
中文摘要
背景：头颈部鳞状细胞癌（HNSCC）需要鉴定新的治疗靶点以改善治疗效果。本研究探讨了微管亲和调节激酶（MARKs）的生物学意义，这是一类调节微管相关蛋白（MAPs）的丝氨酸-苏氨酸激酶家族。由于MARK活性影响细胞结构、胞内运输和增殖，这些激酶可能成为有前景的治疗靶点。
材料与方法：在20株人HNSCC细胞系中评估了MARK家族基因的蛋白表达水平。通过在CA9-22和YD-8细胞中分别将顺铂和多西他赛浓度逐步递增至4 μM和15 nM，建立了顺铂和多西他赛耐药模型。采用WST法测定细胞活力。使用CRISPR/Cas9基因组编辑技术构建MARK2/3/4敲除细胞系。此外，还评估了若干新研发的MARK2/3/4先导化合物的抗癌疗效。
结果：与野生型对照相比，MARK2/3/4敲除细胞表现出显著降低的增殖能力和受损的matrigel侵袭能力。MARK激酶缺失与MAPK和PI3K-AKT信号通路的抑制相关。由于已知MARK蛋白通过磷酸化依赖机制干扰MST/SAV和LATS复合物的形成，我们进一步表明MARK4耗竭减少了YAP/TAZ的核定位，并降低了YAP/TEAD转录靶标的表达。协同筛选在先导化合物阶段鉴定出多个MARK特异性抑制的候选分子；值得注意的是，一个先导化合物在HNSCC细胞（包括顺铂耐药模型）中表现出强效的抗增殖活性。
结论：MARK蛋白作为HIPPO激酶级联的负调节因子，从而增强HNSCC中YAP/TAZ的致癌活性。MARK功能缺失降低了致瘤表型，支持MARKs作为未来药物开发的有前景的治疗靶点。
查看英文原文 English abstract
Background: Head and neck squamous cell carcinoma (HNSCC) requires the identification of new therapeutic targets to improve treatment outcomes. This study investigated the biological significance of Microtubule Affinity-Regulating Kinases (MARKs), a family of serine-threonine kinases that regulate microtubule-associated proteins (MAPs). Because MARK activity influences cell structure, intracellular transport, and proliferation, these kinases may represent promising therapeutic targets.
Materials and Methods: Protein expression levels of MARK family genes were evaluated in 20 human HNSCC cell lines. Cisplatin and docetaxel-resistant models were established by gradually escalating cisplatin and docetaxel concentrations up to 4 μM and 15 nM, respectively in CA9-22 and YD-8 cells. Cell viability was determined using WST assays. CRISPR/Cas9 genome editing was employed to generate MARK2/3/4 knockout cell lines. In addition, the anticancer efficacy of several newly developed MARK2/3/4 hits was assessed.
Results: MARK2/3/4-knock-out cells demonstrated significantly reduced proliferation and impaired matrigel invasion compared with wild-type controls. MARK kinase loss was associated with suppression of the MAPK- and PI3K-AKT signaling pathways. Because MARK proteins are known to interfere with MST/SAV and LATS complex formation through phosphorylation-dependent mechanisms, we further showed that MARK4 depletion reduced nuclear YAP/TAZ localization and decreased expression of YAP/TEAD transcriptional targets. Collaborative screening identified multiple candidate molecules for MARK-specific inhibition at the hit-compound stage; notably, one lead compound exhibited strong antiproliferative activity in HNSCC cells, including cisplatin-resistant models.
Conclusions: MARK proteins function as negative regulators of the HIPPO kinase cascade, thereby enhancing YAP/TAZ oncogenic activity in HNSCC. Loss of MARK function reduces tumorigenic phenotypes, supporting MARKs as promising therapeutic targets for future drug development.
利益披露 Disclosure
S. Oh, None..
H. Lim, None..
S. Park, None..
S. Han, None..
J. Kim, None..
S. Lee, None..
H. Jeung, None.