Saini Heena, Choudhary Mahima, Sharma Harshita, Chowdhury Shibasish, Mukherjee Sudeshna, Chowdhury Rajdeep
Department of Biological Sciences, Birla Institute of Technology and Science, Pilani Campus, Pilani, Rajasthan, 333031, India.
Mol Biol Rep. 2023 Feb;50(2):1045-1058. doi: 10.1007/s11033-022-08072-y. Epub 2022 Nov 16.
Non-small cell lung carcinoma (NSCLC) is the most common cause of cancer-associated deaths worldwide. Though recent development in targeted therapy has improved NSCLC prognosis, yet there is an unmet need to identify novel causative factors and appropriate therapeutic regimen against NSCLCs.
In this study, we identify key molecular factors de-regulated in NSCLCs. Analyze their expression by real-time PCR and immunoblot; map their localization by immuno-fluorescence microscopy. We further propose an FDA approved drug, chloroquine (CQ) that affects the function of the molecular factors and hence can be repurposed as a therapeutic strategy against NSCLCs. Available NSCLC mutation data reflects a high probabilistic chance of patients harboring a p53 mutation, especially a gain of function (GOF)-R273H mutation. The GOF-P53 mutation enables the P53 protein to potentially interact with non-canonical protein partners facilitating oncogenesis. In this context, analysis of existing transcriptomic data from R273H-P53 expressing cells shows a concomitant up-regulation of Yes-associated protein (YAP) transcriptional targets and its protein partner TEAD1 in NSCLCs, suggesting a possible link between R273H-P53 and YAP. We therefore explored the inter-dependence of R273H-P53 and YAP in NSCLC cells. They were found to co-operatively regulate NSCLC proliferation. Genetic or pharmacological inhibition of YAP and GOF-P53 resulted in sensitization of NSCLC cells. Further analysis of pathways controlled by GOF-P53 and YAP showed that they positively regulate the cellular homeostatic process- autophagy to mediate survival. We hence postulated that a modulation of autophagy might be a potent strategy to curb proliferation. In accordance to above, autophagy inhibition, especially with the FDA-approved drug- chloroquine (CQ) resulted in cytoplasmic accumulation and reduced transcriptional activity of GOF-P53 and YAP, leading to growth arrest of NSCLC cells.
Our study highlights the importance of GOF-P53 and YAP in NSCLC proliferation and proposes autophagy inhibition as an efficient strategy to attenuate NSCLC tumorigenesis.
非小细胞肺癌（NSCLC）是全球癌症相关死亡的最常见原因。尽管靶向治疗的最新进展改善了NSCLC的预后，但仍需要确定新的致病因素和针对NSCLC的合适治疗方案。
在本研究中，我们确定了NSCLC中失调的关键分子因素。通过实时PCR和免疫印迹分析它们的表达；通过免疫荧光显微镜定位它们的位置。我们进一步提出一种FDA批准的药物氯喹（CQ），它会影响这些分子因素的功能，因此可以重新用作针对NSCLC的治疗策略。现有的NSCLC突变数据显示患者携带p53突变的概率很高，尤其是功能获得性（GOF）-R273H突变。GOF-P53突变使P53蛋白能够与非经典蛋白伙伴潜在地相互作用，促进肿瘤发生。在这种情况下，对来自表达R273H-P53细胞的现有转录组数据的分析表明，NSCLC中Yes相关蛋白（YAP）转录靶点及其蛋白伙伴TEAD1同时上调，这表明R273H-P53与YAP之间可能存在联系。因此，我们探讨了NSCLC细胞中R273H-P53与YAP的相互依赖性。发现它们协同调节NSCLC增殖。对YAP和GOF-P5进行基因或药理学抑制导致NSCLC细胞致敏。对由GOF-P53和YAP控制的通路的进一步分析表明，它们正向调节细胞稳态过程——自噬以介导存活。因此，我们推测自噬的调节可能是抑制增殖的有效策略。据此，自噬抑制，尤其是使用FDA批准的药物氯喹（CQ），导致GOF-P53和YAP的细胞质积累并降低其转录活性，从而导致NSCLC细胞生长停滞。
我们的研究强调了GOF-P53和YAP在NSCLC增殖中的重要性，并提出自噬抑制是减弱NSCLC肿瘤发生的有效策略。
