Azam Haleema, Veale Colin, Zitzmann Kim, Marcone Simone, Gallagher William M, Prencipe Maria
Cancer Biology and Therapeutics Laboratory, Conway Institute of Biomolecular and Biomedical Research, University College Dublin, Dublin, Ireland.
School of Biomolecular and Biomedical Science, University College Dublin, Dublin, Ireland.
PLoS One. 2024 Dec 13;19(12):e0309491. doi: 10.1371/journal.pone.0309491. eCollection 2024.
The Androgen Receptor (AR) pathway is crucial in driving the progression of prostate cancer (PCa) to an advanced state. Despite the introduction of second-generation AR antagonists, such as enzalutamide, majority of patients develop resistance. Several mechanisms of resistance have been identified, including the constitutive activation of the AR pathway, the emergence of AR spliced variants, and the influence of other signalling pathways. The Serum Response Factor (SRF) was previously identified as a possible player of resistance involved in a crosstalk with the AR signalling pathway. Elevated SRF levels in PCa patients were associated with disease progression and resistance to enzalutamide. However, the molecular mediators of the crosstalk between SRF and AR still need to be elucidated. The objective of this study was to identify common interactors of the AR/SRF crosstalk as therapeutic targets.
Here we used affinity purification mass spectrometry (MS) following immunoprecipitation of SRF and AR, to identify proteins that interact with both SRF and AR. The list of common interactors was expanded using STRING. Four common interactors were functionally validated using MTT assays.
Seven common interactors were identified, including HSP70, HSP0AA1, HSP90AB1, HSAP5, PRDX1 and GAPDH. Pathway analysis revealed that the PI3k/AKT pathway was the most enriched in the AR/SRF network. Moreover, pharmacological inhibition of several proteins in this network, including HSP70, HSP90, PI3k and AKT, significantly decreased cellular viability of PCa cells.
This study identified a list of AR/SRF common interactors that represent a pipeline of druggable targets for the treatment of PCa.
雄激素受体（AR）信号通路在前列腺癌（PCa）进展至晚期过程中起关键作用。尽管已引入第二代AR拮抗剂，如恩杂鲁胺，但大多数患者仍会产生耐药性。已确定多种耐药机制，包括AR信号通路的组成性激活、AR剪接变体的出现以及其他信号通路的影响。血清反应因子（SRF）先前被确定为可能参与与AR信号通路相互作用的耐药相关因子。PCa患者中SRF水平升高与疾病进展和对恩杂鲁胺的耐药性相关。然而，SRF与AR之间相互作用的分子介质仍有待阐明。本研究的目的是确定AR/SRF相互作用的共同蛋白作为治疗靶点。
在此，我们在对SRF和AR进行免疫沉淀后，使用亲和纯化质谱（MS）来鉴定与SRF和AR均相互作用的蛋白质。使用STRING扩展共同相互作用蛋白列表。使用MTT试验对四种共同相互作用蛋白进行功能验证。
鉴定出七种共同相互作用蛋白，包括HSP70、HSP0AA1、HSP90AB1、HSAP5、PRDX1和GAPDH。通路分析显示PI3k/AKT通路在AR/SRF网络中富集程度最高。此外，对该网络中几种蛋白质（包括HSP70、HSP90、PI3k和AKT）的药理学抑制显著降低了PCa细胞的细胞活力。
本研究确定了一系列AR/SRF共同相互作用蛋白，它们代表了用于治疗PCa的可成药靶点。
