Proteomics Lab, National Centre for Cell Science, Ganeshkhind, Pune 411007, MH, India; Savitribai Phule Pune University, Ganeshkhind, Pune 411007, MH, India.
Savitribai Phule Pune University, Ganeshkhind, Pune 411007, MH, India; Cancer Biology and Epigenetics Lab, National Centre for Cell Science, Ganeshkhind, Pune 411007, MH, India.
J Proteomics. 2019 Oct 30;209:103504. doi: 10.1016/j.jprot.2019.103504. Epub 2019 Aug 26.
Among the blood cancers, 13% mortality is caused by Multiple myeloma (MM) type of hematological malignancy. In spite of therapeutic advances in chemotherapy treatment, still MM remains an incurable disease is mainly due to emergence of chemoresistance. At present time, FDA approved bortezomib is the first line drug for MM treatment. However, like other chemotherapy, MM patients are acquiring resistance against bortezomib. The present study aims to identify and validate bortezomib resistant protein targets in MM using iTRAQ and label free quantitative proteomic approaches. 112 differentially expressed proteins were commonly found in both approaches with similar differential expression pattern. Exportin-1 (XPO1) protein was selected for further validation as its significant high expression was observed in both iTRAQ and label free analysis. Bioinformatic analysis of these common differentially expressed proteins showed a clear cluster of proteins such as SMC1A, RCC2, CSE1, NUP88, NUP50, TPR, HSPA14, DYNLL1, RAD21 and RANBP2 being associated with XPO1. Functional studies like cell count assay, flow cytometry assay and soft agar assay proved that XPO1 knock down in RPMI 8226R cell line results in re-sensitization to bortezomib drug. The mass spectrometry data are available via ProteomeXchange with identifier PXD013859. BIOLOGICAL SIGNIFICANCE: Multiple myeloma (MM) is a type of hematological malignancy which constitutes about 13% of all blood cell related malignancies. Chemoresistance is one of the major obstacles for the successful treatment for MM. Bortezomib is a first proteasome inhibitor drug, widely used in MM treatment. The present study aims to identify and validate bortezomib resistant protein targets in MM. Here, we identified 112 candidate proteins to be associated with bortezomib resistance using global quantitative proteomic analysis. Among these candidate proteins, we show that XPO1 plays crucial role in emerging bortezomib resistance using functional studies like cell count assay, flow cytometry assay and soft agar assay. XPO1 could be a potential therapeutic target for MM and development of inhibitors of XPO1 might help to cure MM.
在血液癌症中,13%的死亡率是由多发性骨髓瘤(MM)型血液恶性肿瘤引起的。尽管化疗治疗取得了治疗进展,但多发性骨髓瘤仍然是一种无法治愈的疾病,主要是由于出现了化疗耐药性。目前,美国食品和药物管理局批准的硼替佐米是治疗多发性骨髓瘤的一线药物。然而,与其他化疗药物一样,多发性骨髓瘤患者对硼替佐米产生了耐药性。本研究旨在使用 iTRAQ 和无标记定量蛋白质组学方法鉴定和验证多发性骨髓瘤中的硼替佐米耐药蛋白靶标。两种方法都发现了 112 个差异表达蛋白,其差异表达模式相似。由于其在 iTRAQ 和无标记分析中都观察到明显的高表达,因此选择了输出蛋白 1(XPO1)蛋白进行进一步验证。这些常见差异表达蛋白的生物信息学分析显示,SMC1A、RCC2、CSE1、NUP88、NUP50、TPR、HSPA14、DYNLL1、RAD21 和 RANBP2 等与 XPO1 相关的蛋白质明显聚类。细胞计数测定、流式细胞术测定和软琼脂测定等功能研究证明,在 RPMI 8226R 细胞系中敲低 XPO1 可导致对硼替佐米药物重新敏感。质谱数据可通过 ProteomeXchange 获得,标识符为 PXD013859。生物学意义:多发性骨髓瘤(MM)是一种血液恶性肿瘤,占所有与血细胞相关的恶性肿瘤的 13%左右。化疗耐药性是多发性骨髓瘤成功治疗的主要障碍之一。硼替佐米是一种广泛用于 MM 治疗的第一代蛋白酶体抑制剂药物。本研究旨在鉴定和验证多发性骨髓瘤中的硼替佐米耐药蛋白靶标。在这里,我们使用全局定量蛋白质组学分析鉴定了 112 种候选蛋白与硼替佐米耐药性相关。在这些候选蛋白中,我们通过细胞计数测定、流式细胞术测定和软琼脂测定等功能研究表明,XPO1 在出现硼替佐米耐药性方面起着至关重要的作用。XPO1 可能是多发性骨髓瘤的一个潜在治疗靶点,开发 XPO1 的抑制剂可能有助于治疗多发性骨髓瘤。