Hossen Md Nazir, Elechalawar Chandra Kumar, Sjoelund Virginie, Moore Kathleen, Mannel Robert, Bhattacharya Resham, Mukherjee Priyabrata
Peggy and Charles Stephenson Cancer Laboratory Research, Oklahoma Stanton L. Young Biomedical Research Center, University of Oklahoma Health Sciences Center, 975 N.E., Suite # 1409 10th Street, Oklahoma City, OK 73104 USA.
Department of Pathology, University of Oklahoma Health Science Center, Oklahoma City, OK USA.
Cancer Nanotechnol. 2021;12(1):1. doi: 10.1186/s12645-020-00071-7. Epub 2021 Jan 6.
Ovarian cancer is one of the deadliest gynecological malignancies. While the overall survival of ovarian cancer patients has slightly improved in recent years in the developed world, it remains clinically challenging due to its frequent late diagnosis and the lack of reliable diagnostic and/or prognostic markers. The aim of this study was to identify potential new molecular target proteins (NMTPs) responsible for the poor outcomes. When nanoparticles (NP) are exposed to biological fluids, a protein coat, termed the protein corona (PC), forms around the NP, and the PC represents a tool to identify NMTPs. This study investigates the influence of pre-processing conditions, such as lysis conditions and serum/plasma treatment, on the PC composition and the resulting identification of NMTPs.
Using gel electrophoresis, pre-processing conditions, including cell-lysis techniques and enrichment of low-abundance proteins (LAPs) by immunocentrifugation of serum/plasma, were shown to alter the relative amounts and compositions of proteins. PCs formed when 20 nm gold-NPs (GNPs) were incubated with lysate proteins from either RIPA- or urea lysis. Proteomic analysis of these PCs showed 2-22-fold enrichment of NMTPs in PCs from urea lysates as compared to RIPA lysates. Enriched NMTPs were then classified as cellular components, biological and molecular functions-associated proteins. The impact of enriched LAPs (eLAPs) on both PC composition and NMTP identification was shown by comparative proteomic analysis of original plasma, eLAPs, and PCs derived from eLAPs; eLAPs-PCs enhanced the abundance of NMTPs approximately 13%. Several NMTPs, including gasdermin-B, dermcidin, and kallistatin, were identified by this method demonstrating the potential use of this PC approach for molecular target discovery.
The current study showed that the pre-processing conditions modulate PC composition and can be used to enhance identification of NMTPs.
卵巢癌是最致命的妇科恶性肿瘤之一。尽管近年来发达国家卵巢癌患者的总体生存率略有提高，但由于其经常出现晚期诊断且缺乏可靠的诊断和/或预后标志物，临床上仍然具有挑战性。本研究的目的是确定导致不良预后的潜在新分子靶蛋白（NMTPs）。当纳米颗粒（NP）暴露于生物流体时，会在NP周围形成一层蛋白质外壳，称为蛋白质冠（PC），而PC是识别NMTPs的一种工具。本研究调查了预处理条件，如裂解条件和血清/血浆处理，对PC组成以及由此产生的NMTPs识别的影响。
使用凝胶电泳显示，预处理条件，包括细胞裂解技术和通过血清/血浆免疫离心富集低丰度蛋白（LAPs），会改变蛋白质的相对含量和组成。当20纳米金纳米颗粒（GNPs）与来自RIPA裂解或尿素裂解的裂解蛋白孵育时形成了PC。对这些PC的蛋白质组学分析表明，与RIPA裂解物相比，尿素裂解物中PCs中NMTPs的富集倍数为2至22倍。然后将富集的NMTPs分类为细胞成分、生物和分子功能相关蛋白。通过对原始血浆、eLAPs以及源自eLAPs的PC进行比较蛋白质组学分析，显示了富集的LAPs（eLAPs）对PC组成和NMTP识别的影响；eLAPs-PCs使NMTPs的丰度提高了约13%。通过该方法鉴定了几种NMTPs，包括gasdermin-B、dermcidin和kallistatin，证明了这种PC方法在分子靶点发现中的潜在用途。
当前研究表明，预处理条件可调节PC组成，并可用于增强对NMTPs的识别。
