Expanding the Use of an SS18-SSX Antibody for Molecular Assays in Synovial Sarcoma.
Expanding the Use of an SS18-SSX Antibody for Molecular Assays in Synovial Sarcoma.
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DOI：
10.1097/pai.0000000000001049
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发表时间：
2022-09-01
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影响因子：
--
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中科院分区：
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滑膜肉瘤是一种侵袭性的恶性肿瘤，通常影响青少年和年轻人，以高复发率和转移率为特征，10年生存率约为50%。融合癌蛋白SS18-SSX是致病原染色体易位t(X；18)的产物，是该肉瘤的致癌驱动因素，通过广泛的表观遗传失调扰乱分化。SS18-SSX生物学的实验研究一直受到缺乏特异性检测内源性融合癌蛋白的抗体的限制，而不是其天然的SS18或SSX成分。最近，研制出一种兔抗SS18-SSX融合连接位点表位的抗体，该抗体可特异性地检测至少95%的滑膜肉瘤中SS18-SSX的融合连接位点表位。在这里，我们描述了一套使用这种新抗体的分子生物学分析，既证实了现有的，也报道了新的应用。通过对滑膜肉瘤、组织芯片和全面部切片的免疫组织化学阳性染色，我们证明了该方法对滑膜肉瘤诊断的高度敏感性和特异性。此外，我们还展示了人类SS18-SSX蛋白在滑膜肉瘤基因工程小鼠模型中表达的检测结果。我们还用免疫荧光法证明了SS18-SSX在滑膜肉瘤细胞中的核染色，并通过邻近连接实验观察了SS18-SSX与BAF复合体成员BRG1的相互作用。最后，我们通过染色质免疫沉淀证实了SS18-SSX与目的基因启动子区域的相互作用。该抗体代表了肉瘤研究的一项突破，在扩大滑膜肉瘤生物学知识方面具有广泛的应用价值。
Synovial sarcoma is an aggressive malignancy that generally affects adolescents and young adults and is characterized by high rates of recurrence and metastasis, with a 10-year survival rate of about 50%. The fusion oncoprotein SS18-SSX, the product of a pathognomonic chromosomal translocation t(X;18), is the oncogenic driver of this sarcoma, disrupting differentiation through widespread epigenetic dysregulation. Experimental research into SS18-SSX biology has been limited by the lack of an antibody that specifically detects the endogenous fusion oncoprotein as opposed to its native SS18 or SSX components. Recently, a rabbit monoclonal antibody was developed and made commercially available, which specifically detects the fusion junction site epitope of SS18-SSX as found in at least 95% of synovial sarcomas. Here, we characterize a suite of molecular biology assays using this new antibody, both confirming existing and reporting on novel applications. We demonstrate its high sensitivity and specificity for synovial sarcoma diagnosis on patient samples through positive immunohistochemical staining on synovial sarcoma, tissue microarray, and full face sections. In addition, we demonstrate detection of the human SS18-SSX protein when expressed in a genetically engineered mouse model of synovial sarcoma. We also demonstrate nuclear staining of SS18-SSX in synovial sarcoma cells using immunofluorescence, and visualize the interaction between SS18-SSX and the BAF complex member BRG1 through a proximity ligation assay. Lastly, we confirm the interaction between SS18-SSX and promoter regions of target genes through chromatin immunoprecipitation. This antibody represents a breakthrough in sarcoma research and has value in multiple applications to expand the knowledge of synovial sarcoma biology.
影响因子：
8.8
作者：
Tamborini E;Agus V;Mezzelani A;Riva C;Sozzi G;Azzarelli A;Pierotti MA;Pilotti S
通讯作者：
Pilotti S