Barber H, Tofias A, Lander B, Daniels A, Gong J, Ren Y, Ren X, Liang Y, White P, Kurian K M
Brain Tumour Research Centre, Bristol Medical School, University of Bristol, Bristol, UK.
NanoString Technologies, Inc., Seattle, WA, UK.
J Oncol. 2021 Feb 24;2021:8819702. doi: 10.1155/2021/8819702. eCollection 2021.
Glioblastoma (GBM) is the most common primary adult brain tumour with a median overall survival (OS) of 12-15 months. Molecular characterization of multiple immunooncology targets in GBM may help target novel immunotherapeutic strategies. We used NanoString GeoMx® Digital Spatial Profiling (DSP) to assess multiple immunooncology protein targets in methylated versus unmethylated IDH-wild-type glioblastoma.
NanoString GeoMx® DSP technology uses multiple primary antibodies conjugated to indexing DNA oligos with a UV photocleavable linker. Tissue regions of interest (ROIs) are selected with bound fluorescent antibodies; oligos are released via a UV-mediated linker and quantitated. We used DSP multiplex analysis of 31 immunooncology proteins and controls (CD4, CD14, CD68, CD8A, B7-H3, PD-L1, CD19, FOXP3, CD44, STAT3 (phospho Y705), CD45, Pan Cytokeratin, MS4A1/CD20, CD45RO, PD1, CD3, beta-2 microglobulin, VISTA, Bcl2, GZMB, PTEN, beta-catenin, CD56, Ki-67, STAT3, AKT, p-Akt, S6, Histone H3, IgG Rabbit control, and Mouse IgG control) from ROIs in a cohort of 10 IDH-wild-type glioblastomas (5 methylated and 5 unmethylated). An nCounter platform allowed quantitative comparisons of antibodies between ROIs in MGMT methylated and unmethylated tumours. Mean protein expression counts between methylated and unmethylated GBM were compared using technical and biological replicates.
The analysis showed 10/27 immunooncology target proteins were significantly increased in methylated versus unmethylated IDH-wild-type glioblastoma tumour core (false discovery rate (FDR) <0.1 by Benjamini-Hochberg procedure).
NanoString GeoMx® DSP was used to analyse multiple immunooncology protein target expression in methylated versus unmethylated IDH-wild-type glioblastoma. In this small study, there was a statistical increase in CD4, CD14, CD68, CD8A, B7-H3, PDL-1, CD19, FOXP3, CD44, and STAT3 protein expression in methylated versus unmethylated GBM tumour core; however, this requires larger cohort validation. Advanced multiplex immunooncological biomarker analysis may be useful in identifying biomarkers for novel immunotherapeutic agents in GBMs.
胶质母细胞瘤（GBM）是最常见的原发性成人脑肿瘤，中位总生存期（OS）为12 - 15个月。对GBM中多个免疫肿瘤学靶点进行分子特征分析可能有助于确定新的免疫治疗策略。我们使用NanoString GeoMx®数字空间分析（DSP）来评估甲基化与未甲基化的异柠檬酸脱氢酶（IDH）野生型胶质母细胞瘤中的多个免疫肿瘤学蛋白靶点。
NanoString GeoMx® DSP技术使用与带有紫外线可裂解接头的索引DNA寡核苷酸偶联的多种一抗。用结合的荧光抗体选择感兴趣的组织区域（ROIs）；寡核苷酸通过紫外线介导的接头释放并进行定量。我们对10例IDH野生型胶质母细胞瘤（5例甲基化和5例未甲基化）的ROIs进行了31种免疫肿瘤学蛋白和对照（CD4、CD14、CD68、CD8A、B7 - H3、程序性死亡受体配体1（PD - L1）、CD19、叉头框蛋白P3（FOXP3）、CD44、信号转导和转录激活因子3（磷酸化Y705）（STAT3）、CD45、泛细胞角蛋白、跨膜4结构域A1/CD20（MS4A1/CD20）、CD45RO、程序性死亡蛋白1（PD1）、CD3、β2微球蛋白、VISTA、Bcl2、颗粒酶B（GZMB）、第10号染色体缺失的磷酸酶及张力蛋白同源物（PTEN）、β连环蛋白、CD56、Ki - 67、STAT3、蛋白激酶B（AKT）、磷酸化AKT（p - Akt）、核糖体蛋白S6（S6）、组蛋白H3、兔IgG对照和小鼠IgG对照）的DSP多重分析。nCounter平台允许对甲基化和未甲基化肿瘤中ROIs之间的抗体进行定量比较。使用技术和生物学重复比较甲基化和未甲基化GBM之间的平均蛋白表达计数。
分析显示，在甲基化与未甲基化的IDH野生型胶质母细胞瘤肿瘤核心中，10/27种免疫肿瘤学靶点蛋白显著增加（通过Benjamini - Hochberg方法，错误发现率（FDR）<0.1）。
使用NanoString GeoMx® DSP分析甲基化与未甲基化的IDH野生型胶质母细胞瘤中多个免疫肿瘤学蛋白靶点的表达。在这项小型研究中，甲基化与未甲基化的GBM肿瘤核心中，CD4、CD14、CD68、CD8A、B7 - H3、PD - L1、CD19、FOXP3、CD44和STAT3蛋白表达有统计学意义的增加；然而，这需要更大队列的验证。先进的多重免疫肿瘤学生物标志物分析可能有助于识别GBM中新免疫治疗药物的生物标志物。
