DeCoste Ryan, Amemiya Yutaka, Nersesian Sarah, Westhaver Lauren, Lee Stacey N, Carter Michael D, Sapp Heidi L, Stueck Ashley E, Arnason Thomas, Boudreau Jeanette, Seth Arun, Huang Weei-Yuarn
Department of Pathology & Laboratory Medicine, QEII Health Sciences Centre, Nova Scotia Health Authority (Central Zone), Halifax, NS B3H 1V8, Canada.
Department of Pathology, Dalhousie University, Halifax, NS B3H 1V8, Canada.
J Clin Med. 2022 Jul 21;11(14):4241. doi: 10.3390/jcm11144241.
Genomic profiling of pancreatic cancer using small core biopsies has taken an increasingly prominent role in precision medicine. However, if not appropriately preserved, nucleic acids (NA) from pancreatic tissues are known to be susceptible to degradation due to high intrinsic levels of nucleases. PAXgene fixation (PreAnalytix, Switzerland) represents a novel formalin-free tissue preservation method. We sought to compare the NA and histomorphological preservation of pancreatic cancer tissues preserved with PAXgene-fixed paraffin-embedding (PFPE) and formalin-fixed paraffin-embedding (FFPE). Tissues from 19 patients were obtained prospectively from pancreaticoduodenectomy specimens and evaluated by four gastrointestinal pathologists. The extracted NA were quantified by Nanodrop and Qubit and assessed for quality by qPCR, targeted next-generation sequencing (NGS) assay, and RNA-sequencing. Our results demonstrated that, when assessed blindly for morphological quality, the four pathologists deemed the PFPE slides adequate for diagnostic purposes. PFPE tissues enable greater yields of less fragmented and more amplifiable DNA. PFPE tissues demonstrated significantly improved quality control (QC) metrics in a targeted NGS assay including Median Absolute Pair-wise Difference (MAPD) scores. Our results support the use of PAXgene fixative for the processing of specimens from pancreatic cancers with the potential benefits of improved yields for more amplifiable DNA in low-yield biopsy specimens and its ideal use for amplicon-based NGS assays.
使用小核心活检对胰腺癌进行基因组分析在精准医学中发挥着越来越重要的作用。然而,如果保存不当,胰腺组织中的核酸(NA)因核酸酶固有水平高而容易降解。PAXgene固定(瑞士PreAnalytix公司)是一种新型的无甲醛组织保存方法。我们试图比较采用PAXgene固定石蜡包埋(PFPE)和福尔马林固定石蜡包埋(FFPE)保存的胰腺癌组织的核酸和组织形态学保存情况。前瞻性地从19例患者的胰十二指肠切除标本中获取组织,并由四位胃肠病理学家进行评估。提取的核酸通过Nanodrop和Qubit进行定量,并通过qPCR、靶向二代测序(NGS)分析和RNA测序评估质量。我们的结果表明,在对形态质量进行盲法评估时,四位病理学家认为PFPE切片足以用于诊断目的。PFPE组织能获得产量更高、片段化程度更低且更易于扩增的DNA。在包括中位绝对成对差异(MAPD)分数在内的靶向NGS分析中,PFPE组织显示出显著改善的质量控制(QC)指标。我们的结果支持使用PAXgene固定剂处理胰腺癌标本,其潜在益处包括提高低产量活检标本中更易于扩增的DNA的产量,以及其在基于扩增子的NGS分析中的理想用途。