激光显微切割福尔马林固定石蜡包埋（FFPE）组织标本的新一代测序分析

Next-Generation Sequencing Analysis of Laser-Microdissected Formalin-Fixed and Paraffin-Embedded (FFPE) Tissue Specimens.

作者信息

Mägel Lavinia, Bartels Stephan, Lehmann Ulrich

机构信息

Institute of Pathology, Medizinische Hochschule Hannover, Hannover, Germany.

出版信息

Methods Mol Biol. 2018;1723:111-118. doi: 10.1007/978-1-4939-7558-7_5.

DOI:10.1007/978-1-4939-7558-7_5

PMID:29344856

Abstract

In recent years, next-generation sequencing (NGS) became widely used in molecular pathology. Comprehensive mutational profiling improved diagnosis and prognosis, as well as the identification of therapeutically relevant genetic alterations. However, the vast majority of studies analyzing tissue samples use DNA extracted from bulk tissue or only manually microdissected specimens. Laser-assisted microdissection offers the possibility of isolating morphologically defined small tissue compartments (like individual glands) or even of single cells for further molecular analysis. Even formalin-fixed paraffin-embedded (FFPE) tissue specimens can be used for laser-assisted microdissection. Combining these two innovative powerful methodological approaches provides invaluable insights into the genetic profile of any cell type and tissue compartment of interest, contributing to a better understanding of fundamental biological processes and disease-specific mechanisms.In this chapter, a detailed protocol is provided for microdissection of human mammary adenomyoepithelioma tissue specimens and subsequent targeted resequencing of a panel of cancer-related genes using IonTorrent/PGM technology.

摘要

近年来，下一代测序（NGS）在分子病理学中得到广泛应用。全面的突变分析改善了诊断和预后，以及治疗相关基因改变的鉴定。然而，绝大多数分析组织样本的研究使用从大块组织或仅手动显微切割标本中提取的DNA。激光辅助显微切割为分离形态学上定义的小组织区域（如单个腺体）甚至单个细胞以进行进一步分子分析提供了可能性。即使是福尔马林固定石蜡包埋（FFPE）组织标本也可用于激光辅助显微切割。将这两种创新的强大方法学方法结合起来，能够深入了解任何感兴趣的细胞类型和组织区域的基因谱，有助于更好地理解基本生物学过程和疾病特异性机制。在本章中，提供了一份详细的方案，用于对人乳腺腺肌上皮瘤组织标本进行显微切割，并随后使用IonTorrent/PGM技术对一组癌症相关基因进行靶向重测序。