Single-cell analysis of BRAFV600E and NRASQ61R mutation status in melanoma cell lines as method generation for circulating melanoma cells

Joseph W. Po, Yafeng Ma, Alison W. S. Luk, David Lynch, Bavanthi Balakrishnar, Daniel Brungs, Farhad Azimi, Adam Cooper, Erin Saricilar, Vinay Murthy, Paul de Souza, Therese M. Becker

Research output: Chapter in Book / Conference PaperChapter

2 Citations (Scopus)

Abstract

Molecular testing of tumor biopsies allows for the identification of the key mutations driving a patient’s cancer. However, this is limited to singular nodes and may not accurately reflect cancer heterogeneity. Circulating tumor cell (CTC) analyses offer a noninvasive method of interrogating the genomic profile of patient-derived tumor material. To date, molecular analysis of CTCs has relied on the characterization of bulk or pooled CTC lysates, limiting the detection of minor tumorigenic CTC subclones. Here, we show a workflow enabling BRAFV600E/NRASQ61R mutation detection from single cultured melanoma cells by combining micromanipulation and genomic material amplification methods. This workflow can be directly integrated into circulating tumor cell analysis applications.
Original languageEnglish
Title of host publicationMelanoma: Methods and Protocols
EditorsKristian M. Hargadon
Place of PublicationU.S.
PublisherHumana Press
Pages277-286
Number of pages10
ISBN (Electronic)9781071612057
ISBN (Print)9781071612040
DOIs
Publication statusPublished - 2021

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