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

doi:10.1007/978-1-0716-1205-7_21

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 Paper › Chapter

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 language	English
Title of host publication	Melanoma: Methods and Protocols
Editors	Kristian M. Hargadon
Place of Publication	U.S.
Publisher	Humana Press
Pages	277-286
Number of pages	10
ISBN (Electronic)	9781071612057
ISBN (Print)	9781071612040
DOIs	https://doi.org/10.1007/978-1-0716-1205-7_21
Publication status	Published - 2021

Bibliographical note

Publisher Copyright:
© 2021, Springer Science+Business Media, LLC, part of Springer Nature.

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This output contributes to the following UN Sustainable Development Goals (SDGs)

SDG 3 Good Health and Well-being

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10.1007/978-1-0716-1205-7_21

Cite this

Po, J. W., Ma, Y., Luk, A. W. S., Lynch, D., Balakrishnar, B., Brungs, D., Azimi, F., Cooper, A., Saricilar, E., Murthy, V., de Souza, P., & Becker, T. M. (2021). Single-cell analysis of BRAFV600E and NRASQ61R mutation status in melanoma cell lines as method generation for circulating melanoma cells. In K. M. Hargadon (Ed.), Melanoma: Methods and Protocols (pp. 277-286). Humana Press. https://doi.org/10.1007/978-1-0716-1205-7_21

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

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.",

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

note = "Publisher Copyright: {\textcopyright} 2021, Springer Science+Business Media, LLC, part of Springer Nature.",

year = "2021",

doi = "10.1007/978-1-0716-1205-7\_21",

language = "English",

isbn = "9781071612040",

pages = "277--286",

editor = "Hargadon, \{Kristian M.\}",

booktitle = "Melanoma: Methods and Protocols",

publisher = "Humana Press",

}

Po, JW, Ma, Y, Luk, AWS, Lynch, D, Balakrishnar, B, Brungs, D, Azimi, F, Cooper, A, Saricilar, E, Murthy, V, de Souza, P & Becker, TM 2021, Single-cell analysis of BRAFV600E and NRASQ61R mutation status in melanoma cell lines as method generation for circulating melanoma cells. in KM Hargadon (ed.), Melanoma: Methods and Protocols. Humana Press, U.S., pp. 277-286. https://doi.org/10.1007/978-1-0716-1205-7_21

TY - CHAP

T1 - Single-cell analysis of BRAFV600E and NRASQ61R mutation status in melanoma cell lines as method generation for circulating melanoma cells

AU - Po, Joseph W.

AU - Ma, Yafeng

AU - Luk, Alison W. S.

AU - Lynch, David

AU - Balakrishnar, Bavanthi

AU - Brungs, Daniel

AU - Azimi, Farhad

AU - Cooper, Adam

AU - Saricilar, Erin

AU - Murthy, Vinay

AU - de Souza, Paul

AU - Becker, Therese M.

PY - 2021

Y1 - 2021

N2 - 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.

AB - 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.

UR - https://hdl.handle.net/1959.7/uws:68251

U2 - 10.1007/978-1-0716-1205-7_21

DO - 10.1007/978-1-0716-1205-7_21

M3 - Chapter

C2 - 33704722

SN - 9781071612040

SP - 277

EP - 286

BT - Melanoma: Methods and Protocols

A2 - Hargadon, Kristian M.

PB - Humana Press

CY - U.S.

ER -

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

Abstract

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