Gene Expression Profiling of Mycosis Fungoides in Early and Tumor Stage—A Proof-of-Concept Study Using Laser Capture/Single Cell Microdissection and NanoString Analysis

A subset of patients with mycosis fungoides (MF) progress to the tumor stage, which correlates with a worse clinical outcome. The molecular events driving this progression are not well-understood. To identify the key molecular drivers, we performed gene expression profiling (GEP) using NanoString. T...

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Autores principales:	Justine Lai, Jing Li, Robert Gniadecki, Raymond Lai
Formato:	article
Lenguaje:	EN
Publicado:	MDPI AG 2021
Materias:	mycosis fungoides single cell microdissection NanoString gene expression profiling immunohistochemistry Biology (General) QH301-705.5
Acceso en línea:	https://doaj.org/article/968cc05b83694ea4910b9db88a6ac8bf
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spelling	oai:doaj.org-article:968cc05b83694ea4910b9db88a6ac8bf2021-11-25T17:12:39ZGene Expression Profiling of Mycosis Fungoides in Early and Tumor Stage—A Proof-of-Concept Study Using Laser Capture/Single Cell Microdissection and NanoString Analysis10.3390/cells101131902073-4409https://doaj.org/article/968cc05b83694ea4910b9db88a6ac8bf2021-11-01T00:00:00Zhttps://www.mdpi.com/2073-4409/10/11/3190https://doaj.org/toc/2073-4409A subset of patients with mycosis fungoides (MF) progress to the tumor stage, which correlates with a worse clinical outcome. The molecular events driving this progression are not well-understood. To identify the key molecular drivers, we performed gene expression profiling (GEP) using NanoString. Ten formalin-fixed/paraffin-embedded skin biopsies from six patients (six non-tumor and four tumor MF) were included; non-tumor and tumor samples were available in three patients. Laser capture/single cell microdissection of epidermotropic MF cells was used for non-tumor cases. We found that the RNA extracted from 700–800 single cells was consistently sufficient for GEP, provided that multiplexed target enrichment amplification was used. An un-supervised/hierarchical analysis revealed clustering of non-tumor and tumor cases. Many of the most upregulated or downregulated genes are implicated in the PI3K, RAS, cell cycle/apoptosis and MAPK pathways. Two of the targets, HMGA1 and PTPN11 (encodes SHP2), were validated using immunohistochemistry. HMGA1 was positive in six out of six non-tumor MF samples and negative in five out of five tumor MF samples. An opposite pattern was seen with SHP2. Our study has provided a proof-of-concept that single-cell microdissection/GEP can be applied to archival tissues. Some of our identified gene targets might be key drivers of the disease progression of MF.Justine LaiJing LiRobert GniadeckiRaymond LaiMDPI AGarticlemycosis fungoidessingle cell microdissectionNanoStringgene expression profilingimmunohistochemistryBiology (General)QH301-705.5ENCells, Vol 10, Iss 3190, p 3190 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	mycosis fungoides single cell microdissection NanoString gene expression profiling immunohistochemistry Biology (General) QH301-705.5
spellingShingle	mycosis fungoides single cell microdissection NanoString gene expression profiling immunohistochemistry Biology (General) QH301-705.5 Justine Lai Jing Li Robert Gniadecki Raymond Lai Gene Expression Profiling of Mycosis Fungoides in Early and Tumor Stage—A Proof-of-Concept Study Using Laser Capture/Single Cell Microdissection and NanoString Analysis
description	A subset of patients with mycosis fungoides (MF) progress to the tumor stage, which correlates with a worse clinical outcome. The molecular events driving this progression are not well-understood. To identify the key molecular drivers, we performed gene expression profiling (GEP) using NanoString. Ten formalin-fixed/paraffin-embedded skin biopsies from six patients (six non-tumor and four tumor MF) were included; non-tumor and tumor samples were available in three patients. Laser capture/single cell microdissection of epidermotropic MF cells was used for non-tumor cases. We found that the RNA extracted from 700–800 single cells was consistently sufficient for GEP, provided that multiplexed target enrichment amplification was used. An un-supervised/hierarchical analysis revealed clustering of non-tumor and tumor cases. Many of the most upregulated or downregulated genes are implicated in the PI3K, RAS, cell cycle/apoptosis and MAPK pathways. Two of the targets, HMGA1 and PTPN11 (encodes SHP2), were validated using immunohistochemistry. HMGA1 was positive in six out of six non-tumor MF samples and negative in five out of five tumor MF samples. An opposite pattern was seen with SHP2. Our study has provided a proof-of-concept that single-cell microdissection/GEP can be applied to archival tissues. Some of our identified gene targets might be key drivers of the disease progression of MF.
format	article
author	Justine Lai Jing Li Robert Gniadecki Raymond Lai
author_facet	Justine Lai Jing Li Robert Gniadecki Raymond Lai
author_sort	Justine Lai
title	Gene Expression Profiling of Mycosis Fungoides in Early and Tumor Stage—A Proof-of-Concept Study Using Laser Capture/Single Cell Microdissection and NanoString Analysis
title_short	Gene Expression Profiling of Mycosis Fungoides in Early and Tumor Stage—A Proof-of-Concept Study Using Laser Capture/Single Cell Microdissection and NanoString Analysis
title_full	Gene Expression Profiling of Mycosis Fungoides in Early and Tumor Stage—A Proof-of-Concept Study Using Laser Capture/Single Cell Microdissection and NanoString Analysis
title_fullStr	Gene Expression Profiling of Mycosis Fungoides in Early and Tumor Stage—A Proof-of-Concept Study Using Laser Capture/Single Cell Microdissection and NanoString Analysis
title_full_unstemmed	Gene Expression Profiling of Mycosis Fungoides in Early and Tumor Stage—A Proof-of-Concept Study Using Laser Capture/Single Cell Microdissection and NanoString Analysis
title_sort	gene expression profiling of mycosis fungoides in early and tumor stage—a proof-of-concept study using laser capture/single cell microdissection and nanostring analysis
publisher	MDPI AG
publishDate	2021
url	https://doaj.org/article/968cc05b83694ea4910b9db88a6ac8bf
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Gene Expression Profiling of Mycosis Fungoides in Early and Tumor Stage—A Proof-of-Concept Study Using Laser Capture/Single Cell Microdissection and NanoString Analysis

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