Quantitative Detection of Disseminated Melanoma Cells by Trp-1 Transcript Analysis Reveals Stochastic Distribution of Pulmonary Metastases

A better understanding of the process of melanoma metastasis is required to underpin the development of novel therapies that will improve patient outcomes. The use of appropriate animal models is indispensable for investigating the mechanisms of melanoma metastasis. However, reliable and practicable...

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Autores principales:	Lenka Kyjacova, Rafael Saup, Melanie Rothley, Anja Schmaus, Tabea Wagner, Anja Boßerhoff, Boyan K. Garvalov, Wilko Thiele, Jonathan P. Sleeman
Formato:	article
Lenguaje:	EN
Publicado:	MDPI AG 2021
Materias:	Trp-1 metastasis melanoma animal models lung mice Medicine R
Acceso en línea:	https://doaj.org/article/5efdfbfd7d5b4c978300e2058a145c84
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spelling	oai:doaj.org-article:5efdfbfd7d5b4c978300e2058a145c842021-11-25T18:03:03ZQuantitative Detection of Disseminated Melanoma Cells by Trp-1 Transcript Analysis Reveals Stochastic Distribution of Pulmonary Metastases10.3390/jcm102254592077-0383https://doaj.org/article/5efdfbfd7d5b4c978300e2058a145c842021-11-01T00:00:00Zhttps://www.mdpi.com/2077-0383/10/22/5459https://doaj.org/toc/2077-0383A better understanding of the process of melanoma metastasis is required to underpin the development of novel therapies that will improve patient outcomes. The use of appropriate animal models is indispensable for investigating the mechanisms of melanoma metastasis. However, reliable and practicable quantification of metastases in experimental mice remains a challenge, particularly if the metastatic burden is low. Here, we describe a qRT-PCR-based protocol that employs the melanocytic marker Trp-1 for the sensitive quantification of melanoma metastases in the murine lung. Using this protocol, we were able to detect the presence of as few as 100 disseminated melanoma cells in lung tissue. This allowed us to quantify metastatic burden in a spontaneous syngeneic B16-F10 metastasis model, even in the absence of visible metastases, as well as in the autochthonous Tg(<i>Grm1</i>)/<i>Cyld<sup>−/−</sup></i> melanoma model. Importantly, we also observed an uneven distribution of disseminated melanoma cells amongst the five lobes of the murine lung, which varied considerably from animal to animal. Together, our findings demonstrate that the qRT-PCR-based detection of Trp-1 allows the quantification of low pulmonary metastatic burden in both transplantable and autochthonous murine melanoma models, and show that the analysis of lung metastasis in such models needs to take into account the stochastic distribution of metastatic lesions amongst the lung lobes.Lenka KyjacovaRafael SaupMelanie RothleyAnja SchmausTabea WagnerAnja BoßerhoffBoyan K. GarvalovWilko ThieleJonathan P. SleemanMDPI AGarticleTrp-1metastasismelanomaanimal modelslungmiceMedicineRENJournal of Clinical Medicine, Vol 10, Iss 5459, p 5459 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Trp-1 metastasis melanoma animal models lung mice Medicine R
spellingShingle	Trp-1 metastasis melanoma animal models lung mice Medicine R Lenka Kyjacova Rafael Saup Melanie Rothley Anja Schmaus Tabea Wagner Anja Boßerhoff Boyan K. Garvalov Wilko Thiele Jonathan P. Sleeman Quantitative Detection of Disseminated Melanoma Cells by Trp-1 Transcript Analysis Reveals Stochastic Distribution of Pulmonary Metastases
description	A better understanding of the process of melanoma metastasis is required to underpin the development of novel therapies that will improve patient outcomes. The use of appropriate animal models is indispensable for investigating the mechanisms of melanoma metastasis. However, reliable and practicable quantification of metastases in experimental mice remains a challenge, particularly if the metastatic burden is low. Here, we describe a qRT-PCR-based protocol that employs the melanocytic marker Trp-1 for the sensitive quantification of melanoma metastases in the murine lung. Using this protocol, we were able to detect the presence of as few as 100 disseminated melanoma cells in lung tissue. This allowed us to quantify metastatic burden in a spontaneous syngeneic B16-F10 metastasis model, even in the absence of visible metastases, as well as in the autochthonous Tg(<i>Grm1</i>)/<i>Cyld<sup>−/−</sup></i> melanoma model. Importantly, we also observed an uneven distribution of disseminated melanoma cells amongst the five lobes of the murine lung, which varied considerably from animal to animal. Together, our findings demonstrate that the qRT-PCR-based detection of Trp-1 allows the quantification of low pulmonary metastatic burden in both transplantable and autochthonous murine melanoma models, and show that the analysis of lung metastasis in such models needs to take into account the stochastic distribution of metastatic lesions amongst the lung lobes.
format	article
author	Lenka Kyjacova Rafael Saup Melanie Rothley Anja Schmaus Tabea Wagner Anja Boßerhoff Boyan K. Garvalov Wilko Thiele Jonathan P. Sleeman
author_facet	Lenka Kyjacova Rafael Saup Melanie Rothley Anja Schmaus Tabea Wagner Anja Boßerhoff Boyan K. Garvalov Wilko Thiele Jonathan P. Sleeman
author_sort	Lenka Kyjacova
title	Quantitative Detection of Disseminated Melanoma Cells by Trp-1 Transcript Analysis Reveals Stochastic Distribution of Pulmonary Metastases
title_short	Quantitative Detection of Disseminated Melanoma Cells by Trp-1 Transcript Analysis Reveals Stochastic Distribution of Pulmonary Metastases
title_full	Quantitative Detection of Disseminated Melanoma Cells by Trp-1 Transcript Analysis Reveals Stochastic Distribution of Pulmonary Metastases
title_fullStr	Quantitative Detection of Disseminated Melanoma Cells by Trp-1 Transcript Analysis Reveals Stochastic Distribution of Pulmonary Metastases
title_full_unstemmed	Quantitative Detection of Disseminated Melanoma Cells by Trp-1 Transcript Analysis Reveals Stochastic Distribution of Pulmonary Metastases
title_sort	quantitative detection of disseminated melanoma cells by trp-1 transcript analysis reveals stochastic distribution of pulmonary metastases
publisher	MDPI AG
publishDate	2021
url	https://doaj.org/article/5efdfbfd7d5b4c978300e2058a145c84
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Quantitative Detection of Disseminated Melanoma Cells by Trp-1 Transcript Analysis Reveals Stochastic Distribution of Pulmonary Metastases

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