Preprint Article Version 2 Preserved in Portico This version is not peer-reviewed

Tracking of Melanoma Cell Plasticity by Transcriptional Reporters

Version 1 : Received: 21 December 2021 / Approved: 22 December 2021 / Online: 22 December 2021 (15:04:59 CET)
Version 2 : Received: 12 January 2022 / Approved: 13 January 2022 / Online: 13 January 2022 (12:43:21 CET)

How to cite: Vidal, A.; Redmer, T. Tracking of Melanoma Cell Plasticity by Transcriptional Reporters. Preprints 2021, 2021120373 (doi: 10.20944/preprints202112.0373.v2). Vidal, A.; Redmer, T. Tracking of Melanoma Cell Plasticity by Transcriptional Reporters. Preprints 2021, 2021120373 (doi: 10.20944/preprints202112.0373.v2).

Abstract

Clonal evolution and cellular plasticity are the genetic and non-genetic driving forces of tumor heterogeneity that in turn determines the tumor cell response towards therapeutic drugs. Several lines of evidence suggest that therapeutic interventions foster the selection of drug resistant neural crest stem-like cells (NCSCs) that establish minimal residual disease (MRD) in melanoma. Here we established a dual reporter system enabling the tracking of NGFR expression and mRNA stability, providing insights into the maintenance of NCSC-states. We observed that the transcriptional reporter that contained a 1kb fragment of the human NGFR promoter was activated only in a minor subset (0.72±0.49%, range 0.3-1.5) and ~2-4% of A375 melanoma cells revealed stable NGFR mRNA. The combination of both reporters provided insights into phenotype switching and revealed that both cellular subsets gave rise to cellular heterogeneity. Moreover, whole transcriptome profiling and gene set enrichment analysis (GSEA) of the minor cellular subset revealed hypoxia-associated programs that might serve as potential drivers of an in vitro switching of NGFR-associated phenotypes and relapse of post-BRAF inhibitor treated tumors. Concordantly, we observed that the minor cellular subset increased in response to dabrafenib over time. In summary, our reporter-based approach provided insights into plasticity and identified a cellular subset that might be responsible for the establishment of MRD in melanoma.

Keywords

Transcriptional reporters; Plasticity; Stemness; NGFR

Subject

LIFE SCIENCES, Cell & Developmental Biology

Comments (1)

Comment 1
Received: 13 January 2022
Commenter: Torben Redmer
Commenter's Conflict of Interests: Author
Comment: In the revised version we provide more evidence of a dabrafenib-driven activation of our NGFR-reporter system and investigated genes that are potentially controlled by phenotype switching.
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