Identification of EMT signaling cross-talk and gene regulatory networks by single-cell RNA sequencing

The epithelial-to-mesenchymal transition (EMT) plays a critical role during normal development and in cancer progression. EMT is induced by various signaling pathways, including TGF-β, BMP, Wnt-β-catenin, NOTCH, Shh, and receptor tyrosine kinases. In this study, we performed single-cell RNA sequenci...

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Vydané v:Proceedings of the National Academy of Sciences - PNAS Ročník 118; číslo 19
Hlavní autori: Deshmukh, Abhijeet P, Vasaikar, Suhas V, Tomczak, Katarzyna, Tripathi, Shubham, den Hollander, Petra, Arslan, Emre, Chakraborty, Priyanka, Soundararajan, Rama, Jolly, Mohit Kumar, Rai, Kunal, Levine, Herbert, Mani, Sendurai A
Médium: Journal Article
Jazyk:English
Vydavateľské údaje: United States 11.05.2021
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Abstract The epithelial-to-mesenchymal transition (EMT) plays a critical role during normal development and in cancer progression. EMT is induced by various signaling pathways, including TGF-β, BMP, Wnt-β-catenin, NOTCH, Shh, and receptor tyrosine kinases. In this study, we performed single-cell RNA sequencing on MCF10A cells undergoing EMT by TGF-β1 stimulation. Our comprehensive analysis revealed that cells progress through EMT at different paces. Using pseudotime clustering reconstruction of gene-expression profiles during EMT, we found sequential and parallel activation of EMT signaling pathways. We also observed various transitional cellular states during EMT. We identified regulatory signaling nodes that drive EMT with the expression of important microRNAs and transcription factors. Using a random circuit perturbation methodology, we demonstrate that the NOTCH signaling pathway acts as a key driver of TGF-β-induced EMT. Furthermore, we demonstrate that the gene signatures of pseudotime clusters corresponding to the intermediate hybrid EMT state are associated with poor patient outcome. Overall, this study provides insight into context-specific drivers of cancer progression and highlights the complexities of the EMT process.
AbstractList The epithelial-to-mesenchymal transition (EMT) plays a critical role during normal development and in cancer progression. EMT is induced by various signaling pathways, including TGF-β, BMP, Wnt-β-catenin, NOTCH, Shh, and receptor tyrosine kinases. In this study, we performed single-cell RNA sequencing on MCF10A cells undergoing EMT by TGF-β1 stimulation. Our comprehensive analysis revealed that cells progress through EMT at different paces. Using pseudotime clustering reconstruction of gene-expression profiles during EMT, we found sequential and parallel activation of EMT signaling pathways. We also observed various transitional cellular states during EMT. We identified regulatory signaling nodes that drive EMT with the expression of important microRNAs and transcription factors. Using a random circuit perturbation methodology, we demonstrate that the NOTCH signaling pathway acts as a key driver of TGF-β-induced EMT. Furthermore, we demonstrate that the gene signatures of pseudotime clusters corresponding to the intermediate hybrid EMT state are associated with poor patient outcome. Overall, this study provides insight into context-specific drivers of cancer progression and highlights the complexities of the EMT process.The epithelial-to-mesenchymal transition (EMT) plays a critical role during normal development and in cancer progression. EMT is induced by various signaling pathways, including TGF-β, BMP, Wnt-β-catenin, NOTCH, Shh, and receptor tyrosine kinases. In this study, we performed single-cell RNA sequencing on MCF10A cells undergoing EMT by TGF-β1 stimulation. Our comprehensive analysis revealed that cells progress through EMT at different paces. Using pseudotime clustering reconstruction of gene-expression profiles during EMT, we found sequential and parallel activation of EMT signaling pathways. We also observed various transitional cellular states during EMT. We identified regulatory signaling nodes that drive EMT with the expression of important microRNAs and transcription factors. Using a random circuit perturbation methodology, we demonstrate that the NOTCH signaling pathway acts as a key driver of TGF-β-induced EMT. Furthermore, we demonstrate that the gene signatures of pseudotime clusters corresponding to the intermediate hybrid EMT state are associated with poor patient outcome. Overall, this study provides insight into context-specific drivers of cancer progression and highlights the complexities of the EMT process.
The epithelial-to-mesenchymal transition (EMT) plays a critical role during normal development and in cancer progression. EMT is induced by various signaling pathways, including TGF-β, BMP, Wnt-β-catenin, NOTCH, Shh, and receptor tyrosine kinases. In this study, we performed single-cell RNA sequencing on MCF10A cells undergoing EMT by TGF-β1 stimulation. Our comprehensive analysis revealed that cells progress through EMT at different paces. Using pseudotime clustering reconstruction of gene-expression profiles during EMT, we found sequential and parallel activation of EMT signaling pathways. We also observed various transitional cellular states during EMT. We identified regulatory signaling nodes that drive EMT with the expression of important microRNAs and transcription factors. Using a random circuit perturbation methodology, we demonstrate that the NOTCH signaling pathway acts as a key driver of TGF-β-induced EMT. Furthermore, we demonstrate that the gene signatures of pseudotime clusters corresponding to the intermediate hybrid EMT state are associated with poor patient outcome. Overall, this study provides insight into context-specific drivers of cancer progression and highlights the complexities of the EMT process.
Author Arslan, Emre
Rai, Kunal
Jolly, Mohit Kumar
Mani, Sendurai A
Levine, Herbert
Deshmukh, Abhijeet P
Vasaikar, Suhas V
den Hollander, Petra
Soundararajan, Rama
Tomczak, Katarzyna
Tripathi, Shubham
Chakraborty, Priyanka
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  organization: Department of Translational Molecular Pathology, University of Texas MD Anderson Cancer Center, Houston, TX 77030
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  organization: Department of Translational Molecular Pathology, University of Texas MD Anderson Cancer Center, Houston, TX 77030
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  orcidid: 0000-0002-0141-987X
  surname: Tripathi
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  fullname: den Hollander, Petra
  organization: Department of Translational Molecular Pathology, University of Texas MD Anderson Cancer Center, Houston, TX 77030
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  organization: Department of Genomic Medicine, University of Texas MD Anderson Cancer Center, Houston, TX 77030
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  givenname: Priyanka
  surname: Chakraborty
  fullname: Chakraborty, Priyanka
  organization: Centre for BioSystems Science and Engineering, Indian Institute of Science, 560012 Bangalore, India
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  givenname: Rama
  orcidid: 0000-0001-6976-3784
  surname: Soundararajan
  fullname: Soundararajan, Rama
  organization: Department of Translational Molecular Pathology, University of Texas MD Anderson Cancer Center, Houston, TX 77030
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  givenname: Mohit Kumar
  orcidid: 0000-0002-6631-2109
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  surname: Rai
  fullname: Rai, Kunal
  email: krai@mdanderson.org, h.levine@northeastern.edu, Smani@mdanderson.org
  organization: Department of Genomic Medicine, University of Texas MD Anderson Cancer Center, Houston, TX 77030; krai@mdanderson.org h.levine@northeastern.edu Smani@mdanderson.org
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  surname: Mani
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  organization: Department of Translational Molecular Pathology, University of Texas MD Anderson Cancer Center, Houston, TX 77030; krai@mdanderson.org h.levine@northeastern.edu Smani@mdanderson.org
BackLink https://www.ncbi.nlm.nih.gov/pubmed/33941680$$D View this record in MEDLINE/PubMed
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scRNA-seq
RACIPE
signaling cascade
EMT
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Snippet The epithelial-to-mesenchymal transition (EMT) plays a critical role during normal development and in cancer progression. EMT is induced by various signaling...
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SubjectTerms Cell Line
Epithelial Cells - drug effects
Epithelial Cells - metabolism
Epithelial-Mesenchymal Transition - drug effects
Epithelial-Mesenchymal Transition - genetics
Gene Expression Profiling - methods
Gene Expression Profiling - statistics & numerical data
Gene Regulatory Networks
Humans
Kaplan-Meier Estimate
MicroRNAs - genetics
Neoplasms - classification
Neoplasms - genetics
Prognosis
Proportional Hazards Models
RNA-Seq - methods
Signal Transduction - drug effects
Signal Transduction - genetics
Single-Cell Analysis - methods
Transforming Growth Factor beta1 - metabolism
Transforming Growth Factor beta1 - pharmacology
Title Identification of EMT signaling cross-talk and gene regulatory networks by single-cell RNA sequencing
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