Electrical activity between skin cells regulates melanoma initiation

Oncogenes can only initiate tumors in certain cellular contexts, which is referred to as oncogenic competence. In melanoma, whether cells in the microenvironment can endow such competence remains unclear. Using a combination of zebrafish transgenesis coupled with human tissues, we demonstrate that GABAergic signaling between keratinocytes and melanocytes promotes melanoma initiation by BRAFV600E. GABA is synthesized in melanoma cells, which then acts on GABA-A receptors on keratinocytes. Electron microscopy demonstrates synapse-like structures between keratinocytes and melanoma cells, and multi-electrode array analysis shows that GABA acts to inhibit electrical activity in melanoma/keratinocyte co-cultures. Genetic and pharmacologic perturbation of GABA synthesis abrogates melanoma initiation in vivo. These data suggest that electrical activity across the skin microenvironment determines the ability of oncogenes to initiate melanoma.

[1]  A. Tward,et al.  Human melanocyte development and melanoma dedifferentiation at single-cell resolution , 2021, Nature Cell Biology.

[2]  D. P. Pomeranz Krummel,et al.  Melanoma Cell Intrinsic GABAA Receptor Enhancement Potentiates Radiation and Immune Checkpoint Inhibitor Response by Promoting Direct and T Cell-Mediated Antitumor Activity. , 2020, International journal of radiation oncology, biology, physics.

[3]  R. White,et al.  The Stress-Like Cancer Cell State Is a Consistent Component of Tumorigenesis. , 2020, Cell systems.

[4]  A. Tward,et al.  Human melanocyte development and melanoma dedifferentiation at single cell resolution , 2020, bioRxiv.

[5]  Joshua M. Weiss,et al.  Developmental chromatin programs determine oncogenic competence in melanoma , 2020, bioRxiv.

[6]  S. Simon,et al.  Ca2+ transients in melanocyte dendrites and dendritic spine-like structures evoked by cell-to-cell signaling , 2019, The Journal of cell biology.

[7]  N. Dekker,et al.  Endosomal escape enhancing compounds facilitate functional delivery of extracellular vesicle cargo. , 2019, Nanomedicine.

[8]  Stephen L. Johnson,et al.  Maintenance of Melanocyte Stem Cell Quiescence by GABA-A Signaling in Larval Zebrafish. , 2019, Genetics.

[9]  Stephen L. Johnson,et al.  Maintenance of Melanocyte Stem Cell Quiescence by GABA-A Signaling in Larval Zebrafish. , 2019, Genetics.

[10]  A. Barria,et al.  Dangerous liaisons as tumour cells form synapses with neurons , 2019, Nature.

[11]  T. Kuner,et al.  Glutamatergic synaptic input to glioma cells drives brain tumour progression , 2019, Nature.

[12]  Shawn M. Gillespie,et al.  Electrical and synaptic integration of glioma into neural circuits , 2019, Nature.

[13]  Inti Zlobec,et al.  Synaptic proximity enables NMDAR signaling to promote brain metastasis , 2019, Nature.

[14]  Kornel Labun,et al.  CHOPCHOP v3: expanding the CRISPR web toolbox beyond genome editing , 2019, Nucleic Acids Res..

[15]  Iwei Yeh,et al.  Human tumor genomics and zebrafish modeling identify SPRED1 loss as a driver of mucosal melanoma , 2018, Science.

[16]  R. White,et al.  Cancer modeling by Transgene Electroporation in Adult Zebrafish (TEAZ) , 2018, Disease Models & Mechanisms.

[17]  C. Ceol,et al.  Ligand-activated BMP signaling inhibits cell differentiation and death to promote melanoma , 2018, The Journal of clinical investigation.

[18]  C. Pirker,et al.  FGF5 is expressed in melanoma and enhances malignancy in vitro and in vivo , 2017, Oncotarget.

[19]  R. White,et al.  Microenvironment-derived factors driving metastatic plasticity in melanoma , 2017, Nature Communications.

[20]  Yuri Pritykin,et al.  GuideScan software for improved single and paired CRISPR guide RNA design , 2017, Nature Biotechnology.

[21]  J. Landsberg,et al.  The epidermal polarity protein Par3 is a non–cell autonomous suppressor of malignant melanoma , 2017, The Journal of experimental medicine.

[22]  M. Bosenberg,et al.  The YUMM lines: a series of congenic mouse melanoma cell lines with defined genetic alterations , 2016, Pigment cell & melanoma research.

[23]  K. Öllinger,et al.  Extracellular vesicles are transferred from melanocytes to keratinocytes after UVA irradiation , 2016, Scientific Reports.

[24]  L. Studer,et al.  Feeder-free Derivation of Melanocytes from Human Pluripotent Stem Cells. , 2016, Journal of Visualized Experiments.

[25]  Michelle E. Hung,et al.  A platform for actively loading cargo RNA to elucidate limiting steps in EV-mediated delivery , 2016, Journal of extracellular vesicles.

[26]  J. Rheenen,et al.  Studying extracellular vesicle transfer by a Cre-loxP method , 2015, Nature Protocols.

[27]  Ian Parker,et al.  A comparison of fluorescent Ca²⁺ indicators for imaging local Ca²⁺ signals in cultured cells. , 2015, Cell calcium.

[28]  D. Sprinzak,et al.  Interactions of Melanoma Cells with Distal Keratinocytes Trigger Metastasis via Notch Signaling Inhibition of MITF. , 2015, Molecular cell.

[29]  L. Zon,et al.  A Quantitative System for Studying Metastasis Using Transparent Zebrafish. , 2015, Cancer research.

[30]  Steven J. M. Jones,et al.  Genomic Classification of Cutaneous Melanoma , 2015, Cell.

[31]  Jacco van Rheenen,et al.  In Vivo Imaging Reveals Extracellular Vesicle-Mediated Phenocopying of Metastatic Behavior , 2015, Cell.

[32]  Z. J. Huang,et al.  GAD67 deficiency in parvalbumin interneurons produces deficits in inhibitory transmission and network disinhibition in mouse prefrontal cortex. , 2015, Cerebral cortex.

[33]  Linlin Yin,et al.  Multiplex Conditional Mutagenesis Using Transgenic Expression of Cas9 and sgRNAs , 2015, Genetics.

[34]  Christopher H Contag,et al.  Differential fates of biomolecules delivered to target cells via extracellular vesicles , 2015, Proceedings of the National Academy of Sciences.

[35]  Joseph J. Marlin,et al.  GABA-A Receptor Inhibition of Local Calcium Signaling in Spines and Dendrites , 2014, The Journal of Neuroscience.

[36]  W. Huber,et al.  Moderated estimation of fold change and dispersion for RNA-seq data with DESeq2 , 2014, Genome Biology.

[37]  M. Levin,et al.  Endogenous Voltage Potentials and the Microenvironment: Bioelectric Signals that Reveal, Induce and Normalize Cancer. , 2014, Journal of clinical & experimental oncology.

[38]  P. Altevogt,et al.  Extracellular Vesicle-Mediated Transfer of Genetic Information between the Hematopoietic System and the Brain in Response to Inflammation , 2014, Journal of Neuroimmunology.

[39]  Björn Usadel,et al.  Trimmomatic: a flexible trimmer for Illumina sequence data , 2014, Bioinform..

[40]  Yosef Yarom,et al.  Cerebellar Inhibitory Input to the Inferior Olive Decreases Electrical Coupling and Blocks Subthreshold Oscillations , 2014, Neuron.

[41]  D. Fisher,et al.  UV signaling pathways within the skin , 2014, The Journal of investigative dermatology.

[42]  G. Johnston Muscimol as an Ionotropic GABA Receptor Agonist , 2014, Neurochemical Research.

[43]  D. Laird,et al.  Connexin43 Reduces Melanoma Growth within a Keratinocyte Microenvironment and during Tumorigenesis in Vivo* , 2013, The Journal of Biological Chemistry.

[44]  A. Bosserhoff,et al.  Impact of LIF (leukemia inhibitory factor) expression in malignant melanoma. , 2013, Experimental and molecular pathology.

[45]  Joshua M. Stuart,et al.  The Cancer Genome Atlas Pan-Cancer analysis project , 2013, Nature Genetics.

[46]  Wei Shi,et al.  featureCounts: an efficient general purpose program for assigning sequence reads to genomic features , 2013, Bioinform..

[47]  E. Oancea,et al.  UV light phototransduction activates transient receptor potential A1 ion channels in human melanocytes , 2013, Proceedings of the National Academy of Sciences.

[48]  M. Ichihashi,et al.  Melanosomes are transferred from melanocytes to keratinocytes through the processes of packaging, release, uptake, and dispersion. , 2012, The Journal of investigative dermatology.

[49]  A. Werdich,et al.  The regenerative capacity of zebrafish reverses cardiac failure caused by genetic cardiomyocyte depletion , 2011, Development.

[50]  David A. Orlando,et al.  The histone methyltransferase SETDB1 is recurrently amplified in melanoma and accelerates its onset , 2011, Nature.

[51]  I. Han,et al.  Keratinocyte-derived Laminin-332 Promotes Adhesion and Migration in Melanocytes and Melanoma* , 2011, The Journal of Biological Chemistry.

[52]  L. Zon,et al.  Ubiquitous transgene expression and Cre-based recombination driven by the ubiquitin promoter in zebrafish , 2011, Development.

[53]  C. McCaig,et al.  Electrical dimensions in cell science , 2009, Journal of Cell Science.

[54]  Paolo Massobrio,et al.  A novel algorithm for precise identification of spikes in extracellularly recorded neuronal signals , 2009, Journal of Neuroscience Methods.

[55]  Petra Schwille,et al.  Ceramide Triggers Budding of Exosome Vesicles into Multivesicular Endosomes , 2008, Science.

[56]  L. Zon,et al.  Transparent adult zebrafish as a tool for in vivo transplantation analysis. , 2008, Cell stem cell.

[57]  Melissa Hardy,et al.  The Tol2kit: A multisite gateway‐based construction kit for Tol2 transposon transgenesis constructs , 2007, Developmental dynamics : an official publication of the American Association of Anatomists.

[58]  M. Korte,et al.  Impaired GABAergic transmission and altered hippocampal synaptic plasticity in collybistin‐deficient mice , 2007, The EMBO journal.

[59]  M. Herlyn,et al.  Microenvironmental influences in melanoma progression , 2007, Journal of cellular biochemistry.

[60]  H. Ueno,et al.  GABA-synthesizing enzyme, GAD67, from dermal fibroblasts: evidence for a new skin function. , 2007, Biochimica et biophysica acta.

[61]  Masaru Ishii,et al.  Spot pattern of leopard Danio is caused by mutation in the zebrafish connexin41.8 gene , 2006, EMBO reports.

[62]  D. Schadendorf,et al.  Metastatic potential of melanomas defined by specific gene expression profiles with no BRAF signature. , 2006, Pigment cell research.

[63]  J. Cidlowski,et al.  Selective Role of Intracellular Chloride in the Regulation of the Intrinsic but Not Extrinsic Pathway of Apoptosis in Jurkat T-cells* , 2006, Journal of Biological Chemistry.

[64]  M. Herlyn,et al.  Normal human melanocyte homeostasis as a paradigm for understanding melanoma. , 2005, The journal of investigative dermatology. Symposium proceedings.

[65]  C. Berking,et al.  Differential expression of melanoma‐associated growth factors in keratinocytes and fibroblasts by ultraviolet A and ultraviolet B radiation , 2005, The British journal of dermatology.

[66]  Pablo Tamayo,et al.  Gene set enrichment analysis: A knowledge-based approach for interpreting genome-wide expression profiles , 2005, Proceedings of the National Academy of Sciences of the United States of America.

[67]  L. Zon,et al.  BRAF Mutations Are Sufficient to Promote Nevi Formation and Cooperate with p53 in the Genesis of Melanoma , 2005, Current Biology.

[68]  T. Hirobe Role of keratinocyte-derived factors involved in regulating the proliferation and differentiation of mammalian epidermal melanocytes. , 2005, Pigment cell research.

[69]  M. Owen,et al.  The GDP-GTP Exchange Factor Collybistin: An Essential Determinant of Neuronal Gephyrin Clustering , 2004, The Journal of Neuroscience.

[70]  F. Szoka,et al.  Chloride Accumulation and Swelling in Endosomes Enhances DNA Transfer by Polyamine-DNA Polyplexes* , 2003, Journal of Biological Chemistry.

[71]  J. Shay,et al.  Bypass of telomere-dependent replicative senescence (M1) upon overexpression of Cdk4 in normal human epithelial cells , 2003, Oncogene.

[72]  Kaori Inoue,et al.  gamma-Aminobutyric acid (A) receptor agonists accelerate cutaneous barrier recovery and prevent epidermal hyperplasia induced by barrier disruption. , 2002, The Journal of investigative dermatology.

[73]  Zhiyuan Gong,et al.  Green fluorescent protein expression in germ‐line transmitted transgenic zebrafish under a stratified epithelial promoter from Keratin8 , 2002, Developmental dynamics : an official publication of the American Association of Anatomists.

[74]  Toshikazu Nakamura,et al.  Keratinocyte expression of transgenic hepatocyte growth factor affects melanocyte development, leading to dermal melanocytosis , 2000, Mechanisms of Development.

[75]  M. Herlyn,et al.  Cadherin repertoire determines partner-specific gap junctional communication during melanoma progression. , 2000, Journal of cell science.

[76]  D. Elder,et al.  E-cadherin expression in melanoma cells restores keratinocyte-mediated growth control and down-regulates expression of invasion-related adhesion receptors. , 2000, The American journal of pathology.

[77]  M. Tuszynski,et al.  Leukemia Inhibitory Factor Augments Neurotrophin Expression and Corticospinal Axon Growth after Adult CNS Injury , 1999, The Journal of Neuroscience.

[78]  Bernhard Lüscher,et al.  Postsynaptic clustering of major GABAA receptor subtypes requires the γ2 subunit and gephyrin , 1998, Nature Neuroscience.

[79]  P. Patterson,et al.  Leukemia inhibitory factor (LIF) mRNA-expressing neuronal subpopulations in adult rat basal forebrain , 1997, Neuroscience Letters.

[80]  T. Yagi,et al.  Cleft palate and decreased brain gamma-aminobutyric acid in mice lacking the 67-kDa isoform of glutamic acid decarboxylase. , 1997, Proceedings of the National Academy of Sciences of the United States of America.

[81]  Austin G Smith,et al.  Essential function of LIF receptor in motor neurons , 1995, Nature.

[82]  D. Elder,et al.  Regulation of Mel-CAM/MUC18 expression on melanocytes of different stages of tumor progression by normal keratinocytes. , 1994, The American journal of pathology.

[83]  R. Halaban Signal transduction in normal and malignant melanocytes. , 1994, Pigment cell research.

[84]  S. Zhang,et al.  GABA-activated chloride channels in secretory nerve endings. , 1993, Science.

[85]  R. Halaban,et al.  Basic fibroblast growth factor from human keratinocytes is a natural mitogen for melanocytes , 1988, The Journal of cell biology.

[86]  J. Hornung,et al.  Normal keratinization in a spontaneously immortalized aneuploid human keratinocyte cell line , 1988, The Journal of cell biology.

[87]  M. Hollstein,et al.  Selective lack of intercellular communication between transformed and nontransformed cells as a common property of chemical and oncogene transformation of BALB/c 3T3 cells. , 1987, Cancer research.

[88]  H. Yamasaki,et al.  Lack of intercellular communication between chemically transformed and surrounding nontransformed BALB/c 3T3 cells. , 1984, Cancer research.

[89]  W. Loewenstein,et al.  Intercellular Communication and the Control of Tissue Growth: Lack of Communication between Cancer Cells , 1966, Nature.

[90]  Thomas R. Gingeras,et al.  STAR: ultrafast universal RNA-seq aligner , 2013, Bioinform..

[91]  D. Elder,et al.  Melanoma: The Wistar Melanoma (WM) Cell Lines , 2002 .

[92]  M. Herlyn,et al.  Dynamics of cell interactions and communications during melanoma development. , 2002, Critical reviews in oral biology and medicine : an official publication of the American Association of Oral Biologists.

[93]  J. Kirsch,et al.  Collybistin, a newly identified brain-specific GEF, induces submembrane clustering of gephyrin , 2000, Nature Neuroscience.

[94]  J. Benson,et al.  Postsynaptic clustering of major GABAA receptor subtypes requires the gamma 2 subunit and gephyrin. , 1998, Nature neuroscience.

[95]  R. Halaban,et al.  Met and hepatocyte growth factor/scatter factor signal transduction in normal melanocytes and melanoma cells. , 1992, Oncogene.

[96]  T. Fitzpatrick,et al.  [THE EPIDERMAL MELANIN UNIT SYSTEM]. , 1963, Dermatologische Wochenschrift.