A Proteomics Approach Identifies RREB1 as a Crucial Molecular Target of Imidazo–Pyrazole Treatment in SKMEL-28 Melanoma Cells

International Journal of Molecular Sciences, Journal Year: 2024, Volume and Issue: 25(12), P. 6760 - 6760

Published: June 20, 2024

Cutaneous melanoma is the most dangerous and deadly form of human skin malignancy. Despite its rarity, it accounts for a staggering 80% deaths attributed to cutaneous cancers overall. Moreover, final stages often exhibit resistance drug treatments, resulting in unfavorable outcomes. Hence, ensuring access novel improved chemotherapeutic agents imperative patients grappling with this severe ailment. Pyrazole fused systems derived thereof are heteroaromatic moieties widely employed medicinal chemistry develop effective drugs various therapeutic areas, including inflammation, pain, oxidation, pathogens, depression, fever. In previous study, we described biochemical properties newly synthesized group imidazo-pyrazole compounds. paper, improve our knowledge pharmacological these molecules, conduct differential proteomic analysis on cell line treated one derivatives. Our results detail changes SKMEL-28 proteome induced by 24, 48, 72 h

Language: Английский

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Fate mapping melanoma persister cells through regression and into recurrent disease in adult zebrafish

Disease Models & Mechanisms, Journal Year: 2022, Volume and Issue: 15(9)

Published: Aug. 5, 2022

Melanoma heterogeneity and plasticity underlie therapy resistance. Some tumour cells possess innate resistance, while others reprogramme during drug exposure survive to form persister cells, a source of potential cancer for recurrent disease. Tracing individual melanoma cell populations through regression into disease remains largely unexplored, in part, because complex animal models are required live imaging over time. Here, we applied tamoxifen-inducible creERt2/loxP lineage tracing zebrafish model MITF-dependent recurrence image trace vivo stages. Using this strategy, show that at the minimal residual site originate from primary tumour. Next, fate mapped rare MITF-independent demonstrate these directly contribute progressive Multiplex immunohistochemistry confirmed give rise Mitfa+ Taken together, our work reveals direct contribution disease, provides resource lineage-tracing methodology adult models.

Language: Английский

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Aldh2 is a lineage-specific metabolic gatekeeper in melanocyte stem cells

Development, Journal Year: 2022, Volume and Issue: 149(10)

Published: April 29, 2022

Melanocyte stem cells (McSCs) in zebrafish serve as an on-demand source of melanocytes during growth and regeneration, but metabolic programs associated with their activation regenerative processes are not well known. Here, using live imaging coupled scRNA-sequencing, we discovered that, quiescent McSCs activate a dormant embryonic neural crest transcriptional program followed by aldehyde dehydrogenase (Aldh) 2 switch to generate progeny. Unexpectedly, although ALDH2 is known for its aldehyde-clearing mechanisms, find regenerating McSCs, Aldh2 activity required formate - the one-carbon (1C) building block nucleotide biosynthesis through formaldehyde metabolism. Consequently, that disrupting 1C cycle low doses methotrexate causes melanocyte regeneration defects. In absence Aldh2, purines end product sufficient activated Together, our work reveals undergo two-step cell state transition reaction products enzymes have tissue-specific functions meet demands regeneration.

Language: Английский

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Specific oncogene activation of the cell of origin in mucosal melanoma

bioRxiv (Cold Spring Harbor Laboratory), Journal Year: 2024, Volume and Issue: unknown

Published: April 26, 2024

Abstract Mucosal melanoma (MM) is a deadly cancer derived from mucosal melanocytes. To test the consequences of MM genetics, we developed zebrafish model in which all melanocytes experienced CCND1 expression and loss PTEN TP53. Surprisingly, only lining internal organs, analogous to location patient MM. We found that MMs had unique chromatin landscape cutaneous melanoma. Internal could be labeled using MM-specific transcriptional enhancer. Normal shared gene signature with MMs. Patient have increased migratory neural crest decreased antigen presentation expression, consistent metastatic behavior immunotherapy sensitivity Our work suggests cell state originating melanocyte influences melanomas. animal phenotypically transcriptionally mimics tumors, allowing this used for therapeutic discovery.

Language: Английский

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SASH1 S519N Variant Links Skin Hyperpigmentation and Premature Hair Graying to Dysfunction of Melanocyte Lineage

Journal of Investigative Dermatology, Journal Year: 2024, Volume and Issue: 145(1), P. 144 - 154.e3

Published: June 6, 2024

A better understanding of human melanocyte (MC) and stem cell (McSC) biology is essential for treating melanocyte-related diseases. This study employed an inherited pigmentation disorder carrying the SASH1S519N variant in a Hispanic family to investigate SASH1 function MC lineage underlying mechanism this disorder. We used multidisciplinary approach, including clinical exams, assays, yeast two-hybrid screening, biochemical techniques. Results linked early hair graying variant, previously unrecognized phenotype hyperpigmentation disorders. In vitro, we identified as regulator McSC maintenance discovered that TNKS2 crucial SASH1's role. Additionally, S519N located one multiple tankyrase-binding motifs alters binding kinetics affinity interaction. summary, links both gain loss same individual, hinting accelerated aging McSC. The findings offer insights into roles molecular mechanisms propose comprehensive evaluation patients with MC-related disorders should include assessment history loss.

Language: Английский

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Dynamic regulation of chromatin accessibility during melanocyte stem cell activation

Pigment Cell & Melanoma Research, Journal Year: 2023, Volume and Issue: 36(6), P. 531 - 541

Published: July 18, 2023

Abstract Melanocyte stem cells (McSCs) of the hair follicle are necessary for pigmentation and can serve as melanoma origin when harboring cancer‐driving mutations. McSCs be released from quiescence, activated, undergo differentiation into pigment‐producing melanocytes during cycle or due to environmental stimuli, such ultraviolet‐B (UVB) exposure. However, our current understanding mechanisms regulating McSC stemness, activation, remains limited. Here, capture differing possible states in which murine exist, we sorted melanocyte nuclei quiescent (telogen) skin, skin actively producing shafts (anagen), exposed UVB. With these nuclei, then utilized single‐nucleus assay transposase‐accessible chromatin with high‐throughput sequencing (snATAC‐seq) characterized three lineages: (qMcSCs), activated (aMcSCs), differentiated (dMCs) that co‐exist all conditions. Furthermore, successfully identified differentially accessible genes enriched transcription factor binding motifs each lineage. Our findings reveal potential gene regulators determine cell provide new insights how aMcSC regulated differently under divergent intrinsic extrinsic cues. We also a publicly available online tool user‐friendly interface explore this comprehensive dataset, will resource further studies on regulation upon natural UVB‐mediated activation.

Language: Английский

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Pigment cell progenitor heterogeneity and reiteration of developmental signaling underlie melanocyte regeneration in zebrafish

eLife, Journal Year: 2023, Volume and Issue: 12

Published: April 6, 2023

Tissue-resident stem and progenitor cells are present in many adult organs, where they important for organ homeostasis repair response to injury. However, the signals that activate these mechanisms governing how renew or differentiate highly context-dependent incompletely understood, particularly non-hematopoietic tissues. In skin, melanocyte responsible replenishing mature pigmented melanocytes. mammals, reside hair follicle bulge bulb niches activated during homeostatic turnover following destruction, as occurs vitiligo other skin hypopigmentation disorders. Recently, we identified progenitors zebrafish skin. To elucidate renewal differentiation analyzed individual transcriptomes from thousands of lineage regeneration process. We transcriptional signatures progenitors, deciphered changes intermediate cell states regeneration, cell–cell signaling discover regeneration. KIT via RAS/MAPK pathway a regulator direct asymmetric division. Our findings show activation different subpopulations mitfa -positive underlies cellular transitions required properly reconstitute pigmentary system

Language: Английский

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Generation of tamoxifen‐inducible Tfap2b‐CreER^T2 mice using CRISPR‐Cas9

genesis, Journal Year: 2023, Volume and Issue: 62(1)

Published: Dec. 8, 2023

Summary Tfap2b, a pivotal transcription factor, plays critical roles within neural crest cells and their derived lineage. To unravel the intricate lineage dynamics contribution of these Tfap2b+ during craniofacial development, we established Tfap2b‐CreER T2 knock‐in transgenic mouse line using CRISPR‐Cas9‐mediated homologous direct repair. By breeding with tdTomato reporter mice initiating Cre activity through tamoxifen induction at distinct developmental time points, show Tfap2b key crest‐derived domains, such as facial mesenchyme, midbrain, cerebellum, spinal cord, limbs. Notably, migratory neurons stemming from dorsal root ganglia are visible subsequent to initiated E8.5. Intriguingly, cells, serving progenitors for limb predominantly commencing E10.5. Across landscape, exhibits widespread presence throughout organs. Here validate its role marker in tooth development have confirmed that this process initiates E12.5. Our study not only validates line, but also provides powerful tool tracing genetic targeting ‐expressing progenitor temporally spatially regulated manner organogenesis.

Language: Английский

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Mgat4b mediated selective N-glycosylation regulates melanocyte development and melanoma progression

bioRxiv (Cold Spring Harbor Laboratory), Journal Year: 2024, Volume and Issue: unknown

Published: Oct. 11, 2024

Abstract Melanocyte development involves key pathways that are often recapitulated during melanoma initiation, highlighting the importance of understanding regulators control these early processes and also contribute to cancer onset. Our study identifies mgat4b , a glycosyl transferase involved in selective N -glycan branching enriched pigment progenitors, as regulator directional melanocyte migration establishment stem cell (McSC) pool development. Single RNA (scRNA) sequencing analysis zebrafish upon targeted disruption reveals, migratory progenitors marked by galectin expression fail persist. Lectin affinity proteomic reveals glycosylation proteins GPNMB, KIT, TYRP1 be under MGAT4B melanocytic cells. Additionally, mislocalization Junctional plakoglobin (JUP) explains observed defects adhesion regulated but not its isozyme MGAT4A. meta-analysis further patients with both BRAF V600E mutation elevated levels have significantly worse survival outcomes compared those only mutation. By leveraging MAZERATI platform model driver vivo we show mutant cells aggregate initiate tumors. profiling transformed melanocytes revealed cell-cell junction, ECM binding probable contributing factors resulted failure tumor Using small-molecule inhibitor demonstrate inhibitory role this complex -glycosylation progression early-stage melanoma. underscores suggesting promising therapeutic target for treatment.

Language: Английский

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An attractor state zone precedes neural crest fate in melanoma initiation

bioRxiv (Cold Spring Harbor Laboratory), Journal Year: 2024, Volume and Issue: unknown

Published: Oct. 25, 2024

The field cancerization theory suggests that a group of cells containing oncogenic mutations are predisposed to transformation 1, 2 . We previously identified single in BRAF V600E ;p53 -/- zebrafish reactivate an embryonic neural crest state before initiating melanoma 3–5 Here we show the fate from within large fields adjacent abnormal melanocytes, which term “cancer precursor zone.” These cancer zone melanocytes have aberrant morphology, dysplastic nuclei, and altered gene expression. Using cell RNA-seq ATAC-seq, defined distinct transcriptional attractor for zones validated stage-specific expression initiation signatures human melanoma. identify driver, ID1, binds TCF12 inhibits downstream targets important maintenance melanocyte morphology cycle control. Examination patient samples revealed expressing often surrounding invasive melanoma, indicating role ID1 early melanomagenesis. This work reveals surprising effect vivo tumors arise morphologically distinct, but clinically covert, precursors with fate. Our studies novel could improve diagnosis prevention

Language: Английский

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