Antimelanoma Effects of Concomitant Inhibition of SIRT1 and SIRT3 in Braf/Pten Mice

Chhabra, Gagan; Singh, Chandra K.; Guzmán-Pérez, Glorimar; Ndiaye, Mary A.; Iczkowski, Kenneth A.; Ahmad, Nihal

doi:10.1016/j.jid.2021.08.434

Cited by 5 publications

(4 citation statements)

References 112 publications

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“…The Tyr-CreER.Braf Ca .Pten fl/fl mice were provided by Martin McMahon of UCSF to Duke Cancer Institute, and induced as previously described ( 40 , 42 ). Briefly, the back skins of 5-7 weeks old male and female animals were shaved and the center of the shaved region was treated with 3 topical applications of 1.5 µl of 5 mM 4-hydroxy-tamoxifen (4-OHT, dissolved in 99% ethanol, Sigma, St Louis, MO, USA) spaced at 1-day intervals ( 43 , 44 ). Pigmented lesions became visible 9-13 days after the first 4-OHT application.…”

Section: Methodsmentioning

confidence: 99%

Co-Treatment of Chloroquine and Trametinib Inhibits Melanoma Cell Proliferation and Decreases Immune Cell Infiltration

Degan

May²,

Jin³

et al. 2022

Front. Oncol.

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Autophagy is characterized as a cytoprotective process and inhibition of autophagy with medicinally active agents, such as chloroquine (CQ) is proposed as a prospective adjuvant therapy for cancer. Here, we examined the preclinical effects of CQ combined with the MEK inhibitor trametinib (TRA) on melanoma. We found that cotreatment of CQ and TRA markedly slowed melanoma growth induced in Tyr-CreER.BrafCa.Ptenfl/fl mice. Immunostaining showed that trametinib decreased Ki-67+ proliferating cells, and increased TUNEL+ apoptotic cells. The combo treatment induced a further decrease of Ki-67+ proliferating cells. Consistent with the in vivo findings, CQ and TRA inhibited melanoma cell proliferation in vitro, which was correlated by decreased cyclin D1 expression. In addition, we found that tissues treated with CQ and TRA had significantly decreased numbers of CD4+ and CD8+ T-lymphocytes and F4/80+ macrophages. Together, these results indicate that cotreatment of CQ and TRA decreases cancer cell proliferation, but also dampens immune cell infiltration. Further study is warranted to understand whether CQ-induced immune suppression inadvertently affects therapeutic benefits.

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Section: Methodsmentioning

confidence: 99%

Co-Treatment of Chloroquine and Trametinib Inhibits Melanoma Cell Proliferation and Decreases Immune Cell Infiltration

Degan

May²,

Jin³

et al. 2022

Front. Oncol.

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“…Primary antibodies against PLK1 (#4513, 1:20), NUMB (#2756, 1:40), NOTCH1 (NICD) (#3608, 1:50), E-cadherin (#3195, 1:20), N-cadherin (#13116, 1:50X), Vimentin (#5741, 1:50), ZEB1 (#3396, 1:100), and SNAIL (#3879, 1:50) were purchased from Cell Signaling Technologies (CST). Immunoblotting was performed by capillary-based Simple Western method using the ProteinSimple Jess system, employing a 12-230 KDa Separation Module and anti-rabbit secondary antibody per manufacturer instructions and as previously described 44 . The primary antibody against pS413-NUMB was purchased from GenScript as a custom antibody development order and was used in traditional western blotting to analyze NUMB phosphorylation at S413 after in-vitro kinase assay with PLK1.…”

Section: Methodsmentioning

confidence: 99%

Role of PLK1/NUMB/NOTCH in epithelial-mesenchymal transition in human melanoma

Chhabra,

Singh,

Ndiaye

et al. 2024

npj Precis. Onc.

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Polo-like kinase 1 (PLK1), a serine/threonine kinase, is overexpressed in melanoma and its expression has been associated with poor disease prognosis. PLK1 has been shown to interact with NUMB, a NOTCH antagonist. However, the exact role of PLK1, NUMB, and NOTCH signaling in epithelial-mesenchymal transition (EMT) in melanoma progression is unclear. In this study, Affymetrix microarray analysis was performed to determine differentially expressed genes following shRNA-mediated knockdown of PLK1 in human melanoma cells that showed significant modulations in EMT and metastasis-related genes. Using multiple PLK1-modulated melanoma cell lines, we found that PLK1 is involved in the regulation of cell migration, invasion, and EMT via its kinase activity and NOTCH activation. In vitro kinase assay and mass spectrometry analysis demonstrated a previously unknown PLK1 phosphorylation site (Ser413) on NUMB. Overexpression of non-phosphorylatable (S413A) and phosphomimetic (S413D) mutants of NUMB in melanoma cells implicated the involvement of NUMB-S413 phosphorylation in cell migration and invasion, which was independent of NOTCH activation. To determine the clinical relevance of these findings, immunohistochemistry was performed using melanoma tissue microarray, which indicated a strong positive correlation between PLK1 and N-cadherin, a protein required for successful EMT. These findings were supported by TCGA analysis, where expression of high PLK1 with low NUMB or high NOTCH or N-cadherin showed a significant decrease in survival of melanoma patients. Overall, these results suggest a potential role of PLK1 in EMT, migration, and invasion of melanoma cells. Our findings support the therapeutic targeting of PLK1, NUMB, and NOTCH for melanoma management.

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“…Protein lysates from two mice were pooled in each group to make three groupings of protein for further analysis. The quantitative immunodetection by ProteinSimple was performed as described recently ( 25 ). Primary antibodies used in ProteinSimple analyses are detailed in Supplementary Table S1 .…”

Section: Methodsmentioning

confidence: 99%

“…described recently (25). Primary antibodies used in ProteinSimple analyses are detailed in Supplementary Table S1.…”

Section: Quantitative Immuno-detection Analyses Using Jess Proteinsimplementioning

confidence: 99%

Protective effects of dietary grape against atopic dermatitis-like skin lesions in NC/NgaTndCrlj mice

Singh

Mintie²,

Ndiaye³

et al. 2023

Front. Immunol.

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Atopic dermatitis (AD) is a chronic inflammatory skin disease with significant health/economic burdens. Existing therapies are not fully effective, necessitating development of new approaches for AD management. Here, we report that dietary grape powder (GP) mitigates AD-like symptoms in 2,4-dinitrofluorobenzene (DNFB)-induced AD in NC/NgaTndCrlj mice. Using prevention and intervention protocols, we tested the efficacy of 3% and 5% GP-fortified diet in a 13-weeks study. We found that GP feeding markedly inhibited development and progression of AD-like skin lesions, and caused reduction in i) epidermal thickness, mast cell infiltration, ulceration, excoriation and acanthosis in dorsal skin, ii) spleen weight, extramedullary hematopoiesis and lymph nodes sizes, and iii) ear weight and IgE levels. We also found significant modulations in 15 AD-associated serum cytokines/chemokines. Next, using quantitative global proteomics, we identified 714 proteins. Of these, 68 (normal control) and 21 (5% GP-prevention) were significantly modulated (≥2-fold) vs AD control (DNFB-treated) group, with many GP-modulated proteins reverting to normal levels. Ingenuity pathway analysis of GP-modulated proteins followed by validation using ProteinSimple identified changes in acute phase response signaling (FGA, FGB, FGG, HP, HPX, LRG1). Overall, GP supplementation inhibited DNFB-induced AD in NC/NgaTndCrlj mice in both prevention and intervention trials, and should be explored further.

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Antimelanoma Effects of Concomitant Inhibition of SIRT1 and SIRT3 in Braf/Pten Mice

Cited by 5 publications

References 112 publications

Co-Treatment of Chloroquine and Trametinib Inhibits Melanoma Cell Proliferation and Decreases Immune Cell Infiltration

Co-Treatment of Chloroquine and Trametinib Inhibits Melanoma Cell Proliferation and Decreases Immune Cell Infiltration

Role of PLK1/NUMB/NOTCH in epithelial-mesenchymal transition in human melanoma

Protective effects of dietary grape against atopic dermatitis-like skin lesions in NC/NgaTndCrlj mice

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