Phospho-Specific Flow Cytometry Reveals Signaling Heterogeneity in T-Cell Acute Lymphoblastic Leukemia Cell Lines

Perbellini, Omar; Cavallini, Chiara; Chignola, Roberto; Galasso, Marilisa; Scupoli, Maria Teresa

doi:10.3390/cells11132072

Cited by 5 publications

(3 citation statements)

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“…For functional validation of promoter activity in CD4 + cells, we cloned putative CXCR6 promoter regions (chr3:45942118-45946481, promoter 1; chr3:45939898-45941054, promoter 2; all GRCh38/hg38) ( Figure S2a ) into a pGL3-basic reporter vector. Using a luciferase reporter assay, we tested the activity of constructs in an experimental model of T helper cells, the PMA-treated human T-lymphoblastoid Jurkat cell line [ 30 , 31 , 32 ]. Promoter 1, located in the active chromatin region, showed high activity, while promoter 2 was significantly less active ( Figure S2b ).…”

Section: Resultsmentioning

confidence: 99%

rs71327024 Associated with COVID-19 Hospitalization Reduces CXCR6 Promoter Activity in Human CD4+ T Cells via Disruption of c-Myb Binding

Uvarova,

Stasevich,

Ustiugova

et al. 2023

IJMS

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Single-nucleotide polymorphism rs71327024 located in the human 3p21.31 locus has been associated with an elevated risk of hospitalization upon SARS-CoV-2 infection. The 3p21.31 locus contains several genes encoding chemokine receptors potentially relevant to severe COVID-19. In particular, CXCR6, which is prominently expressed in T lymphocytes, NK, and NKT cells, has been shown to be involved in the recruitment of immune cells to non-lymphoid organs in chronic inflammatory and respiratory diseases. In COVID-19, CXCR6 expression is reduced in lung resident memory T cells from patients with severe disease as compared to the control cohort with moderate symptoms. We demonstrate here that rs71327024 is located within an active enhancer that augments the activity of the CXCR6 promoter in human CD4+ T lymphocytes. The common rs71327024(G) variant makes a functional binding site for the c-Myb transcription factor, while the risk rs71327024(T) variant disrupts c-Myb binding and reduces the enhancer activity. Concordantly, c-Myb knockdown in PMA-treated Jurkat cells negates rs71327024’s allele-specific effect on CXCR6 promoter activity. We conclude that a disrupted c-Myb binding site may decrease CXCR6 expression in T helper cells of individuals carrying the minor rs71327024(T) allele and thus may promote the progression of severe COVID-19 and other inflammatory pathologies.

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Section: Resultsmentioning

confidence: 99%

rs71327024 Associated with COVID-19 Hospitalization Reduces CXCR6 Promoter Activity in Human CD4+ T Cells via Disruption of c-Myb Binding

Uvarova,

Stasevich,

Ustiugova

et al. 2023

IJMS

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“…Earlier studies of multiple cancers have documented discrete molecular profiles of cell lines extracted from the same tumor type, suggesting their dissimilar ability to represent the primary tumors (Jiang et al, 2016;Yu et al, 2019). Notably, analysis of the phosphorylation status of ten signaling pathway proteins with phosopho-specific flow cytometry revealed a higher variability in these proteins across three different T-ALL cell lines under both basal and modulated conditions (Perbellini et al, 2022). Subsequently, T-ALL cell lines, despite their widespread use in cancer biology, differ in their tumorigenic potentials.…”

Section: Introductionmentioning

confidence: 96%

New Mechanisms for Anti-Cancer Drugs

2024

Frontiers Research Topics

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Frontiers is more than just an open access publisher of scholarly articles: it is a pioneering approach to the world of academia, radically improving the way scholarly research is managed. The grand vision of Frontiers is a world where all people have an equal opportunity to seek, share and generate knowledge. Frontiers provides immediate and permanent online open access to all its publications, but this alone is not enough to realize our grand goals. Frontiers journal seriesThe Frontiers journal series is a multi-tier and interdisciplinary set of openaccess, online journals, promising a paradigm shift from the current review, selection and dissemination processes in academic publishing. All Frontiers journals are driven by researchers for researchers; therefore, they constitute a service to the scholarly community. At the same time, the Frontiers journal series operates on a revolutionary invention, the tiered publishing system, initially addressing specific communities of scholars, and gradually climbing up to broader public understanding, thus serving the interests of the lay society, too. Dedication to qualityEach Frontiers article is a landmark of the highest quality, thanks to genuinely collaborative interactions between authors and review editors, who include some of the world's best academicians. Research must be certified by peers before entering a stream of knowledge that may eventually reach the public -and shape society; therefore, Frontiers only applies the most rigorous and unbiased reviews. Frontiers revolutionizes research publishing by freely delivering the most outstanding research, evaluated with no bias from both the academic and social point of view. By applying the most advanced information technologies, Frontiers is catapulting scholarly publishing into a new generation. What are Frontiers Research Topics?Frontiers Research Topics are very popular trademarks of the Frontiers journals series: they are collections of at least ten articles, all centered on a particular subject. With their unique mix of varied contributions from Original Research to Review Articles, Frontiers Research Topics unify the most influential researchers, the latest key findings and historical advances in a hot research area.

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“…Earlier studies of multiple cancers have documented discrete molecular profiles of cell lines extracted from the same tumor type, suggesting their dissimilar ability to represent the primary tumors ( Jiang et al, 2016 ; Yu et al, 2019 ). Notably, analysis of the phosphorylation status of ten signaling pathway proteins with phosopho-specific flow cytometry revealed a higher variability in these proteins across three different T-ALL cell lines under both basal and modulated conditions ( Perbellini et al, 2022 ). Subsequently, T-ALL cell lines, despite their widespread use in cancer biology, differ in their tumorigenic potentials.…”

Section: Introductionmentioning

confidence: 99%

CCR9 overexpression promotes T-ALL progression by enhancing cholesterol biosynthesis

Jamal,

Lei,

et al. 2023

Front. Pharmacol.

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Introduction: T-cell acute lymphoblastic leukemia (T-ALL) is an aggressive hematological malignancy of the lymphoid progenitor cells, contributing to ∼ 20% of the total ALL cases, with a higher prevalence in adults than children. Despite the important role of human T-ALL cell lines in understanding the pathobiology of the disease, a detailed comparison of the tumorigenic potentials of two commonly used T-ALL cell lines, MOLT4 and JURKAT cells, is still lacking.Methodology: In the present study, NOD-PrkdcscidIL2rgdull (NTG) mice were intravenously injected with MOLT4, JURKAT cells, and PBS as a control. The leukemiac cell homing/infiltration into the bone marrow, blood, liver and spleen was investigated for bioluminescence imaging, flow cytometry, and immunohistochemistry staining. Gene expression profiling of the two cell lines was performed via RNA-seq to identify the differentially expressed genes (DEGs). CCR9 identified as a DEG, was further screened for its role in invasion and metastasis in both cell lines in vitro. Moreover, a JURKAT cell line with overexpressed CCR9 (Jurkat-OeCCR9) was investigated for T-ALL formation in the NTG mice as compared to the GFP control. Jurkat-OeCCR9 cells were then subjected to transcriptome analysis to identify the genes and pathways associated with the upregulation of CCR9 leading to enhanced tumirogenesis. The DEGs of the CCR9-associated upregulation were validated both at mRNA and protein levels. Simvastatin was used to assess the effect of cholesterol biosynthesis inhibition on the aggressiveness of T-ALL cells.Results: Comparison of the leukemogenic potentials of the two T-ALL cell lines showed the relatively higher leukemogenic potential of MOLT4 cells, characterized by their enhanced tissue infiltration in NOD-PrkdcscidIL2rgdull (NTG) mice. Transcriptmoe analysis of the two cell lines revealed numerous DEGs, including CCR9, enriched in vital signaling pathways associated with growth and proliferation. Notably, the upregulation of CCR9 also promoted the tissue infiltration of JURKAT cells in vitro and in NTG mice. Transcriptome analysis revealed that CCR9 overexpression facilitated cholesterol production by upregulating the expression of the transcriptional factor SREBF2, and the downstream genes: MSMO1, MVD, HMGCS1, and HMGCR, which was then corroborated at the protein levels. Notably, simvastatin treatment reduced the migration of the CCR9-overexpressing JURKAT cells, suggesting the importance of cholesterol in T-ALL progression.Conclusions: This study highlights the distinct tumorigenic potentials of two T-ALL cell lines and reveals CCR9-regulated enhanced cholesterol biosynthesis in T-ALL.

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Phospho-Specific Flow Cytometry Reveals Signaling Heterogeneity in T-Cell Acute Lymphoblastic Leukemia Cell Lines

Cited by 5 publications

References 82 publications

rs71327024 Associated with COVID-19 Hospitalization Reduces CXCR6 Promoter Activity in Human CD4+ T Cells via Disruption of c-Myb Binding

rs71327024 Associated with COVID-19 Hospitalization Reduces CXCR6 Promoter Activity in Human CD4+ T Cells via Disruption of c-Myb Binding

New Mechanisms for Anti-Cancer Drugs

CCR9 overexpression promotes T-ALL progression by enhancing cholesterol biosynthesis

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