Toll-like receptor 9 signaling in chronic lymphocytic leukemia cell lines
Ontology highlight
ABSTRACT:
ORGANISM(S): Homo sapiens
PROVIDER: GSE233595 | GEO | 2023/09/19
REPOSITORIES: GEO
Similar Datasets
Project description:Chronic Lymphocytic Leukemia (CLL) is strictly dependent on the complex interplay between the intrinsic features of the leukemic cells and microenvironmental stimulations including inflammatory stimuli by Toll Like Receptors (TLR) which protect CLL cells from drug induced apoptosis by upregulating NFKBIZ, an atypical co-transcription factor. To face the challenge of targeting transcription factors, we exploited the capacity of Dimethyl Itaconate (DI), an electrophilic synthetic derivative of Itaconate, to block NFKBIZ acting as anti-inflammatory agent. Primary CLL cells isolated from the peripheral blood of patients, leukemic splenocytes isolated from TCL1 transgenic mice, and circulating leukocytes isolated from healthy donors were treated in vitro with increasing concentrations of DI either alone or in combination with the TLR ligands. DI abrogated the induction of NFKBIZ as well as its target genes IL6 and IL10. Remarkably, DI reduced metabolic activation and cell viability of leukemic cells even if added after a robust TLR pre-stimulation; in contrast, no toxic effect was evident in normal leukocytes including T- and B-lymphocytes. DI induced apoptosis of malignant cells by reducing BCL-XL and MCL1 levels while inducing PARP cleavage. RNA sequencing highlighted that DI abrogated the TLR9-mediated upregulation of transcripts related to the metabolism of rRNAs, interferon and cytokine signaling. Notably, in addition to the expected electrophilic stress signature observed after DI treatment, novel pathways emerged including the downregulation of distinct MHC class II complex genes. In conclusion, DI not only abrogated the pro-inflammatory effects of TLR stimulation but also targeted a specific vulnerability in CLL cells.
2023-09-19 | GSE226904 | GEO
Project description:Chronic Lymphocytic Leukemia (CLL) is strictly dependent on the complex interplay between the intrinsic features of the leukemic cells and microenvironmental stimulations including inflammatory stimuli by Toll Like Receptors (TLR) which protect CLL cells from drug induced apoptosis by upregulating NFKBIZ, an atypical co-transcription factor. To face the challenge of targeting transcription factors, we exploited the capacity of Dimethyl Itaconate (DI), an electrophilic synthetic derivative of Itaconate, to block NFKBIZ acting as anti-inflammatory agent. Primary CLL cells isolated from the peripheral blood of patients, leukemic splenocytes isolated from TCL1 transgenic mice, and circulating leukocytes isolated from healthy donors were treated in vitro with increasing concentrations of DI either alone or in combination with the TLR ligands. DI abrogated the induction of NFKBIZ as well as its target genes IL6 and IL10. Remarkably, DI reduced metabolic activation and cell viability of leukemic cells even if added after a robust TLR pre-stimulation; in contrast, no toxic effect was evident in normal leukocytes including T- and B-lymphocytes. DI induced apoptosis of malignant cells by reducing BCL-XL and MCL1 levels while inducing PARP cleavage. RNA sequencing highlighted that DI abrogated the TLR9-mediated upregulation of transcripts related to the metabolism of rRNAs, interferon and cytokine signaling. Notably, in addition to the expected electrophilic stress signature observed after DI treatment, novel pathways emerged including the downregulation of distinct MHC class II complex genes. In conclusion, DI not only abrogated the pro-inflammatory effects of TLR stimulation but also targeted a specific vulnerability in CLL cells.
2023-09-19 | GSE210972 | GEO
Project description:Transcription profiling of human bone marrow mesenchymal stem cells treated with exosomes isolated from chronic lymphocytic leukemia cell line MEC-1 supernatant. Cells were left untreated or treated with CLL-exosomes for 6h at 37C.
2015-06-21 | E-MTAB-2765 | biostudies-arrayexpress
Project description:To engineer novel SF3B1 mutant cell lines, we developed a methodology called PRECIS to install the pathogenic K700E mutation. We used this method to engineer CLL cell lines HG-3 and MEC-1 with the SF3B1 K700E mutation and then carried out RNA-seq.
2024-08-28 | GSE231387 | GEO
Project description:This work shows that signaling-lymphocytic-activation-molecule-1 (SLAMF1), a co-stimulatory molecule and a microbial sensor, is expressed by normal CD19+/CD5+ B-lymphocytes. Its expression is lost in a subset of patients with chronic lymphocytic leukemia (CLL) characterized by an aggressive form of the disease, with shorter time to first treatment and overall survival. Silencing of SLAMF1 in the CLL-like Mec-1 cell line (constitutively SLAMF1+) modulated pathways connected to cell migration, cytoskeletal organization and intracellular vesicle formation/recirculation. Loss of SLAMF1 was associated to increased expression of CXCR4, CD38 and CD44, positively affecting chemotactic responses to CXCL12. Ligation of SLAMF1 with an agonistic monoclonal antibody promoted the autophagic flux, by increasing reactive oxygen species (ROS) accumulation and inducing phosphorylation of p38, JNK1/2 and bcl-2. The direct consequence was the assembly of the autophagy macro-complex that included SLAMF1, the scaffold protein Beclin1 and the enzyme Vps34. Consistently, SLAMF1-silenced cells or SLAMF1low primary CLL cells were resistant to autophagy-activating therapeutic agents, such as fludarabine or the BH3 mimetic ABT-737. These results indicate that loss of SLAMF1 expression modulates genetic pathways regulating chemotaxis and autophagy and potentially affecting drug responses, thus providing a likely explanation for the unfavorable clinical outcome experienced by this patient subset. Microarrays were performed comparing genetic signature of CLL-like Mec-1 cell line (constitutively SLAMF1+), transfected with a non-effective (scrambled) shRNA cassette (TR30013) used as control, and a Mec-1 variant generated silencing SLAMF1 gene using pGFP-V-RS construct for SLAMF1 shRNA (TG309422). For each cell line variants, grown in vitro in complete medium, three biological replicates were included in the chip.
2015-05-23 | E-GEOD-69165 | biostudies-arrayexpress
Project description:In the activated B-cell-like (ABC) subtype of diffuse large B cell lymphoma (DLBCL), the most frequent gain-of-function mutations target MyD88, a signaling adapter for Tolllike receptors (TLRs). The most prevalent oncogenic mutant, MyD88 L265P, occurs in 29% of cases and is the most active in engaging the NF-kappaB pathway. Here we show that MyD88 mutants do not function autonomously, but rather require TLR7, TLR9, and to a lesser extent, TLR4 to promote the survival of ABC DLBCL cells. Unlike wild type MyD88, MyD88 mutants associate constitutively with TLR7 and TLR9 in ABC DLBCL cells. Like ligand-induced TLR7/9 signaling in normal immune cells, the survival of ABC DLBCL cell lines depends upon translocation of TLR7 and TLR9 to acidic endolysosomes, where proteolytic processing of their ligand binding ectodomains is required for their oncogenic signaling. ABC DLBCL viability also depends upon CD14, a co-receptor for TLR7 and TLR9 that promotes engagement of nucleic acid ligands by these receptors. Point mutations in the TLR7 or TLR9 ectodomains that abrogate ligand binding and/or signaling were incapable of sustaining ABC DLBCL survival. An inhibitory oligonucleotide that suppresses TLR9 responses in normal B cells blocked NF-kappaB signaling and survival of ABC DLBCL lines. Together, these data suggest that an endogenous TLR ligand may play a pathogenic role in ABC DLBCL and provide a rationale for targeting TLR signaling to improve therapy of this aggressive lymphoma. Gene expression was analyzed using Agilent human 2-color 4X44K oligo gene expression arrays. Cell line, TMD8 ABC-DLBCL, was infected with control (shControl, Cy3), shLTR7 (Cy5) or shLTR9 (Cy5) and changes in gene expression were monitored on day 1 and day 2 after induction of the shRNA with doxycycline, co-hybridizing control and experimental samples (Cy3+Cy5), for a total of 4 arrays.
2014-05-21 | E-GEOD-47147 | biostudies-arrayexpress
Project description:In the activated B-cell-like (ABC) subtype of diffuse large B cell lymphoma (DLBCL), the most frequent gain-of-function mutations target MyD88, a signaling adapter for Tolllike receptors (TLRs). The most prevalent oncogenic mutant, MyD88 L265P, occurs in 29% of cases and is the most active in engaging the NF-kappaB pathway. Here we show that MyD88 mutants do not function autonomously, but rather require TLR7, TLR9, and to a lesser extent, TLR4 to promote the survival of ABC DLBCL cells. Unlike wild type MyD88, MyD88 mutants associate constitutively with TLR7 and TLR9 in ABC DLBCL cells. Like ligand-induced TLR7/9 signaling in normal immune cells, the survival of ABC DLBCL cell lines depends upon translocation of TLR7 and TLR9 to acidic endolysosomes, where proteolytic processing of their ligand binding ectodomains is required for their oncogenic signaling. ABC DLBCL viability also depends upon CD14, a co-receptor for TLR7 and TLR9 that promotes engagement of nucleic acid ligands by these receptors. Point mutations in the TLR7 or TLR9 ectodomains that abrogate ligand binding and/or signaling were incapable of sustaining ABC DLBCL survival. An inhibitory oligonucleotide that suppresses TLR9 responses in normal B cells blocked NF-kappaB signaling and survival of ABC DLBCL lines. Together, these data suggest that an endogenous TLR ligand may play a pathogenic role in ABC DLBCL and provide a rationale for targeting TLR signaling to improve therapy of this aggressive lymphoma.
2014-05-21 | GSE47147 | GEO
Project description:Purpose: Nucleolar and spindle associated protein 1 (NUSAP1), serves as a microtubule binding protein in chromosome separation, spindle assembly, and plays significant role to ensure normal regulation of cell cycle as well. This experiment aimed to document the NUSAP1 expression and functions in chronic lymphocytic leukemia (CLL). Methods: Lentivirus vectors either encoding shNUSAP1 or empty lentiviral vector were stably transfected into MEC-1 cells. 3 shNUSAP1 transfected and 3 shControl transfected MEC-1 cell samples were performed RNA sequencing (RNA-seq) analysis, functional enrichment analyses of gene ontology (GO) and gene set enrichment analysis (GSEA). Results: Silencing NUSAP1 inhibited the cell proliferation, promoted cell apoptosis and G0/G1 phase arrest. Mechanistically, high expression of NUSAP1 strengthened DNA damage repairing with RAD51 engagement. Our results also indicated that NUSAP1 knockdown suppressed the growth of CLL cells in vivo. Conclusion: Our research investigates the mechanism by which NUSAP1 enhances chemoresistance via DNA damage repair signaling by stabilizing RAD51 in CLL cells. Hence, NUSAP1 may be expected to be a perspective target for the treatment of CLL with chemotherapy resistance.
2021-10-08 | GSE185467 | GEO
Project description:Genomic studies have recently identified RPS15 as a new driver gene in aggressive and chemorefractory cases of chronic lymphocytic leukemia (CLL). RPS15 encodes a ribosomal protein whose conserved C-terminal domain extends into the decoding center of the ribosome. We demonstrate that mutations in highly conserved residues of this domain affect both protein stability —by increasing its ubiquitin-mediated degradation— and cell proliferation rates. On the other hand, we show that mutated RPS15 can be loaded into the ribosomes, directly impacting on global protein synthesis and/or translational fidelity in a mutation-specific manner. Quantitative mass-spectrometry analyses suggest that RPS15 variants may induce additional alterations in the translational machinery as well as a metabolic shift at the proteome level in both HEK293T and MEC-1 cells. These results indicate that CLL-related RPS15 mutations might act following patterns known for other ribosomal diseases, likely switching from a hypo- to a hyperproliferative phenotype driven by mutated ribosomes. In this scenario, loss of translational fidelity causing altered cell proteostasis can be proposed as a new molecular mechanism involved in CLL pathobiology.
2019-09-09 | PXD010924 | Pride
Project description:B cell chronic lymphocytic leukemia - A model with immune response
Seema Nanda 1, , Lisette dePillis 2, and Ami Radunskaya 3,
1.
Tata Institute of Fundamental Research, Centre for Applicable Mathematics, Bangalore 560065, India
2.
Department of Mathematics, Harvey Mudd College, Claremont, CA 91711
3.
Department of Mathematics, Pomona College, Claremont, CA, 91711, United States
Abstract
B cell chronic lymphocytic leukemia (B-CLL) is known to have substantial clinical heterogeneity. There is no cure, but treatments allow for disease management. However, the wide range of clinical courses experienced by B-CLL patients makes prognosis and hence treatment a significant challenge. In an attempt to study disease progression across different patients via a unified yet flexible approach, we present a mathematical model of B-CLL with immune response, that can capture both rapid and slow disease progression. This model includes four different cell populations in the peripheral blood of humans: B-CLL cells, NK cells, cytotoxic T cells and helper T cells. We analyze existing data in the medical literature, determine ranges of values for parameters of the model, and compare our model outcomes to clinical patient data. The goal of this work is to provide a tool that may shed light on factors affecting the course of disease progression in patients. This modeling tool can serve as a foundation upon which future treatments can be based.
Keywords: NK cell, chronic lymphocytic leukemia, mathematical model, T cell., B-CLL.
2020-01-09 | MODEL2001090002 | BioModels