RESUMO
EBF1 and PAX5 mutations are associated with the development of B progenitor acute lymphoblastic leukemia (B-ALL) in humans. To understand the molecular networks driving leukemia in the Ebf1+/-Pax5+/- (dHet) mouse model for B-ALL, we interrogated the transcriptional profiles and chromatin status of leukemic cells, preleukemic dHet pro-B, and wild-type pro-B cells with the corresponding EBF1 and Pax5 cistromes. In dHet B-ALL cells, many EBF1 and Pax5 target genes encoding pre-BCR signaling components and transcription factors were down-regulated, whereas Myc and genes downstream from IL-7 signaling or associated with the folate pathway were up-regulated. We show that blockade of IL-7 signaling in vivo and methotrexate treatment of leukemic cells in vitro attenuate the expansion of leukemic cells. Single-cell RNA-sequencing revealed heterogeneity of leukemic cells and identified a subset of wild-type pro-B cells with reduced Ebf1 and enhanced Myc expression that show hallmarks of dHet B-ALL cells. Thus, EBF1 and Pax5 may safeguard early stage B cells from transformation to B-ALL by limiting IL-7 signaling, folate metabolism and Myc expression.
Assuntos
Ácido Fólico/metabolismo , Interleucina-7/fisiologia , Fator de Transcrição PAX5/metabolismo , Leucemia-Linfoma Linfoblástico de Células Precursoras/fisiopatologia , Proteínas Proto-Oncogênicas c-myc/genética , Transdução de Sinais/genética , Transativadores/metabolismo , Animais , Carbono/metabolismo , Sobrevivência Celular/genética , Transformação Celular Neoplásica/genética , Modelos Animais de Doenças , Regulação Neoplásica da Expressão Gênica/genética , Redes Reguladoras de Genes/genética , Camundongos , Fator de Transcrição PAX5/genética , Leucemia-Linfoma Linfoblástico de Células Precursoras/genética , Células Precursoras de Linfócitos B/patologia , Ligação Proteica , Análise de Célula Única , Transativadores/genéticaRESUMO
While extended loop extrusion across the entire Igh locus controls VH -DJH recombination, local regulatory sequences, such as the PAIR elements, may also activate VH gene recombination in pro-B-cells. Here, we show that PAIR-associated VH 8 genes contain a conserved putative regulatory element (V8E) in their downstream sequences. To investigate the function of PAIR4 and its V8.7E, we deleted 890 kb containing all 14 PAIRs in the Igh 5' region, which reduced distal VH gene recombination over a 100-kb distance on either side of the deletion. Reconstitution by insertion of PAIR4-V8.7E strongly activated distal VH gene recombination. PAIR4 alone resulted in lower induction of recombination, indicating that PAIR4 and V8.7E function as one regulatory unit. The pro-B-cell-specific activity of PAIR4 depends on CTCF, as mutation of its CTCF-binding site led to sustained PAIR4 activity in pre-B and immature B-cells and to PAIR4 activation in T-cells. Notably, insertion of V8.8E was sufficient to activate VH gene recombination. Hence, enhancers of the PAIR4-V8.7E module and V8.8E element activate distal VH gene recombination and thus contribute to the diversification of the BCR repertoire in the context of loop extrusion.
Assuntos
Células Precursoras de Linfócitos B , Sequências Reguladoras de Ácido Nucleico , Sequências Reguladoras de Ácido Nucleico/genética , Sítios de Ligação , Recombinação GenéticaRESUMO
Humoral immunity depends on the germinal center (GC) reaction where B cells are tightly controlled for class-switch recombination and somatic hypermutation and finally generated into plasma and memory B cells. However, how protein SUMOylation regulates the process of the GC reaction remains largely unknown. Here, we show that the expression of SUMO-specific protease 1 (SENP1) is up-regulated in GC B cells. Selective ablation of SENP1 in GC B cells results in impaired GC dark and light zone organization and reduced IgG1-switched GC B cells, leading to diminished production of class-switched antibodies with high-affinity in response to a TD antigen challenge. Mechanistically, SENP1 directly binds to Paired box protein 5 (PAX5) to mediate PAX5 deSUMOylation, sustaining PAX5 protein stability to promote the transcription of activation-induced cytidine deaminase. In summary, our study uncovers SUMOylation as an important posttranslational mechanism regulating GC B cell response.
Assuntos
Linfócitos B , Cisteína Endopeptidases , Centro Germinativo , Fator de Transcrição PAX5 , Sumoilação , Centro Germinativo/imunologia , Centro Germinativo/metabolismo , Fator de Transcrição PAX5/metabolismo , Fator de Transcrição PAX5/genética , Animais , Linfócitos B/imunologia , Linfócitos B/metabolismo , Cisteína Endopeptidases/metabolismo , Cisteína Endopeptidases/genética , Camundongos , Switching de Imunoglobulina , Humanos , Citidina Desaminase/metabolismo , Citidina Desaminase/genética , Imunidade Humoral , Camundongos Endogâmicos C57BLRESUMO
While PAX5 is an important tumor suppressor gene in B-cell acute lymphoblastic leukemia (B-ALL), it is also involved in oncogenic translocations coding for diverse PAX5 fusion proteins. PAX5-JAK2 encodes a protein consisting of the PAX5 DNA-binding region fused to the constitutively active JAK2 kinase domain. Here, we studied the oncogenic function of the PAX5-JAK2 fusion protein in a mouse model expressing it from the endogenous Pax5 locus, resulting in inactivation of one of the two Pax5 alleles. Pax5Jak2/+ mice rapidly developed an aggressive B-ALL in the absence of another cooperating exogenous gene mutation. The DNA-binding function and kinase activity of Pax5-Jak2 as well as IL-7 signaling contributed to leukemia development. Interestingly, all Pax5Jak2/+ tumors lost the remaining wild-type Pax5 allele, allowing efficient DNA-binding of Pax5-Jak2. While we could not find evidence for a nuclear role of Pax5-Jak2 as an epigenetic regulator, high levels of active phosphorylated STAT5 and increased expression of STAT5 target genes were seen in Pax5Jak2/+ B-ALL tumors, implying that nuclear Pax5-Jak2 phosphorylates STAT5. Together, these data reveal Pax5-Jak2 as an important nuclear driver of leukemogenesis by maintaining phosphorylated STAT5 levels in the nucleus.
Assuntos
Janus Quinase 2 , Leucemia de Células B , Fator de Transcrição PAX5 , Fator de Transcrição STAT5 , Animais , Janus Quinase 2/genética , Leucemia de Células B/genética , Camundongos , Mutação , Fator de Transcrição PAX5/genética , Fator de Transcrição STAT5/genética , Translocação GenéticaRESUMO
As a hallmark of senescent cells, the derepression of Long Interspersed Elements 1 (LINE1) transcription results in accumulated LINE1 cDNA, which triggers the secretion of the senescence-associated secretory phenotype (SASP) and paracrine senescence in a cGAS-STING pathway-dependent manner. However, transcription factors that govern senescence-associated LINE1 reactivation remain ill-defined. Here, we predict several transcription factors that bind to human LINE1 elements to regulate their transcription by analyzing the conserved binding motifs in the 5'-untranslated regions (UTR) of the commonly upregulated LINE1 elements in different types of senescent cells. Further analysis reveals that PAX5 directly binds to LINE1 5'-UTR and the binding is enhanced in senescent cells. The enrichment of PAX5 at the 5'-UTR promotes cellular senescence and SASP by activating LINE1. We also demonstrate that the longevity gene SIRT6 suppresses PAX5 transcription by directly binding to the PAX5 promoter, and overexpressing PAX5 abrogates the suppressive effect of SIRT6 on stress-dependent cellular senescence. Our work suggests that PAX5 could serve as a potential target for drug development aiming to suppress LINE1 activation and treat senescence-associated diseases.
Assuntos
Senescência Celular , Elementos Nucleotídeos Longos e Dispersos , Fator de Transcrição PAX5 , Humanos , Regiões 5' não Traduzidas/genética , Regulação da Expressão Gênica , Fator de Transcrição PAX5/genética , Fator de Transcrição PAX5/metabolismo , Regiões Promotoras Genéticas , Ligação Proteica , Retroelementos/genética , Fenótipo Secretor Associado à Senescência/genéticaRESUMO
B-cell fate determination requires the action of transcription factors that operate in a regulatory network to activate B-lineage genes and repress lineage-inappropriate genes. However, the dynamics and hierarchy of events in B-cell programming remain obscure. To uncouple the dynamics of transcription factor expression from functional consequences, we generated induction systems in developmentally arrested Ebf1-/- pre-pro-B cells to allow precise experimental control of EBF1 expression in the genomic context of progenitor cells. Consistent with the described role of EBF1 as a pioneer transcription factor, we show in a time-resolved analysis that EBF1 occupancy coincides with EBF1 expression and precedes the formation of chromatin accessibility. We observed dynamic patterns of EBF1 target gene expression and sequential up-regulation of transcription factors that expand the regulatory network at the pro-B-cell stage. A continuous EBF1 function was found to be required for Cd79a promoter activity and for the maintenance of an accessible chromatin domain that is permissive for binding of other transcription factors. Notably, transient EBF1 occupancy was detected at lineage-inappropriate genes prior to their silencing in pro-B cells. Thus, persistent and transient functions of EBF1 allow for an ordered sequence of epigenetic and transcriptional events in B-cell programming.
Assuntos
Linfócitos B/metabolismo , Epigênese Genética , Células-Tronco/metabolismo , Transativadores/metabolismo , Transcrição Gênica , Animais , Antígenos CD79/genética , Linhagem da Célula/genética , Células Cultivadas , Cromatina/metabolismo , DNA/metabolismo , Camundongos , Fator de Transcrição PAX5/metabolismo , Regiões Promotoras Genéticas , Proteínas Proto-Oncogênicas/metabolismo , Fatores de Transcrição/metabolismoRESUMO
Earlier studies have identified transcription factors that specify B-cell fate, but the underlying mechanisms remain to be revealed. Two new studies by Miyai and colleagues (pp. 112-126) and Li and colleagues (pp. 96-111) in this issue of Genes & Development provide new and unprecedented insights into the genetic and epigenetic mechanisms that establish B-cell identity.
Assuntos
Linfócitos B/imunologia , Transativadores , Diferenciação Celular , Ativação Linfocitária , Fatores de TranscriçãoRESUMO
BACKGROUND: Existing studies have found that circular RNAs (circRNAs) act as sponges for micro RNAs (miRNAs) to control downstream genes. However, the specific functionalities and mechanisms of circRNAs in human clear cell renal cell carcinoma (ccRCC) have yet to be thoroughly investigated. METHODS: Patient cohorts from online databases were used to screen candidate circRNAs, while another cohort from our hospital was obtained for validation. CircSOD2 was identified as a potential oncogenic target, and its relevant characteristics were investigated during ccRCC progression through various assays. A positive feedback loop containing downstream miRNA and its target gene were identified using bioinformatics and validated by luciferase reporter assays, RNA pull-down, and high-throughput sequencing. RESULTS: CircSOD2 expression was elevated in tumor samples and significantly correlated with overall survival (OS) and the tumor stage of ccRCC patients, which appeared in the enhanced proliferation, invasion, and migration of tumor cells. Through competitive binding to circSOD2, miR-532-3p can promote the expression of PAX5 and the progression of ccRCC, and such regulation can be salvaged by miR-532-3p inhibitor. CONCLUSION: A novel positive feedback loop, PAX5/circSOD2/miR-532-3p/PAX5 was identified in the study, indicating that the loop may play an important role in the diagnosis and prognostic prediction in ccRCC patients.
Assuntos
Carcinoma de Células Renais , Proliferação de Células , Retroalimentação Fisiológica , Regulação Neoplásica da Expressão Gênica , Neoplasias Renais , MicroRNAs , RNA Circular , Humanos , Carcinoma de Células Renais/genética , Carcinoma de Células Renais/patologia , Carcinoma de Células Renais/metabolismo , RNA Circular/genética , RNA Circular/metabolismo , Neoplasias Renais/genética , Neoplasias Renais/patologia , Neoplasias Renais/metabolismo , MicroRNAs/genética , MicroRNAs/metabolismo , Linhagem Celular Tumoral , Proliferação de Células/genética , Feminino , Pessoa de Meia-Idade , Masculino , Carcinogênese/genética , Carcinogênese/patologia , Movimento Celular/genética , Fator de Transcrição PAX5/metabolismo , Fator de Transcrição PAX5/genética , Oncogenes/genética , Sequência de Bases , Progressão da Doença , Invasividade Neoplásica , Reprodutibilidade dos TestesRESUMO
PAX5, a member of the paired box gene family of transcription factors, is a B-cell-specific activator protein that plays important roles during B lymphopoiesis. Two putative PAX5 binding sites in the human GINS1 promoter region were identified. EMSA, ChIP and luciferase assay showed that PAX5 functions as a positive transcription factor for GINS1 expression. Furthermore, coordinated expression of PAX5 and GINS1 was observed in mice B cells under physiological conditions and LPS stimulation situations. A similar pattern was also observed in human DLBCL cell lines under differentiation-inducing conditions. In addition, both PAX5 and GINS1 were highly expressed and significantly correlated in DLBCL specimens and cell lines. These findings suggested that dysregulation of PAX5 played an extremely important role in controlling the universal phenomenon of tumor progression through increased expression of GINS1 in DLBCL. In addition, circ1857 that was generated using back splicing of PAX5 pre-mRNA could further stabilize GINS1 mRNA, modulate GINS1 expression and promote lymphoma progression. To the best of our knowledge, this report is the first to demonstrate the role of GINS1 in DLBCL progression, and the mechanism of GINS1 upregulation using both circ1857 and PAX5 in DLBCL was revealed. Our results suggested that GINS1 may be a possible therapeutic target for DLBCL.
Assuntos
Fator de Transcrição PAX5 , Fatores de Transcrição , Camundongos , Animais , Humanos , Fator de Transcrição PAX5/genética , Fator de Transcrição PAX5/metabolismo , Fatores de Transcrição/genética , Regulação da Expressão Gênica , Linhagem Celular , Proliferação de Células/genética , Proteínas de Ligação a DNA/genéticaRESUMO
BACKGROUND: Breast cancer is one of the most common malignancies occurred in female around the globe. Recent studies have revealed the crucial characters of miRNA and genes, as well as the essential roles of epigenetic regulation in breast cancer initiation and progression. In our previous study, miR-142-3p was identified as a tumor suppressor and led to G2/M arrest through targeting CDC25C. However, the specific mechanism is still uncertain. METHODS: We identified PAX5 as the upstream regulator of miR-142-5p/3p through ALGGEN website and verified by series of assays in vitro and in vivo. The expression of PAX5 in breast cancer was detected by qRT-PCR and western blot. Besides, bioinformatics analysis and BSP sequencing were performed to analyze the methylation of PAX5 promoter region. Finally, the binding sites of miR-142 on DNMT1 and ZEB1 were predicted by JASPAR, and proved by luciferase reporter assay, ChIP analysis and co-IP. RESULTS: PAX5 functioned as a tumor suppressor by positive regulation of miR-142-5p/3p both in vitro and in vivo. The expression of PAX5 was regulated by the methylation of its promoter region induced by DNMT1 and ZEB1. In addition, miR-142-5p/3p could regulate the expression of DNMT1 and ZEB1 through binding with their 3'UTR region, respectively. CONCLUSION: In summary, PAX5-miR-142-DNMT1/ZEB1 constructed a negative feedback loop to regulate the progression of breast cancer, which provided emerging strategies for breast cancer therapy.
Assuntos
Neoplasias da Mama , MicroRNAs , Humanos , Feminino , Linhagem Celular Tumoral , Retroalimentação , Neoplasias da Mama/patologia , Apoptose/genética , Epigênese Genética , Pontos de Checagem da Fase G2 do Ciclo Celular , MicroRNAs/genética , MicroRNAs/metabolismo , Proliferação de Células/genética , Regulação Neoplásica da Expressão Gênica , Movimento Celular/genética , Homeobox 1 de Ligação a E-box em Dedo de Zinco/genética , Homeobox 1 de Ligação a E-box em Dedo de Zinco/metabolismo , Fator de Transcrição PAX5/genética , Fator de Transcrição PAX5/metabolismoRESUMO
Objective: In this study, we investigate the molecular rearrangement of FOXO1 in alveolar rhabdomyosarcoma (ARHS) in Saudi pediatric patients. Method: We performed a molecular detection of molecular translocation in 30 pediatric cases of ARHS using FOXO1 dual color break-apart FISH probe (ZytoLight®, 13q14.11) and PAX5 dual color break-apart FISH probe (ZytoLight®, 9p13.2). Results: All analyzable cases of ARHS demonstrated FOXO1 translocation whereas PAX5 translocation was not detected in any case. Conclusion: Although the testing for PAX5 rearrangement was based on protein-protein network analysis, our study showed that PAX5 translocation is not conspicuous in ARHS. PAX7/3::FOXO1 fusion genes feature ARMS, rendering crossreactivity between PAX7 and PAX3 a possible explanation. Nevertheless, PAX5 immunoreactivity and molecular translocation could be an adjunctive pathway that is confined to aggressive ARMS.
Assuntos
Rabdomiossarcoma Alveolar , Rabdomiossarcoma , Humanos , Rabdomiossarcoma Alveolar/diagnóstico , Rabdomiossarcoma Alveolar/genética , Arábia Saudita , Hibridização in Situ Fluorescente , Translocação Genética , Proteínas de Fusão Oncogênica/genética , Proteína Forkhead Box O1/genética , Fator de Transcrição PAX5/genéticaRESUMO
PAX5 is a transcription factor associated with abnormal posterior midbrain and cerebellum development in mice. PAX5 is highly loss-of-function intolerant and missense constrained, and has been identified as a candidate gene for autism spectrum disorder (ASD). We describe 16 individuals from 12 families who carry deletions involving PAX5 and surrounding genes, de novo frameshift variants that are likely to trigger nonsense-mediated mRNA decay, a rare stop-gain variant, or missense variants that affect conserved amino acid residues. Four of these individuals were published previously but without detailed clinical descriptions. All these individuals have been diagnosed with one or more neurodevelopmental phenotypes including delayed developmental milestones (DD), intellectual disability (ID), and/or ASD. Seizures were documented in four individuals. No recurrent patterns of brain magnetic resonance imaging (MRI) findings, structural birth defects, or dysmorphic features were observed. Our findings suggest that PAX5 haploinsufficiency causes a neurodevelopmental disorder whose cardinal features include DD, variable ID, and/or ASD.
Assuntos
Transtorno do Espectro Autista , Deficiência Intelectual , Transtornos do Neurodesenvolvimento , Animais , Transtorno do Espectro Autista/genética , Haploinsuficiência , Humanos , Deficiência Intelectual/diagnóstico , Deficiência Intelectual/genética , Camundongos , Transtornos do Neurodesenvolvimento/genética , Transtornos do Neurodesenvolvimento/patologia , Fator de Transcrição PAX5 , FenótipoRESUMO
Lineage switch is a rare event at leukemic relapse. While mostly known to occur in KMT2A-rearranged infant leukemia, the underlying mechanism is yet to be depicted. This case report describes a female infant who achieved remission of KMT2A-MLLT3-rearranged acute monocytic leukemia, but 6 months thereafter, relapsed as KMT2A-MLLT3-rearranged acute lymphocytic leukemia. Whole exome sequencing of the bone marrow obtained pre-post lineage switch revealed two somatic mutations of PAX5 in the relapse sample. These two PAX5 alterations were suggested to be loss of function, thus to have played the driver role in the lineage switch from acute monocytic leukemia to acute lymphocytic leukemia.
Assuntos
Leucemia Monocítica Aguda , Leucemia Mieloide Aguda , Leucemia-Linfoma Linfoblástico de Células Precursoras , Medula Óssea , Criança , Feminino , Histona-Lisina N-Metiltransferase , Humanos , Lactente , Leucemia Monocítica Aguda/genética , Leucemia Mieloide Aguda/genética , Proteína de Leucina Linfoide-Mieloide/genética , Fator de Transcrição PAX5 , Leucemia-Linfoma Linfoblástico de Células Precursoras/genética , RecidivaRESUMO
Postmenopausal osteoporosis (OPO) is one of the most common types of primary osteoporosis. There is currently lack of a plasma biomarker for sensitive and early diagnosis of OPO. Here we aimed to explore the potential of early B cell factor 1 (EBF1) as a new plasma biomarker of OPO. Quantitative real-time PCR was used to measure the plasma EBF1 levels. Absorptiometry markers, such as lumbar spine (LS) bone mineral density (BMD) and LS T score were obtained after X-ray scans. Biochemical analyses used to measure osteopontin (OPN), ß-isomerized C-terminal telopeptides and total N-terminal procollagen of type-I collagen levels of patients with osteopenia (OPE, nâ¯=â¯81), osteoporosis (OPO, nâ¯=â¯98) as well as healthy subjects (NC, nâ¯=â¯110). Quantitative real-time PCR was used to measure the plasma levels of PAX5 and GSTP1, which are target genes of EBF1. EBF1 was downregulated in OPO patients. Levels of EBF1 were positively correlated to clinicopathological characteristics, including LS BMD and LS T scores, and negatively correlated to OPN and total N-terminal procollagen of type-I collagen levels. Increased PAX5 and GSTP1 levels also demonstrated strong correlations with higher EBF1, LS BMD and LS T score. Anti-osteoporotic treatment resulted in significant upregulation of EBF1, PAX5 and GSTP1 at 6 mo after treatment. Our study suggests that plasma EBF1 is a potential biomarker for diagnosing and assessing treatment outcome of OPO.
Assuntos
Osteoporose Pós-Menopausa , Osteoporose , Transativadores , Absorciometria de Fóton , Biomarcadores/sangue , Densidade Óssea/fisiologia , Colágeno Tipo I/genética , Feminino , Humanos , Vértebras Lombares , Osteoporose/tratamento farmacológico , Osteoporose Pós-Menopausa/diagnóstico por imagem , Pró-Colágeno , Transativadores/sangueRESUMO
BACKGROUND: Hodgkin's lymphoma (HL) is among the most prevalent lymphomas worldwide. PAX5 has a great value in separating this entity from other T cell lymphoma; however, it is weakly expressed in neoplastic cells. Polyclonal PAX8 was positive in a variety of lymphoid neoplasms in several previous studies and the staining paralleled that of PAX5 but monoclonal PAX8 was negative in the same neoplasms. The aim of this study was to compare immunohistochemical patterns of monoclonal PAX8 with PAX5 in Classical and NLPHL samples. MATERIAL AND METHODS: This retrospective study was conducted on 89 formalin-fixed paraffin embedded blocks from HL patients (69 Classical and 20 NLPHL) admitted to Imam Khomeini and Dr. Shariati hospitals, Tehran, Iran during 2016-2020. Diagnoses were confirmed by reviewing previous immunohistochemistry (IHC) studies, including PAX5. All samples were stained for PAX8 (clone MRQ-50). Expression intensity scoring was made for both antibodies in neoplastic and background cells based on nuclear staining percentage. RESULTS: PAX8 was positive in neoplastic and background B lymphocytes of all classical and NLPHL samples. PAX8 Expression intensity was significantly higher in neoplastic and background cells compared to PAX5 in classical HL samples (P = 0.001). PAX5 expression intensity in neoplastic cells was significantly higher in NLPHL samples compared to classical HL (P = 0.040); however, no significant difference in PAX8 expression between neoplastic cells of NLPHL and HL was seen. PAX8 expression intensity was not significantly correlated with gender, histologic subtype, tumor location, and relapse. CONCLUSIONS: PAX8 monoclonal antibody (clone MRQ-50) showed strong nuclear reactivity in neoplastic and background cells of classical HL and NLPHL samples. Therefore, this marker can be utilized as a valuable alternative for PAX5 in differentiating HL from other T cell lymphoma in challenging cases.
Assuntos
Doença de Hodgkin , Linfoma de Células T , Anticorpos Monoclonais , Formaldeído , Doença de Hodgkin/metabolismo , Humanos , Irã (Geográfico) , Recidiva Local de Neoplasia , Fator de Transcrição PAX5 , Fator de Transcrição PAX8 , Estudos Retrospectivos , Coloração e RotulagemRESUMO
Loss-of-function mutations in hematopoietic transcription factors including PAX5 occur in most cases of B-progenitor acute lymphoblastic leukemia (B-ALL), a disease characterized by the accumulation of undifferentiated lymphoblasts. Although PAX5 mutation is a critical driver of B-ALL development in mice and humans, it remains unclear how its loss contributes to leukemogenesis and whether ongoing PAX5 deficiency is required for B-ALL maintenance. Here we used transgenic RNAi to reversibly suppress endogenous Pax5 expression in the hematopoietic compartment of mice, which cooperates with activated signal transducer and activator of transcription 5 (STAT5) to induce B-ALL. In this model, restoring endogenous Pax5 expression in established B-ALL triggers immunophenotypic maturation and durable disease remission by engaging a transcriptional program reminiscent of normal B-cell differentiation. Notably, even brief Pax5 restoration in B-ALL cells causes rapid cell cycle exit and disables their leukemia-initiating capacity. These and similar findings in human B-ALL cell lines establish that Pax5 hypomorphism promotes B-ALL self-renewal by impairing a differentiation program that can be re-engaged despite the presence of additional oncogenic lesions. Our results establish a causal relationship between the hallmark genetic and phenotypic features of B-ALL and suggest that engaging the latent differentiation potential of B-ALL cells may provide new therapeutic entry points.
Assuntos
Diferenciação Celular/genética , Fator de Transcrição PAX5/genética , Leucemia-Linfoma Linfoblástico de Células Precursoras/genética , Leucemia-Linfoma Linfoblástico de Células Precursoras/patologia , Células Precursoras de Linfócitos B/citologia , Animais , Linhagem Celular Tumoral , Células Cultivadas , Regulação Neoplásica da Expressão Gênica , Técnicas de Silenciamento de Genes , Genes myc/genética , Humanos , Camundongos , Camundongos Transgênicos , Fator de Transcrição PAX5/metabolismo , Fator de Transcrição STAT5/metabolismo , Transdução de SinaisRESUMO
PAX5, a member of the Paired Box (PAX) transcription factor family, is an essential factor for B-lineage identity during lymphoid differentiation. Mechanistically, PAX5 controls gene expression profiles, which are pivotal to cellular processes such as viability, proliferation, and differentiation. Given its crucial function in B-cell development, PAX5 aberrant expression also correlates with hallmark cancer processes leading to hematological and other types of cancer lesions. Despite the well-established association of PAX5 in the development, maintenance, and progression of cancer disease, the use of PAX5 as a cancer biomarker or therapeutic target has yet to be implemented. This may be partly due to the assortment of PAX5 expressed products, which layers the complexity of their function and role in various regulatory networks and biological processes. In this review, we provide an overview of the reported data describing PAX5 products, their regulation, and function in cellular processes, cellular biology, and neoplasm.
Assuntos
Fator de Transcrição PAX5 , Proteínas , Diferenciação Celular/genética , Hematopoese , Fator de Transcrição PAX5/genética , Fator de Transcrição PAX5/metabolismoRESUMO
An 82-year-old Japanese male patient was initially diagnosed with lymphocytosis. His complete blood count revealed a white blood cell count of 30.9×109/l with 81% abnormal lymphocytes. The abnormal lymphocytes included monoclonal clones of CD38+ and CD138+cytoplasmic κ+ and IgG-κ M-protein, which led to the final diagnosis of plasma cell leukemia (PCL). Bortezomib and dexamethasone therapy was initiated, but the patient succumbed to the disease on the 8th day of hospitalization. A cytogenetic examination revealed a t (9;14)(p13;q32) translocation and the Western blotting confirmed high PAX5 expression. Similar to our present case, PCL cases with "lymphocytosis" have been widely reported, which some speculating the involvement of PAX5 overexpression in the pathogenesis. Such cases, including ours, may be classified as a unique group of disorders (PCL presenting as "lymphocytosis"), which requires accurate differential diagnosis and subsequent urgent multidisciplinary intensive treatment.
Assuntos
Leucemia Plasmocitária , Linfocitose , Idoso de 80 Anos ou mais , Humanos , Masculino , Leucemia Plasmocitária/diagnóstico , Linfócitos/metabolismo , Linfocitose/diagnóstico , Fator de Transcrição PAX5/genética , Translocação GenéticaRESUMO
For many years, immortalized tumor cell lines have been used as reliable tools to understand the function of oncogenes and tumor suppressor genes. Today, we know that tumors can comprise subclones with common and with subclone-specific genetic alterations. We sequenced DNA and RNA of sequential sister cell lines obtained from patients with pre-B acute lymphoblastic leukemia at different phases of the disease. All five pairs of cell lines carry alterations that are typical for this disease: loss of tumor suppressors (CDKN2A, CDKN2B), expression of fusion genes (ETV6-RUNX1, BCR-ABL1, MEF2D-BCL9) or of genes targeted by point mutations (KRAS A146T, NRAS G12C, PAX5 R38H). MEF2D-BCL9 and PAX R38H mutations in cell lines have hitherto been undescribed, suggesting that YCUB-4 (MEF2D-BCL9), PC-53 (PAX R38H) and their sister cell lines will be useful models to elucidate the function of these genes. All aberrations mentioned above occur in both sister cell lines, demonstrating that the sisters derive from a common ancestor. However, we also found mutations that are specific for one sister cell line only, pointing to individual subclones of the primary tumor as originating cells. Our data show that sequential sister cell lines can be used to study the clonal development of tumors and to elucidate the function of common and clone-specific mutations.
Assuntos
Biomarcadores Tumorais , Linhagem Celular Tumoral , Predisposição Genética para Doença , Leucemia-Linfoma Linfoblástico de Células Precursoras B/genética , Alelos , Aberrações Cromossômicas , Análise Mutacional de DNA , Progressão da Doença , Humanos , Mutação , Leucemia-Linfoma Linfoblástico de Células Precursoras B/patologia , RNA-Seq , Sequenciamento do ExomaRESUMO
PAX5 is a tumor suppressor in B-ALL, while the role of PAX5 fusion proteins in B-ALL development is largely unknown. Here, we studied the function of PAX5-ETV6 and PAX5-FOXP1 in mice expressing these proteins from the Pax5 locus. Both proteins arrested B-lymphopoiesis at the pro-B to pre-B-cell transition and, contrary to their proposed dominant-negative role, did not interfere with the expression of most regulated Pax5 target genes. Pax5-Etv6, but not Pax5-Foxp1, cooperated with loss of the Cdkna2a/b tumor suppressors in promoting B-ALL development. Regulated Pax5-Etv6 target genes identified in these B-ALLs encode proteins implicated in pre-B-cell receptor (BCR) signaling and migration/adhesion, which could contribute to the proliferation, survival, and tissue infiltration of leukemic B cells. Together with similar observations made in human PAX5-ETV6+ B-ALLs, these data identified PAX5-ETV6 as a potent oncoprotein that drives B-cell leukemia development.