RESUMEN
The ability of activation-induced cytidine deaminase (AID) to efficiently mediate class-switch recombination (CSR) is dependent on its phosphorylation at Ser38; however, the trigger that induces AID phosphorylation and the mechanism by which phosphorylated AID drives CSR have not been elucidated. Here we found that phosphorylation of AID at Ser38 was induced by DNA breaks. Conversely, in the absence of AID phosphorylation, DNA breaks were not efficiently generated at switch (S) regions in the immunoglobulin heavy-chain locus (Igh), consistent with a failure of AID to interact with the endonuclease APE1. Additionally, deficiency in the DNA-damage sensor ATM impaired the phosphorylation of AID at Ser38 and the interaction of AID with APE1. Our results identify a positive feedback loop for the amplification of DNA breaks at S regions through the phosphorylation- and ATM-dependent interaction of AID with APE1.
Asunto(s)
Linfocitos B/inmunología , Citidina Desaminasa/inmunología , ADN-(Sitio Apurínico o Apirimidínico) Liasa/inmunología , Retroalimentación Fisiológica , Cambio de Clase de Inmunoglobulina , Cadenas Pesadas de Inmunoglobulina/inmunología , Animales , Proteínas de la Ataxia Telangiectasia Mutada/genética , Proteínas de la Ataxia Telangiectasia Mutada/inmunología , Linfocitos B/citología , Citidina Desaminasa/genética , Roturas del ADN de Doble Cadena , ADN-(Sitio Apurínico o Apirimidínico) Liasa/genética , Regulación de la Expresión Génica , Cadenas Pesadas de Inmunoglobulina/genética , Ratones , Fosforilación , Unión Proteica , Serina/inmunología , Serina/metabolismo , Transducción de SeñalRESUMEN
Somatic hypermutation and class-switch recombination of the immunoglobulin (Ig) genes occur in germinal center (GC) B cells and are initiated through deamination of cytidine to uracil by activation-induced cytidine deaminase (AID). Resulting uracil-guanine mismatches are processed by uracil DNA glycosylase (UNG)-mediated base-excision repair and MSH2-mediated mismatch repair (MMR) to yield mutations and DNA strand lesions. Although off-target AID activity also contributes to oncogenic point mutations and chromosome translocations associated with GC and post-GC B-cell lymphomas, the role of downstream AID-associated DNA repair pathways in the pathogenesis of lymphoma is unknown. Here, we show that simultaneous deficiency of UNG and MSH2 or MSH2 alone causes genomic instability and a shorter latency to the development of BCL6-driven diffuse large B-cell lymphoma (DLBCL) in a murine model. The additional development of several BCL6-independent malignancies in these mice underscores the critical role of MMR in maintaining general genomic stability. In contrast, absence of UNG alone is highly protective and prevents the development of BCL6-driven DLBCL. We further demonstrate that clonal and nonclonal mutations arise within non-Ig AID target genes in the combined absence of UNG and MSH2 and that DNA strand lesions arise in an UNG-dependent manner but are offset by MSH2. These findings lend insight into a complex interplay whereby potentially deleterious UNG activity and general genomic instability are opposed by the protective influence of MSH2, producing a net protective effect that promotes immune diversification while simultaneously attenuating malignant transformation of GC B cells.
Asunto(s)
Transformación Celular Neoplásica/patología , Citidina Desaminasa/metabolismo , Reparación del ADN/genética , Proteínas de Unión al ADN/metabolismo , Linfoma de Células B Grandes Difuso/genética , Linfoma de Células B Grandes Difuso/patología , Proteína 2 Homóloga a MutS/fisiología , Uracil-ADN Glicosidasa/fisiología , Animales , Linfocitos B/metabolismo , Linfocitos B/patología , Transformación Celular Neoplásica/genética , Proteínas de Unión al ADN/genética , Femenino , Citometría de Flujo , Perfilación de la Expresión Génica , Centro Germinal , Técnicas para Inmunoenzimas , Cambio de Clase de Inmunoglobulina/genética , Masculino , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Mutación , Proteínas Proto-Oncogénicas c-bcl-6 , ARN Mensajero/genética , Reacción en Cadena en Tiempo Real de la Polimerasa , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Hipermutación Somática de Inmunoglobulina/genética , Cariotipificación Espectral , Células Tumorales CultivadasRESUMEN
In this issue of Blood, complementary studies by Amin et al and Linley et al demonstrate that sugar moieties linked to surface immunoglobulin (sIg) of follicular lymphoma (FL) cells directly interact with endogenous lectins within the lymphoma niche and lead to activation of downstream B-cell receptor (BCR) signaling pathways. In addition to providing further insight into the role of the microenvironment in lymphomagenesis, these findings expose a unique molecular interaction that may represent a viable target for therapeutic intervention.
Asunto(s)
Linfocitos B/metabolismo , Señalización del Calcio , Moléculas de Adhesión Celular/inmunología , Moléculas de Adhesión Celular/metabolismo , Regulación de la Expresión Génica/inmunología , Inmunoglobulina M/inmunología , Región Variable de Inmunoglobulina/metabolismo , Lectinas Tipo C/inmunología , Lectinas Tipo C/metabolismo , Linfoma Folicular/inmunología , Linfoma Folicular/metabolismo , Sistema de Señalización de MAP Quinasas , Macrófagos/inmunología , Receptores de Antígenos de Linfocitos B/inmunología , Receptores de Superficie Celular/inmunología , Receptores de Superficie Celular/metabolismo , Transducción de Señal/inmunología , Femenino , Humanos , MasculinoRESUMEN
Tumor microenvironment (TME) is commonly implicated in regulating the growth of tumors, but whether it can directly alter the genetics of tumors is not known. Genomic instability and dendritic cell (DC) infiltration are common features of several cancers, including multiple myeloma (MM). Mechanisms underlying genomic instability in MM are largely unknown. Here, we show that interaction between myeloma and DCs, but not monocytes, leads to rapid induction of the genomic mutator activation-induced cytidine deaminase (AID) and AID-dependent DNA double-strand breaks (DSBs) in myeloma cell lines as well as primary MM cells. Both myeloid as well as plasmacytoid DCs have the capacity to induce AID in tumor cells. The induction of AID and DSBs in tumor cells by DCs requires DC-tumor contact and is inhibited by blockade of receptor activator of NF-κB/receptor activator of NF-κB ligand (RANKL) interactions. AID-mediated genomic damage led to altered tumorigenicity and indolent behavior of tumor cells in vivo. These data show a novel pathway for the capacity of DCs in the TME to regulate genomic integrity. DC-mediated induction of AID and resultant genomic damage may therefore serve as a double-edged sword and be targeted by approaches such as RANKL inhibition already in the clinic.
Asunto(s)
Citidina Desaminasa/genética , Células Dendríticas/metabolismo , Inestabilidad Genómica , Mieloma Múltiple/genética , Animales , Western Blotting , Línea Celular Tumoral , Supervivencia Celular/genética , Células Cultivadas , Técnicas de Cocultivo , Citidina Desaminasa/metabolismo , Roturas del ADN de Doble Cadena , Femenino , Regulación Enzimológica de la Expresión Génica , Regulación Neoplásica de la Expresión Génica , Humanos , Subunidad gamma Común de Receptores de Interleucina/deficiencia , Subunidad gamma Común de Receptores de Interleucina/genética , Ratones , Ratones Endogámicos NOD , Ratones Noqueados , Ratones SCID , Mieloma Múltiple/metabolismo , Mieloma Múltiple/patología , FN-kappa B/metabolismo , Ligando RANK/metabolismo , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Trasplante Heterólogo , Células Tumorales CultivadasRESUMEN
PURPOSE OF REVIEW: Activation-induced cytidine deaminase (AID) is expressed in germinal center B cells and initiates the events that lead to somatic hypermutation and class switch recombination of immunoglobulin genes. In addition to this fundamental role in immune diversification, aberrant targeting of AID activity contributes to point mutations and translocations of oncogenes associated with B-cell lymphoma. This review discusses recent findings on the role of AID in lymphomagenesis. RECENT FINDINGS: AID is expressed in many malignancies of mature B-cell origin and contributes to the development of lymphoma in several mouse models. The mechanism that guides AID to its genetic target is unknown and may be relatively nonspecific, as numerous nonimmunoglobulin genes appear to be targeted by AID in both normal and neoplastic B cells. Indeed, AID binds to genes on every chromosome throughout the genome and can induce double-stranded DNA breaks that lead to chromosome translocations at these sites. SUMMARY: Emerging evidence supports a key role of AID in lymphomagenesis through genome-wide off-target induction of point mutations and chromosome translocations. Additional work is needed to further define the full scope and consequences of off-target AID activity in human lymphoma as well as to understand the protective mechanisms that break down during the development and progression of disease.
Asunto(s)
Transformación Celular Neoplásica/genética , Citidina Desaminasa/fisiología , Linfoma de Células B/enzimología , Animales , Humanos , Cambio de Clase de Inmunoglobulina/genética , Linfoma de Células B/genética , Ratones , Hipermutación Somática de Inmunoglobulina/genética , Translocación GenéticaRESUMEN
Activation-induced cytidine deaminase (AID) initiates somatic hypermutation and class switch recombination of immunoglobulin genes in B cells, whereas off-targeted AID activity contributes to oncogenic mutations and chromosomal translocations associated with B cell malignancies. Paradoxically, only a minority of AID is allowed to access the nuclear genome, but the majority of AID is retained in the cytoplasm. It is unknown whether cytoplasmic AID can access and target the mitochondrial genome [mitochondrial DNA (mtDNA)]. To address this issue, we developed high-fidelity differential DNA denaturation PCR, which allowed the enrichment of genuine mtDNA mutations and therefore the identification of endogenous mtDNA mutation signatures in vitro. With this approach, we showed that AID targeting to mtDNA is a rare event in AID-expressing lymphoma lines. Further biochemical and microscopic analysis revealed that a fraction of cytosol AID is associated with the outer membrane of mitochondria but unable to access the mitochondrial matrix. Together, our data suggested that the mitochondrial genome is protected from AID-mediated mutagenesis by physical segregation of AID from accessing mtDNA within the mitochondrial matrix.
Asunto(s)
Citidina Desaminasa/genética , ADN Mitocondrial/genética , Mitocondrias/metabolismo , Reacción en Cadena de la Polimerasa , Células Cultivadas , Citidina Desaminasa/metabolismo , ADN Mitocondrial/metabolismo , HumanosRESUMEN
We previously identified a rearrangement of mixed-lineage leukemia (MLL) gene (also known as ALL-1, HRX, and HTRX1), consisting of an in-frame partial tandem duplication (PTD) of exons 5 through 11 in the absence of a partner gene, occurring in approximately 4%-7% of patients with acute myeloid leukemia (AML) and normal cytogenetics, and associated with a poor prognosis. The mechanism by which the MLL PTD contributes to aberrant hematopoiesis and/or leukemia is unknown. To examine this, we generated a mouse knockin model in which exons 5 through 11 of the murine Mll gene were targeted to intron 4 of the endogenous Mll locus. Mll(PTD/WT) mice exhibit an alteration in the boundaries of normal homeobox (Hox) gene expression during embryogenesis, resulting in axial skeletal defects and increased numbers of hematopoietic progenitor cells. Mll(PTD/WT) mice overexpress Hoxa7, Hoxa9, and Hoxa10 in spleen, BM, and blood. An increase in histone H3/H4 acetylation and histone H3 lysine 4 (Lys4) methylation within the Hoxa7 and Hoxa9 promoters provides an epigenetic mechanism by which this overexpression occurs in vivo and an etiologic role for MLL PTD gain of function in the genesis of AML.
Asunto(s)
Epigénesis Genética/genética , Duplicación de Gen , Expresión Génica/genética , Proteínas de Homeodominio/genética , Proteína de la Leucemia Mieloide-Linfoide/genética , Secuencias Repetidas en Tándem/genética , Animales , Apoptosis/genética , Proliferación Celular , Transformación Celular Neoplásica/genética , Reordenamiento Génico/genética , Genotipo , Hematopoyesis/genética , Células Madre Hematopoyéticas/metabolismo , Células Madre Hematopoyéticas/patología , N-Metiltransferasa de Histona-Lisina , Histonas/metabolismo , Proteínas Homeobox A10 , Proteínas de Homeodominio/metabolismo , Ratones , Ratones Endogámicos C57BL , Ratones Transgénicos , Proteínas de Neoplasias/genética , Fenotipo , Regiones Promotoras Genéticas/genética , Costillas/anomalías , Costillas/metabolismoRESUMEN
Chronic lymphocytic leukemia (CLL) is a clonal malignancy of mature B cells that displays immense clinical heterogeneity as reflected by the observation that many patients have an indolent disease that will not require intervention for many years while others will present with an aggressive and symptomatic leukemia requiring immediate treatment. Although there is no cure for CLL, the disease is treatable and current standard chemotherapy regimens have been shown to prolong survival. There is no obvious survival advantage to early treatment versus observation but the timing as to when a patient will require treatment is highly unpredictable. Thus, there has been great interest in identifying prognostic markers that can be used to distinguish those patients who may have an aggressive form of CLL and might benefit from early intervention. While clinical staging systems have been used to stratify patients into risk categories, they lack the ability to predict disease progression or response to therapy. Recent advances in our understanding of the biology of CLL have led to the identification of numerous cellular and molecular markers with potential prognostic and therapeutic significance. This review provides a concise overview of prognostic markers in CLL and a discussion of how those markers have impacted the clinical management of the disease.
Asunto(s)
Leucemia Linfocítica Crónica de Células B/diagnóstico , Leucemia Linfocítica Crónica de Células B/terapia , Biomarcadores de Tumor/sangre , Humanos , Cadenas Pesadas de Inmunoglobulina/genética , Región Variable de Inmunoglobulina/genética , Leucemia Linfocítica Crónica de Células B/sangre , Leucemia Linfocítica Crónica de Células B/genética , MicroARNs/genética , PronósticoRESUMEN
Although the majority of published cases of lead poisoning come from occupational exposures, some traditional remedies may also contain toxic amounts of lead. Ayurveda is a system of traditional medicine that is native to India and is used in many parts of world as an alternative to standard treatment regimens. Here, we report the case of a 58-year-old woman who presented with abdominal pain, anemia, liver function abnormalities, and an elevated blood lead level. The patient was found to have been taking the Ayurvedic medicine Jambrulin prior to presentation. Chemical analysis of the medication showed high levels of lead. Following treatment with an oral chelating agent, the patient's symptoms resolved and laboratory abnormalities normalized. This case highlights the need for increased awareness that some Ayurvedic medicines may contain potentially harmful levels of heavy metals and people who use them are at risk of developing associated toxicities.
Asunto(s)
Intoxicación por Plomo/etiología , Preparaciones de Plantas/efectos adversos , Quelantes/uso terapéutico , Contaminación de Medicamentos , Femenino , Humanos , Intoxicación por Plomo/diagnóstico , Intoxicación por Plomo/tratamiento farmacológico , Intoxicación por Plomo/patología , Medicina Ayurvédica , Persona de Mediana Edad , Preparaciones de Plantas/química , Succímero/uso terapéuticoRESUMEN
BACKGROUND: A 39-year-old woman with no relevant medical or family history was admitted to hospital with episodic fever, which persisted despite antibiotic therapy. Other notable findings at admission were splenomegaly, pancytopenia, hyponatremia, elevated levels of liver enzymes, hyperferritinemia and hypofibrinogenemia. INVESTIGATIONS: Physical examination, laboratory tests, rheumatic marker serology, pathogen detection assays, complete blood counts, measurement of levels of ferritin, fibrinogen, triglycerides and soluble CD25, natural killer cell functional studies, PRF1 mutation analysis, renal biopsy, bone marrow biopsy, CT imaging of the chest and abdomen. DIAGNOSIS: Idiopathic, atypical hemophagocytic lymphohistiocytosis. MANAGEMENT: Initial treatment with antibiotics was followed by immunosuppressive therapy (including intravenous immunoglobulin, ciclosporin, infliximab, corticosteroids and etoposide). Remission was achieved by treatment with the anti-CD52 monoclonal antibody, alemtuzumab, after which allogeneic stem-cell transplantation (with reduced-intensity conditioning treatment and graft-versus-host disease prophylaxis) resulted in a definitive cure.
Asunto(s)
Anticuerpos Monoclonales/uso terapéutico , Anticuerpos Antineoplásicos/uso terapéutico , Antineoplásicos/uso terapéutico , Linfohistiocitosis Hemofagocítica/terapia , Trasplante de Células Madre , Adulto , Alemtuzumab , Anticuerpos Monoclonales Humanizados , Terapia Combinada , Femenino , Humanos , Inmunosupresores/uso terapéutico , Linfohistiocitosis Hemofagocítica/diagnóstico , Inducción de Remisión , Trasplante Homólogo , Resultado del TratamientoRESUMEN
Very little is known about how acquired oncogenic mutations arise. In this issue of Cancer Cell, Klemm and colleagues present evidence supporting a role for the antibody diversification enzyme activation-induced deaminase (AID) in the generation of mutations associated with disease progression and drug resistance in chronic myeloid leukemia.
Asunto(s)
Crisis Blástica/tratamiento farmacológico , Citidina Desaminasa/metabolismo , Resistencia a Antineoplásicos/genética , Leucemia Mielógena Crónica BCR-ABL Positiva/tratamiento farmacológico , Mutación , Piperazinas/uso terapéutico , Pirimidinas/uso terapéutico , Antineoplásicos/uso terapéutico , Benzamidas , Línea Celular Tumoral , Progresión de la Enfermedad , Humanos , Mesilato de Imatinib , Leucemia Mielógena Crónica BCR-ABL Positiva/genética , Leucemia Mielógena Crónica BCR-ABL Positiva/patología , Modelos BiológicosRESUMEN
MLL (ALL-1) chimeric fusions and MLL partial tandem duplications (PTD) may have mechanistically distinct contributions to leukemogenesis. Acute myeloid leukemia (AML) blasts with the t(9;11)(p22; q23) express MLL-AF9 and MLL wild-type (WT) transcripts, while normal karyotype AML blasts with the MLL(PTD/WT) genotype express MLL PTD but not the MLL WT. Silencing of MLL WT in MLL(PTD/WT) blasts was reversed by DNA methyltransferase (DNMT) and histone deacetylase (HDAC) inhibitors, and MLL WT induction was associated with selective sensitivity to cell death. Reduction of MLL PTD expression induced MLL WT and reduced blast colony-forming units, supporting opposing functions for MLL PTD and MLL WT whereby the MLL PTD contributes to the leukemic phenotype via a recessive gain-of-function. The coincident suppression of the MLL WT allele with the expression of the MLL PTD allele, along with the functional data presented here, supports the hypothesis that loss of WT MLL function via monoallelic repression contributes to the leukemic phenotype by the remaining mutant allele. These data from primary AML and the pharmacologic reversal of MLL WT silencing associated with a favorable alteration in the threshold for apoptosis suggest that these patients with poor prognosis may benefit from demethylating or histone deacetylase inhibitor therapy, or both.