RESUMO
Classical myeloproliferative neoplasms (MPNs) are characterized by distinct clinical phenotypes. The discovery of driver mutations in JAK2, CALR and MPL genes provided new insights into their pathogenesis. Next-generation sequencing (NGS) identified additional somatic mutations, most frequently in epigenetic modulator genes. In this study, a cohort of 95 MPN patients was genetically characterized using targeted NGS. Clonal hierarchies of detected mutations were subsequently analysed using colony forming progenitor assays derived from single cells to study mutation acquisition. Further, the hierarchy of mutations within distinct cell lineages was evaluated. NGS revealed mutations in three epigenetic modulator genes (TET2, DNMT3A, ASXL1) as most common co-mutations to the classical driver mutations. JAK2V617F as well as DNMT3A and TET2 mutations were detected as primary events in disease formation and most cases presented with a linear mutation pattern. Mutations appear mostly in the myeloid lineages but can also appear in lymphoid subpopulations. In one case with a double mutant MPL gene, mutations exclusively appeared in the monocyte lineage. Overall, this study confirms the mutational heterogeneity of classical MPNs and highlights the role of JAK2V617F and epigenetic modifier genes as early events in hematologic disease formation.
Assuntos
Transtornos Mieloproliferativos , Neoplasias , Humanos , Transtornos Mieloproliferativos/genética , Transtornos Mieloproliferativos/patologia , Janus Quinase 2/genética , Janus Quinase 2/metabolismo , Evolução Clonal/genética , Fenótipo , Mutação , Calreticulina/genéticaAssuntos
Antineoplásicos/administração & dosagem , Medula Óssea/efeitos dos fármacos , Leucemia Neutrofílica Crônica/genética , Pirazóis/administração & dosagem , Pirazóis/uso terapêutico , Receptores de Fator Estimulador de Colônias/genética , Idoso , Medula Óssea/patologia , Dasatinibe/uso terapêutico , Sequenciamento de Nucleotídeos em Larga Escala , Humanos , Leucemia Neutrofílica Crônica/tratamento farmacológico , Leucemia Neutrofílica Crônica/fisiopatologia , Masculino , Mutação , Nitrilas , Pirazóis/efeitos adversos , PirimidinasRESUMO
OBJECTIVES: The principal aim of this study was to assess the safety and effectiveness of the middle ear implant Vibrant Soundbridge (VSB) in patients with moderate-to-severe sensorineural hearing loss up to a mean (± standard deviation) duration of 11.1 ± 2.1 years (min. = 8.2, max. = 13.9, n = 16) after the intervention. DESIGN: This was a retrospective, single-subject repeated-measurements study over a long-term period. A total of 104 German-speaking adults (for 122 implants) were included in this study (54 male, 50 female). The mean age at implantation was 54.5 years (min. = 19.0, max. = 80.4). Audiological outcome and speech intelligibility were assessed in all VSB patients at different time points in non-overlapping groups. RESULTS: Bone conduction (BC) thresholds were preserved after the implantation and no indication was found of an increase over time of the small air-bone gaps introduced by the implantation. BC and air conduction thresholds worsened similarly in both implanted and non-implanted ears over time. The decrease in audiological benefit provided by the VSB was moderate and the Word Recognition Score in quiet conditions at 65 dB SPL was still largely improved with the VSB in the longest observed group. CONCLUSIONS: These results confirm that the VSB does not affect the integrity of the inner/middle ear and is still beneficial in long-term follow-up.
Assuntos
Bigorna/cirurgia , Prótese Ossicular , Substituição Ossicular/métodos , Adulto , Idoso , Idoso de 80 Anos ou mais , Audiometria de Tons Puros , Condução Óssea , Feminino , Perda Auditiva Neurossensorial , Humanos , Estudos Longitudinais , Masculino , Pessoa de Meia-Idade , Estudos Retrospectivos , Índice de Gravidade de Doença , Percepção da Fala , Resultado do Tratamento , Adulto JovemRESUMO
OBJECTIVE: To compare audiological outcomes in mild-to-moderate mixed hearing loss patients treated with a bone-anchored hearing aid or an active middle-ear implant. Analysis aimed to refine criteria used in preoperative selection of implant type. DESIGN: Retrospective comparative analysis of audiological data. Follow-up time ranged between 0.55 and 8.8 years. STUDY SAMPLE: For detailed comparative analysis, 12 patients (six in each group) with comparable bone conduction thresholds and similar clinical characteristics were selected. A larger cohort of 48 patient files were used to evaluate overall audiological indication criteria (24 per group). RESULTS: In free-field tone audiometry, Baha patients showed mean aided thresholds between 40-48 dB, whereas hearing thresholds for VSB patients were 25-43 dB. Baha and VSB users had mean WRS of 56% and 82%, respectively, at 65 dB. Better speech understanding in noise was seen with the VSB. CONCLUSION: Analysis of the main cohort (n = 48) showed that treatment with round window vibroplasty leads to better hearing performance than treatment with a bone-anchored hearing device, if the bone conduction pure-tone average (0.5 to 4 kHz) is poorer than 35 dB HL. Audiological analysis in the smaller comparative analysis showed similar findings.
Assuntos
Estimulação Acústica/métodos , Correção de Deficiência Auditiva/métodos , Auxiliares de Audição , Perda Auditiva Condutiva-Neurossensorial Mista/reabilitação , Prótese Ossicular , Estimulação Acústica/instrumentação , Adulto , Idoso , Audiometria , Limiar Auditivo , Condução Óssea/fisiologia , Correção de Deficiência Auditiva/instrumentação , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Ruído , Estudos Retrospectivos , Janela da Cóclea/fisiopatologia , Percepção da Fala , Adulto JovemRESUMO
AIMS: Clonal haematopoiesis of indeterminate potential (CHIP)-associated mutation is a risk factor for the development of ischaemic cardiomyopathy (ICM), but its association with non-ischaemic dilated cardiomyopathy (DCM) remains unclear. We aimed to determine the prevalence of CHIP in patients with DCM and define its risk for disease progression. METHODS AND RESULTS: Next-generation sequencing targeting 54 common CHIP-associated genes was performed in 48 ICM and 52 DCM patients. The patients were monitored for a median of 3.1 years, and a COX proportional hazards model was used to examine the association between CHIP and adverse clinical outcome with regard to all-cause death or all-cause hospitalization. Overall, the prevalence of CHIP mutations was 19% and 13% in DCM and ICM, respectively. Seventeen per cent of ICM patients over 75 years were CHIP carriers. In DCM cohort, mutation event had already been observed in the patients who were under the age of 45 (13%). Among 54 genes analysed, DNMT3A had the highest mutation frequency, followed by TET2 and CUX1. Kaplan-Meier curve over a median of 3.1 year tracking period showed a trend towards poor clinical outcome in the DCM patients who carried DNMT3A or TET2 mutation; however, such association was not statistically significant. CONCLUSIONS: The prevalence of CHIP is detected at a young age in DCM, and accumulation of mutational frequency in DCM patients is independent of age. However, a larger patient cohort is required to validate the association between CHIP and clinical progression in the DCM patients.
Assuntos
Cardiomiopatia Dilatada , Isquemia Miocárdica , Humanos , Hematopoiese Clonal/genética , Cardiomiopatia Dilatada/epidemiologia , Cardiomiopatia Dilatada/genética , Cardiomiopatia Dilatada/complicações , Isquemia Miocárdica/complicações , Isquemia Miocárdica/epidemiologia , Isquemia Miocárdica/genética , Fatores de Risco , MutaçãoRESUMO
Gene mutations independent of BCR::ABL1 have been identified in newly diagnosed patients with chronic myeloid leukemia (CML) in chronic phase, whereby mutations in epigenetic modifier genes were most common. These findings prompted the systematic analysis of prevalence, dynamics, and prognostic significance of such mutations, in a clinically well-characterized patient population of 222 CML patients from the TIGER study (CML-V) by targeted next-generation sequencing covering 54 myeloid leukemia-associated genes. In total, 53/222 CML patients (24%) carried 60 mutations at diagnosis with ASXL1 being most commonly affected (n = 20). To study mutation dynamics, longitudinal deep sequencing analysis of serial samples was performed in 100 patients after 12, 24, and 36 months of therapy. Typical patterns of clonal evolution included eradication, persistence, and emergence of mutated clones. Patients carrying an ASXL1 mutation at diagnosis showed a less favorable molecular response to nilotinib treatment, as a major molecular response (MMR) was achieved less frequently at month 12, 18, and 24 compared to all other patients. Patients with ASXL1 mutations were also younger and more frequently found in the high risk category, suggesting a central role of clonal evolution associated with ASXL1 mutations in CML pathogenesis.
Assuntos
Proteínas de Fusão bcr-abl , Leucemia Mielogênica Crônica BCR-ABL Positiva , Humanos , Mutação , Inibidores de Proteínas Quinases , Pirimidinas , Proteínas RepressorasRESUMO
Clonal hematopoiesis is frequently observed in elderly people. To investigate the prevalence and dynamics of genetic alterations among healthy elderly individuals, a cohort of 50 people >80 years was genotyped for commonly mutated leukemia-associated genes by targeted deep next-generation sequencing. A total of 16 somatic mutations were identified in 13/50 (26%) individuals. Mutations occurred at low variant allele frequencies (median 11.7%) and remained virtually stable over 3 years without development of hematologic malignancies in affected individuals. With DNMT3A mutations most frequently detected, another cohort of 160 healthy people spanning all age groups was sequenced specifically for DNMT3A revealing an overall mutation rate of 6.2% (13/210) and an age-dependent increase of mutation prevalence. A significant difference (p = 0.017) in the DNMT3A expression pattern was detected between younger and healthy elderly people as determined by qRT-PCR. To evaluate the selection of clonal hematopoietic stem cells (HSCs), bone marrow of two healthy individuals with mutant DNMT3A was transplanted in a humanized mouse model. Xenografts displayed stable kinetics of DNMT3A mutations over 8 months. These findings indicate that the appearance of low-level clones with leukemia-associated mutations is a common age-associated phenomenon, but insufficient to initiate clonal selection and expansion without the additional influence of other factors.
Assuntos
DNA (Citosina-5-)-Metiltransferases/genética , Hematopoese/genética , Leucemia/genética , Mutação , Fatores Etários , Idoso , Idoso de 80 Anos ou mais , Animais , Células Clonais , DNA Metiltransferase 3A , Feminino , Humanos , Masculino , Camundongos , Prevalência , RNA Mensageiro/análiseRESUMO
An amendment to this paper has been published and can be accessed via a link at the top of the paper.
RESUMO
To date, only one subtype of acute myeloid leukemia (AML), acute promyelocytic leukemia (APL) can be effectively treated by differentiation therapy utilizing all-trans retinoic acid (ATRA). Non-APL AMLs are resistant to ATRA. Here we demonstrate that the acetyltransferase GCN5 contributes to ATRA resistance in non-APL AML via aberrant acetylation of histone 3 lysine 9 (H3K9ac) residues maintaining the expression of stemness and leukemia associated genes. We show that inhibition of GCN5 unlocks an ATRA-driven therapeutic response. This response is potentiated by coinhibition of the lysine demethylase LSD1, leading to differentiation in most non-APL AML. Induction of differentiation was not correlated to a specific AML subtype, cytogenetic, or mutational status. Our study shows a previously uncharacterized role of GCN5 in maintaining the immature state of leukemic blasts and identifies GCN5 as a therapeutic target in AML. The high efficacy of the combined epigenetic treatment with GCN5 and LSD1 inhibitors may enable the use of ATRA for differentiation therapy of non-APL AML. Furthermore, it supports a strategy of combined targeting of epigenetic factors to improve treatment, a concept potentially applicable for a broad range of malignancies.
Assuntos
Resistencia a Medicamentos Antineoplásicos , Leucemia Mieloide Aguda/tratamento farmacológico , Leucemia Promielocítica Aguda/tratamento farmacológico , Tretinoína/farmacologia , Fatores de Transcrição de p300-CBP/metabolismo , Apoptose , Medula Óssea/metabolismo , Diferenciação Celular , Linhagem Celular Tumoral , Membrana Celular/metabolismo , Epigênese Genética , Genótipo , Células HEK293 , Células HL-60 , Histona Desmetilases/antagonistas & inibidores , Histonas/química , Humanos , Leucócitos Mononucleares/citologiaRESUMO
OBJECTIVE: To present power stapes, stapedotomy, and middle ear implantation with Vibrant SoundBridge (VSB) performed in a one-step surgery as an alternative option for hearing rehabilitation in patients with osteogenesis imperfecta (OI). STUDY DESIGN: Retrospective case series. SETTING: Tertiary referral ear center. PATIENTS: A family with genetically proven OI Type I. INTERVENTIONS: Two patients, mother and son, with severe to profound mixed hearing loss underwent 3 power stapes, 1 unilateral and 1 bilateral sequential. MAIN OUTCOME MEASURES: Thorough audiological diagnostic batteries including aided and unaided pure-tone and free-field audiometry and Freiburg monosyllabic word test were used to assess the preoperative status and the postoperative hearing outcome. High-resolution computed tomography of the temporal bones was performed as well. Surgical procedure and any special considerations were analyzed in detail. RESULTS: The hearing outcome was favorable in all cases, showing in comparison to the preoperative values an average improvement of 36.8 dB. Severe intraoperative bleeding of the middle ear mucosa was the only complication and could be easily controlled by allowing short time intervals. Inner ear trauma did not occur in any case. CONCLUSION: Power stapes represents a safe and promising procedure for treating hearing loss in selected patients with OI. Furthermore, it introduces a new, advantageous VSB application in cases of mixed hearing loss with severe otosclerosis and increased bone-conduction thresholds.