RESUMO
Certain subtypes of acute myeloid leukemia (AML) in children have inferior outcome, such as AML with translocation t(7;12)(q36;p13) leading to an MNX1::ETV6 fusion along with high expression of MNX1. We have identified the transforming event in this AML and possible ways of treatment. Retroviral expression of MNX1 was able to induce AML in mice, with similar gene expression and pathway enrichment to t(7;12) AML patient data. Importantly, this leukemia was only induced in immune incompetent mice using fetal but not adult hematopoietic stem and progenitor cells. The restriction in transforming capacity to cells from fetal liver is in alignment with t(7;12)(q36;p13) AML being mostly seen in infants. Expression of MNX1 led to increased histone 3 lysine 4 mono-, di- and trimethylation, reduction in H3K27me3, accompanied with changes in genome-wide chromatin accessibility and genome expression, likely mediated through MNX1 interaction with the methionine cycle and methyltransferases. MNX1 expression increased DNA damage, depletion of the Lin-/Sca1+/c-Kit+ population and skewing toward the myeloid lineage. These effects, together with leukemia development, were prevented by pre-treatment with the S-adenosylmethionine analog Sinefungin. In conclusion, we have shown the importance of MNX1 in development of AML with t(7;12), supporting a rationale for targeting MNX1 and downstream pathways.
Assuntos
Histonas , Leucemia Mieloide Aguda , Criança , Lactente , Humanos , Animais , Camundongos , Metiltransferases , Cromatina , S-Adenosilmetionina , Leucemia Mieloide Aguda/genética , Metilação , Fatores de Transcrição , Proteínas de Homeodomínio/genéticaRESUMO
Acute myeloid leukemia (AML) results from aberrant hematopoietic processes and these changes are frequently initiated by chromosomal translocations. One particular subtype, AML with translocation t(7;12)(q36;p13), is found in children diagnosed before 2 years of age. The mechanisms for leukemogenesis induced by t(7;12) is not understood, in part because of the lack of efficient methods to reconstruct the leukemia-associated genetic aberration with correct genomic architecture and regulatory elements. We therefore created induced pluripotent stem cell (iPSC) lines that carry the translocation t(7;12) using CRISPR/Cas9. These t(7;12) iPSC showed propensity to differentiate into all three germ layers, confirming retained stem cell properties. The potential for differentiation into hematopoietic stem and progenitor cells (HSPC) was shown by expression of CD34, CD43 and CD45. Compared with the parental iPSC line, a significant decrease in cells expressing CD235a and CD41a was seen in the t(7;12) iPSC-derived HSPC (iHSPC), suggesting a block in differentiation. Moreover, colony formation assay showed an accumulation of cells at the erythroid and myeloid progenitor stages. Gene expression analysis revealed significant down-regulation of genes associated with megakaryocyte differentiation and up-regulation of genes associated with myeloid pathways but also genes typically seen in AML cases with t(7;12). Thus, this iPSC t(7;12) leukemia model of the t(7;12) AML subtype constitutes a valuable tool for further studies of the mechanisms for leukemia development and to find new treatment options.
Assuntos
Diferenciação Celular , Proteínas de Homeodomínio , Células-Tronco Pluripotentes Induzidas , Leucemia Mieloide Aguda , Células Progenitoras de Megacariócitos e Eritrócitos , Fatores de Transcrição , Diferenciação Celular/genética , Criança , Expressão Gênica/genética , Expressão Gênica/fisiologia , Perfilação da Expressão Gênica , Células-Tronco Hematopoéticas/fisiologia , Proteínas de Homeodomínio/genética , Humanos , Células-Tronco Pluripotentes Induzidas/metabolismo , Leucemia Mieloide Aguda/genética , Células Progenitoras de Megacariócitos e Eritrócitos/fisiologia , Megacariócitos/fisiologia , Fatores de Transcrição/genética , Translocação GenéticaRESUMO
The aims of the study were to investigate healthy subjects' performance on a clinical test of high-level language (HLL) and how it is related to demographic characteristics and verbal working memory (VWM). One hundred healthy subjects (20-79 years old) were assessed with the Swedish BeSS test (Laakso, Brunnegård, Hartelius, & Ahlsén, 2000) and two digit span tasks. Relationships between the demographic variables, VWM and BeSS were investigated both with bivariate correlations and multiple regression analysis. The results present the norms for BeSS. The correlations and multiple regression analysis show that demographic variables had limited influence on test performance. Measures of VWM were moderately related to total BeSS score and weakly to moderately correlated with five of the seven subtests. To conclude, education has an influence on the test as a whole but measures of VWM stood out as the most robust predictor of HLL.
Assuntos
Idioma , Memória de Curto Prazo/fisiologia , Adulto , Idoso , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Testes Neuropsicológicos/estatística & dados numéricosRESUMO
OBJECTIVE: Acceptance and commitment therapy (ACT) is a transdiagnostic, behavioral treatment focusing on common processes behind different diagnoses. Internet-based treatment programs can improve access to treatment and easily be integrated into the individual's life. METHOD: This single-arm, pre-post-follow-up pilot study examined the acceptability, safety, and effectiveness of internet-based ACT (iACT) for participants with comorbid posttraumatic stress disorder (PTSD) and chronic pain treated at a tertiary pain clinic. All participants (N = 10) received the iACT program over a period of 10 weeks and were assessed pre- and postintervention and at a 3-month follow-up. RESULTS: This study provides preliminary evidence for the acceptability, safety, and effectiveness of iACT for comorbid PTSD and chronic pain. The program showed clinically significant improvement in relation to the primary outcomes PTSD symptom severity and pain interference for this patient group with complex, comorbid symptoms, with sustained effects at the 3-month follow-up. CONCLUSIONS: These results add to previous research where face-to-face ACT has been shown to be of benefit to individuals with either chronic pain or PTSD. The findings also suggest that iACT can be delivered to individuals with comorbid PTSD and chronic pain with an effectiveness that is comparable to other cognitive behavioral therapy-based treatments for somatic and psychiatric disorders. (PsycInfo Database Record (c) 2024 APA, all rights reserved).
RESUMO
Acute myeloid leukemia (AML) with the t(7;12)(q36;p13) translocation occurs only in very young children and has a poor clinical outcome. The expected oncofusion between breakpoint partners (MNX1 and ETV6) has only been reported in a subset of cases. However, a universal feature is the strong transcript and protein expression of MNX1, a homeobox transcription factor that is normally not expressed in hematopoietic cells. Here, we map the translocation breakpoints on chromosomes 7 and 12 in affected patients to a region proximal to MNX1 and either introns 1 or 2 of ETV6. The frequency of MNX1 overexpression in pediatric AML (n=1556, own and published data) is 2.4% and occurs predominantly in t(7;12)(q36;p13) AML. Chromatin interaction assays in a t(7;12)(q36;p13) iPSC cell line model unravel an enhancer-hijacking event that explains MNX1 overexpression in hematopoietic cells. Our data suggest that enhancer-hijacking may be a more widespread consequence of translocations where no oncofusion product was identified, including e.g. t(1;3) or t(4;12) AML.
Assuntos
Leucemia Mielogênica Crônica BCR-ABL Positiva/patologia , NADPH Oxidase 2/fisiologia , Animais , Células da Medula Óssea/metabolismo , Transformação Celular Neoplásica/patologia , Modelos Animais de Doenças , Proteínas de Fusão bcr-abl/metabolismo , Técnicas de Silenciamento de Genes , Camundongos Endogâmicos C57BL , NADPH Oxidase 2/deficiência , Transplante de Neoplasias , Análise de SobrevidaRESUMO
In the last decades, a well-established equine wound model has been used to study fibroproliferative wound healing disorders. The aim of this study was to characterize the degree of discomfort of wounding and sampling in an equine excisional wound model by evaluating systemic and local inflammatory responses and signs of pain. A total of 12 cutaneous wounds, three on each shoulder and each metatarsus, were created in a standing surgical procedure. Wounds were biopsied on days 2, 4, 7, 14, 21, and 28 after surgery. Clinical parameters (rectal temperature, heart rate, respiratory frequency) and blood levels of white blood cell, serum amyloid A, fibrinogen, and iron were monitored to evaluate the systemic inflammatory response. Local signs of inflammation (swelling, heat, pain) were subjectively assessed, the limb circumference recorded, and temperature of the wound measured by thermometry. Pain was evaluated by a composite measure pain scale (CMPS). The results demonstrated that the wounding procedure elicits an inflammatory response. Day 1 after surgery, two horses scored 2 and 7 units (of 27 units), respectively, on the CMPS, and day 8 after surgery, one horse scored 3 units. The biopsy procedure did not elicit local or systemic signs of inflammation. Based on these findings, it appears that the equine experimental wound model causes mild discomfort and pain manifestations. This information is important for researchers, who consider using the model. To justify the use of an animal model, it should be demonstrated that the expected benefits of the research outweigh the discomfort imposed to the animal.
RESUMO
Endothelin receptor type A (EDNRA) is known as a mediator of cell proliferation and survival. Aberrant regulation of EDNRA has been shown to play a role in tumor growth and metastasis. Using a global gene expression screen, we found that expression of Ednra was upregulated in murine leukemia inducing cells co-expressing Hoxa9 and Meis1 compared to cells only expressing Hoxa9. The aim of this study was to explore the role of Ednra in leukemogenesis further. In a murine bone marrow transplantation model, mice transplanted with cells overexpressing Ednra and Hoxa9 succumbed to leukemia significantly earlier than mice transplanted with cells overexpressing Hoxa9 only. Furthermore, overexpression of Ednra led to increased proliferation and resistance to apoptosis of bone marrow cells in vitro. We could also show that Meis1 binds to the Ednra promoter region, suggesting a regulatory role for Meis1 in Ednra expression. Taken together, our results suggest a role for Ednra in Hoxa9/Meis1-driven leukemogenesis.
Assuntos
Regulação Leucêmica da Expressão Gênica/fisiologia , Proteínas de Homeodomínio/metabolismo , Leucemia Mieloide Aguda/metabolismo , Proteína Meis1/metabolismo , Receptor de Endotelina A/metabolismo , Animais , Humanos , Leucemia Mieloide Aguda/genética , Leucemia Mieloide Aguda/patologia , Camundongos , Camundongos Endogâmicos C57BLRESUMO
OBJECTIVE: We investigated the potential role of ADAMTS-1 (a disintegrin and metalloprotease with thrombospondin motif type I) in atherogenesis. METHODS AND RESULTS: ADAMTS-1 is expressed at the highest levels in the aorta when compared with other human tissues examined. Immunolocalization studies in human aorta and coronary artery indicate that ADAMTS-1 expression is mainly seen at low levels in the medial layer, but upregulated in the intima when plaque is present. We found that ADAMTS-1 mRNA levels are significantly higher in proliferating/migrating cultured primary aortic vascular smooth muscle cells (VSMCs) compared with resting/confluent cells. Using the mouse carotid artery flow cessation model, we show that there are differences in vessel remodeling in ADAMTS-1 transgenic/apoE-deficient mice compared with apoE deficiency alone, particularly a significant increase in intimal hyperplasia. We show that ADAMTS-1 can cleave the large versican containing proteoglycan population purified from cultured human aortic VSMCs. Finally, using versican peptide substrates, we show data suggesting that ADAMTS-1 cleaves versican at multiple sites. CONCLUSIONS: We hypothesize that ADAMTS-1 may promote atherogenesis by cleaving extracellular matrix proteins such as versican and promoting VSMC migration.
Assuntos
Arteriosclerose/patologia , Artéria Carótida Primitiva/patologia , Proteoglicanas de Sulfatos de Condroitina/metabolismo , Desintegrinas/fisiologia , Imuno-Histoquímica/métodos , Metaloendopeptidases/fisiologia , Peptídeo Hidrolases/metabolismo , Proteínas ADAM , Proteína ADAMTS1 , Adolescente , Animais , Arteriosclerose/metabolismo , Artéria Carótida Primitiva/química , Artéria Carótida Primitiva/metabolismo , Artéria Carótida Primitiva/cirurgia , Linhagem Celular , Modelos Animais de Doenças , Desintegrinas/biossíntese , Desintegrinas/imunologia , Desintegrinas/metabolismo , Humanos , Hidrólise , Lectinas Tipo C , Ligadura/métodos , Masculino , Metaloendopeptidases/biossíntese , Metaloendopeptidases/imunologia , Metaloendopeptidases/metabolismo , Camundongos , Camundongos Endogâmicos C57BL , Pessoa de Meia-Idade , Músculo Liso Vascular/química , Músculo Liso Vascular/metabolismo , Músculo Liso Vascular/patologia , Miócitos de Músculo Liso/química , Miócitos de Músculo Liso/metabolismo , Miócitos de Músculo Liso/patologia , Neovascularização Patológica/patologia , Reação em Cadeia da Polimerase Via Transcriptase Reversa/métodos , VersicanasRESUMO
BACKGROUND: Acute myeloid leukemia (AML) carrying nucleophosmin 1 (NPM1) mutations (NPMc(+)) is regarded as a separate entity of myeloid neoplasms due to its distinct biological and clinical features. However, NPMc(+) alone displays low leukemogenic activity and cooperating events appear crucial for AML to develop. Dysregulation of homeobox genes, such as HOXA9 and MEIS1, is a common transcriptional signature of NPMc(+) AML. Furthermore, the pathogenic role for NPMc(+) in AML remains incompletely understood. AIM: To elucidate if NPMc(+) collaborates with Meis1 or Hoxa9 in the evolvement of AML. METHODS: Murine bone marrow cells were genetically engineered to express mutated NPM1 variant A in combination with overexpression of Meis1 or Hoxa9. The capacity of the transduced cells to transform in vitro and to cause leukemia in vivo was then assessed. FINDINGS AND CONCLUSION: There was no synergy between NPMc(+) and Meis1 or Hoxa9 in causing leukemogenic transformation of murine bone marrow cells, or in inducing AML in a transplantation model. Hence, overexpression of Meis1 or Hoxa9 in combination with NPMc(+) expression was not sufficient to generate an NPMc(+) AML mouse model.
RESUMO
Sexual selection can lead to rapid divergence in reproductive characters. Recent studies have indicated that postmating events, such as sperm precedence, may play a key role in speciation. Here, we stress that other components of postmating sexual selection may be involved in the evolution of reproductive isolation. One of these is the reproductive investment made by females after mating (i.e., differential allocation). We performed an experiment designed to assess genetic divergence in the effects of mating on female reproductive performance in flour beetles, Tribolium castaneum. Females were mated to males of three different wild-type genotypes at two different frequencies, in all possible reciprocal combinations. Male genotype affected all aspects of female reproduction, through its effects on female longevity, total offspring production, reproductive rate, mating rate, and fertility. Moreover, male and female genotype interacted in their effects on offspring production and reproductive rate. We use the pattern of these interactions to discuss the evolutionary process of divergence and suggest that the pattern is most consistent with that expected if divergence was driven by sexually antagonistic coevolution. In particular, the fact that females exhibited a relatively weak response to males with which they were coevolved suggests that females have evolved resistance to male gonadotropic signals/stimuli.
Assuntos
Besouros/fisiologia , Reprodução , Comportamento Sexual Animal , Animais , Feminino , MasculinoRESUMO
The transactivating function of the oriPI-EBNA1 complex is essential for activation of the Epstein-Barr virus (EBV) C promoter (Cp) in lymphoblastoid cell lines expressing the viral growth programme. Furthermore, the oriPI-EBNA1 complex is believed to play an important role during promoter switching upon primary infection of B-lymphocytes and establishment of latent infection in vivo. Previously, it was shown that six EBNA1-binding sites within oriPI were required for transactivation of the heterologous thymidine kinase promoter. Here, we define the number of EBNA1-binding sites within oriPI necessary for its biological function as EBNA1-dependent Cp enhancer. We show that four EBNA1-binding sites within oriPI lead to significant upregulation of Cp in response to EBNA1 and eight or more to full activation. Thus, multiple EBNA1 homodimers at oriPI are required for the formation of a transcriptionally active Cp complex, a process that involves EBNA1-induced changes in the chromatin structure including DNA looping and nucleosome destabilization.
Assuntos
Antígenos Nucleares do Vírus Epstein-Barr/metabolismo , Herpesvirus Humano 4/genética , Regiões Promotoras Genéticas/genética , Origem de Replicação , Ativação Transcricional , Sítios de Ligação , Elementos Facilitadores Genéticos/genética , Antígenos Nucleares do Vírus Epstein-Barr/genéticaRESUMO
Retinoic acid receptor-related orphan receptor-alpha (RORalpha) (NR1F1) is an orphan nuclear receptor with a potential role in metabolism. Previous studies have shown that RORalpha regulates transcription of the murine Apolipoprotein AI gene and human Apolipoprotein CIII genes. In the present study, we present evidence that RORalpha also induces transcription of the human Apolipoprotein AV gene, a recently identified apolipoprotein associated with triglyceride levels. Adenovirus-mediated overexpression of RORalpha increased the endogenous expression of ApoAV in HepG2 cells and RORalpha also enhanced the activity of an ApoAV promoter construct in transiently transfected HepG2 cells. Deletion and mutation studies identified three AGGTCA motifs in the ApoAV promoter that mediate RORalpha transactivation, one of which overlaps with a previously identified binding site for PPARalpha. Together, these results suggest a novel mechanism whereby RORalpha modulates lipid metabolism and implies RORalpha as a potential target for the treatment of dyslipidemia and atherosclerosis.