Role of FOXM1 and FOXO3a in B-lymphoblastic leukaemia progression and glucocorticoid responsiveness

B-cell acute lymphoblastic leukaemia (B-ALL) is one of the most common paediatric disorders characterized by an accumulation of B-cell blasts reminiscent of normal stages of differentiation and by infiltration of various extramedullary sites. B-ALL patients that respond poorly to glucocorticoid therapy when diagnosed are usually predicted to undergo relapse. Therefore understanding the biological mechanisms underlying this poor responsiveness is crucial for the development of more effective therapies. FOXM1 belongs to the Forkhead Box (FOX) superfamily of transcriptional factors and is a major regulator of cell proliferation and cell cycle progression. Its expression increases at the entry of cell cycle S-phase, remains stable during G2/M phase, and finally, is degraded at the mitotic exit. FOXM1 is a bona fide proto-oncogene and its up-regulation is often associated with a high proliferation rate, resistance to drugs and poor prognosis of many tumours, as well as with the development and progression of many malignancies. Therefore, downregulation of FOXM1 expression and activity might be a key strategy to inhibit tumour progression. In the first study we investigated the potential role of FOXM1 in B-ALL cell proliferation, aiming to understand if FOXM1 can be targeted to either increase the efficiency of chemotherapeutic treatment, or to overcome drug resistance in this haematological malignancy. We observed that FOXM1 protein and mRNA levels are higher in B-ALL patients and cell lines when compared to lymphocytes of healthy donors (peripheral blood mononuclear cells, PBMCs). We then downregulated FOXM1 in REH and NALM-6 cell lines using a specific FOXM1 siRNA or thiostrepton treatment: there was a significant reduction in cell proliferation, cell cycle arrest in G2/M and an increase in sub-G1 cells. Importantly, we observed that thiostrepton acts in synergy with conventional chemotherapeutic agents, increasing their efficiency and partially reversing the glucocorticoid resistance in REH cells. Altogether, our results suggest that FOXM1 inhibition could be a useful strategy to increase the efficacy of existing therapeutics for B-ALL and to overcome drug resistance. In the second study we focused our attention on another FOX family member: the transcription factor FOXO3a. Although FOXO3a is mainly described as a bona fide tumour suppressor, literature reveals that it might also play a role as a proto-oncogene, by inducing cancer development and drug resistance according to the cellular context. FOXO3a transcriptional activity is regulated by a complex array of post-translational modifications. Amongst there, phosphorylation and acetylation are the major regulators of FOXO3a subcellular localization and transcriptional function. In this work, we investigated whether FOXO3a is involved in dexamethasone responsiveness of B-lymphoblastic leukaemia (B-ALL). Western blot and immunostaining data showed that FOXO3a becomes active only in sensitive cells following dexamethasone treatment. In dexamethasone sensitive cells (RS4;11 and SUP-B15), FOXO3a translocates from the cytoplasm to the nucleus, where it induces the expression of BIM, p27 and the activation of apoptosis signature through caspase-3 cleavage. Moreover knockdown experiments clearly reveal that FOXO3a expression and function is fundamental for dexamethasone mediated cytotoxicity. We then performed a series of experiments to clarify if its activity is regulated by phosphorylation and/or acetylation. More importantly, two post-translational modifications, phosphorylation on Ser7 and acetylation on Lys242 and Lys245, appeared to be crucial for dexamethasone-mediated FOXO3a activation in sensitive cells. Clarifying which are the biological processes that prevent these post-translational modifications from occurring in dexamethasone-resistant REH cells, might pave the way to overcoming glucocorticoid resistance in B-AL

The Chilean Spanish version of the Juvenile Arthritis Multidimensional Assessment Report (JAMAR)

The Juvenile Arthritis Multidimensional Assessment Report (JAMAR) is a new parent/patient reported outcome measure that enables a thorough assessment of the disease status in children with juvenile idiopathic arthritis (JIA). We report the results of the cross-cultural adaptation and validation of the parent and patient versions of the JAMAR in the Chilean Spanish language. The reading comprehension of the questionnaire was tested in ten JIA parents and patients. Each participating centre was asked to collect demographic, clinical data and the JAMAR in 100 consecutive JIA patients or all consecutive patients seen in a 6-month period and to administer the JAMAR to 100 healthy children and their parents. The statistical validation phase explored descriptive statistics and the psychometric issues of the JAMAR: the three Likert assumptions, floor/ceiling effects, internal consistency, Cronbach\u2019s alpha, interscale correlations, and construct validity (convergent and discriminant validity). A total of 49 JIA patients (12.2% systemic, 24.5% oligoarticular, 22.5% RF-negative polyarthritis, 40.8% other categories) and 70 healthy children, were enrolled. The JAMAR components discriminated well healthy subjects from JIA patients. All JAMAR components revealed good psychometric performances. In conclusion, the Chilean Spanish version of the JAMAR is a valid tool for the assessment of children with JIA and is suitable for use both in routine clinical practice and clinical research

The Estonian version of the Juvenile Arthritis Multidimensional Assessment Report (JAMAR)

The Juvenile Arthritis Multidimensional Assessment Report (JAMAR) is a new parent/patient-reported outcome measure that enables a thorough assessment of the disease status in children with juvenile idiopathic arthritis (JIA). We report the results of the cross-cultural adaptation and validation of the parent and patient versions of the JAMAR in the Estonian language. The reading comprehension of the questionnaire was tested in 10 JIA parents and patients. Each participating centre was asked to collect demographic, clinical data and the JAMAR in 100 consecutive JIA patients or all consecutive patients seen in a 6-month period and to administer the JAMAR to 100 healthy children and their parents. The statistical validation phase explored descriptive statistics and the psychometric issues of the JAMAR: the three Likert assumptions, floor/ceiling effects, internal consistency, Cronbach\u2019s alpha, interscale correlations, test\u2013retest reliability, and construct validity (convergent and discriminant validity). A total of 110 JIA patients (71.8% oligoarticular, 18.2% RF-negative polyarthritis, 10% other categories) and 98 healthy children were enrolled in one paediatric rheumatology centre. Notably, none of the enrolled JIA patients is affected with systemic JIA. The JAMAR components discriminated healthy subjects from JIA patients, except for the Paediatric Rheumatology Quality of Life (HRQoL) Psychosocial Health (PsH) subscales and for the satisfaction with current health status. All JAMAR components revealed good psychometric performances. In conclusion, the Estonian version of the JAMAR is a valid tool for the assessment of children with JIA and is suitable for use both in routine clinical practice and clinical research

The Georgian version of the Juvenile Arthritis Multidimensional Assessment Report (JAMAR)

The Juvenile Arthritis Multidimensional Assessment Report (JAMAR) is a new parent/patient reported outcome measure that enables a thorough assessment of the disease status in children with juvenile idiopathic arthritis (JIA). We report the results of the cross-cultural adaptation and validation of the parent and patient versions of the JAMAR in the Georgian language. The reading comprehension of the questionnaire was tested in 10 JIA parents and patients. Each participating centre was asked to collect demographic, clinical data and the JAMAR in 100 consecutive JIA patients or all consecutive patients seen in a 6-month period and to administer the JAMAR to 100 healthy children and their parents. The statistical validation phase explored descriptive statistics and the psychometric issues of the JAMAR: the 3 Likert assumptions, floor/ceiling effects, internal consistency, Cronbach\u2019s alpha, interscale correlations, test\u2013retest reliability, and construct validity (convergent and discriminant validity). A total of 100 JIA patients (26% systemic, 57% oligoarticular, 16% RF negative polyarthritis, and 1% RF positive polyarthritis) and 100 healthy children, were enrolled at two paediatric rheumatology centre. Notably, none of the enrolled JIA patients is affected with psoriatic arthritis or with enthesitis-related arthritis or with undifferentiated arthritis. The JAMAR components discriminated healthy subjects from JIA patients, except for the school-related problems variable. All JAMAR components revealed good psychometric performances. In conclusion, the Georgian version of the JAMAR is a valid tool for the assessment of children with JIA and is suitable for use both in routine clinical practice and clinical research

The Ukrainian version of the Juvenile Arthritis Multidimensional Assessment Report (JAMAR)

The Juvenile Arthritis Multidimensional Assessment Report (JAMAR) is a new parent/patient-reported outcome measure that enables a thorough assessment of the disease status in children with juvenile idiopathic arthritis (JIA). We report the results of the cross-cultural adaptation and validation of the parent and patient versions of the JAMAR in the Ukrainian language. The reading comprehension of the questionnaire was tested in 10 JIA parents and patients. Each participating centre was asked to collect demographic, clinical data and the JAMAR in 100 consecutive JIA patients or all consecutive patients seen in a 6-month period and to administer the JAMAR to 100 healthy children and their parents. The statistical validation phase explored descriptive statistics and the psychometric issues of the JAMAR: the three Likert assumptions, floor/ceiling effects, internal consistency, Cronbach\u2019s alpha, interscale correlations, test\u2013retest reliability, and construct validity (convergent and discriminant validity). A total of 100 JIA patients (2% systemic, 44% oligoarticular, 20% RF-negative polyarthritis, 34% other categories) and 100 healthy children were enrolled at the paediatric rheumatology centre of the Western Ukrainian Specialised Children\u2019s Medical Centre. The JAMAR components discriminated well between healthy subjects and JIA patients. Notably, there was no significant difference between healthy subjects and their affected peers in the school-related problems variable. All JAMAR components revealed good psychometric performances. In conclusion, the Ukrainian version of the JAMAR is a valid tool for the assessment of children with JIA and is suitable for use both in routine clinical practice and clinical research

The Swedish version of the Juvenile Arthritis Multidimensional Assessment Report (JAMAR)

The Juvenile Arthritis Multidimensional Assessment Report (JAMAR) is a new parent/patient-reported outcome measure that enables a thorough assessment of the disease status in children with juvenile idiopathic arthritis (JIA). We report the results of the cross-cultural adaptation and validation of the parent and patient versions of the JAMAR in the Swedish language. The reading comprehension of the questionnaire was tested in 10 JIA parents and patients. Each participating centre was asked to collect demographic, clinical data and the JAMAR in 100 consecutive JIA patients or all consecutive patients seen in a 6-month period and to administer the JAMAR to 100 healthy children and their parents. The statistical validation phase explored descriptive statistics and the psychometric issues of the JAMAR: the 3 Likert assumptions, floor/ceiling effects, internal consistency, Cronbach\u2019s alpha, interscale correlations, test\u2013retest reliability and construct validity (convergent and discriminant validity). A total of 68 JIA patients (8.8% systemic, 44.1% oligoarticular, 13.2% RF negative polyarthritis, 33.9% other categories) and 76 healthy children, were enrolled in two centres. The JAMAR components discriminated well healthy subjects from JIA patients. All JAMAR components revealed good psychometric performances. In conclusion, the Swedish version of the JAMAR is a valid tool for the assessment of children with JIA and is suitable for use both in routine clinical practice and clinical research

FOXM1 is overexpressed in B-acute lymphoblastic leukemia (B-ALL) and its inhibition sensitizes B-ALL cells to chemotherapeutic drugs

The Forkhead box protein M1 (FOXM1) is a transcription factor that plays a central role in the regulation of cell cycle, proliferation, DNA repair, and apoptosis. FOXM1 is overexpressed in many human tumors and its upregulation has been linked to high proliferation rates and poor prognosis. We therefore studied the role of FOXM1 in B-lymphoblastic leukemia (B-ALL) in order to understand whether FOXM1 could be a key target for leukemia therapy. RT-PCR and western blot analysis were carried out in a small cohort of pediatric B-ALL patients to evaluate FOXM1 levels. To assess its biological relevance, its expression was down-modulated by transient RNA interference in B-ALL cell lines (REH and NALM-6). Our results show that FOXM1 expression is higher in both B-ALL patients and cell lines when compared to PBMC or normal B-cells (CD19+) from healthy donors. Furthermore, blocking FOXM1 activity in two B-ALL cell lines, by either knockdown or treatment with the FOXM1 inhibitor thiostrepton, causes significant decrease in their cell proliferation. This decrease in cell proliferation was coupled with both an induction of the G2/M cell cycle arrest and with a reduction in the S phase population. Finally, we noted how thiostrepton synergises with chemotherapeutic agents commonly used in B-ALL therapy, thus increasing their efficiency. Therefore our results suggest that FOXM1 is highly expressed in both patients and B-ALL cell lines, and that targeting FOXM1 could be an attractive strategy for leukemia therapy and for overcoming drug resistance

IL-1 Inhibition in Systemic Juvenile Idiopathic Arthritis

Systemic juvenile idiopathic arthritis (sJIA) is the form of childhood arthritis whose treatment is most challenging. The demonstration of the prominent involvement of interleukin (IL)-1 in disease pathogenesis has provided the rationale for the treatment with biologic medications that antagonize this cytokine. The three IL-1 blockers that have been tested so far (anakinra, canakinumab, and rilonacept) have all been proven effective and safe, although only canakinumab is currently approved for use in sJIA. The studies on IL-1 inhibition in sJIA published in the past few years suggest that children with fewer affected joints, higher neutrophil count, younger age at disease onset, shorter disease duration, or, possibly, higher ferritin level may respond better to anti-IL-1 treatment. In addition, it has been postulated that use of IL-1 blockade as first-line therapy may take advantage of a window of opportunity, in which disease pathophysiology can be altered to prevent the occurrence of chronic arthritis. In this review, we analyze the published literature on IL-1 inhibitors in sJIA and discuss the rationale underlying the use of these medications, the results of therapeutic studies, and the controversial issues

The British English version of the Juvenile Arthritis Multidimensional Assessment Report (JAMAR)

The Juvenile Arthritis Multidimensional Assessment Report (JAMAR) is a new parent/patient-reported outcome measure that enables a thorough assessment of the disease status in children with juvenile idiopathic arthritis (JIA). We report the results of the cross-cultural adaptation and validation of the parent and patient versions of the JAMAR in the British English language. The reading comprehension of the questionnaire was tested in ten JIA parents and patients. Each participating centre was asked to collect demographic, clinical data and the JAMAR in 100 consecutive JIA patients or all consecutive patients seen in a 6-month period and to administer the JAMAR to 100 healthy children and their parents. The statistical validation phase explored descriptive statistics and the psychometric issues of the JAMAR: the three Likert assumptions, floor/ceiling effects, internal consistency, Cronbach\u2019s alpha, interscale correlations, test\u2013retest reliability, and construct validity (convergent and discriminant validity). A total of 100 JIA patients (7.0% systemic, 38.0% oligoarticular, 27.0% RF negative polyarthritis, 28% other categories) and 100 healthy children, were enrolled at the Royal Hospital for Sick Children in Glasgow. The JAMAR components discriminated well healthy subjects from JIA patients. All JAMAR components revealed good psychometric performances. In conclusion, the British English version of the JAMAR is a valid tool for the assessment of children with JIA and is suitable for use both in routine clinical practice and clinical research