Pharmacogenetics of pediatric acute lymphoblastic leukemia in Uruguay: adverse events related to induction phase drugs.

In Uruguay, the pediatric acute lymphoblastic leukemia (ALL) cure rate is 82.2%, similar to those reported in developed countries. However, many patients suffer adverse effects that could be attributed, in part, to genetic variability. This study aims to identify genetic variants related to drugs administered during the induction phase and analyze their contribution to adverse effects, considering individual genetic ancestry. Ten polymorphisms in five genes (ABCB1, CYP3A5, CEP72, ASNS, and GRIA1) related to prednisone, vincristine, and L-asparaginase were genotyped in 200 patients. Ancestry was determined using 45 ancestry informative markers (AIMs). The sample ancestry was 69.2% European, 20.1% Native American, and 10.7% African, but with high heterogeneity. Mucositis, Cushing syndrome, and neurotoxicity were the only adverse effects linked with genetic variants and ancestry. Mucositis was significantly associated with ASNS (rs3832526; 3R/3R vs. 2R carriers; OR: = 6.88 [1.88-25.14], p = 0.004) and CYP3A5 (non-expressors vs. expressors; OR: 4.55 [1.01-20.15], p = 0.049) genes. Regarding Cushing syndrome, patients with the TA genotype (rs1049674, ASNS) had a higher risk of developing Cushing syndrome than those with the TT genotype (OR: 2.60 [1.23-5.51], p = 0.012). Neurotoxicity was significantly associated with ABCB1 (rs9282564; TC vs. TT; OR: 4.25 [1.47-12.29], p = 0.007). Moreover, patients with <20% Native American ancestry had a lower risk of developing neurotoxicity than those with ≥20% (OR: 0.312 [0.120-0.812], p = 0.017). This study shows the importance of knowing individual genetics to improve the efficacy and safety of acute lymphoblastic leukemia.

COVID-19 in children and the influence on the employment activity of their female caregivers: A cross sectional gender perspective study.

Introduction: During the COVID-19 pandemic, women disproportionately assume more unpaid activities, affecting their employment. Objective: Describe the influence of COVID-19 on the employment of caregivers of children and adolescents from a gender perspective. Methods: Cross-sectional study in three high-complexity hospitals in Bogotá, Colombia from April 2020 to June 2021. A subsample of the FARA cohort was taken, including those patients with a positive test for SARS-COV2. We took as our analysis category children older than 8 years and younger than 18 years who had a positive SARS-COV2 test, as well as, caregivers of all children with a positive SARS-COV2 test. This subsample was drawn from the FARA cohort. A survey was applied to them. We carried out a descriptive and stratified analysis by age group, educational, and socioeconomic level. Results: We included 60 surveys of caregivers and 10 surveys of children. The main caregiver in 94.8% of the cases was a female. At the beginning of the pandemic, 63.3% of the caregivers were employed, and 78.9% of those lost their employment. The vast majority of these caregiver were women (96.6%, n = 29). A predominance of loss of work activity was documented in caregivers of children in early childhood 66.6% (n = 20), with lower education 66.6% (n = 20), and from lower strata 56.6% (n = 17). Conclusion: Caregivers of children with COVID-19 with low educational levels and lower socioeconomic conditions, as well as those with children under 5 years showed greater likelihood of employment loss between the interviewed subsample.

Respuesta con enfoque de derechos de la niñez frente a la pandemia por COVID-19 en Chile, Colombia y Perú

[RESUMEN]. Objetivo. Describir las estrategias que fueron establecidas por Chile, Colombia y Perú durante el primer año de la pandemia por COVID-19 y compararlas desde el enfoque de derechos de la niñez. Métodos. Se realizó un estudio cualitativo de análisis comparado de políticas públicas, tomando como eje siete categorías construidas por el Capítulo Latinoamericano de la International-Society-for-Social-Pediatrics-and-Child-Health a partir de la Convención de Derechos de la Niñez (CDN). La selección de los documentos de los países se realizó por conveniencia y su análisis en diálogos deliberativos. Resultados. Se revisaron 173 documentos de los tres países. Destaca como convergencia la priorización de la prevención de la transmisión comunitaria del virus, por sobre la promoción del ejercicio de derechos de la niñez, la falta de participación de niños, niñas y adolescentes (NNA) en el proceso de elaboración de las políticas públicas, y la falta de avance en el reconocimiento y protección del ejercicio de todos sus derechos. No hubo mayores divergencias más allá de brechas de desigualdad identificadas con base a la realidad de cada país. Conclusión. La pandemia ha afectado el funcionamiento de los sistemas económicos, sociales, de salud, educación, medioambiente y gobernanza de estos tres países. Si bien este estudio muestra un avance en la inclusión del enfoque de derechos de NNA en las políticas formuladas, su comprensión como sujetos sociales y políticos titulares de derecho podría permitir la construcción de alternativas colectivas que garanticen la salud y el bienestar para todas las personas en el curso de vida.

[ABSTRACT]. Objective. Describe the strategies established by Chile, Colombia, and Peru during the first year of the COVID-19 pandemic and compare them from a children's rights perspective. Methods. A qualitative study with comparative analysis of public policies was conducted around seven categories constructed by the Latin American Chapter of the International Society for Social Pediatrics and Child Health, based on the Convention on the Rights of the Child. Country documents were selected based on convenience sampling and were analyzed in deliberative dialogues. Results. 173 documents from the three countries were reviewed. There was convergence around: prioritization of prevention of community transmission of the virus over promotion of the exercise of children's rights; lack of participation of children and adolescents in the process of developing public policies; and lack of progress in the recognition and protection of the exercise of children’s rights overall. There were no major divergences beyond identified inequality gaps grounded in the reality of each country. Conclusion. The pandemic has affected the functioning of the economic, social, health, education, environmental, and governance systems in these three countries. While this study shows progress in the inclusion of the children's rights approach in formulated policies, the recognition of children and adolescents as holders of social and political rights could allow the construction of collective alternatives that guarantee health and well-being for all people throughout the life course.

[RESUMO]. Objetivo. Descrever as estratégias que foram utilizadas por Chile, Colômbia e Peru durante o primeiro ano da pandemia de COVID-19 e compará-las a partir da perspectiva dos direitos da criança. Métodos. Realizou-se um estudo qualitativo de análise comparativa de políticas públicas, tomando como eixo sete categorias construídas pelo Capítulo Latino-americano da International Society for Social Pediatrics & Child Health a partir da Convenção sobre os Direitos da Criança (CDC). A seleção de documentos dos países foi feita por conveniência, e sua análise ocorreu mediante diálogos deliberativos. Resultados. Foram revisados 173 documentos dos três países. A prioridade da prevenção da transmissão comunitária do vírus destaca-se por sua convergência, em detrimento da promoção do exercício dos direitos da criança, da falta de participação de crianças e adolescentes no processo de formulação de políticas públicas e da falta de progresso no reconhecimento e proteção da efetivação de todos os seus direitos. Não houve grandes divergências além das lacunas de desigualdade identificadas com base na realidade de cada país. Conclusões. A pandemia afetou o funcionamento dos sistemas econômico, social, de saúde, educação, meio ambiente e governança desses três países. Embora este estudo mostre avanços na inclusão da perspectiva dos direitos da criança e do adolescente nas políticas formuladas, a compreensão deles como sujeitos sociais e políticos titulares de direitos pode permitir a construção de alternativas coletivas que garantam saúde e bem-estar a todas as pessoas ao longo do curso da vida.

[Response to the COVID-19 Pandemic in Chile, Colombia, and Peru from a children's rights perspectiveResposta à pandemia de COVID-19 no Chile, Colômbia e Peru com foco nos direitos da criança]. / Respuesta con enfoque de derechos de la niñez frente a la pandemia por COVID-19 en Chile, Colombia y Perú.

OBJECTIVE: Describe the strategies established by Chile, Colombia, and Peru during the first year of the COVID-19 pandemic and compare them from a children's rights perspective. METHODS: A qualitative study with comparative analysis of public policies was conducted around seven cat-egories constructed by the Latin American Chapter of the International Society for Social Pediatrics and Child Health, based on the Convention on the Rights of the Child. Country documents were selected based on convenience sampling and were analyzed in deliberative dialogues. RESULTS: 173 documents from the three countries were reviewed. There was convergence around: prioritization of prevention of community transmission of the virus over promotion of the exercise of children's rights; lack of participation of children and adolescents in the process of developing public policies; and lack of progress in the recognition and protection of the exercise of children's rights overall. There were no major divergences beyond identified inequality gaps grounded in the reality of each country. CONCLUSION: The pandemic has affected the functioning of the economic, social, health, education, environmental, and governance systems in these three countries. While this study shows progress in the inclusion of the children's rights approach in formulated policies, the recognition of children and adolescents as holders of social and political rights could allow the construction of collective alternatives that guarantee health and well-being for all people throughout the life course.

OBJETIVO: Descrever as estratégias que foram utilizadas por Chile, Colômbia e Peru durante o primeiro ano da pandemia de COVID-19 e compará-las a partir da perspectiva dos direitos da criança. MÉTODOS: Realizou-se um estudo qualitativo de análise comparativa de políticas públicas, tomando como eixo sete categorias construídas pelo Capítulo Latino-americano da International Society for Social Pediatrics & Child Health a partir da Convenção sobre os Direitos da Criança (CDC). A seleção de documentos dos países foi feita por conveniência, e sua análise ocorreu mediante diálogos deliberativos. RESULTADOS: Foram revisados 173 documentos dos três países. A prioridade da prevenção da transmissão comunitária do vírus destaca-se por sua convergência, em detrimento da promoção do exercício dos direitos da criança, da falta de participação de crianças e adolescentes no processo de formulação de políticas públicas e da falta de progresso no reconhecimento e proteção da efetivação de todos os seus direitos. Não houve grandes divergências além das lacunas de desigualdade identificadas com base na realidade de cada país. CONCLUSÕES: A pandemia afetou o funcionamento dos sistemas econômico, social, de saúde, educação, meio ambiente e governança desses três países. Embora este estudo mostre avanços na inclusão da perspectiva dos direitos da criança e do adolescente nas políticas formuladas, a compreensão deles como sujeitos sociais e políticos titulares de direitos pode permitir a construção de alternativas coletivas que garantam saúde e bem-estar a todas as pessoas ao longo do curso da vida.

Ancestry and TPMT-VNTR Polymorphism: Relationship with Hematological Toxicity in Uruguayan Patients with Acute Lymphoblastic Leukemia.

6-Mercaptopurine (6-MP) is a thiopurine drug widely used in childhood acute lymphoblastic leukemia (ALL) therapy. Genes such as TPMT and NUDT15 have an outstanding role in 6-MP metabolism. Mutations in both genes explain a significant portion of hematological toxicities suffered by ALL Uruguayan pediatric patients. A variable number tandem repeat in the TPMT promoter (TPMT-VNTR) has been associated with TPMT expression. This VNTR has a conservative architecture (AnBmC). To explore new causes of hematological toxicities related to ALL therapy, we genotyped the TPMT-VNTR of 130 Uruguayan pediatric patients. Additionally, individual genetic ancestry was estimated by 45 ancestry-informative markers (AIMs). Hematological toxicity was measured as the number of leukopenia events and 6-MP dose along the maintenance phase. As previously reported, we found TPMT*2 and TPMT*3C alleles were associated to TPMT-VNTR A2BC and AB2C, respectively. However, contrasting with other reports, TPMT*3A allele was found in a heterogeneous genetic background in linkage equilibrium. Patients carrying more than 5 A repeats present a significant higher number of leukopenia events among patients without TPMT and/or NUDT15 variants. Native American ancestry and the number of A repeats were significantly correlated with the number of leukopenia events. However, the correlation between Native American ancestry and the number of leukopenia events was lost when the number of A repeats was considered as covariate. This suggests that TPMT-VNTR alleles are more relevant than Native American ancestry in the hematological toxicity. Our results emphasize that TPMT-VNTR may be used as a pharmacogenetic biomarker to predict 6-MP-related hematological toxicity in ALL childhood therapy.

GM2 gangliosidoses in Spain: analysis of the HEXA and HEXB genes in 34 Tay-Sachs and 14 Sandhoff patients.

The GM2 gangliosidoses are autosomal recessive lysosomal storage diseases caused by a deficiency of the ß-hexosaminidase A enzyme. This enzyme is composed of two polypeptide chains designated the α- and ß- subunits and it interacts with the GM2 activator protein. The HEXA and HEXB genes encode the α-subunit and the ß-subunit, respectively. Mutations in these genes are causative of Tay-Sachs disease (HEXA) and Sandhoff disease (HEXB). We analyzed the complete HEXA gene in 34 Spanish patients with Tay-Sachs disease and the HEXB gene in 14 Spanish patients with Sandhoff disease. We identified 27 different mutations, 14 of which were novel, in the HEXA gene and 14 different mutations, 8 of which unreported until now, in the HEXB gene, and we attempted to correlate these mutations with the clinical presentation of the patients. We found a high frequency of c.459+5G>A (IVS4+5G>A) mutation in HEXA affected patients, 22 of 68 alleles, which represent the 32.4%. This is the highest percentage found of this mutation in a population. All patients homozygous for mutation c.459+5G>A presented with the infantile form of the disease and, as previously reported, patients carrying mutation p.R178H in at least one of the alleles presented with a milder form. In HEXB affected patients, the novel deletion c.171delG accounts for 21.4% of the mutant alleles (6/28). All patients with this deletion showed the infantile form of the disease. The Spanish GM2 gangliosidoses affected patients show a great mutational heterogeneity as seen in other inherited lisosomal diseases in this country.

The p38 MAPK-MK2 axis regulates E2F1 and FOXM1 expression after epirubicin treatment.

E2F1 is responsible for the regulation of FOXM1 expression, which plays a key role in epirubicin resistance. Here, we examined the role and regulation of E2F1 in response to epirubicin in cancer cells. We first showed that E2F1 plays a key role in promoting FOXM1 expression, cell survival, and epirubicin resistance as its depletion by siRNA attenuated FOXM1 induction and cell viability in response to epirubicin. We also found that the p38-MAPK activity mirrors the expression patterns of E2F1 and FOXM1 in both epirubicin-sensitive and -resistant MCF-7 breast cancer cells, suggesting that p38 has a role in regulating E2F1 expression and epirubicin resistance. Consistently, studies using pharmacologic inhibitors, siRNA knockdown, and knockout mouse embryonic fibroblasts (MEF) revealed that p38 mediates the E2F1 induction by epirubicin and that the induction of E2F1 by p38 is, in turn, mediated through its downstream kinase MK2 [mitogen-activated protein kinase (MAPK)-activated protein kinase 2; MAPKAPK2]. In agreement, in vitro phosphorylation assays showed that MK2 can directly phosphorylate E2F1 at Ser-364. Transfection assays also showed that E2F1 phosphorylation at Ser-364 participates in its induction by epirubicin but also suggests that other phosphorylation events are also involved. In addition, the p38-MK2 axis can also limit c-jun-NH(2)-kinase (JNK) induction by epirubicin and, notably, JNK represses FOXM1 expression. Collectively, these findings underscore the importance of p38-MK2 signaling in the control of E2F1 and FOXM1 expression as well as epirubicin sensitivity.

Phosphorylation of FOXO3a on Ser-7 by p38 promotes its nuclear localization in response to doxorubicin.

FOXO3a is a forkhead transcription factor that regulates a multitude of important cellular processes, including proliferation, apoptosis, differentiation, and metabolism. Doxorubicin treatment of MCF-7 breast carcinoma cells results in FOXO3a nuclear relocation and the induction of the stress-activated kinase p38 MAPK. Here, we studied the potential regulation of FOXO3a by p38 in response to doxorubicin. Co-immunoprecipitation studies in MCF-7 cells demonstrated a direct interaction between p38 and FOXO3a. We also showed that p38 can bind and phosphorylate a recombinant FOXO3a directly in vitro. HPLC-coupled phosphopeptide mapping and mass spectrometric analyses identified serine 7 as a major site for p38 phosphorylation. Using a phosphorylated Ser-7 FOXO3a antibody, we demonstrated that FOXO3a is phosphorylated on Ser-7 in response to doxorubicin. Immunofluorescence staining studies showed that upon doxorubicin treatment, the wild-type FOXO3a relocalized to the nucleus, whereas the phosphorylation-defective FOXO3a (Ala-7) mutant remained largely in the cytoplasm. Treatment with SB202190 also inhibits the doxorubicin-induced FOXO3a Ser-7 phosphorylation and nuclear accumulation in MCF-7 cells. In addition, doxorubicin caused the nuclear translocation of FOXO3a in wild-type but not p38-depleted mouse fibroblasts. Together, our results suggest that p38 phosphorylation of FOXO3a on Ser-7 is essential for its nuclear relocalization in response to doxorubicin.

ATM and p53 regulate FOXM1 expression via E2F in breast cancer epirubicin treatment and resistance.

In this report, we investigated the role and regulation of forkhead box M1 (FOXM1) in breast cancer and epirubicin resistance. We generated epirubicin-resistant MCF-7 breast carcinoma (MCF-7-EPI(R)) cells and found FOXM1 protein levels to be higher in MCF-7-EPI(R) than in MCF-7 cells and that FOXM1 expression is downregulated by epirubicin in MCF-7 but not in MCF-7-EPI(R) cells. We also established that there is a loss of p53 function in MCF-7-EPI(R) cells and that epirubicin represses FOXM1 expression at transcription and gene promoter levels through activation of p53 and repression of E2F activity in MCF-7 cells. Using p53(-/-) mouse embryo fibroblasts, we showed that p53 is important for epirubicin sensitivity. Moreover, transient promoter transfection assays showed that epirubicin and its cellular effectors p53 and E2F1 modulate FOXM1 transcription through an E2F-binding site located within the proximal promoter region. Chromatin immunoprecipitation analysis also revealed that epirubicin treatment increases pRB (retinoblastoma protein) and decreases E2F1 recruitment to the FOXM1 promoter region containing the E2F site. We also found ataxia-telangiectasia mutated (ATM) protein and mRNA to be overexpressed in the resistant MCF-7-EPI(R) cells compared with MCF-7 cells and that epirubicin could activate ATM to promote E2F activity and FOXM1 expression. Furthermore, inhibition of ATM in U2OS cells with caffeine or depletion of ATM in MCF-7-EPI(R) with short interfering RNAs can resensitize these resistant cells to epirubicin, resulting in downregulation of E2F1 and FOXM1 expression and cell death. In summary, our data show that ATM and p53 coordinately regulate FOXM1 via E2F to modulate epirubicin response and resistance in breast cancer.

Characterization of Drosophila melanogaster JmjC+N histone demethylases.

In this article, we characterize histone demethylase activity of the entire family of JmjC+N proteins of Drosophila melanogaster. Our results show that Lid (little imaginal discs), which is structurally homologous to JARID1, demethylates H3K4me3. However, contrary to what would be inferred from its demethylase activity, lid contributes to the establishment of transcriptionally competent chromatin states as: (i) is required for histone H3 acetylation; (ii) contributes to expression of the homoeotic gene Ultrabithorax (Ubx); and (iii) antagonizes heterochromatin-mediated gene silencing (PEV). These results, which are consistent with the identification of lid as a trithorax group (trxG) gene, are discussed in the context of current models for the contribution of H3K4me3 to the regulation of gene expression. Here, we also show that the two Drosophila JMJD2 homologues, dJMJD2(1)/CG15835 and dJMJD2(2)/CG33182, are capable of demethylating both H3K9me3 and H3K36me3. dJMJD2(1)/CG15835 regulates heterochromatin organization, as its over-expression induces spreading of HP1, out of heterochromatin, into euchromatin, without affecting the actual pattern of histone modifications of heterochromatin. dJMJD2(1)/CG15835 is excluded from heterochromatin and localizes to multiple euchromatic sites, where it regulates H3K36 methylation. These results indicate that dJMJD2(1)/CG15835 contributes to delimit hetero- and euchromatic territories through the regulation of H3K36 methylation in euchromatin. On the other hand, dJARID2/CG3654 shows no demethylase activity on H3K4me3, H3K9me3, H3K27me3, H3K36me3 and H4K20me3.