The clinical phenotype of germline RUNX1 mutations in relation to the accompanying somatic variants and RUNX1 isoform expression.

Germline RUNX1 mutations lead to familial platelet disorder with associated myeloid malignancy (FPDMM), characterized by thrombocytopenia, abnormal bleeding, and an elevated risk of developing myelodysplastic neoplasia (MDS) and acute myeloid leukemia (AML) at young age. However, it is not known why or how germline carriers of RUNX1 mutations have a particular propensity to develop myeloid hematologic malignancies, but the acquisition and composition of somatic mutations are believed to initiate and determine disease progression. We present a novel family pedigree that shares a common germline RUNX1R204* variant and exhibits a spectrum of somatic mutations and related myeloid malignancies (MM). RUNX1 mutations are associated with inferior clinical outcome; however, the proband of this family developed MDS with ring sideroblasts (MDS-RS), classified as a low-risk MDS subgroup. His relatively indolent clinical course is likely due to a specific somatic mutation in the SF3B1 gene. While the three main RUNX1 isoforms have been ascribed various roles in normal hematopoiesis, they are now being increasingly recognized as involved in myeloid disease. We investigated the RUNX1 transcript isoform patterns in the proband and his sister, who carries the same germline RUNX1R204* variant, and has FPDMM but no MM. We demonstrate a RUNX1a increase in MDS-RS, as previously reported in MM. Interestingly, we identify a striking unbalance of RUNX1b and -c in FPDMM. In conclusion, this report reinforces the relevance of somatic variants on the clinical phenotypic heterogeneity in families with germline RUNX1 deficiency and investigates a potential new role for RUNX1 isoform disequilibrium as a mechanism for development of MM.

Erythroid Differentiation Enhances RNA Mis-Splicing in SF3B1-Mutant Myelodysplastic Syndromes with Ring Sideroblasts.

Myelodysplastic syndromes with ring sideroblasts (MDS-RS) commonly develop from hematopoietic stem cells (HSC) bearing mutations in the splicing factor SF3B1 (SF3B1mt). Direct studies into MDS-RS pathobiology have been limited by a lack of model systems that fully recapitulate erythroid biology and RS development and the inability to isolate viable human RS. Here, we combined successful direct RS isolation from patient samples, high-throughput multiomics analysis of cells encompassing the SF3B1mt stem-erythroid continuum, and functional assays to investigate the impact of SF3B1mt on erythropoiesis and RS accumulation. The isolated RS differentiated, egressed into the blood, escaped traditional nonsense-mediated decay (NMD) mechanisms, and leveraged stress-survival pathways that hinder wild-type hematopoiesis through pathogenic GDF15 overexpression. Importantly, RS constituted a contaminant of magnetically enriched CD34+ cells, skewing bulk transcriptomic data. Mis-splicing in SF3B1mt cells was intensified by erythroid differentiation through accelerated RNA splicing and decreased NMD activity, and SF3B1mt led to truncations in several MDS-implicated genes. Finally, RNA mis-splicing induced an uncoupling of RNA and protein expression, leading to critical abnormalities in proapoptotic p53 pathway genes. Overall, this characterization of erythropoiesis in SF3B1mt RS provides a resource for studying MDS-RS and uncovers insights into the unexpectedly active biology of the "dead-end" RS. SIGNIFICANCE: Ring sideroblast isolation combined with state-of-the-art multiomics identifies survival mechanisms underlying SF3B1-mutant erythropoiesis and establishes an active role for erythroid differentiation and ring sideroblasts themselves in SF3B1-mutant myelodysplastic syndrome pathogenesis.

Integrated Genomic and Transcriptomic Analysis Improves Disease Classification and Risk Stratification of MDS with Ring Sideroblasts.

PURPOSE: Ring sideroblasts (RS) define the low-risk myelodysplastic neoplasm (MDS) subgroup with RS but may also reflect erythroid dysplasia in higher risk myeloid neoplasm. The benign behavior of MDS with RS (MDSRS+) is limited to SF3B1-mutated cases without additional high-risk genetic events, but one third of MDSRS+ carry no SF3B1 mutation, suggesting that different molecular mechanisms may underlie RS formation. We integrated genomic and transcriptomic analyses to evaluate whether transcriptome profiles may improve current risk stratification. EXPERIMENTAL DESIGN: We studied a prospective cohort of MDSRS+ patients irrespective of World Health Organization (WHO) class with regard to somatic mutations, copy-number alterations, and bone marrow CD34+ cell transcriptomes to assess whether transcriptome profiles add to prognostication and provide input on disease classification. RESULTS: SF3B1, SRSF2, or TP53 multihit mutations were found in 89% of MDSRS+ cases, and each mutation category was associated with distinct clinical outcome, gene expression, and alternative splicing profiles. Unsupervised clustering analysis identified three clusters with distinct hemopoietic stem and progenitor (HSPC) composition, which only partially overlapped with mutation groups. IPSS-M and the transcriptome-defined proportion of megakaryocyte/erythroid progenitors (MEP) independently predicted survival in multivariable analysis. CONCLUSIONS: These results provide essential input on the molecular basis of SF3B1-unmutated MDSRS+ and propose HSPC quantification as a prognostic marker in myeloid neoplasms with RS.

Análise dos indicadores das metas do ODS 6 e sua relação com indicadores socioeconômicos para os municípios do estado de São Paulo / Analysis of SDG 6 targets indicators and their relationship with socioeconomic indicator for municipalities in the State of São Paulo

A falta de acesso a sistemas água potável e esgotamento adequados afeta todos os aspectos da vida humana, tendo os maiores impactos sobre localidades menos desenvolvidas e comunidades marginalizadas. A proposição do ODS6 destacou a importância do desenvolvimento inclusivo e o compromisso, dos países membros da ONU no acesso universal e equitativo a esses serviços, segundo a premissa de "não deixar ninguém para trás". As condições e dados atuais de mensuração para o ODS 6, medindo apenas as taxas de cobertura desses serviços, projetam o Brasil, e especialmente São Paulo, em níveis de atendimento altos, mas não permitem demonstrar as desigualdades no acesso de determinados grupos, visando alcançar a redução progressiva e eliminação das desigualdades entre grupos populacionais. Diante deste cenário, este trabalho visou analisar a evolução temporal e dos déficits, no período de 2010 a 2019 dos indicadores de acesso a serviços adequados de água e esgoto para o Estado de São Paulo, coletados do Painel ODS6, elaborado pela equipe do NARA-FSP/USP, e identificar o perfil socioeconômico dos grupos mais afetados pela falta de acesso a esses serviços, por meio da relação com os dados autônomos do Censo Demográfico 2010. Os resultados permitiram observar que não obstante a evolução quantitativa dos indicadores de água e esgotamento sanitário, as regiões com maiores desigualdades de renda, com maior presença de comunidades tradicionais e de geografia naturalmente frágil são as que permanecem com menores índices de atendimento à população. Da mesma forma, os municípios com maior proporção de domicílios rurais, chefiados por mulheres e com rendimento mensal de até 2 salários-mínimos tem menor proporção de acesso a serviços adequados de abastecimento de água e coleta de esgoto. Regiões historicamente impactadas pela miséria e pobreza e grupos populacionais socialmente vulneráveis não são considerados nas principais bases de dados do país. Devido a esse viés, que não considera essa parcela da população, não há acesso a informações relevantes para a elaboração e monitoramento de políticas públicas. O acesso universal e equitativo, em 2030, apenas será possível se medidas afirmativas forem adotadas, dando preferência a certos grupos e indivíduos, a fim de corrigir a discriminação que se mantêm até os dias de hoje.

The lack of access to adequate water and sanitation systems affects all aspects of human life, having the greatest impacts on less developed localities and marginalized communities. The proposition of SDG6 highlighted the importance of inclusive development and the commitment of UN member countries to universal and equitable access to these services, according to the premise of "leaving no one behind". The current measurement conditions and data for SDG 6, measuring only the coverage rates of these services, indicate high service levels for Brazil, and mainly São Paulo, but it is not able to identify the inequalities in access of certain groups, in order to reach the progressive reduction and elimination of inequalities between population groups. In view of this scenario, this work aimed to analyze the evolution over time and the deficits, between 2010 and 2019, of the indicators of access to adequate water and sewage services for the State of São Paulo, collected from the SDG 6 Panel, prepared by the NARA- FSP/USP, and to identify the socioeconomic profile of the groups that are the most affected by the lack of access to these services, through the relation with the autonomous data from the 2010 Demographic Census. The results show that, despite the quantitative evolution of water and sanitation income indicators, regions with greater population inequalities, presence of traditional communities and naturally fragile geography remain with the lowest rates of these services. Likewise, municipalities with a higher proportion of rural households headed by women and with a monthly income of up to 2 minimum salaries have a lower proportion of access to adequate water supply and sewage collection services. Regions historically impacted by misery, poverty and socially considered population groups are not present in the country's main databases. Due to this bias, that does not consider this population portion, there is no access to relevant information for the development and monitoring of public policies. Universal and equitable access to these services, in 2030, will only be possible if affirmative measures are adopted, focusing on certain groups and individuals, in order to abolish the social discrimination that remains to the present days.