FIND: the importance of early diagnosis in mucopolyssacharidoses

As Mucopolissacaridoses (MPS) são um sub-grupo das Doenças Lisossomais de Sobrecarga, causadas por deficiências em enzimas lisossomais, que catalisam a degradação dos glicosaminoglicanos (também designados GAGs ou mucopolissacarídeos), que se acumulam nos lisossomas de diferentes órgãos e tecidos. As MPS têm apresentação multissistémica, com diferentes graus de gravidade, e evolução variável. Sendo patologias de apresentação heterogénea e consequentemente de diagnóstico difícil, o objetivo do estudo FIND, que resulta de uma parceria entre a Secção de Doenças Hereditárias do Metabolismo da Sociedade Portuguesa de Pediatria e o Instituto Nacional de Saúde Doutor Ricardo Jorge, é alertar os clínicos para sinais e sintomas de risco ao mesmo tempo que disponibiliza uma ferramenta de diagnóstico. Este diagnóstico é efetuado através da determinação enzimática em sangue colhido em cartão, sendo possível a identificação da enzima deficiente, em sete tipos de MPS. O estudo FIND, devido à forma fácil e económica de obtenção de amostra, associada à baixa quantidade necessária para a análise, coloca à disposição dos clínicos, um ótimo meio para a identificação e caraterização de casos sintomáticos de MPS em idade pediátrica.Mucopolyssacharidoses (MPS) are a sub-goup of Lysosomal Storage Disorders, caused by the impairment of lysosomal enzymes, that are responsible for the degradation of Glycosaminoglycans (also known as GAGs or mucopolyssacharides), which are stored in the lysosome. MPS are multissystemic disorders, with dif ferent degrees of severity and evolution. Since they have a wide spectrum of presentation, becoming the diagnosis very dif ficult, it appears the Project FIND, that results from the association between Secção de Doenças Hereditárias do Metabolismo da Sociedade Por tuguesa de Pediatria and Instituto Nacional de Saúde Dr. Ricardo Jorge, in order to claim awareness to red flags of MPS and to provide a useful tool to diagnose it. This diagnosis is done by measuring lysosomal enzymatic activities in dried blood spots, in order to detect the deficient one. Project FIND, due to the small amount, inexpensive and easy way to collect the sample, make available to the physicians a good way to identify and characterize symptomatic MPS patients at pediatric age.info:eu-repo/semantics/publishedVersio

FIND: a importância de um diagnóstico

O Projeto FIND é um estudo de colaboração entre a SDHM da SPP e o INSA e permite disponibilizar uma ferramenta de diagnóstico aos médicos que sigam um doente com suspeita clínica de mucopolissacaridose (MPS).N/

Public preferences for involvement in the governance of health data

Public involvement in the governance of epidemiological and public health studies can foster needs-driven research, enhance participants' recruitment, reduce attrition and improve the quality of and ethics in research and surveillance. However, it can also reinforce health inequalities if it fails to ensure public representation across socioeconomic gradients. This study aimed to assess patients' and carers' preferences for involvement in collective health data governance, and its associated factors, to strengthen the evidence base for policy development

Benefits and risks of sharing genomic data for research: comparing the views of rare disease patients, informal carers and healthcare professionals

Assessing public and patients’ expectations and concerns about genomic data sharing is essential to promote adequate data governance and engagement in rare diseases genomics research. This cross-sectional study compared the views of 159 rare disease patients, 478 informal carers and 63 healthcare professionals in Northern Portugal about the benefits and risks of sharing genomic data for research, and its associated factors. The three participant groups expressed significantly different views. The majority of patients (84.3%) and informal carers (87.4%) selected the discovery of a cure for untreatable diseases as the most important benefit. In contrast, most healthcare professionals revealed a preference for the development of new drugs and treatments (71.4%), which was the second most selected benefit by carers (48.3%), especially by the more educated (OR (95% CI): 1.58 (1.07–2.34)). Lack of security and control over information access and the extraction of information exceeding research objectives were the two most often selected risks by patients (72.6% and 50.3%, respectively) and carers (60.0% and 60.6%, respectively). Conversely, professionals were concerned with genomic data being used to discriminate citizens (68.3%), followed by the extraction of information exceeding research objectives (54.0%). The latter risk was more frequently expressed by more educated carers (OR (95% CI): 1.60 (1.06–2.41)) and less by those with blue-collar (OR (95% CI): 0.44 (0.25–0.77) and other occupations (OR (95% CI): 0.44 (0.26–0.74)). Developing communication strategies and consent approaches tailored to participants’ expectations and needs can benefit the inclusiveness of genomics research that is key for patient-centred care.FEDER through the Operational Programme for Competitiveness and Internationalisation and national funding from the Foundation for Science and Technology—FCT (Portuguese Ministry of Science, Technology and Higher Education) (Ref. POCI-01–0145-FEDER-032194), under the project Public and patient involvement in health data governance: a people-centred approach to data protection in genetic diseases (Ref. FCT PTDC/SOC-SOC/32194/2017) and the Unidade de Investigação em Epidemiologia—Instituto de Saúde Pública da Universidade do Porto (EPIUnit) (Ref. UIDB/04750/2020), Laboratório para a Investigação Integrativa e Translacional em Saúde Populacional (ITR) LA/P/0064/2020), the individual contract grant L57/2016/CP1336/CT0001 (C.d.F)

Trimetilaminúria (Síndroma de odor a peixe) uma doença subestimada: espectro mutacional da população portuguesa

Pretende-se com este trabalho divulgar a capacidade instalada de estudo desta doença, esclarecer a etiologia de casos investigados e tentar correlacionar o genótipo/fenótipo da doença, minimizando os impactos psicossociais que esta patologia acarreta. Por outro lado, pretende-se também alertar para a necessidade do estudo desta patologia de uma forma integrada com a farmacogenética, uma vez que certos genótipos poderão condicionar a atuação de um determinado fármaco

Public and patient involvement in health data governance (DATAGov): protocol of a people-centred, mixed-methods study on data use and sharing for rare diseases care and research.

INTRODUCTION: International policy imperatives for the public and patient involvement in the governance of health data coexist with conflicting cross-border policies on data sharing. This can challenge the planning and implementation of participatory data governance in healthcare services locally. Engaging with local stakeholders and understanding how their needs, values and preferences for governing health data can be articulated with policies made at the supranational level is crucial. This paper describes a protocol for a project that aims to coproduce a people-centred model for involving patients and the public in decision-making processes about the use and sharing of health data for rare diseases care and research. METHODS AND ANALYSIS: This multidisciplinary project draws on an explanatory sequential mixed-methods study. A hospital-based survey with patients, informal carers, health professionals and technical staff recruited at two reference centres for rare diseases in Portugal will be conducted first. The qualitative study will follow consisting of semi-structured interviews and scenario-based workshops with a subsample of the participant groups recruited at baseline. Quantitative data will be analysed using descriptive and inferential statistics. Inductive and deductive approaches will be combined to analyse the qualitative interviews. Data from scenario-based workshops will be iteratively compared using the constant comparison method to identify cross-cutting themes and categories. ETHICS AND DISSEMINATION: The Ethics Committee for Health from the University Hospital Centre São João/Faculty of Medicine of University of Porto approved the study protocol (Ref. 99/19). Research findings will be disseminated at academic conferences and science promotion events, and through public meetings involving patient representatives, practitioners, policy-makers and students, a project website and peer-reviewed journal publications

NPC1 silent variant induces skipping of exon 11 (p.V562V) and unfolded protein response was found in a specific Niemann‐Pick type C patient

Background: Niemann-Pick type C (NPC, MIM #257220) is a neuro-visceral disease, caused predominantly by pathogenic variants in the NPC1 gene. Here we studied patients with clinical diagnosis of NPC but inconclusive results regarding the molecular analysis. Methods: We used a Next-Generation Sequencing (NGS)-panel followed by cDNA analysis. Latter, we used massively parallel single-cell RNA-seq (MARS-Seq) to address gene profiling changes and finally the effect of different variants on the protein and cellular levels. Results: We identified novel variants and cDNA analysis allowed us to establish the functional effect of a silent variant, previously reported as a polymorphism. We demonstrated that this variant induces the skipping of exon 11 leading to a premature stop codon and identified it in NPC patients from two unrelated families. MARS-Seq analysis showed that a number of upregulated genes were related to the unfolded protein response (UPR) and endoplasmic reticulum (ER) stress in one specific patient. Also, for all analyzed variants, the NPC1 protein was partially retained in the ER. Conclusion: We showed that the NPC1 silent polymorphism (p.V562V) is a disease-causing variant in NPC and that the UPR is upregulated in an NPC patient.info:eu-repo/semantics/publishedVersio

Advances in the diagnosis of mitochondrial diseases by next generation sequencing

O recente desenvolvimento da tecnologia de sequenciação de nova geração (NGS) revolucionou o diagnóstico molecular das doenças genéticas raras, de difícil diagnóstico, tais como as doenças mitocondriais. O estudo destas patologias foi implementado em 1993 pelo nosso grupo e até à data foram investigados mais de 2500 doentes portugueses. Muitos destes doentes ainda não dispõem de diagnóstico molecular, pelo que foi desenvolvida uma estratégia de NGS para a identificação da mutação causal. A sequenciação de um painel de 209 genes nucleares associados a doenças mitocondriais e do DNA mitocondrial completo por NGS, foi realizada num sequenciador MiSeq (Illumina). O estudo de 145 doentes permitiu identificar 41 mutações causais e caraterizar 35 doentes. Esta investigação contribuiu para esclarecer a etiologia molecular destes doentes (35/145; 24%), ii) alargar o espetro mutacional destas patologias e, iii) oferecer um aconselhamento genético e um eventual diagnóstico pré-natal aos casais em risco. O desenvolvimento de um painel, específico para estas patologias, tem um caráter inovador e reforça o nosso Centro como laboratório nacional para o estudo e investigação de doenças mitocondriais.Recent development of high throughput, next-generation sequencing (NGS) technology has revolutionized the research and molecular diagnosis of hardto- diagnose genetic disorders such as mitochondrial disorders. The study of these diseases was implemented in 1993 by our group and to date more than 2,500 Portuguese patients have been investigated. As many of these patients do not yet have molecular diagnosis, an NGS strategy was developed to identify the causal mutation. NGS was performed in a MiSeq Illumina instrument using a custom mitochondrial gene panel with around 209 genes involved in mitochondria metabolism and the entire human mitochondrial genome. The study of 145 patients allowed the identification of 41 causal mutations and the molecular characterization of 35 patients. This investigation contributed to i) identify the pathogenic mutations in the studied patients (35/145; 24%), ii) expand the mutational spectrum in the etiology of these disorders, and iii) propose an accurate genetic counseling. Custom design panels have been widely used for molecular heterogeneous disorders however, the development of this panel will be innovative in our country strengthening our Center as a national reference for the study and research of mitochondrial disorders.Estudo financiado por: Fundação da Ciência e Tecnologia (PTDC/DTP-PIC/2220/2014, Genetic Defects of Mitochondrial Diseases: a Next Generation Sequencing Approach) - implementação da tecnologia de NGS e o desenho de um painel de genes nucleares aplicado ao diagnóstico das doenças mitocondriais; Programa NORTE 2020 (NORTE-01-0246-FEDER-000014, DESVENDAR “DEScobrir, VENcer as Doenças rARas”) - aquisição de equipamentos para a realização da sequenciação de nova geração.info:eu-repo/semantics/publishedVersio

Leukocyte Imbalances in Mucopolysaccharidoses Patients

(This article belongs to the Special Issue Inherited Metabolic Disorders: From Bench to Bedside)Mucopolysaccharidoses (MPSs) are rare inherited lysosomal storage diseases (LSDs) caused by deficient activity in one of the enzymes responsible for glycosaminoglycans lysosomal degradation. MPS II is caused by pathogenic mutations in the IDS gene, leading to deficient activity of the enzyme iduronate-2-sulfatase, which causes dermatan and heparan sulfate storage in the lysosomes. In MPS VI, there is dermatan sulfate lysosomal accumulation due to pathogenic mutations in the ARSB gene, leading to arylsulfatase B deficiency. Alterations in the immune system of MPS mouse models have already been described, but data concerning MPSs patients is still scarce. Herein, we study different leukocyte populations in MPS II and VI disease patients. MPS VI, but not MPS II patients, have a decrease percentage of natural killer (NK) cells and monocytes when compared with controls. No alterations were identified in the percentage of T, invariant NKT, and B cells in both groups of MPS disease patients. However, we discovered alterations in the naïve versus memory status of both helper and cytotoxic T cells in MPS VI disease patients compared to control group. Indeed, MPS VI disease patients have a higher frequency of naïve T cells and, consequently, lower memory T cell frequency than control subjects. Altogether, these results reveal MPS VI disease-specific alterations in some leukocyte populations, suggesting that the type of substrate accumulated and/or enzyme deficiency in the lysosome may have a particular effect on the normal cellular composition of the immune system.This work was funded by National Funds through FCT—Fundação para a Ciência e a Tecnologia, I.P., under the project UIDB/04293/2020.info:eu-repo/semantics/publishedVersio

Comissão Coordenadora do Tratamento das Doenças Lisossomais de Sobrecarga − Relatório de actividades 2018

Relatório de atividades da Comissão Coordenadora do Tratamento das Doenças Lisossomais de Sobrecarga (CCTDLS) relativo ao ano de 2018. A CCTDLS funciona no âmbito do INSA, sendo constituída por profissionais designados pelo INSA, pelos Centros de Referência nesta área, pela Administração Central do Sistema de Saúde, pela Direção-Geral da Saúde e pela Autoridade Nacional do Medicamento e Produtos de Saúde (Infarmed). As Doenças Lisossomais de Sobrecarga (DLS) constituem um grupo de patologias raras, progressivas, com elevada morbilidade, que engloba, atualmente, mais de 60 patologias, cuja apresentação clínica é extremamente variável, podendo ir desde doença neurológica grave até a casos menos graves. Em Portugal, a prevalência deste tipo de patologias em recém-nascidos é de 25 por cada 100.000 nados vivos. Nos últimos anos, várias terapêuticas de substituição enzimática e de redução do substrato têm sido desenvolvidas possibilitando o tratamento dos doentes com algumas destas patologias, nomeadamente, a Doença de Gaucher, a Doença de Fabry, a Doença de Pompe, as Mucopolissacaridoses tipo I, II, IVA e VI, a Deficiência de lipase ácida lisossomal e a Doença de Niemann Pick tipo C. Do relatório anual da CCTDLS, relativo ao ano 2018, destaca-se a realização das seguintes atividades: i) Discussão e avaliação de 47 casos clínicos com emissão de 40 pareceres relativos a pedidos de início de tratamento (19), ajuste de dose (12), alteração do tratamento (8) e transferência de hospital (1). Foram emitidos pareceres para a Doença de Gaucher, Doença de Fabry, Mucopolissacaridoses tipo I, II e VI, Doença de Pompe e Doença de Niemann Pick tipo C; ii) Definição de critérios gerais de início e de suspensão de terapêuticas; iii) Monitorização da evolução clínica dos doentes com terapias em curso. Foi efetuada a monitorização de 29 doentes com Doença de Fabry (1 do Centro de Referência Centro Hospitalar Universitário S. João e 28 do Centro de Referência Hospital Senhora da Oliveira – Guimarães; iv) Avaliação e elaboração de protocolos de avaliação e seguimento das diferentes patologias de acordo com regras de anonimização; v) Atualização de dados referentes ao número total de doentes com DLS em tratamento no território continental, por centro hospitalar, por patologia e terapêutica; vi) Desenvolvimento de folhetos informativos para o doente.info:eu-repo/semantics/publishedVersio