Inflammation and lipoperoxidation in mucopolysaccharidoses type II patients at diagnosis and post-hematopoietic stem cell transplantation.

INTRODUCTION: Mucopolysaccharidosis type II (MPS II) is caused by deficiency of the enzyme iduronate-2-sulfatase; one possible therapy for MPS II is hematopoietic stem cell transplantation (HSCT). It is established that there is excessive production of reactive species in MPS II patients, which can trigger several processes, such as the inflammatory cascade. OBJECTIVES: Our aim was to outline an inflammatory profile and lipoperoxidation of MPS II patients for a better understanding of disease and possible benefits that HSCT can bring in these processes. MATERIALS AND METHODS: We investigate oxidative damage to lipids by 15-F2t-isoprostane urinary concentrations and plasma pro-and anti-inflammatory cytokine concentrations in MPS II patients at diagnosis, MPS II post-HSCT patients, and controls. RESULTS: Interleukin (IL)-1ß and IL-17a concentrations were significantly increased and a tendency toward increased IL-6 production in the diagnosis group was verified. We found significant decrease in IL-4 and increase in 15-F2t-isoprostane concentrations in the diagnosis group, while IL-1ß, IL-6, IL-17a and 15-F2t-isoprostane concentrations were similar between control and post-HSCT groups. CONCLUSIONS: Our study demonstrated that MPS II patients at diagnosis are in a pro-inflammatory state, bringing a novel result showing increased production of IL-17a, an osteoclastogenic cytokine, as well as demonstrating that these patients have oxidative damage to lipids. Furthermore, evidence suggests that HSCT can reduce inflammation and lipoperoxidation in MPS II patients.

Ectasia coronariana difusa: anticoagulação pode prevenir infarto precoce? / Diffuse coronary ectasia: can anticoagulation prevent early infarction?

INTRODUÇÃO: Conceitua-se a ectasia de coronária (EC) como uma dilatação de pelo menos 1,5 vezes o tamanho da coronária adjacente normal. É documentada em 5% das coronariografias, sendo difusa ou localizada. A EC isolada aumenta o risco de isquemia miocárdica, e associada à doença arterial coronariana constitui uma combinação perigosa para eventos cardiovasculares. DESCRIÇÃO DO CASO: Homem, 44 anos, obeso, em uso de dupla antiagregação plaquetária (DAPT), estatina e antianginosos por infarto agudo do miocárdio sem supradesnivelamento de ST há um ano. História familiar de dois irmãos com infarto precoce aos 45 e 55 anos. Chega ao pronto socorro queixando de dor torácica retroesternal atípica iniciada há três horas da admissão. O exame físico não apresentava alterações e o eletrocardiograma demonstrava área elétrica inativa em parede inferior. Troponina de 3362ng/L (VR>1,5ng/L). Realizado cateterismo cardíaco (CATE) que evidenciou ectasia difusa em coronária esquerda e seus ramos, bem como coronária direita (CD) ectasiada e ocluída em seu óstio, com imagem sugestiva de trombo e acinesia em parede inferior. A ressonância cardíaca evidenciou disfunção biventricular (FEVE 40% e FEVD 26%), infarto extenso em ventrículo direito, e ausência de viabilidade em parede inferior e inferosseptal médio-basal de ventrículo esquerdo, achados corroborados pela cintilografia miocárdica com Tálio. CATE realizado há 1 ano também evidenciou CD ocluída e EC, portanto, visto a baixa probabilidade de sucesso através do tratamento invasivo percutâneo devido à anatomia desfavorável relacionado às ectasias, optado por manutenção do tratamento clínico com medicações que diminuem mortalidade na insuficiência cardíaca e associação de anticoagulação com Rivaroxabana à terapia com DAPT prévia. Iniciada investigação para trombofilias como fator desencadeante do quadro, corroborado pelo histórico familiar. Segue assintomático em reabilitação cardiovascular e acompanhamento ambulatorial. CONCLUSÕES: Descrevemos um caso de EC com a presença de trombo em paciente com história familiar de infarto precoce, apontando para possível etiologia trombótica. Levanta-se a possibilidade de se prevenir o segundo evento coronariano e a evolução para disfunção ventricular em caso de instituída a precoce anticoagulação, embora tal quadro tenha poucos relatos na literatura. Ratificamos ainda a importância da multimodalidade da imagem na construção de alicerces para decisão terapêutica.

Um caso de estenose de artéria renal por displasia fibromuscular: devemos resistir à tentação de revascularizar estes pacientes? / A case of renal artery stenosis due to fibromuscular dysplasia: should we resist the temptation to revascularize these patients?

INTRODUÇÃO: A hipertensão renovascular (HRV) é uma das principais causas de hipertensão arterial (HA) secundária, 90% das vezes por aterosclerose. A displasia fibromuscular (DFM) é uma doença vascular sistêmica rara, com incidência predominante em mulheres abaixo dos 50 anos e responsável por 10% dos casos de HRV. O padrão-ouro no tratamento da HRV por DFM é a angioplastia percutânea por balão (APB), com relatos de taxa de cura da HA de 35 a 80%. RELATO DE CASO: Mulher de 50 anos, com diagnóstico de HA aos 28 anos, relato de pré-eclâmpsia com dois abortos e mantendo quadro de HA refratária (PA média na MAPA de 24h: PA181x112 mmHg) em uso diário de olmesartana 40mg, clortalidona 25mg, metildopa 1500mg, anlodipino 10mg, espironolactona 50mg, hidralazina 100mg e metoprolol 100mg, foi submetida a rastreio de HA secundária. Observou-se no doppler de artérias renais (AR) estenose de AR direita maior que 60%, confirmada e melhor caracterizada na angiotomografia como múltiplas estenoses sugestivas de DFM. Demais exames complementares: aldosterona 8 ng/dL, atividade de renina plasmática: 0,8 ng/ml/h e creatinina: 0,8 mg/dL. A pesquisa de lesão de órgão-alvo revelou retinopatia grau 3 de Keith Wagener Barker; microalbuminúria (137 mg/g) hipertrofia ventricular esquerda (massa indexada de VE: 159g/m2). Foi submetida à APB com stent da AR direita, com resultado angiográfico satisfatório e sem intercorrências clínicas. PA na alta 160x108 mmHg e creatinina mantida em 0,8 mg/dL. Manteve-se hipertensa apesar das 7 classes medicamentosas prescritas e MAPA de 24h realizado 15 dias após o procedimento revelou PA média: 182x109 mmHg. CONCLUSÃO: Relatamos um caso de HA refratária associada à estenose de AR direita por DFM, submetida a APB e implante de stent, sem benefício clínico associado, mantendo-se a refratariedade do quadro hipertensivo ao longo do seguimento ambulatorial. A APB figura na literatura como o padrão-ouro no tratamento da DFM. Estudos observacionais reportam taxas de controle pressórico em torno de 70% e de cura da HA em aproximadamente 50% dos pacientes. Todavia, o caráter observacional desses estudos limita suas conclusões e os maiores estudos clínicos randomizados de revascularização arterial renal excluíram indivíduos com DFM. Este caso se soma ao conjunto de evidências contrárias à revascularização rotineira da AR na DFM. O tratamento precisa ser individualizado com base em novos preditores clínico-laboratoriais de sucesso.

"Ring-like" à ressonância cardíaca: intervenção precoce com cardiodesfibrilador implantável / "Ring-like" cardiac resonance: early intervention with implantable cardioverter defibrillator

INTRODUÇÃO: O padrão de realce tardio miocárdico (RT) do ventrículo esquerdo, denominado "ring-like", localiza-se no meso-epicárdio e abrange pelo menos três segmentos contíguos no mesmo corte de eixo curto, na ressonância cardíaca (RMC). Este padrão tem sido associado a uma maior incidência de eventos arrítmicos potencialmente fatais. Descreve-se um caso de intervenção precoce com a indicação de implantação de cardiodesfibrilador implantável (CDI), considerando o achado de "Ring like" com uma possível associação com laminopatia. RELATO DE CASO: Homem de 51 anos iniciou quadro de dispneia paroxística noturna, ortopneia e dispneia aos mínimos esforços em 2022. O ecocardiograma evidenciou disfunção ventricular com Fração de Ejeção (FE) de 32% às custas de hipocinesia difusa. Angiotomografia de coronárias sem lesões. A RMC demonstrou dilatação de todas câmaras (Ventrículo Esquerdo 78 mm x 75 mm), disfunção sistólica biventricular (FE de ventrículo esquerdo 12% e FE de ventrículo direito 17%), com RT de padrão não coronariano mesocárdico em todo o septo e nos segmentos anterior e inferior mediobasal. Os achados de RMC são compatíveis com cardiomiopatia dilatada com fibrose padrão ring-like (Figuras 1 e 2). Iniciado terapia médica com terapia quádrupla, com evolução satisfatória. Ao Holter de 24h, alta densidade de ectopias ventriculares, com episódios de Taquicardia Ventricular Polimórfica Não Sustentada. Apesar do teste genético não confirmatório, os achados de imagem com extensa quantidade de fibrose sugeriram uma cardiomiopatia dilatada por provável laminopatia. Considerando a FEVE < %, o sexo masculino e as arritmias ventriculares no Holter, caso foi discutido em reunião e indicado CDI como profilaxia primária. Evoluiu assintomático, sem novas intercorrências. CONCLUSÕES: Este relato de caso destaca a importância da RMC na avaliação e manejo da cardiomiopatia dilatada com padrão "ring-like", com papel fundamental na caracterização da extensão da fibrose mesocárdica, fornecendo informações adicionais que corroboram na indicação de CDI por profilaxia primária. O sexo masculino, as arritmias ventriculares não sustentadas no Holter e a fração de ejeção inferior a 45% foram identificados como fatores de risco para morte súbita nas séries de casos descritas de laminopatias até o momento.

Hematoma pericárdico organizado promovendo pericardite crônica constritiva: um relato de caso / Organized pericardial hematoma causing chronic constrictive pericarditis: a case report

INTRODUÇÃO: Diversas causas podem provocar inflamação do pericárdio, sendo as mais comuns as de origem tuberculosa e viral. A inflamação do pericárdio, assim como o processo de reparação subsequente que envolve fibrose e aumento da espessura, estão associados a condições de constrição e insuficiência cardíaca. Apresentamos um caso em que a causa da pericardite constritiva (PC) foi atípica, resultante de trauma cardíaco. DESCRIÇÃO DO CASO: Homem, 69 anos, agricultor, ex-tabagista, sem outras comorbidades. Há 3 meses passou a apresentar dispneia aos moderados esforços e edema de membros inferiores. À avaliação, apresentava sinais de congestão sistêmica, como turgência jugular e ascite, além de pulso paradoxal e sinal de kussmaul. Negou febre, perda de peso, sudorese noturna ou uso de medicações. Na radiografia do tórax, evidenciou-se área cardíaca normal e calcificação por todo o pericárdio. Ecocardiograma transtorácico evidenciou trombo em átrio direito e pericárdio espesso, associado à imagem hiperrefringente sugestiva de "massa" com sinais de compressão extrínseca do ventrículo direito e rechaçamento em direção ao ventrículo esquerdo (VE), com retificação do septo interventricular e diminuição da cavidade do VE, resultando em uma disfunção diastólica acentuada, mantendo função sistólica biventricular preservada. Realizado estudo tomográfico, que confirmou intensa calcificação pericárdica com imagem de "pseudo-tumor" de contornos irregulares, gerando intensa constricção e confirmando o diagnóstico de PC. Paciente foi submetido à pericardiectomia, que evidenciou grande quantidade de trombo calcificado no interior do "pseudo-tumor", com posterior resolução do quadro clínico. Após exclusão de múltiplas etiologias de pericardite e revisar seu histórico clínico, o paciente relatou trauma torácico contundente por cabeçada bovina há cerca de 10 anos, que cursou com dor torácica e dispneia por meses, sem atendimento médico na ocasião, sendo o hematoma pericárdico derivado do trauma, à provável etiologia do quadro. CONCLUSÕES: A pericardite constritiva, uma condição diagnóstica infrequente, está associada a significativa morbimortalidade e pode ser atribuída a uma variedade de acometimentos do pericárdio, incluindo trauma torácico. Portanto, é crucial identificar e gerenciar situações que possam levar à pericardite aguda e, potencialmente, à PC.

Partial Shoulder Arthroplasty Guided by Three-dimensional Prototyping.

Three-dimensional (3D) printing technology is a reality in medicine. In Orthopedics and Traumatology, 3D printing guides a precise and tailored surgical treatment. Understanding and disseminating its applicability, use, and outcomes can foster academicism and improve patient care. This is a report of a rare case of a female young adult patient with osteonecrosis of the humeral head due to avascular necrosis developed in early childhood. The treatment was tailored and optimized with 3D printing, which helped determine the steps for partial humeral arthroplasty.

Laronidase-loaded liposomes reach the brain and other hard-to-treat organs after noninvasive nasal administration.

Mucopolysaccharidosis type I (MPS I) is caused by a lack of the lysosomal enzyme α-L-iduronidase (IDUA), responsible for the degradation of the glycosaminoglycans (GAGs) dermatan and heparan sulfate, leading to multisystemic signs and symptoms. Enzyme replacement therapy (ERT) is a treatment that consists of weekly intravenous administrations of laronidase, a recombinant version of IDUA. However, ERT has limited access to certain tissues, such as bone, cartilage, and brain, and laronidase fails to trespass the BBB. In this sense, this study reports the development and characterization of laronidase-loaded liposomes for the treatment of MPS I mice. Liposomal complexes were obtained by the thin film formation method followed by microfluidization. The main characterization results showed mean vesicle size of 103.0 ± 3.3 nm, monodisperse populations of vesicles, zeta potential around + 30.0 ± 2.1 mV, and mucoadhesion strength of 5.69 ± 0.14 mN. Treatment of MPS I mice fibroblasts showed significant increase in enzyme activity. Nasal administration of complexes to MPS I mice resulted in significant increase in laronidase activity in the brain cortex, heart, lungs, kidneys, eyes, and serum. The overall results demonstrate the feasibility of nasal administration of laronidase-loaded liposomes to deliver enzyme in difficult-to-reach tissues, circumventing ERT issues and bringing hope as a potential treatment for MPS I.

Ectasia de coronária: anticoagulação pode prevenir infarto precoce? / Coronary ectasia: can anticoagulation prevent early heart attack?

INTRODUÇÃO: Conceitua-se a ectasia de coronária EC como uma dilatação de pelo menos 1,5 vezes o tamanho da coronária adjacente normal. É documentada em 5% das coronariografias, sendo difusa ou localizada. A EC isolada aumenta o risco de isquemia miocárdica, e associada à doença arterial coronariana constitui uma combinação perigosa para eventos cardiovasculares. DESCRIÇÃO DO CASO: Paciente de 44 anos, masculino, obeso, em uso de AAS, clopidogrel, estatina e antianginosos por infarto agudo do miocárdio sem supradesnivelamento de ST há um ano. História familiar de dois primos com infarto precoce aos 45 e 55 anos. Deu entrada no pronto socorro com dor torácica retroesternal em pontada iniciada há três horas da admissão. O eletrocardiograma demonstrava área elétrica inativa na parede inferior. Troponina de 3362ng/L (VR>1,5ng/L). Realizado cateterismo (CATE) que evidenciou ectasia difusa em coronária esquerda e seus ramos, bem como coronária direita (CD) ectasiada e ocluída em seu óstio, com imagem sugestiva de trombo e acinesia em parede inferior. A ressonância cardíaca evidenciou disfunção biventricular (FEVE 40% e FEVD 26%), infarto extenso em ventrículo direito, e ausência de viabilidade em parede inferior e inferosseptal médio-basal de ventrículo esquerdo, achados corroborados pela cintilografia com Tálio. Como cate realizado há 1 ano também evidenciou CD ocluída e EC, foi optado por tratamento clínico supracitado e foi associado metoprolol, sacubitril-valsartana, espironolactona, dapaglifozina, e rivaroxabana. Iniciada investigação para trombofilias como fator desencadeante do quadro, corroborado pelo histórico familiar. Segue assintomático em reabiltação cardiovascular e acompanhamento ambulatorial. DISCUSSÃO/CONCLUSÃO: Descrevemos um caso de EC com a presença de trombo em paciente com história familiar de infarto precoce, apontando para possível etiologia trombótica. Levanta-se a possibilidade de se prevenir o segundo evento coronariano e a evolução para disfunção ventricular em caso de instituída a precoce da anticoagulação, embora tal quadro tenha poucos relatos na literatura. Ratificamos ainda a importância da multimodalidade da imagem na construção de alicerces para decisão terapêutica.

Disturbances in mitochondrial bioenergetics and control quality and unbalanced redox homeostasis in the liver of a mouse model of mucopolysaccharidosis type II.

Mucopolysaccharidosis type II (MPS II; Hunter syndrome) is a lysosomal storage disease caused by mutations in the gene encoding the enzyme iduronate 2-sulfatase (IDS) and biochemically characterized by the accumulation of glycosaminoglycans (GAGs) in different tissues. It is a multisystemic disorder that presents liver abnormalities, the pathophysiology of which is not yet established. In the present study, we evaluated bioenergetics, redox homeostasis, and mitochondrial dynamics in the liver of 6-month-old MPS II mice (IDS-). Our findings show a decrease in the activity of α-ketoglutarate dehydrogenase and an increase in the activities of succinate dehydrogenase and malate dehydrogenase. The activity of mitochondrial complex I was also increased whereas the other complex activities were not affected. In contrast, mitochondrial respiration, membrane potential, ATP production, and calcium retention capacity were not altered. Furthermore, malondialdehyde levels and 2',7'-dichlorofluorescein oxidation were increased in the liver of MPS II mice, indicating lipid peroxidation and increased ROS levels, respectively. Sulfhydryl and reduced glutathione levels, as well as glutathione S-transferase, glutathione peroxidase (GPx), superoxide dismutase, and catalase activities were also increased. Finally, the levels of proteins involved in mitochondrial mass and dynamics were decreased in knockout mice liver. Taken together, these data suggest that alterations in energy metabolism, redox homeostasis, and mitochondrial dynamics can be involved in the pathophysiology of liver abnormalities observed in MPS II.

Disease progression in Sanfilippo type B: Case series of Brazilian patients.

Mucopolysaccharidosis type IIIB (MPS IIIB) is caused by deficiency of alpha-N-acetylglucosaminidase, leading to storage of heparan sulphate. The disease is characterized by intellectual disability and hyperactivity, among other neurological and somatic features. Here we studied retrospective data from a total of 19 MPS IIIB patients from Brazil, aiming to evaluate disease progression. Mean age at diagnosis was 7.2 years. Speech delay was one of the first symptoms to be identified, around 2-3 years of age. Behavioral alterations include hyperactivity and aggressiveness, starting around age four. By the end of the first decade, patients lost acquired abilities such as speech and ability to walk. Furthermore, as disease progresses, respiratory, cardiovascular and joint abnormalities were found in more than 50% of the patients, along with organomegaly. Most common cause of death was respiratory problems. The disease progression was characterized in multiple systems, and hopefully these data will help the design of appropriate clinical trials and clinical management guidelines.

Effect of genistein and coenzyme Q10 in oxidative damage and mitochondrial membrane potential in an attenuated type II mucopolysaccharidosis cellular model.

Mucopolysaccharidosis type II (MPS II) is an inborn error of the metabolism resulting from several possible mutations in the gene coding for iduronate-2-sulfatase (IDS), which leads to a great clinical heterogeneity presented by these patients. Many studies demonstrate the involvement of oxidative stress in the pathogenesis of inborn errors of metabolism, and mitochondrial dysfunction and oxidative stress can be related since most of reactive oxygen species come from mitochondria. Cellular models have been used to study different diseases and are useful in biochemical research to investigate them in a new promising way. The aim of this study is to develop a heterozygous cellular model for MPS II and analyze parameters of oxidative stress and mitochondrial dysfunction and investigate the in vitro effect of genistein and coenzyme Q10 on these parameters for a better understanding of the pathophysiology of this disease. The HP18 cells (heterozygous c.261_266del6/c.259_261del3) showed almost null results in the activity of the IDS enzyme and presented accumulation of glycosaminoglycans (GAGs), allowing the characterization of this knockout cellular model by MPS II gene editing. An increase in the production of reactive species was demonstrated (p < .05 compared with WT vehicle group) and genistein at concentrations of 25 and 50 µm decreased in vitro its production (p < .05 compared with HP18 vehicle group), but there was no effect of coenzyme Q10 in this parameter. There was a tendency for lysosomal pH change in HP18 cells in comparison to WT group and none of the antioxidants tested demonstrated any effect on this parameter. There was no increase in the activity of the antioxidant enzymes superoxide dismutase and catalase and oxidative damage to DNA in HP18 cells in comparison to WT group and neither genistein nor coenzyme q10 had any effect on these parameters. Regarding mitochondrial membrane potential, genistein induced mitochondrial depolarization in both concentrations tested (p < .05 compared with HP18 vehicle group and compared with WT vehicle group) and incubation with coenzyme Q10 demonstrated no effect on this parameter. In conclusion, it is hypothesized that our cellular model could be compared with a milder MPS II phenotype, given that the accumulation of GAGs in lysosomes is not as expressive as another cellular model for MPS II presented in the literature. Therefore, it is reasonable to expect that there is no mitochondrial depolarization and no DNA damage, since there is less lysosomal impairment, as well as less redox imbalance.

Regulatory Framework, Challenges, and Initial Strategic Planning for Advanced Therapy Products (PTAs) Development in Brazil.

Advanced Therapies are a class of innovative complex biological products used for therapeutic purposes, encompassing cell therapy, tissue engineering, and gene therapy products. These are promising therapeutic strategies for several complex diseases with low or non-existent therapeutic alternatives. The proper transposition of basic research in this area into medicinal products must comply with regulatory requirements. Here we review the main regulatory recommendations, emphasizing on the Brazilian regulation. The critical points are the manufacturing process, challenges in characterizing the product, development of non-clinical trials, lack of adequate animal models representative of the clinical situation, and absence of valid and measurable therapeutic endpoints. Based on that, we propose a framework for strategic planning of pre-clinical studies in this field. The detailed example involves producing a nonviral vector-based gene editing product, but the regulations and methods may be extrapolated for developing different types of advanced therapies.

Partial Shoulder Arthroplasty Guided by Three-dimensional Prototyping / Artroplastia parcial de ombro guiada por prototipagem tridimensional

Abstract Three-dimensional (3D) printing technology is a reality in medicine. In Orthopedics and Traumatology, 3D printing guides a precise and tailored surgical treatment. Understanding and disseminating its applicability, use, and outcomes can foster academicism and improve patient care. This is a report of a rare case of a female young adult patient with osteonecrosis of the humeral head due to avascular necrosis developed in early childhood. The treatment was tailored and optimized with 3D printing, which helped determine the steps for partial humeral arthroplasty.

Resumo A tecnologia de impressão tridimensional (3D) é uma realidade no âmbito da medicina. Na Ortopedia e Traumatologia, a impressão 3D direciona um tratamento cirúrgico preciso e individualizado. Compreender e difundir suas aplicações, utilidades e resultados pode fomentar o academicismo e melhorar o cuidado ao paciente. Trata-se de relato de um caso raro de uma paciente adulta jovem com osteonecrose da cabeça umeral por necrose avascular desenvolvida na primeira infância. O tratamento foi individualizado e otimizado com a impressão 3D, que ajudou a determinar os passos para o procedimento de artroplastia parcial de úmero.

Characterization of heart disease in mucopolysaccharidosis type II mice.

Mucopolysaccharidosis type II (MPSII) is a progressive lysosomal storage disease caused by mutations in the IDS gene, that leads to iduronate 2-sulfatase (IDS) enzyme deficiency. The enzyme catalyzes the first step of degradation of two glycosaminoglycans (GAGs), heparan sulfate (HS) and dermatan sulfate (DS). The consequences of MPSII are progressively harmful and can lead to death by cardiac failure. The aim of this study was to characterize the cardiovascular disease in MPSII mice. Thus, we evaluated the cardiovascular function of MPSII male mice at 6, 8, and 10 months of age, through functional, histological, and biochemical analyzes. Echocardiographic analyses showed a progressive loss in cardiac function, observed through parameters such as reduction in ejection fraction (46% in control versus 28% in MPS II at 10 months, P < .01) and fractional area change (31% versus 23%, P < .05). Similar results were found in parameters of vascular competence, obtained by echo Doppler. Both aortic dilatation and an increase in pulmonary resistance were observed at all time points in MPSII mice. The histological analyses showed an increase in the thickness of the heart valves (2-fold thicker than control values at 10 months). Biochemical analyzes confirmed GAG storage in these tissues, with a massive elevation of DS in the myocardium. Furthermore, an important increase in the activity of proteases such as cathepsin S and B (up to 5-fold control values) was found and could be related to the progressive loss of cardiac function observed in MPSII mice. In this work, we demonstrated that loss of cardiac function in MPSII mice started at 6 months of age, although its global cardiac capacity was still preserved at this time. Disease progressed at later time points leading to heart failure. The MPSII mice at later times reproduce many of the cardiovascular events found in patients with Hunter's disease.

Genome Editing Tools for Lysosomal Storage Disorders.

Genome editing has multiple applications in the biomedical field. They can be used to modify genomes at specific locations, being able to either delete, reduce, or even enhance gene transcription and protein expression. Here, we summarize applications of genome editing used in the field of lysosomal disorders. We focus on the development of cell lines for study of disease pathogenesis, drug discovery, and pathogenicity of specific variants. Furthermore, we highlight the main studies that use gene editing as a gene therapy platform for these disorders, both in preclinical and clinical studies. We conclude that gene editing has been able to change quickly the scenario of these disorders, allowing the development of new therapies and improving the knowledge on disease pathogenesis. Should they confirm their hype, the first gene editing-based products for lysosomal disorders could be available in the next years.

Ex vivo gene therapy for lysosomal storage disorders: future perspectives.

INTRODUCTION: Lysosomal storage disorders (LSD) are a group of monogenic rare diseases caused by pathogenic variants in genes that encode proteins related to lysosomal function. These disorders are good candidates for gene therapy for different reasons: they are monogenic, most of lysosomal proteins are enzymes that can be secreted and cross-correct neighboring cells, and small quantities of these proteins are able to produce clinical benefits in many cases. Ex vivo gene therapy allows for autologous transplant of modified cells from different sources, including stem cells and hematopoietic precursors. AREAS COVERED: Here, we summarize the main gene therapy and genome editing strategies that are currently being used as ex vivo gene therapy approaches for lysosomal disorders, highlighting important characteristics, such as vectors used, strategies, types of cells that are modified and main results in different disorders. EXPERT OPINION: Clinical trials are already ongoing, and soon approved therapies for LSD based on ex vivo gene therapy approaches should reach the market.

Inflammatory process and oxidative/nitrative stress: in vivo study in mucopolysaccharidosis type IV A patients under long-term enzyme replacement therapy.

Mucopolysaccharidosis type IV A (MPS IVA) is an inborn error of the metabolism (IEM) caused by a deficiency of the enzyme N-acetylgalactosamine 6-sulfate sulfatase (GALNS). Since 2014, enzyme replacement therapy (ERT) is the recommended treatment for these patients. It is known that the inflammatory response is closely related to antioxidant defenses and oxidative stress, and literature shows involvement of oxidative stress in the pathogenesis of IEM. The aim of this study is to investigate the mechanisms of oxidative/nitrative stress and inflammation in patients with MPS IVA under long-term ERT. In the present work we investigate parameters of oxidative/nitrative stress in plasma and urine of MPS IVA patients under long-term ERT and controls, such as plasmatic nitrate/nitrite levels using the LDH Method, urinary di-tyrosine levels by fluorometric method, plasmatic content of sulfhydryl groups, urinary oxidized guanine species by ELISA kit and the plasmatic total antioxidant status. We next evaluated the plasmatic pro and anti-inflammatory cytokines concentration (IL-1ß, IL-2, IL-4, IL-6, IL-8, IL-10, TNF-α) and the expression of factors and enzymes Nrf-2, NF-κß and HO-1, main mediators between inflammation and oxidative stress. In concern to the oxidative/nitrative stress parameters, there was no significant difference between the groups MPS IVA patients under long-term ERT and controls, showing that there is no overproducing of RNS, no protein damage, no DNA/RNA oxidative damage and no modification in the non-enzymatic antioxidant capacity of a tissue to prevent the damage associated to free radical processes in these patients. It was also verified no significant difference between the MPS IVA patients under long-term ERT and controls groups regarding the production of proinflammatory cytokines. About anti-inflammatory cytokines, IL 10 was shown to be elevated in MPS IVA patients under long-term ERT in comparison to the control group. We next evaluated the genic expression of Nrf-2, NF-κß and HO-1and there was no significant difference between the MPS IVA patients under long-term ERT and control groups. In conclusion, MPS IVA patients under long term ERT are not in an inflammatory state and there is no alteration in genic expression in the genes analyzed which are involved in oxidative stress and inflammatory pathways. It is,however, important to consider that absence of imbalance of antioxidant defenses in MPS IVA patients under long term ERT is so far preliminary it is supported by methodologies that are not highly sensitive nor very accurate. Further experiments in future using state-of-the-art methodologies will corroborate these findings. Nevertheless, our results demonstrated the protective effect of the treatment in relation to the parameters studied and the importance of starting treatment in the early stages of the disease.

Copy number variations in SPAST and ATL1 are rare among Brazilians.

Copy number variations (CNV) may represent a significant proportion of SPG4 and SPG3A diagnosis, the most frequent autosomal dominant subtypes of hereditary spastic paraplegias (HSP). We aimed to assess the frequency of CNVs in SPAST and ATL1 and to update the molecular epidemiology of HSP families in southern Brazil. A cohort study that included 95 Brazilian index cases with clinical suspicion of HSP was conducted between April 2011 and September 2022. Multiplex Ligation Dependent Probe Amplification (MLPA) was performed in 41 cases without defined diagnosis by different massive parallel sequencing techniques (MPS). Diagnosis was obtained in 57/95 (60%) index cases, 15/57 (26.3%) being SPG4. Most frequent autosomal recessive HSP subtypes were SPG7 followed by SPG11, SPG76 and cerebrotendinous xanthomatosis. No CNVs in SPAST and ATL1 were found. Copy number variations are rare among SPG4 and SPG3A families in Brazil. Considering the possibility of CNVs detection by specific algorithms with MPS data, we consider that this is likely the most cost-effective approach to investigate CNVs in these genes in low-risk populations, with MLPA being reserved as an orthogonal confirmatory test.