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Papers by Francyne Kubaski

Research paper thumbnail of Review Establishment of Glycosaminoglycan Assays

Research paper thumbnail of Review Establishment of Glycosaminoglycan Assays

Research paper thumbnail of MPS Brazil Network: A summary of all mucopolysaccharidosis type IIIB patients

Molecular Genetics and Metabolism

Research paper thumbnail of Newborn screening for six lysosomal diseases in Brazil: Pilot study update

Molecular Genetics and Metabolism

Research paper thumbnail of Difficulties in the Diagnosis of Gaucher Disease in a Low-Income Country: A Case Report from Mozambique

Research paper thumbnail of Evaluation of 3-O-methyldopa as a biomarker for aromatic L-amino acid decarboxylase deficiency in 7 Brazilian cases

Molecular Genetics and Metabolism Reports

Research paper thumbnail of Precision Medicine for Lysosomal Disorders

Biomolecules

Precision medicine (PM) is an emerging approach for disease treatment and prevention that account... more Precision medicine (PM) is an emerging approach for disease treatment and prevention that accounts for the individual variability in the genes, environment, and lifestyle of each person. Lysosomal diseases (LDs) are a group of genetic metabolic disorders that include approximately 70 monogenic conditions caused by a defect in lysosomal function. LDs may result from primary lysosomal enzyme deficiencies or impairments in membrane-associated proteins, lysosomal enzyme activators, or modifiers that affect lysosomal function. LDs are heterogeneous disorders, and the phenotype of the affected individual depends on the type of substrate and where it accumulates, which may be impacted by the type of genetic change and residual enzymatic activity. LDs are individually rare, with a combined incidence of approximately 1:4000 individuals. Specific therapies are already available for several LDs, and many more are in development. Early identification may enable disease course prediction and a s...

Research paper thumbnail of Prevalence of thrombophilia-associated genetic risk factors in blood donors of a regional hospital in southern Brazil

Hematology, Transfusion and Cell Therapy

Research paper thumbnail of Newborn screening of mucopolysaccharidoses: past, present, and future

Journal of Human Genetics

Mucopolysaccharidoses (MPS) are a subtype of lysosomal storage disorders (LSDs) characterized by ... more Mucopolysaccharidoses (MPS) are a subtype of lysosomal storage disorders (LSDs) characterized by the deficiency of the enzyme involved in the breakdown of glycosaminoglycans (GAGs). Mucopolysaccharidosis type I (MPS I, Hurler Syndrome) was endorsed by the U.S. Secretary of the Department of Health and Human Services for universal newborn screening (NBS) in February 2016. Its endorsement exemplifies the need to enhance the accuracy of diagnostic testing for disorders that are considered for NBS. The progression of MPS disorders typically incudes irreversible CNS involvement, severe bone dysplasia, and cardiac and respiratory issues. Patients with MPS have a significantly decreased quality of life if untreated and require timely diagnosis and management for optimal outcomes. NBS provides the opportunity to diagnose and initiate treatment plans for MPS patients as early as possible. Most newborns with MPS are asymptomatic at birth; therefore, it is crucial to have biomarkers that can be identified in the newborn. At present, there are tiered methods and different instrumentation available for this purpose. The screening of quick, cost-effective, sensitive, and specific biomarkers in patients with MPS at birth is important. Rapid newborn diagnosis enables treatments to maximize therapeutic efficacy and to introduce immune tolerance during the neonatal period. Currently, newborn screening for MPS I and II has been implemented and/or in pilot testing in several countries. In this review article, historical aspects of NBS for MPS and the prospect of newborn screening for MPS are described, including the potential tiers of screening.

Research paper thumbnail of Neonatal Screening for MPS Disorders in Latin America: A Survey of Pilot Initiatives

International Journal of Neonatal Screening

Newborn screening enables the diagnosis of treatable disorders at the early stages, and because o... more Newborn screening enables the diagnosis of treatable disorders at the early stages, and because of its countless benefits, conditions have been continuously added to screening panels, allowing early intervention, aiming for the prevention of irreversible manifestations and even premature death. Mucopolysaccharidoses (MPS) are lysosomal storage disorders than can benefit from an early diagnosis, and thus are being recommended for newborn screening. They are multisystemic progressive disorders, with treatment options already available for several MPS types. MPS I was the first MPS disorder enrolled in the newborn screening (NBS) panel in the USA and a few other countries, and other MPS types are expected to be added. Very few studies about NBS for MPS in Latin America have been published so far. In this review, we report the results of pilot studies performed in Mexico and Brazil using different methodologies: tandem mass spectrometry, molecular analysis, digital microfluidics, and fl...

Research paper thumbnail of Newborn screening for lysosomal disorders in Brazil: A pilot study using customized fluorimetric assays

Genetics and Molecular Biology

Research paper thumbnail of Mucopolysaccharidosis Type I

Diagnostics

Mucopolysaccharidosis type I (MPS I) is caused by the deficiency of α-l-iduronidase, leading to t... more Mucopolysaccharidosis type I (MPS I) is caused by the deficiency of α-l-iduronidase, leading to the storage of dermatan and heparan sulfate. There is a broad phenotypical spectrum with the presence or absence of neurological impairment. The classical form is known as Hurler syndrome, the intermediate form as Hurler–Scheie, and the most attenuated form is known as Scheie syndrome. Phenotype seems to be largely influenced by genotype. Patients usually develop several somatic symptoms such as abdominal hernias, extensive dermal melanocytosis, thoracolumbar kyphosis odontoid dysplasia, arthropathy, coxa valga and genu valgum, coarse facial features, respiratory and cardiac impairment. The diagnosis is based on the quantification of α-l-iduronidase coupled with glycosaminoglycan analysis and gene sequencing. Guidelines for treatment recommend hematopoietic stem cell transplantation for young Hurler patients (usually at less than 30 months of age). Intravenous enzyme replacement is approv...

Research paper thumbnail of Diagnosis of Mucopolysaccharidoses

Diagnostics

The mucopolysaccharidoses (MPSs) include 11 different conditions caused by specific enzyme defici... more The mucopolysaccharidoses (MPSs) include 11 different conditions caused by specific enzyme deficiencies in the degradation pathway of glycosaminoglycans (GAGs). Although most MPS types present increased levels of GAGs in tissues, including blood and urine, diagnosis is challenging as specific enzyme assays are needed for the correct diagnosis. Enzyme assays are usually performed in blood, with some samples (as leukocytes) providing a final diagnosis, while others (such as dried blood spots) still being considered as screening methods. The identification of variants in the specific genes that encode each MPS-related enzyme is helpful for diagnosis confirmation (when needed), carrier detection, genetic counseling, prenatal diagnosis (preferably in combination with enzyme assays) and phenotype prediction. Although the usual diagnostic flow in high-risk patients starts with the measurement of urinary GAGs, it continues with specific enzyme assays and is completed with mutation identific...

Research paper thumbnail of Quantification of glycosaminoglycans by liquid chromatography tandem mass spectrometry is a useful tool for screening of GlcNAc-phosphotransferase deficient patients

Molecular Genetics and Metabolism

Research paper thumbnail of Quantification of glycosaminoglycan species in patients with multiple sulfatase deficiency by liquid chromatography tandem mass spectrometry: A potential biomarker for this condition

Molecular Genetics and Metabolism

Research paper thumbnail of Newborn screening for six lysosomal diseases: Pilot study in Brazil

Molecular Genetics and Metabolism

INTRODUCTION: Lysosomal storage disorders (LSDs) are inborn errors of metabolism caused by excess... more INTRODUCTION: Lysosomal storage disorders (LSDs) are inborn errors of metabolism caused by excessive accumulation of undegraded metabolites due to the deficiency of soluble lysosomal hydrolases, membrane proteins or accessory proteins that lead to an impaired turnover of complex macromolecules, including glycosaminoglycans, proteins and lipids. Many LSDs already have specific therapies, and in most cases the earlier introduction of therapy provides better outcomes. However, patients are usually diagnosed only after a long “diagnostic odyssey”, with therapies introduced when irreversible manifestations are already present. OBJECTIVE: This project aims to evaluate the feasibility of newborn screening (NBS) for selected LSDs in Brazil, using a tandem mass spectrometry (MS/MS) platform with a 6-Plex kit (supplied by PerkinElmer). MATERIALS AND METHODS: The study includes the screening for Gaucher, Fabry, Pompe, Krabbe, Niemann-Pick A/B and Mucopolysaccharidosis I. This is a prospective study in 20,000 unselected newborns from the state of Bahia, Brazil. The newborns with low enzyme activity are further evaluated by biochemical and molecular genetics methods until the diagnosis is confirmed and are referred for treatment as appropriate. All lysosomal enzymes were analyzed with NeoLSD MS/MS kit (Perkin Elmer) on a Waters Xevo TQ-S Micro. RESULTS: Validation of the method was conducted in dried blood spots provided by the supplier and from unselected newborns. Instrument optimization was conducted in order to increase the signal and to decrease the in source fragmentation. Initial cutoffs were established as percentage of median in nmol/h/mL, as 0.8 (Gaucher), 1 (Fabry), 1.6 (Pompe), 0.8 (MPS I), 0.46 (Krabbe) and 0.9 (Niemann-Pick A/B). CONCLUSIONS: Further positive samples will be included in order to confirm the cutoffs. This validation of the MS/MS method enabled the beginning of a pilot study, which, when completed, will include 20,000 newborns and will provide important information about the feasibility of a NBS for LSDs in Brazil.

Research paper thumbnail of Quantitation of glycosaminoglycans in amniotic fluid by liquid chromatography tandem mass spectrometry: A potential tool for the rapid prenatal identification of MPS in pregnancies at risk

Molecular Genetics and Metabolism

Research paper thumbnail of Lysosomal diseases: Overview on current diagnosis and treatment

Genetics and Molecular Biology

Research paper thumbnail of Population medical genetics: translating science to the community

Genetics and Molecular Biology

Research paper thumbnail of Neurological manifestations of lysosomal disorders and emerging therapies targeting the CNS

The Lancet Child & Adolescent Health

Lysosomal disorders have been an area of interest since intravenous enzyme replacement therapy wa... more Lysosomal disorders have been an area of interest since intravenous enzyme replacement therapy was successfully introduced for the treatment of Gaucher's disease in the early 1990s. This treatment approach has also been developed for several other lysosomal disorders, including Fabry's disease, Pompe's disease, lysosomal acid lipase deficiency, and five types of mucopolysaccharidosis. Despite the benefits of enzyme replacement therapy, it has limitations-most importantly, its ineffectiveness in treating the neurological components of lysosomal disorders, as only a small proportion of recombinant enzymes can cross the blood-brain barrier. Development of strategies to improve drug delivery to the CNS is now the primary focus in lysosomal disorder research. This Review discusses the neurological manifestations and emerging therapies for the CNS component of these diseases. The therapies in development (which are now in phase 1 or phase 2 clinical trials) might be for specific lysosomal disorders (enzyme replacement therapy via intrathecal or intracerebroventricular routes or with fusion proteins, or gene therapy) or applicable to more than one lysosomal disorder (haemopoietic stem cell transplantation, pharmacological chaperones, substrate reduction therapy, or stop codon readthrough). The combination of early diagnosis with effective therapies should change the outlook for patients with lysosomal disorders with neurological involvement in the next 5-10 years.

Research paper thumbnail of Review Establishment of Glycosaminoglycan Assays

Research paper thumbnail of Review Establishment of Glycosaminoglycan Assays

Research paper thumbnail of MPS Brazil Network: A summary of all mucopolysaccharidosis type IIIB patients

Molecular Genetics and Metabolism

Research paper thumbnail of Newborn screening for six lysosomal diseases in Brazil: Pilot study update

Molecular Genetics and Metabolism

Research paper thumbnail of Difficulties in the Diagnosis of Gaucher Disease in a Low-Income Country: A Case Report from Mozambique

Research paper thumbnail of Evaluation of 3-O-methyldopa as a biomarker for aromatic L-amino acid decarboxylase deficiency in 7 Brazilian cases

Molecular Genetics and Metabolism Reports

Research paper thumbnail of Precision Medicine for Lysosomal Disorders

Biomolecules

Precision medicine (PM) is an emerging approach for disease treatment and prevention that account... more Precision medicine (PM) is an emerging approach for disease treatment and prevention that accounts for the individual variability in the genes, environment, and lifestyle of each person. Lysosomal diseases (LDs) are a group of genetic metabolic disorders that include approximately 70 monogenic conditions caused by a defect in lysosomal function. LDs may result from primary lysosomal enzyme deficiencies or impairments in membrane-associated proteins, lysosomal enzyme activators, or modifiers that affect lysosomal function. LDs are heterogeneous disorders, and the phenotype of the affected individual depends on the type of substrate and where it accumulates, which may be impacted by the type of genetic change and residual enzymatic activity. LDs are individually rare, with a combined incidence of approximately 1:4000 individuals. Specific therapies are already available for several LDs, and many more are in development. Early identification may enable disease course prediction and a s...

Research paper thumbnail of Prevalence of thrombophilia-associated genetic risk factors in blood donors of a regional hospital in southern Brazil

Hematology, Transfusion and Cell Therapy

Research paper thumbnail of Newborn screening of mucopolysaccharidoses: past, present, and future

Journal of Human Genetics

Mucopolysaccharidoses (MPS) are a subtype of lysosomal storage disorders (LSDs) characterized by ... more Mucopolysaccharidoses (MPS) are a subtype of lysosomal storage disorders (LSDs) characterized by the deficiency of the enzyme involved in the breakdown of glycosaminoglycans (GAGs). Mucopolysaccharidosis type I (MPS I, Hurler Syndrome) was endorsed by the U.S. Secretary of the Department of Health and Human Services for universal newborn screening (NBS) in February 2016. Its endorsement exemplifies the need to enhance the accuracy of diagnostic testing for disorders that are considered for NBS. The progression of MPS disorders typically incudes irreversible CNS involvement, severe bone dysplasia, and cardiac and respiratory issues. Patients with MPS have a significantly decreased quality of life if untreated and require timely diagnosis and management for optimal outcomes. NBS provides the opportunity to diagnose and initiate treatment plans for MPS patients as early as possible. Most newborns with MPS are asymptomatic at birth; therefore, it is crucial to have biomarkers that can be identified in the newborn. At present, there are tiered methods and different instrumentation available for this purpose. The screening of quick, cost-effective, sensitive, and specific biomarkers in patients with MPS at birth is important. Rapid newborn diagnosis enables treatments to maximize therapeutic efficacy and to introduce immune tolerance during the neonatal period. Currently, newborn screening for MPS I and II has been implemented and/or in pilot testing in several countries. In this review article, historical aspects of NBS for MPS and the prospect of newborn screening for MPS are described, including the potential tiers of screening.

Research paper thumbnail of Neonatal Screening for MPS Disorders in Latin America: A Survey of Pilot Initiatives

International Journal of Neonatal Screening

Newborn screening enables the diagnosis of treatable disorders at the early stages, and because o... more Newborn screening enables the diagnosis of treatable disorders at the early stages, and because of its countless benefits, conditions have been continuously added to screening panels, allowing early intervention, aiming for the prevention of irreversible manifestations and even premature death. Mucopolysaccharidoses (MPS) are lysosomal storage disorders than can benefit from an early diagnosis, and thus are being recommended for newborn screening. They are multisystemic progressive disorders, with treatment options already available for several MPS types. MPS I was the first MPS disorder enrolled in the newborn screening (NBS) panel in the USA and a few other countries, and other MPS types are expected to be added. Very few studies about NBS for MPS in Latin America have been published so far. In this review, we report the results of pilot studies performed in Mexico and Brazil using different methodologies: tandem mass spectrometry, molecular analysis, digital microfluidics, and fl...

Research paper thumbnail of Newborn screening for lysosomal disorders in Brazil: A pilot study using customized fluorimetric assays

Genetics and Molecular Biology

Research paper thumbnail of Mucopolysaccharidosis Type I

Diagnostics

Mucopolysaccharidosis type I (MPS I) is caused by the deficiency of α-l-iduronidase, leading to t... more Mucopolysaccharidosis type I (MPS I) is caused by the deficiency of α-l-iduronidase, leading to the storage of dermatan and heparan sulfate. There is a broad phenotypical spectrum with the presence or absence of neurological impairment. The classical form is known as Hurler syndrome, the intermediate form as Hurler–Scheie, and the most attenuated form is known as Scheie syndrome. Phenotype seems to be largely influenced by genotype. Patients usually develop several somatic symptoms such as abdominal hernias, extensive dermal melanocytosis, thoracolumbar kyphosis odontoid dysplasia, arthropathy, coxa valga and genu valgum, coarse facial features, respiratory and cardiac impairment. The diagnosis is based on the quantification of α-l-iduronidase coupled with glycosaminoglycan analysis and gene sequencing. Guidelines for treatment recommend hematopoietic stem cell transplantation for young Hurler patients (usually at less than 30 months of age). Intravenous enzyme replacement is approv...

Research paper thumbnail of Diagnosis of Mucopolysaccharidoses

Diagnostics

The mucopolysaccharidoses (MPSs) include 11 different conditions caused by specific enzyme defici... more The mucopolysaccharidoses (MPSs) include 11 different conditions caused by specific enzyme deficiencies in the degradation pathway of glycosaminoglycans (GAGs). Although most MPS types present increased levels of GAGs in tissues, including blood and urine, diagnosis is challenging as specific enzyme assays are needed for the correct diagnosis. Enzyme assays are usually performed in blood, with some samples (as leukocytes) providing a final diagnosis, while others (such as dried blood spots) still being considered as screening methods. The identification of variants in the specific genes that encode each MPS-related enzyme is helpful for diagnosis confirmation (when needed), carrier detection, genetic counseling, prenatal diagnosis (preferably in combination with enzyme assays) and phenotype prediction. Although the usual diagnostic flow in high-risk patients starts with the measurement of urinary GAGs, it continues with specific enzyme assays and is completed with mutation identific...

Research paper thumbnail of Quantification of glycosaminoglycans by liquid chromatography tandem mass spectrometry is a useful tool for screening of GlcNAc-phosphotransferase deficient patients

Molecular Genetics and Metabolism

Research paper thumbnail of Quantification of glycosaminoglycan species in patients with multiple sulfatase deficiency by liquid chromatography tandem mass spectrometry: A potential biomarker for this condition

Molecular Genetics and Metabolism

Research paper thumbnail of Newborn screening for six lysosomal diseases: Pilot study in Brazil

Molecular Genetics and Metabolism

INTRODUCTION: Lysosomal storage disorders (LSDs) are inborn errors of metabolism caused by excess... more INTRODUCTION: Lysosomal storage disorders (LSDs) are inborn errors of metabolism caused by excessive accumulation of undegraded metabolites due to the deficiency of soluble lysosomal hydrolases, membrane proteins or accessory proteins that lead to an impaired turnover of complex macromolecules, including glycosaminoglycans, proteins and lipids. Many LSDs already have specific therapies, and in most cases the earlier introduction of therapy provides better outcomes. However, patients are usually diagnosed only after a long “diagnostic odyssey”, with therapies introduced when irreversible manifestations are already present. OBJECTIVE: This project aims to evaluate the feasibility of newborn screening (NBS) for selected LSDs in Brazil, using a tandem mass spectrometry (MS/MS) platform with a 6-Plex kit (supplied by PerkinElmer). MATERIALS AND METHODS: The study includes the screening for Gaucher, Fabry, Pompe, Krabbe, Niemann-Pick A/B and Mucopolysaccharidosis I. This is a prospective study in 20,000 unselected newborns from the state of Bahia, Brazil. The newborns with low enzyme activity are further evaluated by biochemical and molecular genetics methods until the diagnosis is confirmed and are referred for treatment as appropriate. All lysosomal enzymes were analyzed with NeoLSD MS/MS kit (Perkin Elmer) on a Waters Xevo TQ-S Micro. RESULTS: Validation of the method was conducted in dried blood spots provided by the supplier and from unselected newborns. Instrument optimization was conducted in order to increase the signal and to decrease the in source fragmentation. Initial cutoffs were established as percentage of median in nmol/h/mL, as 0.8 (Gaucher), 1 (Fabry), 1.6 (Pompe), 0.8 (MPS I), 0.46 (Krabbe) and 0.9 (Niemann-Pick A/B). CONCLUSIONS: Further positive samples will be included in order to confirm the cutoffs. This validation of the MS/MS method enabled the beginning of a pilot study, which, when completed, will include 20,000 newborns and will provide important information about the feasibility of a NBS for LSDs in Brazil.

Research paper thumbnail of Quantitation of glycosaminoglycans in amniotic fluid by liquid chromatography tandem mass spectrometry: A potential tool for the rapid prenatal identification of MPS in pregnancies at risk

Molecular Genetics and Metabolism

Research paper thumbnail of Lysosomal diseases: Overview on current diagnosis and treatment

Genetics and Molecular Biology

Research paper thumbnail of Population medical genetics: translating science to the community

Genetics and Molecular Biology

Research paper thumbnail of Neurological manifestations of lysosomal disorders and emerging therapies targeting the CNS

The Lancet Child & Adolescent Health

Lysosomal disorders have been an area of interest since intravenous enzyme replacement therapy wa... more Lysosomal disorders have been an area of interest since intravenous enzyme replacement therapy was successfully introduced for the treatment of Gaucher's disease in the early 1990s. This treatment approach has also been developed for several other lysosomal disorders, including Fabry's disease, Pompe's disease, lysosomal acid lipase deficiency, and five types of mucopolysaccharidosis. Despite the benefits of enzyme replacement therapy, it has limitations-most importantly, its ineffectiveness in treating the neurological components of lysosomal disorders, as only a small proportion of recombinant enzymes can cross the blood-brain barrier. Development of strategies to improve drug delivery to the CNS is now the primary focus in lysosomal disorder research. This Review discusses the neurological manifestations and emerging therapies for the CNS component of these diseases. The therapies in development (which are now in phase 1 or phase 2 clinical trials) might be for specific lysosomal disorders (enzyme replacement therapy via intrathecal or intracerebroventricular routes or with fusion proteins, or gene therapy) or applicable to more than one lysosomal disorder (haemopoietic stem cell transplantation, pharmacological chaperones, substrate reduction therapy, or stop codon readthrough). The combination of early diagnosis with effective therapies should change the outlook for patients with lysosomal disorders with neurological involvement in the next 5-10 years.