A large deletion together with a point mutation in the GALC gene is a common mutant allele in patients with infantile Krabbe disease

Introduction: Krabbe disease (KD) is an autosomal recessive disorder caused by mutations in the galactocerebrosidase (GALC) gene resulting in neuro-inflammation and defective myelination in the central and peripheral nervous systems. Most infantile patients present with clinical features before six months of age and die before two years of age. The only treatment available for pre-symptomatic or mildly affected individuals is hematopoietic stem cell transplantation (HSCT). In the animal models, combining bone marrow transplantation (BMT) with gene therapy has shown the best results in disease outcome. In this study, we examine the outcome of gene therapy alone. Methods: Twitcher (twi) mice used in the study, have a W339X mutation in the GALC gene. Genotype identification of the mice was performed shortly after birth or post-natal day 1 (PND1), using polymerase chain reaction on the toe clips followed by restriction enzyme digestion and electrophoresis. Eight or nine-day-old affected mice were used for gene therapy treatment alone or combined with BMT. While iv injection of 4 × 1013 gc/kg of body weight of viral vector was used originally, different viral titers were also used without BMT to evaluate their outcomes. Results: When the standard viral dose was increased four- and ten-fold (4X and 10X) without BMT, the lifespans were increased significantly. Without BMT the affected mice were fertile, had the same weight and appearance as wild type mice and had normal strength and gait. The brains showed no staining for CD68, a marker for activated microglia/macrophages, and less astrogliosis than untreated twi mice. Conclusion: Our results demonstrate that, it may be possible to treat human KD patients with high dose AAVrh10 without blood stem cell transplantation which would eliminate the side effects of HSCT.

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A point mutation and large deletion at the candidate avirulence locus AvrMlp7 in the poplar rust fungus correlate with poplar RMlp7 resistance breakdown

Molecular Ecology ◽

10.1111/mec.16294 ◽

2021 ◽

Author(s):

Clémentine Louet ◽

Méline Saubin ◽

Axelle Andrieux ◽

Antoine Persoons ◽

Mathilde Gorse ◽

...

Keyword(s):

Point Mutation ◽

Rust Fungus ◽

Large Deletion ◽

Resistance Breakdown

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Characterization of the GALC Gene in Three Japanese Patients with Adult-Onset Krabbe Disease

Genetic Testing ◽

10.1089/gte.1997.1.217 ◽

1997 ◽

Vol 1 (3) ◽

pp. 217-223 ◽

Cited By ~ 6

Author(s):

YOJI KUKITA ◽

HIROKAZU FURUYA ◽

TAKURO KOBAYASHI ◽

NORIO SAKAI ◽

KENSHI HAYASHI

Keyword(s):

Japanese Patients ◽

Krabbe Disease ◽

Adult Onset ◽

Galc Gene

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Pathogenic Variants in GALC Gene Correlate With Late Onset Krabbe Disease and Vision Loss: Case Series and Review of Literature

Frontiers in Neurology ◽

10.3389/fneur.2020.563724 ◽

2020 ◽

Vol 11 ◽

Author(s):

Nicholas A. Bascou ◽

Maria L. Beltran-Quintero ◽

Maria L. Escolar

Keyword(s):

Vision Loss ◽

Late Onset ◽

Case Series ◽

Krabbe Disease ◽

Review Of Literature ◽

Pathogenic Variants ◽

Galc Gene

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Factor VII Deficiency Remains Post-Puberty Due to Point Mutation in the HNF4 Binding Site in the Coagulant Factor VII Gene.

Blood ◽

10.1182/blood.v106.11.1947.1947 ◽

2005 ◽

Vol 106 (11) ◽

pp. 1947-1947

Author(s):

XingWu Zheng ◽

Theresa M. Tidd ◽

Donna DiMichele ◽

Eleanor S. Pollak

Keyword(s):

Binding Site ◽

Point Mutation ◽

Mutant Allele ◽

Factor Ix ◽

Hemophilia B ◽

Factor Vii ◽

Start Site ◽

Expression Plasmid ◽

Fvii Deficiency ◽

Factor Ix Deficiency

Abstract A novel T to C point mutation at −60 in the gene for coagulant Factor VII results in life-long severe coagulant Factor VII deficiency in post-pubertal twin males. The clinical course of these patients provides an informative in vivo example of the regulation of expression of vitamin K-dependent clotting protein Factor VII. An analogous point mutation in the HNF4 binding site in the Factor IX gene results in the clinical phenotype Hemophilia B Leyden, a sex-linked antigen-negative Factor IX deficiency that resolves post-puberty. The affected Factor VII deficient patients have prolonged prothrombin times (46 and 52 secs), normal aPTTs and decreased FVII levels of FVII:Coagulant activity: < 1% and FVII:Antigen: < 3%. The −60 mutation, ACTTTG → ACTCTG occurs 9 base pairs before the start site of transcription and 59 bps before the before the start site of translation. The twins are compound heterozygotes and also possess a mutation in exon 8 at amino acid 348, a mutation that has previously been reported to cause FVII deficiency. Both affected individuals have recurrent target joint hemorrhage (shoulder, elbow, ankle) requiring replacement therapy 6–12 times/year. Results: Gel mobility shift assays using a radio-labeled probe spanning from −76 to −46 in the FVII promoter region demonstrate the loss of binding of transcription factor HNF-4. Transient transfection assays in HepG2 cells using 186 bps of the mutant and the wildtype promoters (−185 to +1) revealed a loss of expression with the mutant allele. Co-transfection with an HNF4 expression plasmid resulted in an increase in expression of the wildetype construct in HeLa cells, a non-hepatic cell line. However, co-transfection of the HNF4 expression plasmid failed to increase expression with the construct containing the mutant allele sequence. Conclusion: The lack of phenotypic change of the FVII:C in 19 yo twin boys provides dynamic support of the necessity of an overlapping androgen binding site in the homologous Factor IX gene as responsible for the phenotypic resolution of Factor IX deficiency (Hemophilia B Leyden) post-puberty. It is of interest that an increase in FVII:C did not occur with advancing age in FVII deficiency due to this HNF4 binding site mutation.

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Prion-like α-synuclein pathology in the brains of infants: Krabbe disease as a novel seed-competent α-synucleinopathy

10.1101/2021.10.12.463948 ◽

2021 ◽

Author(s):

Christopher Hatton ◽

Simona S. Ghanem ◽

David Koss ◽

Ilham Yahya Abdi ◽

Elizabeth Gibbons ◽

...

Keyword(s):

Lewy Body ◽

Neurodegenerative Disorder ◽

Lewy Bodies ◽

Krabbe Disease ◽

Biological Processes ◽

Pathogenic Variants ◽

Increased Risk ◽

Galc Gene ◽

The Brain ◽

Lewy Body Diseases

Krabbe disease (KD) is an infantile neurodegenerative disorder resulting from pathogenic variants in the GALC gene which causes accumulation of the toxic sphingolipid psychosine. GALC variants are associated with increased risk of Lewy body diseases (LBD), an umbrella term for age-associated neurodegenerative diseases in which the protein α-synuclein aggregates into Lewy bodies. To explore whether α-synuclein in KD has pathological similarities to that in LBD, we compared post-mortem KD tissue to that of infant control cases and identified alterations to α-synuclein localisation and expression of modifications associated with LBD. To determine whether α-synuclein in KD displayed pathogenic properties associated with LBD we evaluated its seeding capacity using the real-time quaking-induced conversion assay. Strikingly, seeded aggregation of α-synuclein resulted in the formation of fibrillar aggregates similar to those observed in LBD, confirming the prion-like capacity of KD-derived α-synuclein. These observations constitute the first report of prion-like α-synuclein in the brain tissue of infants and challenge the putative view that α-synuclein pathology is merely an age-associated phenomenon, instead suggesting it can result from alterations to biological processes such as sphingolipid homeostasis. Our findings have important implications for understanding the mechanisms underlying Lewy body formation in LBD.

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Duchenne muscular dystrophy: A immunohistochemical profile and deletion pattern in dystrophin gene in North Indian population

Asian Journal of Medical Sciences ◽

10.3126/ajms.v8i6.18281 ◽

2017 ◽

Vol 8 (6) ◽

pp. 13-18

Author(s):

Rachna Agarwal ◽

Sujata Chaturvedi ◽

Neelam Chhillar ◽

Ishita Pant ◽

Anuradha Sharma

Keyword(s):

Duchenne Muscular Dystrophy ◽

Muscular Dystrophy ◽

Point Mutation ◽

Dystrophin Gene ◽

Becker Muscular Dystrophy ◽

Large Deletion ◽

Mutation Spectrum ◽

Specific Information ◽

Complete Work ◽

Deletion Pattern

Background: Duchenne muscular dystrophy (DMD), one of the most common X linked muscular disorder, affecting 1 in 3500 male births and is caused by mutation in dystrophin gene. 65% of DMD cases are caused by large deletion of dystrophin gene, followed by duplication (5-10%) or point mutation (25-30%). There is wide mutation spectrum of the mutations in dystrophin gene. Hence, population specific information is needed on mutation spectrum and frequency of common mutations occurring in that particular population for appropriate counseling, prenatal diagnosis and for developing genetic therapy in future. Aims and Objectives: To find out the frequency and distribution of deletion in dystrophin gene in DMD patients along with contribution of pathology and genetic testing in diagnosis of DMD and Becker muscular dystrophy (BMD) in North Indian population.Materials and Methods: Dystrophin gene was screened for deletion by multiplex polymerase chain reaction (PCR). Out of 41 patients, 09 patients underwent muscle biopsy, on which immunohistochemistry was performed for dystrophin, sarcoglycan, dysferlin and merosin.Results: Majority of the deletions were located in distal hotspot region (26/39 ~66.66%) which includes the exons 45-55 and 15.38% of deletions were located at the proximal hotspot region (2- 19 exons).Conclusion: In the present study, 34% patients only showed deletion. Hence complete work up of any muscular dystrophy requires immnohistochemical analysis to see the expression of muscle proteins along with multipleplex PCR test to detect any exon deletion, multiplex ligation-dependent probe amplification (MLPA) to detect point mutation and duplication and western blotting to quantify the dystrophin protein.Asian Journal of Medical Sciences Vol.8(6) 2017 13-18

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