ORCID Profile
0000-0002-4969-3026
Current Organisation
Yale University
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Publisher: MDPI AG
Date: 09-11-2016
DOI: 10.3390/NU8110713
Publisher: Mary Ann Liebert Inc
Date: 03-2021
DOI: 10.1089/HUM.2020.241
Abstract: Clustered regularly interspaced short palindromic repeat (CRISPR)-based technology has been adapted to achieve a wide range of genome modifications, including transcription regulation. The focus of this review is on the application of CRISPR-based platforms such as nuclease-deficient Cas9 and Cas12a, to achieve targeted gene activation. We review studies to date that have used CRISPR-based activation technology for the elucidation of biological mechanism and disease correction, as well as its application in genetic screens as a powerful tool for high-throughput genotype-phenotype mapping. In addition to our synthesis and critical analysis of published studies, we explore key considerations for the potential clinical translation of CRISPR-based activation technology.
Publisher: Elsevier BV
Date: 10-2019
Publisher: Cold Spring Harbor Laboratory
Date: 12-07-2023
DOI: 10.1101/2023.07.12.548370
Abstract: Our inability to interpret the consequences of rare variants is an unappreciated challenge in the diagnosis of rare diseases. We developed a democratized workflow called Saturation Mutagenesis-Reinforced Functional assays (SMuRF) to inspect the direct impact variants have on enzymatic activity. We employed SMuRF to score all possible coding single nucleotide variants (SNVs) of Dystroglycanopathies-related enzyme-coding genes, FKRP and LARGE1 . The utility of SMuRF scores was enhanced through the assignment of confidence scores and orthogonal assays for validation. SMuRF recapitulated and significantly expanded the knowledge gained from clinical reports and population databases, aiding in alleviating ethnic disparity in biomedical databases and improving variant classification. SMuRF expanded the training datasets of the computational predictors with the potential to improve their variant classification capability. SMuRF highlighted the critical regions in the enzyme structure which shed light on different disease mechanisms. SMuRF is the first high-throughput functional workflow to study dystroglycanopathy variants and opens the door for better variant interpretation underlying other rare diseases.
Publisher: Cold Spring Harbor Laboratory
Date: 25-03-2018
DOI: 10.1101/288621
Abstract: Duchenne Muscular Dystrophy (DMD) is a progressive and fatal neuromuscular disease which arises from mutations in the dystrophin gene ( DMD ) that result in the absence or severe reduction of the cytoskeletal protein dystrophin. In addition to the primary dystrophin defect, secondary processes such as inflammation, calcium influx, dysregulated autophagy and fibrosis exacerbate dystrophic pathology and thus increase disease progression. While therapies to restore dystrophin deficiency are being developed, strategies which target these secondary processes could be of benefit to patients. Benfotiamine is a lipid soluble precursor to thiamine that can reduce secondary processes such as inflammation and oxidative stress in diabetic patients. As such we tested it in the mdx mouse model of DMD and found that benfotiamine reduced multiple markers of dystrophic pathology and improved grip strength. In addition, members of the utrophin and dystrophin glycoprotein complexes were significantly increased at the sarcolemma which could improve cell adhesion. We also demonstrated that benfotiamine treatment lowered the expression of macrophage markers and pro-inflammatory cytokines suggesting that benfotiamine is reducing dystrophic pathology by acting on inflammatory processes.
Publisher: Oxford University Press (OUP)
Date: 07-11-2020
DOI: 10.1093/HMG/DDZ266
Abstract: Duchenne muscular dystrophy (DMD) is a lethal muscle wasting disorder caused by mutations in the DMD gene that lead to the absence or severe reduction of dystrophin protein in muscle. The mdx mouse, also dystrophin deficient, is the model most widely used to study the pathology and test potential therapies, but the phenotype is milder than human DMD. This limits the magnitude and range of histological damage parameters and molecular changes that can be measured in pre-clinical drug testing. We used three weeks of voluntary wheel running to exacerbate the mdx phenotype. In mdx mice voluntary exercise increased the amount of damaged necrotic tissue and macrophage infiltration. Global gene expression profiling revealed that exercise induced additional and larger gene expression changes in mdx mice and the pathways most impacted by exercise were all related to immune function or cell-extracellular matrix (ECM) interactions. When we compared the matrisome and inflammation genes that were dysregulated in mdx with those commonly differentially expressed in DMD, we found the exercised mdx molecular signature more closely resembled that of DMD. These gene expression changes in the exercised mdx model thus provide more scope to assess the effects of pre-clinical treatments. Our gene profiling comparisons also highlighted upregulation of extracellular matrix proteins involved in innate immunity pathways, proteases that can release them, and downstream receptors and signalling molecules in exercised mdx and DMD, suggesting that the ECM could be a major source of pro-inflammatory molecules that trigger and maintain the immune response in dystrophic muscle.
Publisher: MDPI AG
Date: 06-02-2021
DOI: 10.3390/MOLECULES26040853
Abstract: Duchenne muscular dystrophy (DMD) is a progressive fatal neuromuscular disorder with no cure. Therapies to restore dystrophin deficiency have been approved in some jurisdictions but long-term effectiveness is yet to be established. There is a need to develop alternative strategies to treat DMD. Resveratrol is a nutraceutical with anti-inflammatory properties. Previous studies have shown high doses (100–400 mg/kg bodyweight/day) benefit mdx mice. We treated 4-week-old mdx and wildtype mice with a lower dose of resveratrol (5 mg/kg bodyweight/day) for 15 weeks. Voluntary exercise was used to test if a lower dosage than previously tested could reduce exercise-induced damage where a greater inflammatory infiltrate is present. We found resveratrol promoted skeletal muscle hypertrophy in wildtype mice. In dystrophic muscle, resveratrol reduced exercise-induced muscle necrosis. Gene expression of immune cell markers, CD86 and CD163 were reduced however, signalling targets associated with resveratrol’s mechanism of action including Sirt1 and NF-κB were unchanged. In conclusion, a lower dose of resveratrol compared to the dosage used by other studies reduced necrosis and gene expression of inflammatory cell markers in dystrophic muscle suggesting it as a therapeutic candidate for treating DMD.
Publisher: Cold Spring Harbor Laboratory
Date: 27-03-2018
DOI: 10.1101/289587
Abstract: Duchenne muscular dystrophy (DMD) is a progressive and fatal neuromuscular disorder for which there is no treatment. Therapies to restore dystrophin deficiency are not ready for clinical use and long-term efficiency is yet to be established. Therefore, there is a need to develop alternative strategies to treat DMD. Resveratrol is a nutraceutical with anti-inflammatory properties and previous studies have shown that high doses can benefit mdx mice. We treated 4-week-old mdx and wildtype mice with low-dose resveratrol (5mg/kg bodyweight/day) for 15 weeks. A voluntary exercise protocol was added to test if low dose resveratrol could reduce exercise-induced damage. We showed that resveratrol promoted skeletal muscle hypertrophy in the wildtype mice. There was no change in markers of pathology in the mdx mice however, the low-dose resveratrol reduced exercised induced damage. Gene expression of immune cell markers such as CD86, CD163 and PCNA was reduced however signalling targets associated with resveratrol’s mechanism of action of action including SIRT1 and NF-κB were unchanged. In conclusion, low-dose resveratrol was not effective in reducing disease pathology however, its ability to promote hypertrophy in wildtype skeletal muscle could have direct applications to the livestock industry or in sports medicine.
Publisher: Cold Spring Harbor Laboratory
Date: 18-05-2023
DOI: 10.1101/2023.05.16.23289881
Abstract: An N-of-1 trial was developed to deliver a dCas9-VP64 transgene designed to upregulate the cortical dystrophin as a custom therapy for a Duchenne muscular dystrophy (DMD) patient. After showing signs of mild cardiac dysfunction and pericardial effusion, the patient acutely decompensated and sustained cardiac arrest six-days after dosing and succumbed two-days later. Post-mortem examination revealed severe acute-respiratory distress syndrome with diffuse alveolar damage. Vector biodistribution data was obtained and revealed minimal expression of transgene in liver. There was no evidence of AAV9 antibodies nor of effector T cell reactivity. These findings demonstrate innate immune signaling with capillary leak as a form of toxicity in an advanced DMD case treated with high-dose rAAV gene therapy.
Publisher: American Association for the Advancement of Science (AAAS)
Date: 25-03-2020
DOI: 10.1126/SCITRANSLMED.AAY0271
Abstract: Genome-wide CRISPR-Cas9 screens identify druggable pathways associated with facioscapulohumeral muscular dystrophy.
Location: Australia
No related grants have been discovered for Keryn Woodman.