A5085271893- Muscle Physiology and Disorders
- Virus-based gene therapy research
- CRISPR and Genetic Engineering
- Tissue Engineering and Regenerative Medicine
- Muscle activation and electromyography studies
- Cardiomyopathy and Myosin Studies
- Adipose Tissue and Metabolism
- Viral Infections and Immunology Research
- Exercise and Physiological Responses
- Muscle metabolism and nutrition
- RNA Interference and Gene Delivery
- Prosthetics and Rehabilitation Robotics
- Viral Infectious Diseases and Gene Expression in Insects
- Human-Animal Interaction Studies
- Advanced Sensor and Energy Harvesting Materials
- Genetic Neurodegenerative Diseases
- Neurogenetic and Muscular Disorders Research
- Silk-based biomaterials and applications
- Connexins and lens biology
- Cytomegalovirus and herpesvirus research
- Genetics, Aging, and Longevity in Model Organisms
- Biotin and Related Studies
- Animal Genetics and Reproduction
- Congenital heart defects research
- Extracellular vesicles in disease
University of Missouri
2015-2025
University of Missouri Hospital
2010-2022
National Center for Advancing Translational Sciences
2016-2021
National Institutes of Health
2019-2021
National Nuclear Security Administration
2021
National Academies of Sciences, Engineering, and Medicine
2021
Missouri College
2019
Editing can help build stronger muscles Much of the controversy surrounding gene-editing technology called CRISPR/Cas9 centers on ethics germline editing human embryos to correct disease-causing mutations. For certain disorders such as muscular dystrophy, it may be possible achieve therapeutic benefit by faulty gene in somatic cells. In proof-of-concept studies, Long et al. , Nelson and Tabebordbar used adeno-associated virus-9 deliver system young mice with a mutation coding for dystrophin,...
Adeno-associated virus (AAV)-mediated CRISPR-Cas9 editing holds promise to treat many diseases. The immune response bacterial-derived Cas9 has been speculated as a hurdle for AAV-CRISPR therapy. However, immunological consequences of AAV-mediated expression have thus far not thoroughly investigated in large mammals. We evaluate Cas9-specific responses canine models Duchenne muscular dystrophy (DMD) following intramuscular and intravenous Treatment results initially robust dystrophin...
Dystrophin deficiency results in lethal Duchenne muscular dystrophy (DMD). Substituting missing dystrophin with abbreviated microdystrophin has dramatically alleviated disease mouse DMD models. Unfortunately, translation of therapy been unsuccessful dystrophic dogs, the only large mammalian model. Approximately 70% dystrophin-coding sequence is removed microdystrophin. Intriguingly, loss ≥50% frequently severe patients. To test whether small gene size constitutes a fundamental design error...
The ultimate goal of muscular dystrophy gene therapy is to treat all muscles in the body. Global delivery was demonstrated dystrophic mice more than a decade ago using adeno-associated virus (AAV). However, translation affected large mammals has been challenging. only reported attempt performed newborn Duchenne (DMD) dogs. Unfortunately, AAV injection resulted growth delay, muscle atrophy and contracture. Here we report safe bodywide juvenile DMD Three ∼2-m-old dogs received intravenous...
Adeno-associated virus–mediated (AAV-mediated) CRISPR editing is a revolutionary approach for treating inherited diseases. Sustained, often life-long mutation correction required these Unfortunately, this has never been demonstrated with AAV therapy. We addressed question in the mdx model of Duchenne muscular dystrophy (DMD). DMD caused by dystrophin gene mutation. Dystrophin deficiency leads to ambulation loss and cardiomyopathy. treated 6-week-old mice intravenously evaluated disease...
The mechanism of force reduction is not completely understood in Duchenne muscular dystrophy (DMD), a dystrophin-deficient lethal disease. Nitric oxide regulates muscle force. Interestingly, neuronal nitric synthase µ (nNOSµ), major source oxide, lost from the sarcolemma DMD muscle. We hypothesize that nNOSµ delocalization contributes to DMD. To test this hypothesis, we generated dystrophin/nNOSµ double knockout mice. Genetic elimination significantly enhanced dystrophin-null Pharmacological...
Dual adeno-associated virus (AAV) technology was developed in 2000 to double the packaging capacity of AAV vector. The proof principle has been demonstrated various mouse models. Yet, pivotal evidence is lacking large animal models human diseases. Here we report expression a 7-kb canine ΔH2–R15 mini-dystrophin gene using pair dual vectors model Duchenne muscular dystrophy (DMD). minigene by far most potent synthetic dystrophin engineered for DMD therapy. We packaged Y731F tyrosine-modified...
CRISPR editing of muscle stem cells (MuSCs) with adeno-associated virus serotype-9 (AAV9) holds promise for sustained gene repair therapy muscular dystrophies. However, conflicting evidence exists on whether AAV9 transduces MuSCs. To rigorously address this question, we used a graft model. The grafted underwent complete necrosis before regenerating from its We injected AAV9.Cre into Ai14 mice. These mice express tdTomato upon Cre-mediated removal floxed stop codon. About 28%–47% and 24%–89%...
Muscle rigidity and myotendinous junction (MTJ) deficiency contribute to immobilization in Duchenne muscular dystrophy (DMD), a lethal disease caused by the absence of dystrophin. However, little is known about muscle passive properties MTJ strength diseased muscle. Here, we hypothesize that dystrophin-deficient pathology renders skeletal stiffer weaker. To test our hypothesis, examined an intact noncontracting muscle-tendon unit mdx mice, mouse model for DMD. The extensor digitorum longus...
Neuronal nitric oxide synthase (nNOS) membrane delocalization contributes to the pathogenesis of Duchenne muscular dystrophy (DMD) by promoting functional muscle ischemia and exacerbating injury during exercise. We have previously shown that supra-physiological expression nNOS-binding mini-dystrophin restores normal blood flow regulation prevents in transgenic mdx mice, a DMD model. A critical next issue is whether systemic dual adeno-associated virus (AAV) gene therapy can restore mitigate...
Body movements are mainly provided by mechanical function of skeletal muscle. Skeletal muscle is composed numerous bundles myofibers that sheathed intramuscular connective tissues. Each myofiber contains many myofibrils run longitudinally along the length myofiber. Myofibrils contractile apparatus and they repeated units known as sarcomeres. A sarcomere unit actin myosin filaments spaced Z discs titin protein. Mechanical defined passive properties The used to characterize amount force...
Dystrophin is a large sub-sarcolemmal protein. Its absence leads to Duchenne muscular dystrophy (DMD). Binding the sarcolemma essential for dystrophin protect muscle from contraction-induced injury. It has long been thought that membrane binding of depends on its cysteine-rich (CR) domain. Here, we provide in vivo evidence suggesting contains three additional membrane-binding domains including spectrin-like repeats (R)1-3, R10-12 and C-terminus (CT). To systematically study binding, split...
Excessive cytosolic calcium accumulation contributes to muscle degeneration in Duchenne muscular dystrophy (DMD). Sarco/endoplasmic reticulum ATPase (SERCA) is a sarcoplasmic (SR) pump that actively transports from the cytosol into SR. We previously showed adeno-associated virus (AAV)-mediated SERCA2a therapy reduced overload and improved heart function murine DMD model. Here, we tested whether AAV could ameliorate disease canine 7.83 × 1013 vector genome particles of were injected extensor...
ABSTRACT Background Adeno‐associated virus (AAV) 8 and 9 are in clinical trials for treating neuromuscular diseases such as Duchenne muscular dystrophy (DMD). Muscle consists of myofibres different types sizes. However, little is known about the fibre type size tropism AAV large mammals. Methods We evaluated type‐ size‐specific transduction properties AAV8 AAV9 17 dogs that received systemic gene transfer (dose 1.94 ± 0.52 × 10 14 vg/kg; injected at 2.86 0.30 months; harvested 20.79 3.30...
ABSTRACT To establish a minimally invasive approach to studying body-wide muscle inflammation in the canine Duchenne muscular dystrophy (DMD) model, we evaluated 13 cytokines/chemokines frozen sera from 90 affected (239 sera) and 73 normal (189 dogs (0.00 45.2 months of age). Linear mixed-effects model analysis suggested that ten were significantly elevated dogs, including interleukin (IL)-2, IL-6, IL-7, IL-8, IL-10, IL-15, IL-18, C-C motif chemokine ligand 2 (CCL2), C-X-C 1 (CXCL1)...
Abstract mdx , mdx3cv and mdx4cv mice are among the most commonly used models for study of Duchenne muscular dystrophy. Their disease is caused by point mutations in dystrophin gene. Despite widespread use these models, genotyping has not always been straightforward. Current methods require multiple polymerase chain reactions (PCRs), post‐PCR manipulations, and/or special equipment/reagents. Herein we report a simple, robust PCR method based on primer competition. This approach could also be...
ABSTRACT Objective: Gap junction channels formed by connexin (Cx) protein subunits enable cell‐to‐cell conduction of vasoactive signals. Given the lack quantitative measurements Cx expression in microvascular endothelial cells (EC) and smooth muscle (SMC), objective was to determine whether differed between EC SMC resistance microvessels for which is well‐characterized. Methods: Cheek pouch arterioles (CPA) retractor feed arteries (RFA) were hand‐dissected dissociated obtain or tubes. In...
Abstract Introduction : Duchenne muscular dystrophy (DMD) is a severe, muscle‐wasting disease caused by mutations in the dystrophin gene. The mdx mouse first and perhaps most commonly used animal model for study of DMD. Both male female mice are used. However, it not completely clear whether gender influences contraction passive mechanical properties skeletal muscle. Methods We compared isometric tetanic forces extensor digitorum longus muscle between mice. Results At age 6 months, showed...
Loss of muscle force is a salient feature Duchenne muscular dystrophy (DMD), fatal disease caused by dystrophin deficiency. Assessment production from single intact has been considered as the gold standard for studying physiological consequences in murine models DMD. Unfortunately, equivalent assays have not established dystrophic dogs. To fill gap, we developed novel situ protocol to measure generated extensor carpi ulnaris (ECU) dog. We also determined length fiber ratio and pennation...
To study Ca(2+) signaling in the endothelium of murine feed arteries, we determined vitro stability endothelial cell (EC) tubes freshly isolated from abdominal muscle arteries male and female C57BL/6 mice (5-9 mo, 25-35 g). We tested hypothesis that intracellular concentration ([Ca(2+)](i)) responses to muscarinic receptor activation would increase with temperature. Intact EC (length: 1-2 mm, width: 65-80 μm) were using gentle enzymatic digestion trituration remove smooth cells. A tube was...
The muscular dystrophies are a group of devastating genetic disorders that affect both skeletal and cardiac muscle. An effective gene therapy for these diseases requires bodywide muscle delivery. Tyrosine mutant adeno-associated virus (AAV) has been considered as class highly potent transfer vectors. Here, we tested the hypothesis systemic delivery tyrosine AAV can result in transduction newborn dogs. Three vectors (Y445F/Y731F AAV-1, Y445F AAV-6, Y731F AAV-9) were evaluated. These expressed...