Publications by authors named "William R Lagor"

Gene therapy with Adeno-Associated Viral (AAV) vectors requires knowledge of their tropism within the body. Here we analyze the tropism of ten naturally occurring AAV serotypes (AAV3B, AAV4, AAV5, AAV6, AAV7, AAV8, AAV9, AAVrh8, AAVrh10 and AAVrh74) following systemic delivery into male and female mice. A transgene expressing ZsGreen and Cre recombinase was used to identify transduction in a cell-dependent manner based on fluorescence.

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  • Adeno-associated virus (AAV) gene therapy is increasingly recognized for its ability to target the central nervous system (CNS), particularly in treating neurological diseases.
  • Recent research focused on a new AAV gene therapy aimed at reducing amyloid aggregation in the brain by using a modified Aβ sequence variant to inhibit fibril formation.
  • The study showed that adjusting DNA plasmid elements—like signal peptides and incorporating a fusion tag—greatly enhanced the release and tracking of therapeutic peptides, ultimately improving peptide production by 10-fold in experimental models.
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  • * Results showed a complete reduction in ventricular arrhythmias in treated mice, both at 6 weeks and 12 months post-injection, indicating the long-term effectiveness of the treatment.
  • * The genome editing was found to be safe, with no negative effects on normal heart function or structure, suggesting a potential therapeutic avenue for CPVT.
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Homology Directed Repair (HDR)-based genome editing is an approach that could permanently correct a broad range of genetic diseases. However, its utility is limited by inefficient and imprecise DNA repair mechanisms in terminally differentiated tissues. Here, we tested "Repair Drive", a novel method for improving targeted gene insertion in the liver by selectively expanding correctly repaired hepatocytes .

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Viruses have evolved the ability to bind and enter cells through interactions with a wide variety of cell macromolecules. We engineered peptide-modified adeno-associated virus (AAV) capsids that transduce the brain through the introduction of de novo interactions with 2 proteins expressed on the mouse blood-brain barrier (BBB), LY6A or LY6C1. The in vivo tropisms of these capsids are predictable as they are dependent on the cell- and strain-specific expression of their target protein.

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  • The study explores the role of lipoprotein(a) (Lp(a)) in atherosclerotic cardiovascular disease by creating transgenic mice that express human apolipoprotein(a) and apoB-100 to achieve higher plasma Lp(a) levels, addressing limitations of previous models.
  • After feeding these mice a high-fat, high-cholesterol diet for 12 weeks, researchers observed significant differences in plaque size and composition, particularly in female Tg(LPA;APOB) mice, who showed increased necrotic core size and more extensive calcification compared to control mice.
  • The findings indicate that elevated Lp(a) correlates with more severe atherosclerosis, as evidenced by greater levels of pro-inflammatory markers and
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The liver is the metabolic center of the body and an ideal target for gene therapy of inherited metabolic disorders (IMDs). Adeno-associated viral (AAV) vectors can deliver transgenes to the liver with high efficiency and specificity and a favorable safety profile. Recombinant AAV vectors contain only the transgene cassette, and their payload is converted to non-integrating circular double-stranded DNA episomes, which can provide stable expression from months to years.

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Glutaric aciduria type I (GA-1) is an inborn error of metabolism with a severe neurological phenotype caused by the deficiency of glutaryl-coenzyme A dehydrogenase (GCDH), the last enzyme of lysine catabolism. Current literature suggests that toxic catabolites in the brain are produced locally and do not cross the blood-brain barrier. In a series of experiments using knockout mice of the lysine catabolic pathway and liver cell transplantation, we uncovered that toxic GA-1 catabolites in the brain originated from the liver.

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Targeting the Kv1.3 potassium channel has proven effective in reducing obesity and the severity of animal models of autoimmune disease. Stichodactyla toxin (ShK), isolated from the sea anemone Stichodactyla helianthus, is a potent blocker of Kv1.

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  • The CRISPR-Cas9 system is a revolutionary gene-editing tool that is being developed for gene therapy, but effective delivery to target cells is still a challenge due to pre-existing anti-vector immunity and off-target edits.
  • Various delivery methods, including both viral and non-viral vectors, are being explored, with adenoviruses showing promise for targeted delivery.
  • Researchers have engineered a simian adenovirus (SAd36) to specifically target myeloid cells and enhance gene editing in vascular endothelium, potentially addressing genetic disorders without triggering immune responses.
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  • Lipoprotein(a) (Lp(a)) is a distinct type of lipoprotein linked to cardiovascular disease risk, and current drugs do not effectively lower its levels.
  • Researchers created a mouse model to study the effects of a CRISPR-Cas9 therapy designed to reduce Lp(a) by removing the apolipoprotein(a) component from circulation.
  • The study showed that the CRISPR-Cas9 method can significantly diminish apo(a) levels, but also revealed unintended genetic changes, emphasizing the need to assess potential side effects of gene editing in complex genomic regions.
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The low-density lipoprotein receptor (Ldlr) and apolipoprotein E (Apoe) germline knockout (KO) models have provided fundamental insights in lipid and atherosclerosis research for decades. However, testing new candidate genes in these models requires extensive breeding, which is highly time and resource consuming. In this chapter, we provide methods for rapidly modeling hypercholesterolemia and atherosclerosis as well as testing new genes in adult mice through somatic gene editing.

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Purpose Of Review: This review examines recent progress in somatic genome editing for cardiovascular disease. We briefly highlight new gene editing approaches, delivery systems, and potential targets in the liver.

Recent Findings: In recent years, new editing and delivery systems have been applied successfully in model organisms to modify genes within hepatocytes.

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Background: The intestine occupies the critical interface between cholesterol absorption and excretion. Surprisingly little is known about the role of de novo cholesterol synthesis in this organ, and its relationship to whole body cholesterol homeostasis. Here, we investigate the physiological importance of this pathway through genetic deletion of the rate-limiting enzyme.

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Ciliogenic components, such as the family of intraflagellar transport (IFT) proteins, are recognized to play key roles in endochondral ossification, a critical process to form most bones. However, the unique functions and roles of each IFT during endochondral ossification remain unclear. Here, we show that IFT20 is required for endochondral ossification in mice.

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Methylmalonic acidemia (MMA) is a metabolic disorder most commonly caused by mutations in the methylmalonyl-CoA mutase () gene. Although adeno-associated viral (AAV) gene therapy has been effective at correcting the disease phenotype in MMA mouse models, clinical translation may be impaired by loss of episomal transgene expression and magnified by the need to treat patients early in life. To achieve permanent correction, we developed a dual AAV strategy to express a codon-optimized transgene from and tested various CRISPR-Cas9 genome-editing vectors in newly developed knockin mouse models of MMA.

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Genome editing in the lung has the potential to provide long-term expression of therapeutic protein to treat lung genetic diseases. Yet efficient delivery of CRISPR to the lung remains a challenge. The NIH Somatic Cell Genome Editing (SCGE) Consortium is developing safe and effective methods for genome editing in disease tissues.

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Clinical application of somatic genome editing requires therapeutics that are generalizable to a broad range of patients. Targeted insertion of promoterless transgenes can ensure that edits are permanent and broadly applicable while minimizing risks of off-target integration. In the liver, the Albumin () locus is currently the only well-characterized site for promoterless transgene insertion.

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Background & Aims: The accumulation of neutral lipids within hepatocytes underlies non-alcoholic fatty liver disease (NAFLD), which affects a quarter of the world's population and is associated with hepatitis, cirrhosis, and hepatocellular carcinoma. Despite insights gained from both human and animal studies, our understanding of NAFLD pathogenesis remains limited. To better study the molecular changes driving the condition we aimed to generate a humanised NAFLD mouse model.

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  • - The NIH Somatic Cell Genome Editing Consortium aims to enhance human health by developing safer and more effective genome editing techniques for treating diseases directly in patients' cells.
  • - The consortium plans to create a toolkit that includes new genome editing technologies, delivery methods, and validated data, which will be shared with the biomedical research community.
  • - By conducting thorough testing and validation, the initiative seeks to accelerate the discovery of new therapies for various health conditions.
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HMG-CoA reductase (Hmgcr) is the rate-limiting enzyme in the mevalonate pathway and is inhibited by statins. In addition to cholesterol, Hmgcr activity is also required for synthesizing nonsterol isoprenoids, such as dolichol, ubiquinone, and farnesylated and geranylgeranylated proteins. Here, we investigated the effects of Hmgcr inhibition on nonsterol isoprenoids in the liver.

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