Abstract Background Tumors are complex tissues containing collections of phenotypically diverse malignant and nonmalignant cells. We know little the mechanisms that govern heterogeneity tumor cells nor role plays in overcoming stresses, such as adaptation to different microenvironments. Osteosarcoma is an ideal model for studying these mechanisms—it exhibits widespread inter- intra-tumoral heterogeneity, predictable patterns metastasis, a lack clear targetable driver mutations. Understanding...

Duchenne muscular dystrophy (DMD) is typically caused by mutations that disrupt the DMD reading frame, but nonsense in 5' part of gene induce utilization an internal ribosomal entry site (IRES) exon 5, driving expression a highly functional N-truncated dystrophin. We have developed AAV9 vector expressing U7 small nuclear RNAs targeting 2 and tested it mouse containing duplication 2, which skipping both copies induces IRES-driven expression, one copy leads to wild-type dystrophin expression....

In a phase 1/2, open-label dose escalation trial, we delivered rAAVrh74.MCK.GALGT2 (also B4GALNT2) bilaterally to the legs of two boys with Duchenne muscular dystrophy using intravascular limb infusion. Subject 1 (age 8.9 years at dosing) received 2.5 × 1013 vector genome (vg)/kg per leg (5 vg/kg total) and subject 2 6.9 5 (1 1014 total). No serious adverse events were observed. Muscle biopsy evaluated 3 or 4 months post treatment versus baseline showed evidence GALGT2 gene expression...

Duchenne muscular dystrophy (DMD) is an X-linked progressive disease characterized by loss of dystrophin protein that typically results from truncating mutations in the DMD gene. Current exon-skipping therapies have sought to treat deletion abolish open reading frame (ORF) skipping adjacent exon, order restore ORF allows translation internally deleted yet partially functional protein, as seen with many patients milder Becker (BMD) phenotype. In contrast approach, one copy a duplicated exon...

Abstract Aims Dystrophin, the protein product of DMD gene, plays a critical role in muscle integrity by stabilising sarcolemma during contraction and relaxation. The gene is vulnerable to variety mutations that may cause complete loss, depletion or truncation protein, leading Duchenne Becker muscular dystrophies. Precise reproducible dystrophin quantification essential characterising evaluating outcome efforts induce through therapies. Immunofluorescence microscopy offers high sensitivity...

Duchenne muscular dystrophy is an X-linked disorder typically caused by out-of-frame mutations in the

Duchenne muscular dystrophy (DMD) is a devastating muscle-wasting disease that arises due to the loss of dystrophin expression, leading progressive motor and cardiorespiratory function. Four exon-skipping approaches using antisense phosphorodiamidate morpholino oligomers (PMOs) have been approved by FDA restore DMD open reading frame, resulting in expression functional but internally deleted protein, patients with single-exon duplications, exon skipping has potential full-length expression....

Anc80L65 is a synthetic, ancestral adeno-associated virus that has high tropism toward retinal photoreceptors after subretinal injection in mice and non-human primates. We characterized, for the first time, post-intravitreal cell-specific transduction profile of compared with AAV9. Here we use AAV9 to intravitreally deliver copy gene encoding GFP into WT C57Bl/6J mice. expression was driven by one two clinically relevant promoters, chicken β actin (CB) or truncated MECP2 (P546). After...

Abstract Purpose For patients with osteosarcoma, disease-related mortality most often results from lung metastasis—a phenomenon shared many solid tumors. While established metastatic lesions behave aggressively, very few of the tumor cells that reach will survive. By identifying mechanisms facilitate survival disseminated cells, we can develop therapeutic strategies prevent and treat metastasis. Methods We analyzed single cell RNA-sequencing (scRNAseq) data murine metastasis-bearing lungs to...

Duchenne muscular dystrophy (DMD) is a progressive X-linked disease caused by mutations in the

Single-stranded DNA (ssDNA) templates along with Cas9 have been used for gene insertion but suffer from low efficiency. Here, we show that ssDNA chemical modifications in 10-17% of internal bases (eDNA) is compatible the homologous recombination machinery. Moreover, eDNA improve by 2-3 fold compared to unmodified and end-modified airway basal stem cells (ABCs), hematopoietic progenitor (HSPCs), T-cells endothelial cells. Over 50% alleles showed three clinically relevant loci (CFTR, HBB,...

Abstract The blood-brain barrier (BBB) serves as a highly intricate and dynamic interface connecting the brain bloodstream, playing vital role in maintaining homeostasis. BBB dysfunction has been associated with multiple neurodegenerative diseases, including amyotrophic lateral sclerosis (ALS); however, of neurodegeneration is understudied. We developed an ALS patient-derived model by using cells derived from 5 patient donors carrying C9ORF72 mutations. Brain microvascular endothelial-like...

Neuroinflammation is a miscreant in accelerating progression of many neurodegenerative diseases, including amyotrophic lateral sclerosis (ALS). However, treatments targeting neuroinflammation alone have led to disappointing results clinical trials. Both neuronal and non-neuronal cell types been implicated the pathogenesis ALS, multiple studies shown correction each type has beneficial effects on disease outcome. Previously, we that AAV9-mediated superoxide dismutase 1 (SOD1) suppression...

Abstract Single-stranded DNA (ssDNA) templates along with Cas9 have been used for knocking-in exogenous sequences in the genome but suffer from low efficiency. Here, we show that ssDNA chemical modifications 12–19% of internal bases, which denote as enhanced (esDNA), improve knock-in (KI) by 2–3-fold compared to end-modified airway basal stem cells (ABCs), CD34 + hematopoietic (CD34 cells), T-cells and endothelial cells. Over 50% alleles showed KI three clinically relevant loci (CFTR, HBB...

ABSTRACT Cystic fibrosis (CF) is caused by mutations in the cystic transmembrane conductance regulator ( CFTR ) gene. Although many people with CF (pwCF) are treated using modulators, some non-responsive due to their genotype or other uncharacterized reasons. Autologous airway stem cell therapies, which cDNA has been replaced, may enable a durable therapy for all pwCF. Previously, CRISPR-Cas9 two AAVs was used sequentially insert halves of and an enrichment cassette into locus. However,...

Cystic fibrosis (CF) is caused by mutations in the cystic transmembrane conductance regulator (

Abstract Background Tumors are complex tissues containing collections of phenotypically diverse malignant and nonmalignant cells. We know little the mechanisms that govern heterogeneity tumor cells nor role plays in overcoming stresses, such as adaptation to different microenvironments. Osteosarcoma is an ideal model for studying these mechanisms—it exhibits widespread inter- intra-tumoral heterogeneity, predictable patterns metastasis, a lack clear targetable driver mutations. Understanding...

Myotonic dystrophy type 1 (DM1) is the most common form of muscular in adults and affects mainly skeletal muscle, heart, brain. DM1 caused by a CTG repeat expansion 3′UTR region DMPK gene that sequesters muscleblind-like proteins, blocking their splicing activity forming nuclear RNA foci . Consequently, many genes have reversed to fetal pattern. There no treatment for DM1, but several approaches been explored, including antisense oligonucleotides (ASOs) aiming knock down expression or bind...