A5086264524- Retinal Development and Disorders
- Virus-based gene therapy research
- Retinal Diseases and Treatments
- Photoreceptor and optogenetics research
- CRISPR and Genetic Engineering
- Mitochondrial Function and Pathology
- RNA Interference and Gene Delivery
- Retinal and Optic Conditions
- interferon and immune responses
- Retinopathy of Prematurity Studies
- RNA regulation and disease
- Glaucoma and retinal disorders
- bioluminescence and chemiluminescence research
- Advanced biosensing and bioanalysis techniques
- Neuroscience and Neural Engineering
- RNA and protein synthesis mechanisms
- Metabolism and Genetic Disorders
- Photochromic and Fluorescence Chemistry
- Advanced Fluorescence Microscopy Techniques
- Ocular Disorders and Treatments
- Connexins and lens biology
- Receptor Mechanisms and Signaling
- Herpesvirus Infections and Treatments
- Retinoids in leukemia and cellular processes
- Retinal and Macular Surgery
University of Florida
2014-2023
Florida College
2012-2022
American Academy of Ophthalmology
2017
University of California, San Francisco
1997-2012
Ophthalmology Associates (United States)
1989-2012
University of North Texas Health Science Center
2012
University of California, San Diego
2012
King Khalid University Hospital
2012
King Saud University
2012
King Faisal Specialist Hospital & Research Centre
2012
Leber congenital amaurosis (LCA) is a group of autosomal recessive blinding retinal diseases that are incurable. One molecular form caused by mutations in the RPE65 (retinal pigment epithelium-specific 65-kDa) gene. A recombinant adeno-associated virus serotype 2 (rAAV2) vector, altered to carry human gene (rAAV2-CBSB-hRPE65), restored vision animal models with deficiency. clinical trial was designed assess safety rAAV2-CBSB-hRPE65 subjects RPE65-LCA. Three young adults (ages 21–24 years)...
The neuromuscular disease myotonic dystrophy (DM) is caused by microsatellite repeat expansions at two different genomic loci. Mutant DM transcripts are retained in the nucleus together with muscleblind (Mbnl) proteins, and these abnormal RNAs somehow interfere pre-mRNA splicing regulation. Here, we show that disruption of mouse Mbnl1 gene leads to muscle, eye, RNA abnormalities characteristic disease. Our results support hypothesis manifestations can result from sequestration specific...
The RPE65 gene encodes the isomerase of retinoid cycle, enzymatic pathway that underlies mammalian vision. Mutations in disrupt cycle and cause a congenital human blindness known as Leber amaurosis (LCA). We used adeno-associated virus-2-based replacement therapy to treat three young adults with RPE65-LCA measured their vision before up 90 days after intervention. All patients showed statistically significant increase visual sensitivity at 30 treatment localized retinal areas had received...
We constructed gfph, a synthetic version of the jellyfish Aequorea victoria green fluorescent protein (gfp) cDNA that is adapted for high-level expression in mammalian cells, especially those human origin. A total 92 base substitutions were made 88 codons order to change codon usage within gfp10 coding sequence so it was more appropriate cells. also describe series versatile recombinant adeno-associated virus and adenovirus vectors delivery genes into cells and, using these vectors,...
The short- and long-term effects of gene therapy using AAV-mediated RPE65 transfer to canine retinal pigment epithelium were investigated in dogs affected with disease caused by deficiency. Results AAV 2/2, 2/1, 2/5 vector pseudotypes, human or cDNA, constitutive tissue-specific promoters similar. Subretinally administered vectors restored function 23 26 eyes, but intravitreal injections consistently did not. Photoreceptoral postreceptoral both rod cone systems improved therapy. In followed...
Significance The first retinal gene therapy in human blindness from RPE65 mutations has focused on safety and efficacy, as defined by improved vision. disease component not studied, however, been the fate of photoreceptors this progressive degeneration. We show that improves vision for at least 3 y, but photoreceptor degeneration progresses unabated humans. In canine model, same result occurs when treatment is stage equivalent to study shows need combinatorial improve short term also slow long term.
We describe a general approach for achieving efficient and cell type-specific expression of exogenous genes in photoreceptor cells the mammalian retina. Recombinant adeno-associated virus (rAAV) vectors were used to transfer bacterial lacZ gene or synthetic green fluorescent protein (gfp) mouse rat retinas after injection into subretinal space. Using proximal murine rod opsin promoter (+86 -385) drive expression, reporter product was found exclusively photoreceptors, not any other retinal...
Vectors derived from adeno-associated viruses (AAVs) have become important gene delivery tools for the treatment of many inherited ocular diseases in well-characterized animal models. Previous studies determined that viral capsid plays an essential role cellular tropism and efficiency transgene expression. Recently, it was shown phosphorylation surface-exposed tyrosine residues AAV2 targets particles ubiquitination proteasome- mediated degradation, mutations these lead to highly efficient...
Two mitochondrial genotypes are shown to exist within one Holstein cow maternal lineage. They were detected by the appearance of an extra Hae III recognition site in genotype. The nucleotide sequence this region has been determined and distinguished adenine/guanine base transition which creates new site. This point mutation occurs open reading frame at third position a glycine codon therefore does not alter amino acid sequence. present pattern lineage demands that multiple shifts between...
Retinal gene therapy for Leber's congenital amaurosis, an autosomal recessive childhood blindness, has been widely considered to be safe and efficacious. Three years after therapy, improvement in vision was maintained, but the rate of loss photoreceptors treated retina same as that untreated retina. Here we describe long-term follow-up data from three patients. Topographic maps visual sensitivity regions, nearly 6 two patients 4.5 third patient, indicate progressive diminution areas improved...
Human gene therapy with rAAV2-vector was performed for the RPE65 form of childhood blindness called Leber congenital amaurosis. In three contemporaneous studies by independent groups, procedure deemed safe and there evidence visual gain in short term. At 12 months after treatment, our young adult subjects remained healthy without vector-related serious adverse events. Results immunological assays to identify reaction AAV serotype 2 capsid were unchanged from baseline measurements. clinical...
A clone of human cells (Detroit 6) latently infected by adeno-associated virus (AAV) has been characterized with regard to the status viral DNA. In both early (9 10) and late (118) passages clone, AAV-DNA was recombined host DNA, at least in some cases as a head-to-tail tandem repeat, via terminal sequences genome. However, it not possible distinguish between integration into chromosomal DNA very large plasmids (< 20 x 10(6) molecular weight) which contain cellular sequences. Although...
Previous work established retinal expression of channelrhodopsin-2 (ChR2), an algal cation channel gated by light, restored physiological and behavioral visual responses in otherwise blind rd1 mice. However, a viable ChR2-based human therapy must meet several key criteria: (i) ChR2 be targeted, robust, long-term, (ii) provide long-term continuous therapeutic efficacy, (iii) both viral vector delivery safe. Here, we demonstrate the development clinically relevant for late stage degeneration...
The P23H mutation within the rhodopsin gene ( RHO ) causes misfolding, endoplasmic reticulum (ER) stress, and activates unfolded protein response (UPR), leading to rod photoreceptor degeneration autosomal dominant retinitis pigmentosa (ADRP). Grp78/BiP is an ER-localized chaperone that induced by UPR signaling in ER stress. We have previously demonstrated BiP mRNA levels are selectively reduced animal models of ADRP arising from expression at ages precede degeneration. now overexpressed test...
Fluorescence of adenine, guanine, cytosine, and uracil at room temperature in neutral aqueous solution has been detected by means a digital signal accumulation technique. Corrected emission excitation spectra are presented compared with low-temperature data. The quantum yields are, respectively, 2.6 x 10(-4), 3.0 0.8 0.5 10(-4) when the bases excited their low-energy absorption maxima.
The successful restoration of visual function with recombinant adeno-associated virus (rAAV)-mediated gene replacement therapy in animals and humans an inherited disease the retinal pigment epithelium has ushered a new era therapeutics. For many disorders, however, targeting therapeutic vectors to mutant rods and/or cones will be required. In this study, primary cone photoreceptor disorder achromatopsia served as ideal translational model develop directed photoreceptors. We demonstrate that...
Hereditary retinal blindness is caused by mutations in genes expressed photoreceptors or pigment epithelium. Gene therapy mouse and dog models of a primary epithelium disease has already been translated to human clinical trials with encouraging results. Treatment for common photoreceptor blindness, however, not yet moved from proof concept the clinic. We evaluated gene augmentation two blinding canine diseases that model X-linked form retinitis pigmentosa GTPase regulator ( RPGR ) gene,...
Vectors based on adeno-associated virus serotype 2 (AAV2) have been used extensively in many gene-delivery applications, including several successful clinical trials for one type of Leber congenital amaurosis the retina. Many studies focused improving AAV2 transduction efficiency and cellular specificity by genetically engineering its capsid. We previously shown that vectors-containing single-point mutations capsid surface tyrosines serotypes AAV2, AAV8, AAV9 displayed significantly...