A5048270390- Virus-based gene therapy research
- Clostridium difficile and Clostridium perfringens research
- Gut microbiota and health
- Antibiotic Resistance in Bacteria
- Hemophilia Treatment and Research
- Viral Infectious Diseases and Gene Expression in Insects
- CRISPR and Genetic Engineering
- CAR-T cell therapy research
- Antibiotics Pharmacokinetics and Efficacy
- Retinal Diseases and Treatments
- RNA Interference and Gene Delivery
- RNA and protein synthesis mechanisms
- Blood Coagulation and Thrombosis Mechanisms
- Chromosomal and Genetic Variations
- Probiotics and Fermented Foods
- Bacterial Infections and Vaccines
- RNA Research and Splicing
- Retinal Development and Disorders
- Angiogenesis and VEGF in Cancer
- Plant Disease Resistance and Genetics
- Plant Virus Research Studies
- Glioma Diagnosis and Treatment
- Influenza Virus Research Studies
- Diet and metabolism studies
- Platelet Disorders and Treatments
Synthetic Biologics (United States)
2014-2020
Dana-Farber Brigham Cancer Center
2015
Graybug Vision (United States)
2013
Harvard University
2012
Novartis (Switzerland)
2002-2005
Research Institute for Genetic and Human Therapy
1995-2004
Novartis (United States)
1999-2004
Advanced Vision Institute
2003-2004
Queen's University
2001
Novartis (Canada)
2001
Polyadenylation of pre-mRNAs requires the conserved hexanucleotide AAUAAA, as well sequences located downstream from poly(A) addition site. The role these in production functional mRNAs was studied by analyzing a series mutants containing deletions or substitutions SV40 early region As expected, both previously defined GU-rich element and an AAUAAA sequence were required for efficient usage wild-type However, when either elements deleted, greatly increased levels SV40-specific RNA detected...
Endostatin, a proteolytic fragment of collagen XVIII, is an endogenous inhibitor tumor angiogenesis that also inhibits choroidal neovascularization. In this study, we assessed the effects increased intraocular expression endostatin on vascular endothelial growth factor (VEGF)-induced changes in retina. After subretinal injection pair gutless adenoviral vectors (AGV) designed to provide tamoxifen-inducible endostatin, diffuse immunoreactivity was induced thoroughout retina by administration...
Our previous studies of the 3'-end processing simian virus 40 late mRNAs indicated existence an essential element (or elements) downstream AAUAAA signal. We report here use transient expression analysis to study a functional which we located within sequence AGGUUUUUU, beginning 59 nucleotides recognized signal AAUAAA. Deletion this resulted in (i) at least 75% drop normal site and (ii) appearance readthrough transcripts with alternate 3' ends. Some flexibility position relative was noted by...
The gut microbiome, composed of the microflora that inhabit gastrointestinal tract and their genomes, make up a complex ecosystem can be disrupted by antibiotic use. ensuing dysbiosis is conducive to emergence opportunistic pathogens such as Clostridium difficile. A novel approach protect microbiome from antibiotic-mediated use beta-lactamase enzymes degrade residual antibiotics in before are harmed. Here we present preclinical development early clinical studies enzymes, P3A, currently...
Gutless adenoviral vectors are devoid of all viral coding regions and display reduced cytotoxicity, diminished immunogenicity, an increased capacity compared with early generation vectors. Using hemophilia A, a deficiency in clotting factor VIII (FVIII), as model disease, we generated evaluated gutless vector encoding human FVIII. The FVIII grew to high titer was reproducibly scaled-up from seed lots. Extensive DNA analyses revealed no rearrangements the genome. A quantitative PCR assay...
Abstract Increased expression of vascular endothelial growth factor (VEGF) in the retina starting after postnatal day (P)7 results neovascularization originating from deep retinal capillaries, but not those superficial capillary bed. Doxycycline was administered P0 to double transgenic mice with inducible VEGF retina. These showed proliferation and dilation indicating that at this stage development, capillaries are sensitive effects VEGF. Angiopoietin‐2 (Ang2) is expressed along surface for...
Abstract Increased expression of vascular endothelial cell growth factor (VEGF) in the retina is sufficient to stimulate sprouting neovascularization from deep capillary bed retina, but not superficial retinal capillaries or choriocapillaris. Coexpression VEGF and angiopoietin 2 (Ang2) results capillaries, However, retina‐derived Ang2 may reach choriocapillaris, because tight junctions between pigmented epithelial (RPE) cells. To eliminate this possible confounding factor, we used human...
Hemophilia A is caused by blood coagulation factor VIII (FVIII) deficiency and an attractive target for gene therapy. However, features of FVIII physiology, such as the instability mRNA protein, have provided obstacles to design a feasible strategy transfer expression human in vivo. We constructed recombinant adenoviral vector, Av1ALH81, that contains cDNA from which B-domain has been deleted (BDD FVIII) extensively characterized this vector vitro In vitro, HepG2, hepatoma cells, transduced...
A region in the adenovirus major late promoter (MLP) containing a CCAAT consensus sequence can direct transcription termination of RNA polymerase II, mechanism that possibly prevents transcriptional interference from upstream genes. Using chimeric plasmid template contains MLP directing expression simian virus 40 early region, we showed an inserted oligonucleotide only 13 base pairs sequences, including box, is capable inducing orientation-dependent, position-independent manner. Point...
An antibody prevents the symptoms of whooping cough in newborn baboons when administered 5 weeks before infection.
Antibiotics can damage the gut microbiome, leading to serious adventitious infections and emergence of antibiotic resistant pathogens. Antibiotic inactivation in GI tract represents a strategy protect colonic microbiota integrity reduce resistance. Clinical utility this approach was established when SYN-004 (ribaxamase), an orally-administered beta-lactamase, demonstrated degrade ceftriaxone preserve microbiome. Ribaxamase degrades penicillins cephalosporin beta-lactams, but not carbapenems....
Adenoviral vectors devoid of all viral coding regions are referred to by many names, including gutless vectors. Gutless display reduced toxicity and immunogenicity, increased duration transgene expression, capacity compared early generation vectors, which contain the majority backbone genes. However, production at a scale purity suitable for clinical use has limited utility this technology. In work we describe optimization We constructed an improved helper virus generated alternative vector...
The enzyme glucokinase (GK) plays a central role in glucose homeostasis. Hepatic GK activity is acutely controlled by the action of regulatory protein (GKRP). In vitro evidence suggests that GKRP reversibly binds to and inhibits its activity; however, less known about vivo function GKRP. To further explore physiological vivo, we used an E1/E2a/E3-deficient adenoviral vector containing cDNA encoding human (Av3hGKRP). High fat diet-induced diabetic mice were administered Av3hGKRP or control...
Hemophilia A results from subnormal levels of blood coagulation factor VIII (FVIII) and is an attractive target for gene therapy. However, progress has been impeded by features FVIII biology such as low mRNA accumulation the instability protein. We have shown previously that a adenoviral vector, Av1ALH81, allowed high-level expression human in mice sustained several weeks. Here, we generated second Av1ALAPH81, which intron was introduced into cassette. Administration Av1ALAPH81 to resulted...
Our previous studies of the 3'-end processing simian virus 40 late mRNAs indicated existence an essential element (or elements) downstream AAUAAA signal. We report here use transient expression analysis to study a functional which we located within sequence AGGUUUUUU, beginning 59 nucleotides recognized signal AAUAAA. Deletion this resulted in (i) at least 75% drop normal site and (ii) appearance readthrough transcripts with alternate 3' ends. Some flexibility position relative was noted by...
To evaluate an antibiotic inactivation strategy to protect the gut microbiome from antibiotic-mediated damage.SYN-004 (ribaxamase) is orally delivered beta-lactamase intended degrade penicillins and cephalosporins within gastrointestinal tract microbiome. Pigs (20 kg, n = 10) were treated with ceftriaxone (CRO) (IV, 50 mg kg-1 , SID) for 7 days a cohort (n 5) received ribaxamase (PO, 75 mg, QID) 9 beginning day before administration. Ceftriaxone serum levels not statistically different in...
The use of adenoviral vectors for gene therapy has been limited due to host immune responses directed toward the vector and/or transgene and toxicity. To decrease immunogenicity toxicity, we attenuated viral expression by eliminating E1, E2a, E3, E4 early genes from backbone. Two highly attenuated, fourth-generation (Av4) E1/E2a/E3/E4-deficient encoding human factor VIII (FVIII) under control a liver-specific albumin promoter were generated. One Av4 (Av4DeltaE4FVIII) was deficient in entire...