A5019745578- Virus-based gene therapy research
- RNA Interference and Gene Delivery
- Viral Infectious Diseases and Gene Expression in Insects
- CRISPR and Genetic Engineering
- CAR-T cell therapy research
- Viral gastroenteritis research and epidemiology
- Spectroscopy and Quantum Chemical Studies
- X-ray Diffraction in Crystallography
- Crystallization and Solubility Studies
- Metabolism and Genetic Disorders
- Nanotechnology research and applications
- Immunotherapy and Immune Responses
- Cancer Research and Treatments
- Herpesvirus Infections and Treatments
- Carbon Nanotubes in Composites
- Graphene research and applications
- Photochemistry and Electron Transfer Studies
- Cholesterol and Lipid Metabolism
- Animal Virus Infections Studies
- Respiratory viral infections research
- Advancements in Semiconductor Devices and Circuit Design
- Lipoproteins and Cardiovascular Health
- Electron and X-Ray Spectroscopy Techniques
- interferon and immune responses
- Spectroscopy and Laser Applications
Baylor College of Medicine
2013-2023
Total (Belgium)
2021
Brain (Germany)
2021
Takeda (United States)
2018
The University of Sydney
2015
Cedars-Sinai Medical Center
2007-2008
University of California, Los Angeles
2008
UCLA Jonsson Comprehensive Cancer Center
2008
University of Florence
2007-2008
CEA Paris-Saclay
2007
Helper-dependent adenoviral vectors (HDAds) are devoid of all viral coding sequences and have demonstrated tremendous potential for gene therapy by providing increased cloning capacity (up to 37 kb) long-term, high-level transgene expression in vivo with negligible toxicity. Currently, the most widely used method producing HDAds is Cre/loxP system developed Graham co-workers. However, two major obstacles currently hinder progress this promising technology: (1) difficulty large-scale vector...
Systemic intravascular delivery of adenoviral (Ad) vectors for liver-directed gene therapy has been widely employed because its simplicity, noninvasiveness, and potential high transduction. For first-generation Ad (FGAd), this results in but transient levels transgene expression long-term hepatotoxicity due to viral from the vector backbone. Furthermore, doses also result an acute innate inflammatory response with potentially lethal consequences. Unlike FGAd, helper-dependent (HDAd) contain...
Adenoviruses are robust gene delivery vectors in vivo, but limited by their propensity to provoke strong innate and adaptive responses. Previous work has demonstrated that polyethylene glycol (PEG) modification of adenovirus can protect the from preexisting immune responses reducing protein–protein interactions. To test whether PEGylation reduce interactions with cells, first-generation (FG-Ad) helper-dependent (HD-Ad) Ad5 were PEGylated SPA-PEG tested vitro vivo. We demonstrate increasing...
Research Article4 February 2013Open Access Gene transfer of master autophagy regulator TFEB results in clearance toxic protein and correction hepatic disease alpha-1-anti-trypsin deficiency Nunzia Pastore Telethon Institute Genetics Medicine, Naples, Italy Search for more papers by this author Keith Blomenkamp Department Pediatrics, Saint Louis University School Cardinal Glennon Children's Medical Center, Louis, MO, USA Fabio Annunziata Pasquale Piccolo Pratibha Mithbaokar Rosa Maria Sepe...
Axon initial segments (AISs) generate action potentials and regulate the polarized distribution of proteins, lipids, organelles in neurons. While mechanisms AIS Na+ K+ channel clustering are understood, molecular that stabilize control neuronal polarity remain obscure. Here, we use proximity biotinylation mass spectrometry to identify proteome. We target biotin-ligase BirA* by generating fusion proteins with NF186, Ndel1, Trim46; these chimeras map organization intracellular membrane,...
Gene transfer of CFTR cDNA to airway epithelia is a promising approach treat cystic fibrosis (CF). Most gene vectors use strong viral promoters even though the endogenous promoter very weak. To learn whether expressing at low level in fraction cells would correct Cl(-) transport, we mixed freshly isolated wild-type and CF epithelial varying proportions generated differentiated epithelia. Epithelia with approximately 20% 70% transepithelial current containing 100% cells. These data were...
As much as 90% of an intravenously (i.v.) injected dose adenovirus serotype 5 (Ad5) is absorbed and destroyed by liver Kupffer cells. Viruses that escape these cells can then transduce hepatocytes after binding factor X (FX). Given interactions with FX are thought to occur on the Ad5 hexon protein, we replaced its exposed hypervariable regions (HVR) those from Ad6. When tested in vivo BALB/c mice hamsters, Ad5/6 chimera mediated >10 times higher transduction liver. This effect was not due...
Helper-dependent adenoviral vectors (HDAd) have been shown to mediate a considerably longer duration of transgene expression than first-generation vectors. We previously that from HDAd-transduced hepatocytes can persist at high levels for up 2.6 years in nonhuman primates following single-vector administration. Because and long-term toxicity are critical risk:benefit assessment, we continued monitor these animals. report here has persisted the entire observation period 7 all animals without...
Previous studies have shown that nitric oxide (NO) supplements may prevent bone loss and fractures in preclinical models of estrogen deficiency. However, the mechanisms by which NO modulates anabolism remain largely unclear. Argininosuccinate lyase (ASL) is only mammalian enzyme capable synthesizing arginine, sole precursor for synthase-dependent (NOS-dependent) synthesis. Moreover, ASL also required channeling extracellular arginine to NOS production. deficiency (ASLD) thus a model study...
Significance Loss-of-function mutations in MECP2 cause the neurological disorder Rett syndrome (RTT), but precise molecular mechanism driving pathogenesis remains unclear. Using an unbiased approach to identify proteins that interact with MeCP2, we identified transcription factor 20 (TCF20) complex and discovered RTT-causing disrupt this interaction. biochemical, morphological, behavioral, transcriptional studies, examined importance of interaction for brain function found TCF20 plays a...
Helper-dependent adenoviral vectors (HDAd) are devoid of all viral coding sequences and thus an improvement over early generation Ad because they can provide long-term transgene expression in vivo without chronic toxicity. However, high vector doses required to achieve efficient hepatic transduction by systemic intravenous injection, this unfortunately results dose-dependent acute To overcome important obstacle, we have developed a minimally invasive method preferentially deliver HDAd into...
The main challenge of gene therapy is to provide long-term, efficient transgene expression. Long-term expression from first generation adenoviral vectors (Advs) delivered the central nervous system (CNS) elicited in animals not previously exposed adenovirus (Ad). However, upon systemic immunization against Ad, a Adv abolished. High-capacity Advs (HC-Advs) sustained very long-term brain, even pre-immunized Ad. In this study, we tested hypothesis that HC-Adv brain would allow for expression,...
Helper-dependent adenoviral vectors (HDAds) are attractive for liver-directed gene therapy because they can mediate long-term, high-level transgene expression without chronic toxicity. However, systemic delivery requires high vector doses efficient hepatic transduction, resulting in dose-dependent acute Clearly, strategies to improve transduction with low needed. In this regard, we have previously shown that hydrodynamic injection of helper-dependent into mice results increased reduced...
The physics of confined water has stimulated extensive research in recent years, particular, regarding the role hydrogen bonding as a significant factor observed dynamics. In this work, two-dimensional infrared spectroscopy was employed to investigate response OH moiety phospholipid membrane samples. results show strong evidence for three distinct motifs (${\mathrm{H}}_{2}\mathrm{O}$ with zero, one, or both moieties bonded), whose relative proportions at interface are estimated.
In vivo gene transfer with adenovirus vectors would significantly benefit from a tight control of the adenovirus-inherent liver tropism. For efficient hepatocyte transduction, need to evade Kupffer cell scavenging while delivery peripheral tissues or tumors could be improved if both by cells and uptake hepatocytes were blocked. Here, we provide evidence that single point mutation in hexon capsomere designed enable defined chemical capsid modifications may permit detargeting targeting evasion...
Genome editing with site-specific endonucleases has implications for basic biomedical research as well gene therapy. We generated helper-dependent, capsid-modified adenovirus (HD-Ad5/35) vectors zinc-finger nuclease (ZFN)– or transcription activator-like effector (TALEN)–mediated genome in human CD34+ hematopoietic stem cells (HSCs) from mobilized adult donors. The production of these required that ZFN and TALEN expression HD-Ad5/35 producer 293-Cre was suppressed. To do this, we developed a...
Atherosclerosis, a disease of blood vessels, is driven by cholesterol accumulation and inflammation. Gene therapy that removes from vessels decreases inflammation promising approach for prevention treatment atherosclerosis. In previous work, we reported helper-dependent adenoviral (HDAd) overexpression apolipoprotein A-I (apoAI) in endothelial cells (ECs) increases efflux vitro reduces atherosclerosis vivo. However, the effect HDAdApoAI on partial. To improve this therapy, considered...
In view of recent serious adverse events and advances in gene therapy technologies, the use regulatable expression systems is becoming recognized as indispensable adjuncts to successful clinical therapy. present work we optimized high-capacity adenoviral (HC-Ad) vectors encoding novel tetracycline-dependent (TetOn)-regulatory elements for efficient rat brain vivo. We constructed two HC-Ad beta-galactosidase (beta-gal) driven by a TetOn system containing rtTAS(s)M2 transactivator tTS(Kid)...
We have evaluated the potential of liver-directed, helper-dependent adenoviral (HDAd) vector-mediated gene therapy in hemophilia B dog. Two dogs were injected intravenously with HDAd (3 × 1012 VP/kg) bearing a liver-restricted canine coagulation factor IX (FIX) expression cassette. After injection, whole blood clotting time for both declined from >60 min to ≤20 at least 604 and 446 days, respectively. Peak FIX activities 34.1 129.2% detected 12×14 days then slowly 2 5% by 120 stabilized...
Helper-dependent adenoviral vectors (HDAds) are attractive for liver-directed gene therapy because they can mediate sustained, high-level transgene expression without chronic toxicity. However, high vector doses required to achieve efficient hepatic transduction by systemic delivery of a nonlinear dose response. Unfortunately, such result in dissemination and dose-dependent acute toxicity with potentially severe lethal consequences. We hypothesize that the threshold may be circumvented...
Femtosecond infrared (IR) two-color pump-probe experiments were used to investigate the nonlinear response of D2O stretching vibration in weakly hydrated dimyristoyl-phosphatidylcholine (DMPC) membrane fragments. The vibrational lifetime is comparable or longer than that bulk and frequency dependent, as it decreases with increasing probe frequency. Also, increases when water content sample lowered. measured lifetimes range between 903 390 fs. A long-lived spectral feature grows following...