97 results on '"Pivirotto P"'
Search Results
2. MR-guided delivery of AAV2-BDNF into the entorhinal cortex of non-human primates
- Author
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Nagahara, Alan H, Wilson, Bayard R, Ivasyk, Iryna, Kovacs, Imre, Rawalji, Saytam, Bringas, John R, Pivirotto, Phillip J, Sebastian, Waldy San, Samaranch, Lluis, Bankiewicz, Krystof S, and Tuszynski, Mark H
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Biomedical and Clinical Sciences ,Dementia ,Alzheimer's Disease including Alzheimer's Disease Related Dementias (AD/ADRD) ,Neurosciences ,Brain Disorders ,Neurodegenerative ,Aging ,Biomedical Imaging ,Alzheimer's Disease ,Acquired Cognitive Impairment ,Neurological ,Mental health ,Animals ,Brain-Derived Neurotrophic Factor ,Contrast Media ,Dependovirus ,Entorhinal Cortex ,Female ,Gadolinium ,Genetic Vectors ,Green Fluorescent Proteins ,Heterocyclic Compounds ,Hippocampus ,Macaca fascicularis ,Macaca mulatta ,Magnetic Resonance Imaging ,Male ,Neurons ,Organometallic Compounds ,Protein Transport ,Biological Sciences ,Medical and Health Sciences ,Biotechnology ,Biological sciences ,Biomedical and clinical sciences ,Health sciences - Abstract
Brain-derived neurotrophic factor (BDNF) gene delivery to the entorhinal cortex is a candidate for treatment of Alzheimer's disease (AD) to reduce neurodegeneration that is associated with memory loss. Accurate targeting of the entorhinal cortex in AD is complex due to the deep and atrophic state of this brain region. Using MRI-guided methods with convection-enhanced delivery, we were able to accurately and consistently target AAV2-BDNF delivery to the entorhinal cortex of non-human primates; 86 ± 3% of transduced cells in the targeted regions co-localized with the neuronal marker NeuN. The volume of AAV2-BDNF (3 × 108 vg/µl) infusion linearly correlated with the number of BDNF labeled cells and the volume (mm3) of BDNF immunoreactivity in the entorhinal cortex. BDNF is normally trafficked to the hippocampus from the entorhinal cortex; in these experiments, we also found that BDNF immunoreactivity was elevated in the hippocampus following therapeutic BDNF vector delivery to the entorhinal cortex, achieving growth factor distribution through key memory circuits. These findings indicate that MRI-guided infusion of AAV2-BDNF to the entorhinal cortex of the non-human primate results in safe and accurate targeting and distribution of BDNF to both the entorhinal cortex and the hippocampus. These methods are adaptable to human clinical trials.
- Published
- 2018
3. MR-guided parenchymal delivery of adeno-associated viral vector serotype 5 in non-human primate brain
- Author
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Samaranch, L, Blits, B, San Sebastian, W, Hadaczek, P, Bringas, J, Sudhakar, V, Macayan, M, Pivirotto, PJ, Petry, H, and Bankiewicz, KS
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Medical Biotechnology ,Biomedical and Clinical Sciences ,Neurosciences ,Genetics ,Gene Therapy ,Neurological ,Animals ,Brain ,Dependovirus ,Gene Transfer Techniques ,Genetic Therapy ,Genetic Vectors ,Green Fluorescent Proteins ,Humans ,Neurons ,Primates ,Putamen ,Spinal Cord ,Biological Sciences ,Medical and Health Sciences ,Biotechnology ,Biological sciences ,Biomedical and clinical sciences ,Health sciences - Abstract
The present study was designed to characterize transduction of non-human primate brain and spinal cord with AAV5 viral vector after parenchymal delivery. AAV5-CAG-GFP (1 × 1013 vector genomes per milliliter (vg ml-1)) was bilaterally infused either into putamen, thalamus or with the combination left putamen and right thalamus. Robust expression of GFP was seen throughout infusion sites and also in other distal nuclei. Interestingly, thalamic infusion of AAV5 resulted in the transduction of the entire corticospinal axis, indicating transport of AAV5 over long distances. Regardless of site of injection, AAV5 transduced both neurons and astrocytes equally. Our data demonstrate that AAV5 is a very powerful vector for the central nervous system and has potential for treatment of a wide range of neurological pathologies with cortical, subcortical and/or spinal cord affection.
- Published
- 2017
4. Cerebellomedullary Cistern Delivery for AAV-Based Gene Therapy: A Technical Note for Nonhuman Primates
- Author
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Samaranch, Lluis, Bringas, John, Pivirotto, Philip, San Sebastian, Waldy, Forsayeth, John, and Bankiewicz, Krystof
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Medical Biotechnology ,Biomedical and Clinical Sciences ,Genetics ,Neurosciences ,Biotechnology ,Gene Therapy ,Animals ,Central Nervous System ,Cisterna Magna ,Dependovirus ,Female ,Gene Transfer Techniques ,Genetic Therapy ,Genetic Vectors ,Humans ,Injections ,Epidural ,Male ,Primates - Abstract
Accessing cerebrospinal fluid (CSF) from the craniocervical junction through the posterior atlanto-occipital membrane via cerebellomedullary injection (also known as cisternal puncture or cisterna magna injection) has become a standard procedure in preclinical studies. Such delivery provides broader coverage to the central and peripheral nervous system unlike local parenchymal delivery alone. As a clinical application, this approach offers a more reliable method for neurological gene replacement delivery in infants, where skull-mounted devices are not indicated. Here we describe a consistent, precise, and safe method for CSF injection with minimal equipment and technical skills.
- Published
- 2016
5. Widespread AAV1- and AAV2-mediated transgene expression in the nonhuman primate brain: implications for Huntington's disease.
- Author
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Bankiewicz, Krzysztof, Hadaczek, P, Stanek, L, Ciesielska, A, Sudhakar, V, Samaranch, L, Pivirotto, P, Bringas, J, O'Riordan, C, Mastis, B, and San, W
- Abstract
Huntington's disease (HD) is caused by a toxic gain-of-function associated with the expression of the mutant huntingtin (htt) protein. Therefore, the use of RNA interference to inhibit Htt expression could represent a disease-modifying therapy. The potenti
- Published
- 2016
6. SAFETY AND TOLERABILITY OF MRI-GUIDED INFUSION OF AAV2-hAADC INTO THE MID-BRAIN OF NON-HUMAN PRIMATE.
- Author
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Samaranch, Lluis, Hadaczek, Piotr, Ciesielska, Agnieszka, Macayan, Michael, Pivirotto, Phillip, Forsayeth, John, Osborne, Sheryl, Wright, J, Green, Foad, Heller, Gregory, San Sebastian, Waldy, Kells, Adrian, Bankiewicz, Krzysztof, and Bringas, John
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AADC deficiency ,AAV2-hAADC ,MRI-guided ,pressurized infusion - Abstract
Aromatic L-amino acid decarboxylase (AADC) deficiency is a rare, autosomal-recessive neurological disorder caused by mutations in the DDC gene that leads to an inability to synthesize catecholamines and serotonin. As a result, patients suffer compromised development, particularly in motor function. A recent gene replacement clinical trial explored putaminal delivery of recombinant adeno-associated virus serotype 2 vector encoding human AADC (AAV2-hAADC) in AADC-deficient children. Unfortunately, patients presented only modest amelioration of motor symptoms, which authors acknowledged could be due to insufficient transduction of putamen. We hypothesize that, with the development of a highly accurate MRI-guided cannula placement technology, a more effective approach might be to target the affected mid-brain neurons directly. Transduction of AADC-deficient dopaminergic neurons in the substantia nigra and ventral tegmental area with locally infused AAV2-hAADC would be expected to lead to restoration of normal dopamine levels in affected children. The objective of this study was to assess the long-term safety and tolerability of bilateral AAV2-hAADC MRI-guided pressurized infusion into the mid-brain of non-human primates. Animals received either vehicle, low or high AAV2-hAADC vector dose and were euthanized 1, 3 or 9 months after surgery. Our data indicate that effective mid-brain transduction was achieved without untoward effects.
- Published
- 2014
7. SAFETY AND TOLERABILITY OF MRI-GUIDED INFUSION OF AAV2-hAADC INTO THE MID-BRAIN OF NON-HUMAN PRIMATE.
- Author
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San Sebastian, Waldy, Kells, Adrian P, Bringas, John, Samaranch, Lluis, Hadaczek, Piotr, Ciesielska, Agnieszka, Macayan, Michael, Pivirotto, Phillip J, Forsayeth, John, Osborne, Sheryl, Wright, J Fraser, Green, Foad, Heller, Gregory, and Bankiewicz, Krystof S
- Subjects
AADC deficiency ,AAV2-hAADC ,MRI-guided ,pressurized infusion - Abstract
Aromatic L-amino acid decarboxylase (AADC) deficiency is a rare, autosomal-recessive neurological disorder caused by mutations in the DDC gene that leads to an inability to synthesize catecholamines and serotonin. As a result, patients suffer compromised development, particularly in motor function. A recent gene replacement clinical trial explored putaminal delivery of recombinant adeno-associated virus serotype 2 vector encoding human AADC (AAV2-hAADC) in AADC-deficient children. Unfortunately, patients presented only modest amelioration of motor symptoms, which authors acknowledged could be due to insufficient transduction of putamen. We hypothesize that, with the development of a highly accurate MRI-guided cannula placement technology, a more effective approach might be to target the affected mid-brain neurons directly. Transduction of AADC-deficient dopaminergic neurons in the substantia nigra and ventral tegmental area with locally infused AAV2-hAADC would be expected to lead to restoration of normal dopamine levels in affected children. The objective of this study was to assess the long-term safety and tolerability of bilateral AAV2-hAADC MRI-guided pressurized infusion into the mid-brain of non-human primates. Animals received either vehicle, low or high AAV2-hAADC vector dose and were euthanized 1, 3 or 9 months after surgery. Our data indicate that effective mid-brain transduction was achieved without untoward effects.
- Published
- 2014
8. AAV9-mediated Expression of a Non-self Protein in Nonhuman Primate Central Nervous System Triggers Widespread Neuroinflammation Driven by Antigen-presenting Cell Transduction
- Author
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Samaranch, Lluis, San Sebastian, Waldy, Kells, Adrian P, Salegio, Ernesto A, Heller, Gregory, Bringas, John R, Pivirotto, Philip, DeArmond, Stephen, Forsayeth, John, and Bankiewicz, Krystof S
- Subjects
Biomedical and Clinical Sciences ,Immunology ,Neurosciences ,Neurological ,Animals ,Antigen-Presenting Cells ,Central Nervous System ,Corpus Striatum ,Dependovirus ,Gene Expression ,Genes ,Reporter ,Genetic Vectors ,Green Fluorescent Proteins ,Humans ,Inflammation ,Neurons ,Rats ,Transduction ,Genetic ,Transgenes ,Biological Sciences ,Technology ,Medical and Health Sciences ,Biotechnology ,Genetics ,Clinical sciences ,Medical biotechnology - Abstract
Many studies have demonstrated that adeno-associated virus serotype 9 (AAV9) transduces astrocytes and neurons when infused into rat or nonhuman primate (NHP) brain. We previously showed in rats that transduction of antigen-presenting cells (APC) by AAV9 encoding a foreign protein triggered a full neurotoxic immune response. Accordingly, we asked whether this phenomenon occurred in NHP. We performed parenchymal or intrathecal infusion of AAV9 encoding green fluorescent protein (GFP), a non-self protein derived from jellyfish, or human aromatic L-amino acid decarboxylase (hAADC), a self-protein, in separate NHP. Animals receiving AAV9-GFP into cisterna magna (CM) became ataxic, indicating cerebellar pathology, whereas AAV9-hAADC animals remained healthy. In transduced regions, AAV9-GFP elicited inflammation associated with early activation of astrocytic and microglial cells, along with upregulation of major histocompatibility complex class II (MHC-II) in glia. In addition, we found Purkinje neurons lacking calbindin after AAV9-GFP but not after AAV9-hAADC delivery. Our results demonstrate that AAV9-mediated expression of a foreign-protein, but not self-recognized protein, triggers complete immune responses in NHP regardless of the route of administration. Our results warrant caution when contemplating use of serotypes that can transduce APC if the transgene is not syngeneic with the host. This finding has the potential to complicate preclinical toxicology studies in which such vectors encoding human cDNA's are tested in animals.
- Published
- 2014
9. Dynamic hyperpolarized carbon‐13 MR metabolic imaging of nonhuman primate brain
- Author
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Park, Ilwoo, Larson, Peder EZ, Tropp, James L, Carvajal, Lucas, Reed, Galen, Bok, Robert, Robb, Fraser, Bringas, John, Kells, Adrian, Pivirotto, Philip, Bankiewicz, Krystof, Vigneron, Daniel B, and Nelson, Sarah J
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Engineering ,Biomedical Engineering ,Biomedical Imaging ,Neurosciences ,Animals ,Brain ,Carbon Isotopes ,Equipment Design ,Equipment Failure Analysis ,Feasibility Studies ,Female ,Humans ,Lactic Acid ,Macaca fascicularis ,Magnetic Resonance Imaging ,Magnetic Resonance Spectroscopy ,Pyruvic Acid ,Reproducibility of Results ,Sensitivity and Specificity ,Hyperpolarized carbon-13 magnetic resonance spectroscopic imaging ,dynamic nuclear polarization ,pyruvate ,primate brain ,Nuclear Medicine & Medical Imaging ,Biomedical engineering - Abstract
PurposeTo investigate hyperpolarized (13) C metabolic imaging methods in the primate brain that can be translated into future clinical trials for patients with brain cancer.Methods(13) C coils and pulse sequences designed for use in humans were tested in phantoms. Dynamic (13) C data were obtained from a healthy cynomolgus monkey brain using the optimized (13) C coils and pulse sequences. The metabolite kinetics were estimated from two-dimensional localized (13) C dynamic imaging data from the nonhuman primate brain.ResultsPyruvate and lactate signal were observed in both the brain and the surrounding tissues with the maximum signal-to-noise ratio of 218 and 29 for pyruvate and lactate, respectively. Apparent rate constants for the conversion of pyruvate to lactate and the ratio of lactate to pyruvate showed a difference between brain and surrounding tissues.ConclusionThe feasibility of using hyperpolarized [1-(13) C]-pyruvate for assessing in vivo metabolism in a healthy nonhuman primate brain was demonstrated using a hyperpolarized (13) C imaging experimental setup designed for studying patients with brain tumors. The kinetics of the metabolite conversion suggests that this approach may be useful in future studies of human neuropathology.
- Published
- 2014
10. Safety and tolerability of MRI-guided infusion of AAV2-hAADC into the mid-brain of nonhuman primate
- Author
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San Sebastian, Waldy, Kells, Adrian P, Bringas, John, Samaranch, Lluis, Hadaczek, Piotr, Ciesielska, Agnieszka, Macayan, Michael J, Pivirotto, Phillip J, Forsayeth, John, Osborne, Sheryl, Wright, J Fraser, Green, Foad, Heller, Gregory, Bankiewicz, Krystof S, and work, two authors contributed equally to this
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Medical Biotechnology ,Biomedical and Clinical Sciences ,Clinical Research ,Biomedical Imaging ,Gene Therapy ,Genetics ,Neurosciences ,Brain Disorders ,Neurological ,AADC deficiency ,AAV2-hAADC ,MRI-guided ,pressurized infusion ,Medical biotechnology - Abstract
Aromatic L-amino acid decarboxylase (AADC) deficiency is a rare, autosomal-recessive neurological disorder caused by mutations in the DDC gene that leads to an inability to synthesize catecholamines and serotonin. As a result, patients suffer compromised development, particularly in motor function. A recent gene replacement clinical trial explored putaminal delivery of recombinant adeno-associated virus serotype 2 vector encoding human AADC (AAV2-hAADC) in AADC-deficient children. Unfortunately, patients presented only modest amelioration of motor symptoms, which authors acknowledged could be due to insufficient transduction of putamen. We hypothesize that, with the development of a highly accurate MRI-guided cannula placement technology, a more effective approach might be to target the affected mid-brain neurons directly. Transduction of AADC-deficient dopaminergic neurons in the substantia nigra and ventral tegmental area with locally infused AAV2-hAADC would be expected to lead to restoration of normal dopamine levels in affected children. The objective of this study was to assess the long-term safety and tolerability of bilateral AAV2-hAADC MRI-guided pressurized infusion into the mid-brain of non-human primates. Animals received either vehicle, low or high AAV2-hAADC vector dose and were euthanized 1, 3 or 9 months after surgery. Our data indicate that effective mid-brain transduction was achieved without untoward effects.
- Published
- 2014
11. Adeno-associated virus type 6 is retrogradely transported in the non-human primate brain
- Author
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Bankiewicz, Krzysztof, San, W, Samaranch, L, Heller, G, Kells, AP, Bringas, J, Pivirotto, P, Forsayeth, J, and Bankiewicz, KS
- Abstract
We recently demonstrated that axonal transport of adeno-associated virus (AAV) is serotype-dependent. Thus, AAV serotype 2 (AAV2) is anterogradely transported (e.g., from cell bodies to nerve terminals) in both rat and non-human primate (NHP) brain. In con
- Published
- 2013
12. An estimator of first coalescent time reveals selection on young variants and large heterogeneity in rare allele ages among human populations.
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Alexander Platt, Alyssa Pivirotto, Jared Knoblauch, and Jody Hey
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Genetics ,QH426-470 - Abstract
Allele age has long been a focus of population genetic research, primarily because it can be an important clue to the fitness effects of an allele. By virtue of their effects on fitness, alleles under directional selection are expected to be younger than neutral alleles of the same frequency. We developed a new coalescent-based estimator of a close proxy for allele age, the time when a copy of an allele first shares common ancestry with other chromosomes in a sample not carrying that allele. The estimator performs well, including for the very rarest of alleles that occur just once in a sample, with a bias that is typically negative. The estimator is mostly insensitive to population demography and to factors that can arise in population genomic pipelines, including the statistical phasing of chromosomes. Applications to 1000 Genomes Data and UK10K genome data confirm predictions that singleton alleles that alter proteins are significantly younger than those that do not, with a greater difference in the larger UK10K dataset, as expected. The 1000 Genomes populations varied markedly in their distributions for singleton allele ages, suggesting that these distributions can be used to inform models of demographic history, including recent events that are only revealed by their impacts on the ages of very rare alleles.
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- 2019
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13. Slow AAV2 clearance from the brain of nonhuman primates and anti-capsid immune response
- Author
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Samaranch, L, Hadaczek, P, Kells, A P, Bringas, J R, Stockinger, D, San Sebastian, W, Macayan, M, Samineni, S, Pivirotto, P, Forsayeth, J, and Bankiewicz, K S
- Published
- 2016
- Full Text
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14. Widespread AAV1- and AAV2-mediated transgene expression in the nonhuman primate brain: implications for Huntington's disease
- Author
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Piotr Hadaczek, Lisa Stanek, Agnieszka Ciesielska, Vivek Sudhakar, Lluis Samaranch, Philip Pivirotto, John Bringas, Catherine O'Riordan, Bryan Mastis, Waldy San Sebastian, John Forsayeth, Seng H Cheng, Krystof S Bankiewicz, and Lamya S Shihabuddin
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Genetics ,QH426-470 ,Cytology ,QH573-671 - Abstract
Huntington's disease (HD) is caused by a toxic gain-of-function associated with the expression of the mutant huntingtin (htt) protein. Therefore, the use of RNA interference to inhibit Htt expression could represent a disease-modifying therapy. The potential of two recombinant adeno-associated viral vectors (AAV), AAV1 and AAV2, to transduce the cortico-striatal tissues that are predominantly affected in HD was explored. Green fluorescent protein was used as a reporter in each vector to show that both serotypes were broadly distributed in medium spiny neurons in the striatum and cortico-striatal neurons after infusion into the putamen and caudate nucleus of nonhuman primates (NHP), with AAV1-directed expression being slightly more robust than AAV2-driven expression. This study suggests that both serotypes are capable of targeting neurons that degenerate in HD, and it sets the stage for the advanced preclinical evaluation of an RNAi-based therapy for this disease.
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- 2016
- Full Text
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15. Adeno-associated virus type 6 is retrogradely transported in the non-human primate brain
- Author
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San Sebastian, W, Samaranch, L, Heller, G, Kells, A P, Bringas, J, Pivirotto, P, Forsayeth, J, and Bankiewicz, K S
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- 2013
- Full Text
- View/download PDF
16. Safety and tolerability of MRI-guided infusion of AAV2-hAADC into the mid-brain of nonhuman primate
- Author
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Waldy San Sebastian, Adrian P Kells, John Bringas, Lluis Samaranch, Piotr Hadaczek, Agnieszka Ciesielska, Michael J Macayan, Phillip J Pivirotto, John Forsayeth, Sheryl Osborne, J Fraser Wright, Foad Green, Gregory Heller, and Krystof S Bankiewicz
- Subjects
Genetics ,QH426-470 ,Cytology ,QH573-671 - Abstract
Aromatic L-amino acid decarboxylase (AADC) deficiency is a rare, autosomal-recessive neurological disorder caused by mutations in the DDC gene that leads to an inability to synthesize catecholamines and serotonin. As a result, patients suffer compromised development, particularly in motor function. A recent gene replacement clinical trial explored putaminal delivery of recombinant adeno-associated virus serotype 2 vector encoding human AADC (AAV2-hAADC) in AADC-deficient children. Unfortunately, patients presented only modest amelioration of motor symptoms, which authors acknowledged could be due to insufficient transduction of putamen. We hypothesize that, with the development of a highly accurate MRI-guided cannula placement technology, a more effective approach might be to target the affected mid-brain neurons directly. Transduction of AADC-deficient dopaminergic neurons in the substantia nigra and ventral tegmental area with locally infused AAV2-hAADC would be expected to lead to restoration of normal dopamine levels in affected children. The objective of this study was to assess the long-term safety and tolerability of bilateral AAV2-hAADC MRI-guided pressurized infusion into the mid-brain of nonhuman primates. Animals received either vehicle, low or high AAV2-hAADC vector dose and were euthanized 1, 3, or 9 months after surgery. Our data indicate that effective mid-brain transduction was achieved without untoward effects.
- Published
- 2014
- Full Text
- View/download PDF
17. Adeno-associated virus type 6 is retrogradely transported in the non-human primate brain
- Author
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Sebastian, San W, Samaranch, L, Heller, G, Kells, A P, Bringas, J, Pivirotto, P, Forsayeth, J, and Bankiewicz, K S
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- 2013
- Full Text
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18. Long-term clinical improvement in MPTP-lesioned primates after gene therapy with AAV-hAADC: O151
- Author
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Bankiewicz, K., Eberling, J. L., Avramut, M., Pivirotto, P., Bringas, J., and Forsayeth, J.
- Published
- 2006
19. Technique for Bilateral Intracranial Implantation of Cells in Monkeys Using an Automated Delivery System
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Krys S. Bankiewicz M.D., Ph.D., John Bringas, Phillip Pivirotto, Ethan Kutzscher, Dea Nagy, and Marina E. Emborg
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Medicine - Abstract
Intracerebral grafting combined with gene transfer may provide a powerful technique for local delivery of therapeutic agents into the CNS. The present study was undertaken to: (i) develop a reliable and reproducible automated cell implantation system, (ii) determine optimal implantation parameters of cells into the striatum, (iii) determine upper safe limits of cellular implantation into the neostriatum of monkeys. Autologous fibroblasts were infused into six sites of the striatum in nonhuman primates (Macaca mulatta, n = 11). Twenty-six-gauge cannulae were inserted vertically through cortical entry sites into the striatum (two sites in the caudate nucleus and four sites in the putamen) at predefined coordinates based on magnetic resonance imaging (MRI). The cannulae were guided by an electronically operated, hydraulic micropositioner and withdrawn at controlled rates, while cells (5, 10, 20, 40, or 80 μl/site) were infused simultaneously. Varying infusion rates and cell concentrations were also evaluated. Visualization and evaluation of graft placement were performed using contrast MRI at 3–5 days postsurgery. Animals were monitored for signs of clinical complications and sacrificed 2 weeks following surgery. Postimplantation MRI revealed a tissue mass effect of the implant with shifting of midline, edema, and infiltration of the white tracts at 40 and 80 μl/site. In addition, these animals developed transient hemiparesis contralateral to the implant site. MRI of animals grafted with 20 μl/site exhibited columnar-shaped implants and evidence of infiltration into white matter tracts possibly due to a volume effect. No clinical side effects were seen in this group. At 14 days postsurgery, MRI scans showed consistent columnar grafts (measuring approximately 5 mm in height) throughout the striatum in animals implanted with 5 or 10 μl/site. No signs of clinical side effects were associated with these volumes and postmortem histological examination confirmed MRI observations. Optimal surgical parameters for delivery of cells into the striatum consist of a graft volume of 10 μl/site, an infusion rate of 1.6 μl/min, a cell concentration of 2.0 × 10 5 cells/μl, and a cannula withdrawal rate of 0.75 mm/min. These results show that infusion of cells into the striatum can be done in a safe and routine manner.
- Published
- 2000
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20. Yohimbine co-treatment during chronic morphine administration attenuates naloxone-precipitated withdrawal without diminishing tail-flick analgesia in rats
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Taylor, J. R., Lewis, V. O., Elsworth, J. D., Pivirotto, P., Roth, R. H., and Redmond, Jr., D. E.
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- 1991
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21. Factors Associated With an Abnormal Blood Pressure Response During Exercise After Coarctation Repair
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Pivirotto, Mia, Swartz, Michael F., McGreevy, Megan B., Atallah-Yunes, Nader, Cholette, Jill M., Lipshultz, Steven E., and Alfieris, George M.
- Abstract
Background Although resting blood pressures following aortic arch repair or the extended end-to-end anastomosis (EEA) repair for coarctation can be physiologic, factors associated with an abnormal blood pressure response after exercise are unknown. We measured blood pressure gradients following exercise in children who had undergone previous repair in accordance with a surgical selection algorithm and sought to identify factors associated with an abnormal blood pressure response.Methods In accordance with our practice's surgical algorithm for repair of coarctation, infants were stratified to aortic arch repair when the distal transverse arch-to-left carotid artery ratio (DTA:LCA) ≤ 1.0, or when a brachiocephalic trunk or intra-cardiac lesion requiring repair was present. A thoracotomy and EEA were otherwise used. A follow-up exercise stress test (EST) measured the arm:leg blood pressure gradient after exercise, and a gradient ≥ 20 mm Hg was defined as an abnormal blood pressure response.Results Thirty-seven infants who had previously undergone coarctation repair (aortic arch repair-19, EEA-18) completed an EST at 12.3 ± 2.2 years of age. Thirteen (35%) children (aortic arch repair-5, EEA-8; p = .3) exhibited an abnormal blood pressure response. Factors associated with an abnormal blood pressure response included: smaller DTA:LCA ratios prior to repair (1.0 ± .2 vs. 1.2 ± .3; p = .04) and greater body weight at the time of EST (57.5 ± 19.1 vs. 40.9 ± 15.6 kg; p = .03).Conclusion An abnormal blood pressure response following exercise is associated with smaller DTA:LCA ratios at the time of repair and increased weight during follow-up suggesting that patients with these factors warrant close observation.
- Published
- 2022
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22. Application of Gene Therapy for Parkinson's Disease: Nonhuman Primate Experience
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Bankiewicz, K.S., primary, Bringas, J.R., additional, McLaughlin, W., additional, Pivirotto, P., additional, Hundal, R., additional, Yang, B., additional, Emborg, M.E., additional, and Nagy, D., additional
- Published
- 1997
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23. Slow AAV2 clearance from the brain of nonhuman primates and anti-capsid immune response
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Samaranch, L, primary, Hadaczek, P, additional, Kells, A P, additional, Bringas, J R, additional, Stockinger, D, additional, San Sebastian, W, additional, Macayan, M, additional, Samineni, S, additional, Pivirotto, P, additional, Forsayeth, J, additional, and Bankiewicz, K S, additional
- Published
- 2015
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24. Regeneration of the MPTP-Lesioned Dopaminergic System after Convection-Enhanced Delivery of AAV2-GDNF
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Kells, A. P., primary, Eberling, J., additional, Su, X., additional, Pivirotto, P., additional, Bringas, J., additional, Hadaczek, P., additional, Narrow, W. C., additional, Bowers, W. J., additional, Federoff, H. J., additional, Forsayeth, J., additional, and Bankiewicz, K. S., additional
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- 2010
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25. In vivo PET imaging of gene expression in parkinsonian monkeys
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Eberling, J.L., primary, Cunningham, J., additional, Pivirotto, P., additional, Bringas, J., additional, Daadi, M.M., additional, and Bankiewicz, K.S., additional
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- 2003
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26. Grafting Genetically Engineered Cells into the Striatum of Nonhuman Primates.
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Walker, John M., Maral Mouradian, M., Bankiewicz, Krys S., Pivirotto, Phillip, Sanchez-Pernaute, Rosario, and Major, Eugene O.
- Abstract
An emerging new technology based on genetic engineering of viral vectors that can insert genes into the cells of living organisms may play a significant role in treating disorders of the central nervous system (CNS). Most neurodegenerative disorders affect focal regions of the brain. Preventive and/ or palliative treatment strategies need to be targeted only to the diseased parts of the brain without affecting other regions. Administration of therapeutic genes specifically to the disease-affected regions of the brain may be more beneficial than current treatment strategies, which are largely based on systemically administering small molecules. The latter can result not only in peripheral side effects but also CNS side effects since the drugs can affect both targeted and nontargeted brain sites. In addition, many therapeutic agents are prevented from entering the brain by the blood-brain barrier (BBB). For these reasons, many otherwise potentially useful proteins, such as trophic factors, cannot be administered systemically (1) [ABSTRACT FROM AUTHOR]
- Published
- 2001
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27. Tremor Is Associated with PET Measures of Nigrostriatal Dopamine Function in MPTP-Lesioned Monkeys
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Eberling, J.L., primary, Pivirotto, P., additional, Bringas, J., additional, and Bankiewicz, K.S., additional
- Published
- 2000
- Full Text
- View/download PDF
28. Convection-Enhanced Delivery of Adeno-Associated Virus Type 2 (AAV2) into the Striatum and Transport of AAV2 Within Monkey Brain
- Author
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Hadaczek, Piotr, Kohutnicka, Malgorzata, Krauze, Michal T., Bringas, John, Pivirotto, Phil, Cunningham, Janet, and Bankiewicz, Krystof
- Abstract
Adeno-associated virus type 2 (AAV2)-based vectors are promising transgene carriers for experimental gene therapy treatments of brain diseases. However, detailed evaluation of transgene distribution, trafficking, and transport within the brain is of the utmost importance before applying any type of gene therapy in humans. We examined the distribution of AAV2-thymidine kinase (AAV2-TK) and AAV2-aromatic L-amino acid decarboxylase (AAV2-AADC) in monkey brain after convection-enhanced delivery (CED). The AADC group consisted of two 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP)-lesioned monkeys that received unilateral infusions of AAV2-AADC into six sites in the right hemisphere. The TK group consisted of three monkeys that received bilateral CED infusion of AAV2-TK into the putamen; one side in all three monkeys was coinfused with heparin. Six weeks after AAV delivery, the brains were collected and processed for immunohistochemical staining. Volumetric measurement of TK distribution showed that at least 75% of the putamen could be covered by a single infusion of the vector; however, no effects of heparin coadministration were found, most likely because of the already robust gene transfer achieved by CED. Interestingly, TK- and AADCimmunoreactive cells were also present outside the striatum, in the globus pallidus, subthalamic nucleus, thalamus, and substantia nigra. CED proved to be an efficient method for delivery of the AAV2 vector. Detection of the transgenes in brain structures distant from the site of injection emphasizes the potential for gene transport, and the advantages and disadvantages of CED for gene therapy deserve further study.
- Published
- 2006
- Full Text
- View/download PDF
29. Distribution of AAV2-hAADC-transduced cells after 3 years in Parkinsonian monkeys
- Author
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Daadi, Marcel M., Pivirotto, Phillip, Bringas, John, Cunningham, Janet, Forsayeth, John, Eberling, Jamie, and Bankiewicz, Krys S.
- Abstract
The present report describes for the first time, the stability of recombinant adeno-associated virus serotype 2 (AAV2) human aromatic L-amino acid decarboxylase (hAADC) gene transfer after 3-year survival time in a non-human primate model of Parkinson's disease. 1-Methyl-4-phenyl-1,2,3,6-tetrahydropyridine-lesioned monkeys were treated with six injections of 30 μlsite of AAV2-hAADC at a concentration of 2×1012vgml into the caudate and putamen. Stereological analysis revealed a 46.6 increase in the total number of AAV2-hAADC-transduced cells in the striatum between 8 weeks and 3 years after gene transfer survival time. In the 8-week animals, the distribution of the AADC cells was dispersed and heterogeneous, whereas in the 3-year animals it was widespread and homogenous. Confocal analysis demonstrated that approximately 85 of the AADC cells were neuronal nuclei immunoreactive.
- Published
- 2006
30. Technique for Bilateral Intracranial Implantation of Cells in Monkeys Using an Automated Delivery System
- Author
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Bankiewicz, Krys S., Bringas, John, Pivirotto, Phillip, Kutzscher, Ethan, Nagy, Dea, and Emborg, Marina E.
- Abstract
Intracerebral grafting combined with gene transfer may provide a powerful technique for local delivery of therapeutic agents into the CNS. The present study was undertaken to: (i) develop a reliable and reproducible automated cell implantation system, (ii) determine optimal implantation parameters of cells into the striatum, (iii) determine upper safe limits of cellular implantation into the neostriatum of monkeys. Autologous fibroblasts were infused into six sites of the striatum in nonhuman primates (Macaca mulatta, n = 11). Twenty-six-gauge cannulae were inserted vertically through cortical entry sites into the striatum (two sites in the caudate nucleus and four sites in the putamen) at predefined coordinates based on magnetic resonance imaging (MRI). The cannulae were guided by an electronically operated, hydraulic micropositioner and withdrawn at controlled rates, while cells (5, 10, 20, 40, or 80 μl/site) were infused simultaneously. Varying infusion rates and cell concentrations were also evaluated. Visualization and evaluation of graft placement were performed using contrast MRI at 3–5 days postsurgery. Animals were monitored for signs of clinical complications and sacrificed 2 weeks following surgery. Postimplantation MRI revealed a tissue mass effect of the implant with shifting of midline, edema, and infiltration of the white tracts at 40 and 80 μl/site. In addition, these animals developed transient hemiparesis contralateral to the implant site. MRI of animals grafted with 20 μl/site exhibited columnar-shaped implants and evidence of infiltration into white matter tracts possibly due to a volume effect. No clinical side effects were seen in this group. At 14 days postsurgery, MRI scans showed consistent columnar grafts (measuring approximately 5 mm in height) throughout the striatum in animals implanted with 5 or 10 μl/site. No signs of clinical side effects were associated with these volumes and postmortem histological examination confirmed MRI observations. Optimal surgical parameters for delivery of cells into the striatum consist of a graft volume of 10 μl/site, an infusion rate of 1.6 μl/min, a cell concentration of 2.0 × 105cells/μl, and a cannula withdrawal rate of 0.75 mm/min. These results show that infusion of cells into the striatum can be done in a safe and routine manner.
- Published
- 2000
- Full Text
- View/download PDF
31. Convection-Enhanced Delivery of AAV Vector in Parkinsonian Monkeys; In VivoDetection of Gene Expression and Restoration of Dopaminergic Function Using Pro-drug Approach
- Author
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Bankiewicz, Krys S., Eberling, Jamie L., Kohutnicka, Malgorzata, Jagust, William, Pivirotto, Phillip, Bringas, John, Cunningham, Janet, Budinger, Thomas F., and Harvey-White, Judith
- Abstract
Using an approach that combines gene therapy with aromatic l-amino acid decarboxylase (AADC) gene and a pro-drug (l-dopa), dopamine, the neurotransmitter involved in Parkinson's disease, can be synthesized and regulated. Striatal neurons infected with the AADC gene by an adeno-associated viral vector can convert peripheral l-dopa to dopamine and may therefore provide a buffer for unmetabolized l-dopa. This approach to treating Parkinson's disease may reduce the need for l-dopa/carbidopa, thus providing a better clinical response with fewer side effects. In addition, the imbalance in dopamine production between the nigrostriatal and mesolimbic dopaminergic systems can be corrected by using AADC gene delivery to the striatum. We have also demonstrated that a fundamental obstacle in the gene therapy approach to the central nervous system, i.e., the ability to deliver viral vectors in sufficient quantities to the whole brain, can be overcome by using convection-enhanced delivery. Finally, this study demonstrates that positron emission tomography and the AADC tracer, 6-[18F]fluoro-l-m-tyrosine, can be used to monitor gene therapy in vivo.Our therapeutic approach has the potential to restore dopamine production, even late in the disease process, at levels that can be maintained during continued nigrostriatal degeneration.
- Published
- 2000
- Full Text
- View/download PDF
32. Dopamine transporter loss and clinical changes in MPTP-lesioned primates
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Eberling, Jamie L, Bankiewicz, Krzysztof S, Pivirotto, Phillip, Bringas, John, Chen, Karen, Nowotnik, David P, Steiner, Joseph P, Budinger, Thomas F, and Jagust, William J
- Abstract
Single photon emission computed tomography (SPECT) and the dopamine (DA) transporter tracer, 2 beta-carboxymethoxy-3 beta-(4-iodophenyl)tropane ([123I]β-CIT), were used to determine DA transporter density in 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP)-lesioned monkeys with varying degrees of parkinsonism. The clinical stage of parkinsonism corresponded to SPECT measures of striatal DA transporter density suggesting that more severe parkinsonism was associated with a greater degree of dopaminergic terminal degeneration. These findings are similar to those reported earlier using positron emission tomography (PET) and the DA metabolism tracer, 6-[18F]fluoro-l-m-tyrosine (FMT), indicating that both are good methods for evaluating nigrostriatal degeneration in MPTP primate models.
- Published
- 1999
- Full Text
- View/download PDF
33. Clozapine Preferentially Increases Dopamine Release in the Rhesus Monkey Prefrontal Cortex Compared with the Caudate Nucleus
- Author
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Youngren, Kenneth D, Inglis, Fiona M, Pivirotto, Philip J, Jedema, Hank P, Bradberry, Charles W, Goldman-Rakic, Patricia S, Roth, Robert H, and Moghaddam, Bita
- Abstract
Despite substantial differences between species in the organization and elaboration of the cortical dopamine innervation, little is known about the pharmacological response of cortical or striatal sites to antipsychotic medications in nonhuman primates. To examine this issue, rhesus monkeys were chronically implanted with guide cannulae directed at the principal sulcus, medial prefrontal cortex, premotor cortex, and caudate nucleus. Alterations in dopamine release in these discrete brain regions were measured in response to administration of clozapine or haloperidol. Clozapine produced significant and long-lasting increases in dopamine release in the principal sulcus, and to a lesser extent, in the caudate nucleus. Haloperidol did not produce a consistent effect on dopamine release in the principal sulcus, although it increased dopamine release in the caudate. Clozapine’s preferential augmentation of dopamine release in the dorsolateral prefrontal cortex supports the idea that clozapine exerts its therapeutic effects in part by increasing cortical dopamine neurotransmission.
- Published
- 1999
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34. Engraving.
- Author
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Pivirotto, Kalicia
- Subjects
- ENGRAVING (Poem), PIVIROTTO, Kalicia
- Abstract
The article presents the poem "Engraving," by Kalicia Pivirotto. First Line: Mouth, then a frame; Last Line: here the gondolier lost the water.
- Published
- 2006
35. A novel MPTP primate model of Parkinson's disease: neurochemical and clinical changes
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Eberling, J. L., Jagust, W. J., Taylor, S., Bringas, J., Pivirotto, P., VanBrocklin, H. F., and Bankiewicz, K. S.
- Published
- 1998
- Full Text
- View/download PDF
36. Supersonic electrical‐discharge copper vapor laser
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Russell, G.R., Nerheim, N.M., and Pivirotto, T.J.
- Published
- 1972
- Full Text
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37. MRI-guided targeting of entorhinal cortex for Alzheimer's disease.
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Nagahara, Alan Hiroshi, Wilson, Bayard R., Kovacs, Imre, Ivasyk, Iryna, Kells, Adrian P., Bringas, John R., Pivirotto, Phil, Sebastian, Waldy S., Bankiewicz, Krystof, and Tuszynski, Mark H.
- Published
- 2014
- Full Text
- View/download PDF
38. Untitled.
- Author
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Pivirotto, Kalicia
- Subjects
- PIVIROTTO, Kalicia
- Abstract
The article presents an untitled poem by Kalicia Pivirotto. First Line: So the bed began without sleep, folded in; Last Line: did the world seem a place.
- Published
- 2006
39. air/air.
- Author
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Pivirotto, Kalicia
- Subjects
- AIR/AIR (Poem), PIVIROTTO, Kalicia
- Abstract
Presents the poem "Air/Air," by Kalicia Pivirotto. First Line: Inside the silver bones; Last Line: unaware of the sky at its back.
- Published
- 2004
40. Cash-value life insurance expands retirement options.
- Author
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Pivirotto, John
- Abstract
Describes several cash value life insurance methods. Split-dollar; Reverse split-dollar; Salary continuation.
- Published
- 1996
41. Status of robotic mission studies for the Space Exploration Initiative-1991
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Bourke, R. D., Dias, W. C., Golombek, M. P., and Pivirotto, D. L.
- Published
- 1993
- Full Text
- View/download PDF
42. Sex hormones mediate interleukin-1 production by human osteoblastic HOBIT cells
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Pivirotto, L. A., Cissel, D. S., and Keeting, P. E.
- Published
- 1995
- Full Text
- View/download PDF
43. Creative life insurance: It's about death and taxes.
- Author
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Pivirotto, John
- Abstract
Focuses on company funded life-insurance as a means of providing tax-deferred benefits to owners and key managers of closely held companies in the United States. Split-dollar plan and reverse split dollar plan; Salary continuation.
- Published
- 1996
44. Widespread AAV1- and AAV2-mediated transgene expression in the nonhuman primate brain: implications for Huntington's disease.
- Author
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Hadaczek P, Stanek L, Ciesielska A, Sudhakar V, Samaranch L, Pivirotto P, Bringas J, O'Riordan C, Mastis B, San Sebastian W, Forsayeth J, Cheng SH, Bankiewicz KS, and Shihabuddin LS
- Abstract
Huntington's disease (HD) is caused by a toxic gain-of-function associated with the expression of the mutant huntingtin (htt) protein. Therefore, the use of RNA interference to inhibit Htt expression could represent a disease-modifying therapy. The potential of two recombinant adeno-associated viral vectors (AAV), AAV1 and AAV2, to transduce the cortico-striatal tissues that are predominantly affected in HD was explored. Green fluorescent protein was used as a reporter in each vector to show that both serotypes were broadly distributed in medium spiny neurons in the striatum and cortico-striatal neurons after infusion into the putamen and caudate nucleus of nonhuman primates (NHP), with AAV1-directed expression being slightly more robust than AAV2-driven expression. This study suggests that both serotypes are capable of targeting neurons that degenerate in HD, and it sets the stage for the advanced preclinical evaluation of an RNAi-based therapy for this disease.
- Published
- 2016
- Full Text
- View/download PDF
45. Safety and tolerability of magnetic resonance imaging-guided convection-enhanced delivery of AAV2-hAADC with a novel delivery platform in nonhuman primate striatum.
- Author
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San Sebastian W, Richardson RM, Kells AP, Lamarre C, Bringas J, Pivirotto P, Salegio EA, Dearmond SJ, Forsayeth J, and Bankiewicz KS
- Subjects
- Animals, Catheterization, Caudate Nucleus enzymology, Dopa Decarboxylase metabolism, Female, Humans, Macaca mulatta, Magnetic Resonance Imaging, Neurons enzymology, Neurons pathology, Putamen enzymology, Putamen pathology, Stereotaxic Techniques, Transgenes, Corpus Striatum enzymology, Dependovirus genetics, Dopa Decarboxylase genetics, Gene Transfer Techniques
- Abstract
Degeneration of nigrostriatal neurons in Parkinson's disease (PD) causes progressive loss of aromatic l-amino acid decarboxylase (AADC), the enzyme that converts levodopa (l-DOPA) into dopamine in the striatum. Because loss of this enzyme appears to be a major driver of progressive impairment of response to the mainstay drug, l-DOPA, one promising approach has been to use gene therapy to restore AADC activity in the human putamen and thereby restore normal l-DOPA response in patients with PD. An open-label phase I clinical trial of this approach in patients with PD provided encouraging signs of improvement in Unified Parkinson's Disease Rating Scale scores and reductions in antiparkinsonian medications. However, such improvement was modest compared with the results previously reported in parkinsonian rhesus macaques. The reason for this discrepancy may have been that the relatively small volume of vector infused in the clinical study restricted the distribution of AADC expression, such that only about 20% of the postcommissural putamen was covered, as revealed by l-[3-(18)F]-α-methyltyrosine-positron emission tomography. To achieve more quantitative distribution of vector, we have developed a visual guidance system for parenchymal infusion of AAV2. The purpose of the present study was to evaluate the combined magnetic resonance imaging-guided delivery system with AAV2-hAADC under conditions that approximate the intended clinical protocol. Our data indicate that this approach directed accurate cannula placement and effective vector distribution without inducing any untoward effects in nonhuman primates infused with a high dose of AAV2-hAADC.
- Published
- 2012
- Full Text
- View/download PDF
46. Eight years of clinical improvement in MPTP-lesioned primates after gene therapy with AAV2-hAADC.
- Author
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Hadaczek P, Eberling JL, Pivirotto P, Bringas J, Forsayeth J, and Bankiewicz KS
- Subjects
- Animals, Aromatic-L-Amino-Acid Decarboxylases genetics, Brain metabolism, Corpus Striatum metabolism, Dopamine metabolism, Immunohistochemistry, Levodopa metabolism, Male, Microscopy, Fluorescence, Aromatic-L-Amino-Acid Decarboxylases metabolism, Dependovirus genetics, Macaca mulatta metabolism
- Abstract
This study completes the longest known in vivo monitoring of adeno-associated virus (AAV)-mediated gene expression in nonhuman primate (NHP) brain. Although six of the eight parkinsonian NHP originally on study have undergone postmortem analysis, as described previously, we monitored the remaining two animals for a total of 8 years. In this study, NHP received AAV2-human L-amino acid decarboxylase (hAADC) infusions into the MPTP (1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine)-lesioned putamen. Restoration of AADC activity restored normal response to levodopa and gene expression could be quantitated repeatedly over many years by 6-[(18)F]fluoro-meta-tyrosine (FMT)-positron emission tomography (PET) and confirm that AADC transgene expression remained unchanged at the 8-year point. Behavioral assessments confirmed continued, normalized response to levodopa (improvement by 35% over historical controls). Postmortem analysis showed that, although only 5.6 + or - 1% and 6.6 + or - 1% of neurons within the transduced volumes of the striatum were transduced, this still secured robust clinical improvement. Importantly, there were no signs of neuroinflammation or reactive gliosis at the 8-year point, indicative of the safety of this treatment. The present data suggest that the improvement in the L-3,4-dihydroxyphenylalanine (L-Dopa) therapeutic window brought about by AADC gene therapy is pronounced and persistent for many years.
- Published
- 2010
- Full Text
- View/download PDF
47. Real-time MR imaging with Gadoteridol predicts distribution of transgenes after convection-enhanced delivery of AAV2 vectors.
- Author
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Su X, Kells AP, Salegio EA, Salegio EA, Richardson RM, Hadaczek P, Beyer J, Bringas J, Pivirotto P, Forsayeth J, and Bankiewicz KS
- Subjects
- Animals, Aromatic-L-Amino-Acid Decarboxylases genetics, Aromatic-L-Amino-Acid Decarboxylases metabolism, Gadolinium, Genetic Vectors genetics, Humans, Macaca mulatta, Adenoviridae genetics, Contrast Media, Heterocyclic Compounds, Magnetic Resonance Imaging methods, Organometallic Compounds
- Abstract
Gene therapies that utilize convention-enhanced delivery (CED) will require close monitoring of vector infusion in real time and accurate prediction of drug distribution. The magnetic resonance imaging (MRI) contrast agent, Gadoteridol (Gd), was used to monitor CED infusion and to predict the expression pattern of glial cell line-derived neurotrophic factor (GDNF) protein after administration of adeno-associated virus type 2 (AAV2) vector encoding human pre-pro-GDNF complementary DNA. The nonhuman primate (NHP) thalamus was utilized for modeling infusion to allow delivery of volumes more relevant to planned human studies. AAV2 encoding human aromatic L-amino acid decarboxylase (AADC) was coinfused with AAV2-GDNF/Gd to confirm regions of AAV2 transduction versus extracellular GDNF diffusion. There was a close correlation between Gd distribution and GDNF or AADC expression, and the ratios of expression areas of GDNF or AADC versus Gd were both close to 1. Our data support the use of Gd and MRI to monitor AAV2 infusion via CED and to predict the distribution of GDNF protein after AAV2-GDNF administration.
- Published
- 2010
- Full Text
- View/download PDF
48. Safety evaluation of AAV2-GDNF gene transfer into the dopaminergic nigrostriatal pathway in aged and parkinsonian rhesus monkeys.
- Author
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Su X, Kells AP, Huang EJ, Lee HS, Hadaczek P, Beyer J, Bringas J, Pivirotto P, Penticuff J, Eberling J, Federoff HJ, Forsayeth J, and Bankiewicz KS
- Subjects
- 1-Methyl-4-phenyl-1,2,3,6-tetrahydropyridine pharmacology, Adenoviridae genetics, Adenoviridae immunology, Age Factors, Animals, Cell Line, Disease Models, Animal, Dopamine metabolism, Dopamine Agents pharmacology, Glial Cell Line-Derived Neurotrophic Factor metabolism, Humans, Macaca mulatta, Parkinson Disease pathology, Parkinson Disease, Secondary pathology, Gene Transfer Techniques adverse effects, Genetic Therapy, Glial Cell Line-Derived Neurotrophic Factor genetics, Parkinson Disease therapy, Parkinson Disease, Secondary therapy, Substantia Nigra metabolism
- Abstract
We evaluated neuropathological findings in two studies of AAV2-GDNF efficacy and safety in naive aged (>20 years) or MPTP (1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine)-lesioned rhesus macaques. In the first study, a total of 17 animals received one of two doses of AAV2-GDNF into either putamen or substantia nigra (SN). To control for surgical variables, all animals received identical putaminal and nigral infusions in which phosphate-buffered saline was substituted for vector as appropriate. All 17 aged monkeys were studied for 6 months before necropsy. In a separate study, 11 MPTP-lesioned rhesus macaques with extensive lesions in the right SN and mild lesions in the left SN received bilateral infusions of AAV2-GDNF (9.9 x 10(11) vector genomes) or PBS into the putamen and were then studied for up to 14 months. In the current analysis, we addressed safety issues regarding AAV2-GDNF administration. An extensive series of assessments of in-life behavioral and clinical parameters was conducted. No overt histopathology or immune responses were detected in any experimental monkey. However, the delivery of AAV2-GDNF to the SN of aged monkeys caused a marked and significant loss of body weight (-19.4%). No weight loss was observed in the MPTP-lesioned monkeys despite bilateral axonal transport of glial cell line-derived neurotrophic factor (GDNF) to the SN from the putamen. These findings indicate that putaminal administration of AAV2-GDNF by convection-enhanced delivery shows therapeutic promise without any apparent side effects. Importantly, nigral administration of AAV2-GDNF caused significant weight loss that raises substantial concern for clinical application of this approach.
- Published
- 2009
- Full Text
- View/download PDF
49. Real-time MR imaging of adeno-associated viral vector delivery to the primate brain.
- Author
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Fiandaca MS, Varenika V, Eberling J, McKnight T, Bringas J, Pivirotto P, Beyer J, Hadaczek P, Bowers W, Park J, Federoff H, Forsayeth J, and Bankiewicz KS
- Subjects
- Animals, Aromatic-L-Amino-Acid Decarboxylases genetics, Aromatic-L-Amino-Acid Decarboxylases metabolism, Brain physiology, Gadolinium, Green Fluorescent Proteins genetics, Green Fluorescent Proteins metabolism, Humans, Immunohistochemistry, Liposomes, Macaca mulatta, Magnetic Resonance Imaging, Male, Positron-Emission Tomography, Putamen virology, Thalamus virology, Brain diagnostic imaging, Brain virology, Dependovirus genetics, Gene Transfer Techniques, Genetic Vectors
- Abstract
We are developing a method for real-time magnetic resonance imaging (MRI) visualization of convection-enhanced delivery (CED) of adeno-associated viral vectors (AAV) to the primate brain. By including gadolinium-loaded liposomes (GDL) with AAV, we can track the convective movement of viral particles by continuous monitoring of distribution of surrogate GDL. In order to validate this approach, we infused two AAV (AAV1-GFP and AAV2-hAADC) into three different regions of non-human primate brain (corona radiata, putamen, and thalamus). The procedure was tolerated well by all three animals in the study. The distribution of GFP determined by immunohistochemistry in both brain regions correlated closely with distribution of GDL determined by MRI. Co-distribution was weaker with AAV2-hAADC, although in vivo PET scanning with FMT for AADC activity correlated well with immunohistochemistry of AADC. Although this is a relatively small study, it appears that AAV1 correlates better with MRI-monitored delivery than does AAV2. It seems likely that the difference in distribution may be due to differences in tissue specificity of the two serotypes.
- Published
- 2009
- Full Text
- View/download PDF
50. Functional effects of AAV2-GDNF on the dopaminergic nigrostriatal pathway in parkinsonian rhesus monkeys.
- Author
-
Eberling JL, Kells AP, Pivirotto P, Beyer J, Bringas J, Federoff HJ, Forsayeth J, and Bankiewicz KS
- Subjects
- 1-Methyl-4-phenyl-1,2,3,6-tetrahydropyridine pharmacology, Animals, Disease Models, Animal, Female, Humans, Macaca mulatta, Male, Parkinson Disease pathology, Substantia Nigra pathology, Tyrosine 3-Monooxygenase metabolism, Genetic Therapy, Genetic Vectors metabolism, Glial Cell Line-Derived Neurotrophic Factor genetics, Parkinson Disease metabolism, Parkinson Disease therapy, Substantia Nigra metabolism
- Abstract
We investigated the safety and neuroregenerative potential of an adeno-associated virus (AAV2) containing human glial cell line-derived neurotrophic factor (GDNF) in an MPTP primate model of Parkinson's disease. Dopaminergic function was evaluated by positron emission tomography with 6-[(18)F]fluoro-l-m-tyrosine (FMT) before and after AAV2-GDNF or phosphate-buffered saline infusion bilaterally into the putamen. FMT uptake was significantly increased bilaterally in the putamen of AAV2-GDNF but not phosphate-buffered saline-treated animals 6 months after infusion, indicating increased dopaminergic activity in the nigrostriatal pathways. AAV2-GDNF-treated animals also showed clinical improvement without adverse effects. These findings are consistent with our previous report in aged nonhuman primates that showed evidence of enhanced use of striatal dopamine and dopaminergic nigrostriatal innervation. Clinical improvement and evidence of functional recovery in the nigrostriatal pathway, and the absence of adverse effects, support the safety of this approach for the delivery of GDNF over a 6-month period.
- Published
- 2009
- Full Text
- View/download PDF
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