المؤلفون: Nicola Brunetti-Pierri, Alberto Auricchio, Rita Ferla, Virginia Maria Ginocchio

المساهمون: Ginocchio, V. M., Ferla, R., Auricchio, A., Brunetti-Pierri, N.

المصدر: Human gene therapy. 30(10)

مصطلحات موضوعية: Genetic enhancement, Genetic Vectors, inborn errors of metabolism, Gene delivery, Bioinformatics, medicine.disease_cause, Virus, 03 medical and health sciences, Liver disease, adeno-associated virus vector, 0302 clinical medicine, Genome editing, Genetics, Medicine, Animals, Humans, Molecular Biology, Adeno-associated virus, 030304 developmental biology, Gene Editing, 0303 health sciences, Newborn screening, Clinical Trials as Topic, business.industry, Liver Diseases, Lentivirus, Gene Transfer Techniques, AAV, Genetic Therapy, Dependovirus, medicine.disease, gene therapy, Clinical trial, Liver, 030220 oncology & carcinogenesis, Hepatocytes, Molecular Medicine, business, Metabolism, Inborn Errors

الوصف: Inborn errors of metabolism (IEM) are disorders affecting human biochemical pathways and represent attractive targets for gene therapy because of their severity, high overall prevalence, lack of effective treatments, and possibility of early diagnosis through newborn screening. The liver is a central organ involved in several metabolic reactions and is a favorite target for gene therapy in many IEM. Adeno-associated virus (AAV) vectors have emerged in the last years as the preferred vectors for in vivo gene delivery. Gene replacement strategies are aimed either at correcting liver disease or providing a source for production and secretion of the lacking enzyme for cross-correction of other tissues. A number of preclinical studies have been conducted in the last years and, for several diseases, gene therapy has reached the clinical stage, with a growing number of ongoing clinical trials. Moreover, recent applications of genome editing to the field of inherited metabolic diseases have further expanded potential therapeutic possibilities. This review describes relevant clinical gene therapy studies for IEM with particular attention to current obstacles and drawbacks.

الوصول الحر: https://explore.openaire.eu/search/publication?articleId=doi_dedup___::a0c32744e13353462f9455aeff7facd9Test
https://pubmed.ncbi.nlm.nih.gov/31517544Test

المؤلفون: Pasquale Piccolo, Nicola Brunetti-Pierri

المساهمون: Piccolo, P, BRUNETTI PIERRI, Nicola

المصدر: Human Gene Therapy. 26:186-192

مصطلحات موضوعية: Genetics, Clinical Trials as Topic, Liver Diseases, Genetic enhancement, Genetic Vectors, Genetic Therapy, Biology, Bioinformatics, Genetic therapy, Liver metabolism, Metabolic Diseases, Animals, Humans, Molecular Medicine, Molecular Biology

الوصف: Gene therapy is entering the stage of initial clinical development to treat a growing number of inherited metabolic diseases. This review outlines the development of liver-directed gene therapy for diseases caused by deficiencies of enzymes that are primarily expressed in the liver and discusses the disorders that appear most promising for clinical translation.

الوصول الحر: https://explore.openaire.eu/search/publication?articleId=doi_dedup___::cdd4c2c4d7fd71d1e26e635209e58edcTest
https://doi.org/10.1089/hum.2015.029Test

المؤلفون: Nathan C Grove, Pasquale Piccolo, Nicola Brunetti-Pierri, Donna Palmer, Scott V. Dindot

المساهمون: Dindot, S, Piccolo, P, Grove, N, Palmer, D, BRUNETTI PIERRI, Nicola

المصدر: Human Gene Therapy. 22:745-751

مصطلحات موضوعية: Male, Ependymal Cell, Transgene, Genetic enhancement, Genetic Vectors, Central nervous system, Neuroepithelial Cells, Gene Expression, Biology, Adenoviridae, Mice, Transduction (genetics), Cerebrospinal fluid, Transduction, Genetic, Ependyma, Genetics, medicine, Animals, Secretion, Transgenes, Molecular Biology, Injections, Spinal, Neurons, Brief Report, Genetic Therapy, Virology, Cell biology, Mice, Inbred C57BL, Neuroepithelial cell, medicine.anatomical_structure, Molecular Medicine, Helper Viruses

الوصف: Helper-dependent adenoviral (HDAd) vectors are devoid of all viral genes and result in long-term transgene expression in the absence of chronic toxicity. Because of their ability to infect post-mitotic cells, including cells of the central nervous system, HDAd vectors are particularly attractive for brain-directed gene therapy. In this study, we show that intrathecal injection of HDAd results in extensive transduction of ependymal cells and sustained expression of the transgene up to 1 year post-administration. We also demonstrate, for the first time, the ability of HDAd injected by this route of delivery to transduce neuronal cells. The transduced neuroepithelial cells can be potentially used to secrete therapeutic proteins into the cerebrospinal fluid and provide them via cross-correction to nontransduced cells. Targeting of neuronal cells and long-term transgene expression make this approach attractive for the treatment of several neurologic diseases.

الوصول الحر: https://explore.openaire.eu/search/publication?articleId=doi_dedup___::77a80a42cf7ee62a236c807fc03874efTest
https://doi.org/10.1089/hum.2010.147Test

المؤلفون: Donna Palmer, David Dimmock, Nicola Brunetti-Pierri, Arthur L. Beaudet, Philip Ng

المساهمون: Dimmock, D, BRUNETTI PIERRI, Nicola, Palmer, Dj, Beaudet, Al, Ng, P.

المصدر: Human Gene Therapy. 22:483-488

مصطلحات موضوعية: Crigler–Najjar syndrome, Bilirubin, Genetic enhancement, Genetic Vectors, Rats, Gunn, Pharmacology, Biology, medicine.disease_cause, digestive system, Adenoviridae, chemistry.chemical_compound, Transduction, Genetic, Gene Order, Genetics, medicine, Animals, Molecular Biology, Crigler-Najjar Syndrome, Hyperbilirubinemia, Unconjugated hyperbilirubinemia, Genetic Therapy, medicine.disease, Gunn rat, Uridine, Rats, Disease Models, Animal, Phenotype, Liver, chemistry, Injections, Intravenous, Immunology, Hydrodynamics, Molecular Medicine, Female, Brief Reports, Expression cassette

الوصف: Crigler-Najjar syndrome type I is a severe inborn error of bilirubin metabolism caused by a complete deficiency of uridine diphospho-glucuronosyl transferase 1A1 (UGT1A1) and results in life-threatening unconjugated hyperbilirubinemia. Lifelong correction of hyperbilirubinemia by liver-directed gene therapy using a helper-dependent adenoviral (HDAd) vector has been previously reported in the Gunn rat, a model of Crigler-Najjar syndrome, but was only achieved using high doses (≥ 3 × 10(12) viral particles [vp]/kg), which are likely to elicit a severe toxic response in humans. Therefore, in this study, we investigate strategies to achieve correction of hyperbilirubinemia in the Gunn rat using clinically relevant low HDAd doses. We have found that correction of hyperbilirubinemia in the Gunn rat can be achieved with a low dose of 5 × 10(11) vp/kg by using an HDAd vector bearing a more potent UGT1A1 expression cassette. Furthermore, by using hydrodynamic injection of the improved HDAd vector, correction of hyperbilirubinemia in the Gunn rat can be achieved using an even lower dose of 5 × 10(10) vp/kg. Although hydrodynamic injection as performed in rats is not acceptable in humans, clinically attractive, minimally invasive methods have been successfully developed to mimic hydrodynamic injection of HDAd vector in non-human primates. Therefore, using an improved expression cassette combined with a more efficient method of vector delivery permits correction of hyperbilirubinemia in the Gunn rat using clinically relevant low HDAd doses and may thus pave the way to clinical application of HDAd vectors for Crigler-Najjar syndrome gene therapy.

الوصول الحر: https://explore.openaire.eu/search/publication?articleId=doi_dedup___::9adba95e6344486ee51b790b4cf6bdc5Test
https://doi.org/10.1089/hum.2010.167Test

المؤلفون: Margaretha Guenther, Vincenzo Cerullo, Nicola Brunetti-Pierri, Brendan Lee, Christian Clarke, Terry Bertin, Racel Cela, Masataka Suzuki

المصدر: Human Gene Therapy. 21:325-336

مصطلحات موضوعية: Male, Chromatin Immunoprecipitation, Transgene, Blotting, Western, Genetic Vectors, Antigen-Presenting Cells, Enzyme-Linked Immunosorbent Assay, Adaptive Immunity, Biology, medicine.disease_cause, Adenoviridae, Interferon-gamma, Mice, Immune system, Bone Marrow, Genetics, medicine, Animals, Gene silencing, Gene Silencing, RNA, Messenger, Transgenes, Molecular Biology, Cells, Cultured, Research Articles, Mice, Knockout, Innate immune system, Reverse Transcriptase Polymerase Chain Reaction, Macrophages, Fibroblasts, Th1 Cells, Flow Cytometry, Acquired immune system, Immunity, Innate, Toll-Like Receptor 2, Mice, Inbred C57BL, Liver, Helper virus, Myeloid Differentiation Factor 88, Immunology, Cytokines, Molecular Medicine, Signal transduction, Helper Viruses, Spleen

الوصف: Activation of the host innate immune response after systemic administration of adenoviral vectors constitutes a principal impediment to successful clinical gene replacement therapies. Although helper-dependent adenoviruses (HDAds) lack all viral functional genes, systemic administration of a high dose of HDAd still elicits a potent innate immune response in host animals. Toll-like receptors (TLRs) are innate receptors that sense microbial products and trigger the maturation of antigen-presenting cells and cytokine production via MyD88-dependent signaling (except TLR3). Here we show that mice lacking MyD88 exhibit a dramatic reduction in proinflammatory cytokines after intravenous injection of a high dose of HDAd, and show significantly reduced induction of the adaptive immune response when compared with wild-type and TLR2-deficient mice. Importantly, MyD88(-/-) mice also show significantly higher and longer sustained transgene expression than do wild-type mice. Chromatin immunoprecipitation studies using wild-type and MyD88-deficient primary mouse embryonic fibroblasts showed significant MyD88-dependent transcriptional silencing of the HDAd-encoded transgenes. Our results demonstrate that MyD88 signaling, activated by systemic delivery of HDAd, initiates an innate immune response that suppresses transgene expression at the transcriptional level before initiation of the adaptive immune response.

الوصول الحر: https://explore.openaire.eu/search/publication?articleId=doi_dedup___::2adee076c2d41e2bcd62dae4cdfee107Test
https://doi.org/10.1089/hum.2009.155Test

المؤلفون: Yu Zuo, Philip Ng, Rachel Edwards, Nicola Brunetti-Pierri, Donna Palmer, Nathan C Grove, Jun Teruya, Vincenzo Cerullo

المصدر: Human Gene Therapy. 20:479-485

مصطلحات موضوعية: Scientific Articles, Genetic enhancement, Genetic Vectors, Vectors in gene therapy, Hemophilia B, law.invention, Viral vector, Factor IX, Mice, Transduction (genetics), Protein replacement therapy, Therapeutic index, Transduction, Genetic, law, Genetics, Animals, Medicine, Molecular Biology, business.industry, Thrombosis, Genetic Therapy, Protein Structure, Tertiary, Mice, Inbred C57BL, Immunology, Recombinant DNA, Cancer research, Molecular Medicine, business, medicine.drug

الوصف: Although the desire to develop gene therapy for hemophilia B is high, safety remains a concern. Therefore, improving the therapeutic index of gene therapy vectors is an important goal. Thus, we evaluated the use of three bioengineered factor IX (FIX) variants with improved catalytic activity in the context of the helper-dependent adenoviral vector. The first vector expressed R338A-FIX, an FIX variant with the arginine at position 338 changed to an alanine, which resulted in a 2.9-fold higher specific activity (IU/mg) compared with the wild-type FIX. The second vector expressed FIX(VIIEGF1), a variant with the EGF-1 domain replaced with the EGF-1 domain from FVII, which resulted in a 3.4-fold increase in specific activity. The third expressed R338A + FIX(VIIEGF1), a novel variant containing both aforementioned modifications, which resulted in a 12.6-fold increase in specific activity. High-level, long-term, and stable expression of these three variants was observed in hemophilia B mice with no evidence of increased thrombogenicity compared with wild-type FIX. Thus, these bioengineered FIX variants can increase the therapeutic index of gene therapy vectors by permitting administration of lower doses to achieve the same therapeutic outcome. Furthermore, these variants may also be valuable for recombinant FIX protein replacement therapy.

الوصول الحر: https://explore.openaire.eu/search/publication?articleId=doi_dedup___::227acb0e5bf16c4e278b0f436ce8bb7dTest
https://doi.org/10.1089/hum.2008.084Test

المؤلفون: Nicola Brunetti-Pierri, Milton J. Finegold, Thomas Ng, William G. Cioffi, Donna Palmer, Arthur L. Beaudet, K. Dee Carey, David A. Iannitti, Philip Ng

المصدر: Human Gene Therapy. 17:391-404

مصطلحات موضوعية: Transgene, Genetic enhancement, Genetic Vectors, Pharmacology, medicine.disease_cause, Inferior vena cava, Transduction (genetics), Drug Delivery Systems, Hepatic Artery, Transduction, Genetic, biology.animal, Genetics, medicine, Animals, Molecular Biology, Chronic toxicity, biology, Portal Vein, Adenoviruses, Human, Liver Diseases, Genetic Therapy, beta-Galactosidase, Immunohistochemistry, Adenoviridae, Liver, medicine.vein, Toxicity, Immunology, Molecular Medicine, Venae Cavae, Helper Viruses, Papio, Baboon

الوصف: Helper-dependent adenoviral vectors (HDAds) are attractive vectors for liver-directed gene therapy because they can mediate sustained, high-level transgene expression without chronic toxicity. However, high vector doses are required to achieve efficient hepatic transduction by systemic delivery because of a nonlinear dose response. Unfortunately, such high doses result in systemic vector dissemination and dose-dependent acute toxicity with potentially severe and lethal consequences. We hypothesize that the threshold to efficient hepatic transduction may be circumvented by delivering the vector into the surgically isolated liver via the portal vein. Total hepatic isolation was achieved by occluding hepatic inflow from the portal vein and hepatic artery and by occluding hepatic venous outflow at the inferior vena cava. We demonstrate in nonhuman primates that this approach resulted in significantly higher efficiency hepatic transduction with reduced systemic vector dissemination compared with systemic intravascular delivery. This method of delivery was associated with transient acute toxicity, the severity of which was variable. Importantly, stable, high levels of transgene expression were obtained for at least 665 days for one baboon and for at least 560 days for two baboons with no evidence of long-term toxicity.

الوصول الحر: https://explore.openaire.eu/search/publication?articleId=doi_dedup___::5d00cf4f479bddd002ec2bd30c130e65Test
https://doi.org/10.1089/hum.2006.17.391Test

المؤلفون: Elizabeth P. Merricks, Stephanie McCorquodale, Timothy C. Nichols, Nicola Brunetti-Pierri, Donna Palmer, Philip Ng, Arthur L. Beaudet

المصدر: Human Gene Therapy. 16:811-820

مصطلحات موضوعية: Genetic enhancement, Genetic Vectors, medicine.disease_cause, Hemophilia B, Virus, Factor IX, Dogs, Genetics, Coagulopathy, medicine, Animals, Aspartate Aminotransferases, Vector (molecular biology), Creatine Kinase, Molecular Biology, biology, business.industry, Fissipedia, Alanine Transaminase, Genetic Therapy, Dependovirus, medicine.disease, biology.organism_classification, Phenotype, Blood Cell Count, Adenoviridae, Liver, Injections, Intravenous, Immunology, Systemic administration, Molecular Medicine, business, Helper Viruses

الوصف: We have evaluated the potential of liver-directed, helper-dependent adenoviral (HDAd) vector-mediated gene therapy in the hemophilia B dog. Two dogs were injected intravenously with HDAd (3 x 10(12) VP/kg) bearing a liver-restricted canine coagulation factor IX (FIX) expression cassette. After injection, the whole blood clotting time for both dogs declined from60 min to/=20 min for at least 604 and 446 days, respectively. Peak FIX activities of 34.1 and 129.2% were detected at 12x14 days and then slowly declined to 2 to 5% by 120 days and stabilized at these therapeutic levels for at least 418 and 257 days. For one dog, a peak FIX level of 500 ng/ml was achieved and stabilized at170 ng/ml for at least 256 days. For the other dog, a peak FIX level of 1258 ng/ml was achieved and stabilized at400 ng/ml for at least 213 days. Inhibitor formation was not evident in either animal. Importantly, whereas untreated hemophilia B dogs suffer five or six spontaneous bleeds per year, the treated dogs suffered no such bleeds postinjection. Significantly, this study is the first to demonstrate long-term phenotypic correction of a genetic disorder in a large animal with HDAd. Although no evidence of chronic toxicity was observed in either animal, systemic vector administration at 3 x 10(12) VP/kg was accompanied by acute, albeit transient and variable laboratory abnormalities (alanine aminotransferase, aspartate aminotransferase, alkaline phosphatase, creatine phosphokinase, and platelet counts). The results of this study highlight both the potential benefit and the risk associated with systemic intravascular delivery of high-dose HDAd for liver-directed gene therapy.

الوصول الحر: https://explore.openaire.eu/search/publication?articleId=doi_dedup___::578041b3ca7d90252bf0c2460d028552Test
https://doi.org/10.1089/hum.2005.16.811Test

المؤلفون: Donna Palmer, Philip Ng, Nicola Brunetti-Pierri, Milton J. Finegold, K. Dee Carey, Arthur L. Beaudet

المصدر: Human Gene Therapy. 15:35-46

مصطلحات موضوعية: Male, Indoles, Transgene, Genetic enhancement, Genetic Vectors, medicine.disease_cause, Virus, Adenoviridae, Transduction (genetics), Genes, Reporter, Transduction, Genetic, biology.animal, Genetics, medicine, Animals, Transgenes, Lung, Molecular Biology, biology, Galactosides, Genetic Therapy, Acute toxicity, Liver, Toxicity, Immunology, Molecular Medicine, Spleen, Papio, Baboon

الوصف: Systemic intravascular delivery of adenoviral (Ad) vectors for liver-directed gene therapy has been widely employed because of its simplicity, noninvasiveness, and potential for high transduction. For first-generation Ad vectors (FGAd), this results in high but transient levels of transgene expression and long-term hepatotoxicity due to viral gene expression from the vector backbone. Furthermore, high doses also result in an acute innate inflammatory response with potentially lethal consequences. Unlike FGAd, helper-dependent Ad vectors (HDAd) contain no viral genes and can provide sustained, high-level transgene expression with negligible long-term toxicity. However, whether the absence of viral gene expression leads to any decrease of acute toxicity in nonhuman primates has yet to be determined. To address this, we injected one baboon with 5.6 x 10(12) HDAd viral particles (VP)/kg and a second with 1.1 x 10(13) VP/kg. Approximately 50% hepatocyte transduction, accompanied by mild and transient acute toxicity, was observed in the animal receiving the lower dose. In the animal receiving the higher dose, 100% hepatocyte transduction, accompanied by lethal acute toxicity, was observed. These results indicate that systemic delivery of HDAd, like FGAd, results in acute toxicity in baboons consistent with activation of the innate inflammatory response, the severity of which is dose dependent, and confirm the hypothesis that Ad-mediated acute toxicity is independent of viral gene expression.

الوصول الحر: https://explore.openaire.eu/search/publication?articleId=doi_dedup___::a1d234ccd632fa39c5a703345ce60b5dTest
https://doi.org/10.1089/10430340460732445Test

المؤلفون: Mark A. Law, Gary E. Stapleton, Arthur L. Beaudet, Thomas Ng, John P. Breinholt, William G. Cioffi, Karen Rice, Cassondra Bauer, Donna Palmer, David A. Iannitti, Nicola Brunetti-Pierri, Nathan C Grove, Charles E. Mullins, Philip Ng, Milton J. Finegold

المساهمون: BRUNETTI PIERRI, Nicola, Ng, T, Iannitti, D, Cioffi, W, Stapleton, G, Law, M, Breinholt, J, Palmer, D, Grove, N, Rice, K, Bauer, C, Finegold, M, Beaudet, A, Mullins, C, Ng, P.

المصدر: Human gene therapy. 24(8)

مصطلحات موضوعية: Helper dependent adenoviral, Transgene, Genetic Vectors, Gene Expression, Biology, medicine.disease_cause, Adenoviridae, Time, Transduction (genetics), Transduction, Genetic, Gene expression, Genetics, medicine, Animals, Transgenes, Adverse effect, Molecular Biology, Research Articles, Virology, Liver, Helper virus, Immunology, Toxicity, Molecular Medicine, Helper Viruses, Papio

الوصف: Helper-dependent adenoviral vectors (HDAd) have been shown to mediate a considerably longer duration of transgene expression than first-generation adenoviral vectors. We have previously shown that transgene expression from HDAd-transduced hepatocytes can persist at high levels for up to 2.6 years in nonhuman primates following a single-vector administration. Because duration of transgene expression and long-term toxicity are critical for risk:benefit assessment, we have continued to monitor these animals. We report here that transgene expression has persisted for the entire observation period of up to 7 years for all animals without long-term adverse effects. However, in all cases, transgene expression level slowly declined over time to less than 10% of peak values by the end of the observation period but remained 2.3–111-fold above baseline values. These results will provide important information for a more informed risk:benefit assessment before clinical application of HDAd.

الوصول الحر: https://explore.openaire.eu/search/publication?articleId=doi_dedup___::548f9c9641272421fc460ffca3319b5bTest
https://pubmed.ncbi.nlm.nih.gov/23902403Test