Neprilysin Gene Transfer Reduces Human Amyloid Pathology in Transgenic Mice

Marr, Robert A.; Rockenstein, Edward; Mukherjee, Atish; Kindy, Mark S.; Hersh, Louis B.; Gage, Fred H.; Verma, Inder M.; Masliah, Eliezer

doi:10.1523/jneurosci.23-06-01992.2003

Cited by 361 publications

(276 citation statements)

References 27 publications

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“…We produced lentiviral vectors using a fourplasmid transfection system, as previously described. 45 Briefly, 293T cells were transfected with vector and packaging plasmids, the supernatants were collected and vectors were concentrated by centrifugation. The lentiviral vector titers were estimated by measuring the amount of HIV p24 gag antigen with an ELISA kit (Perkin-Elmer, Wellesley, MA, USA) (100 000 transducing units (TU) per nanogram of p24).…”

Section: Lentiviral Vector Productionmentioning

confidence: 99%

“…[46][47][48] A total of 48 ha-synuclein tg mice from line D (12 months old) were injected with 3 ml of the lentiviral preparations (1.5 Â 10 7 TU) into the temporal cortex and hippocampus. Briefly, as previously described, 45 mice were placed under anesthesia on a Koft stereotaxic apparatus and coordinates (AP -1.5 mm, lateral 1 mm, depth 2 mm) were determined as per the Franklin and Paxinos Atlas. The lentiviral vectors were delivered using a 5 ml Hamilton syringe connected to a hydraulic system to inject the solution at a rate of 0.5 ml every 2 min.…”

Section: Transgenic Mouse Lines and Intracerebral Injections Of Lentimentioning

confidence: 99%

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An antiaggregation gene therapy strategy for Lewy body disease utilizing β-synuclein lentivirus in a transgenic model

et al. 2004

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Current experimental gene therapy approaches for Parkinson's disease (PD) and dementia with Lewy bodies (DLB) include the use of viral vectors expressing antiapoptosis genes, neurotrophic factors and dopaminergic system enzymes. However, since increasing evidence favors a role for a-synuclein accumulation in the pathogenesis of these disorders, an alternative therapy might require the transfer of genes that might block a-synuclein accumulation. b-Synuclein, the nonamyloidogenic homologue of a-synuclein, has recently been identified as a potential candidate. Thus, in vivo transfer of genes encoding b-synuclein might provide a novel approach to the development of experimental treatments for PD and DLB. To assess this possibility and to better understand the mechanisms involved, a lentiviral vector expressing human (h) b-synuclein (lenti-b-synuclein) was tested in a transgenic (tg) mouse model of ha-synuclein aggregation. This study showed that unilateral intracerebral injection of lenti-b-synuclein reduced the formation of hasynuclein inclusions and the accumulation of ha-synuclein in synapses and ameliorated the neurodegenerative alterations in the tg mice. Both in vivo and in vitro coimmunoprecipitation and immunoblot experiments show that the mechanisms of b-synuclein neuroprotection involve binding of this molecule to ha-synuclein and Akt, resulting in the decreased aggregation and accumulation of ha-synuclein in the synaptic membrane. Together, these data further support a role for b-synuclein in regulating the conformational state of a-synuclein and suggest that this gene transfer approach might have potential for the development of alternative therapies for PD and DLB.

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Section: Lentiviral Vector Productionmentioning

confidence: 99%

Section: Transgenic Mouse Lines and Intracerebral Injections Of Lentimentioning

confidence: 99%

An antiaggregation gene therapy strategy for Lewy body disease utilizing β-synuclein lentivirus in a transgenic model

et al. 2004

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“…Among many proteases, recent studies have built strong evidence for the ability of two metalloproteases, neprilysin (NEP) and insulin degrading enzyme (IDE), to hydrolyze Aβ and reduce amyloid plaque burden in mice [8,14,22,27]. Similarly, two receptorassociated proteins, low-density lipoprotein receptor-related protein (LRP) and the receptor for advanced glycation end products (RAGE), are instrumental in Aβ transport from the brain to the periphery for degradation [7,16,28,34].…”

Section: Discussionmentioning

confidence: 99%

Spatial and temporal control of age-related APP processing in genomic-based β-secretase transgenic mice

Chiocco

Lamb

2007

Neurobiology of Aging

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Genetic mutations associated with Alzheimer's disease (AD) in the Amyloid Precursor Protein (APP) gene specifically alter the production of the APP processing product, amyloid-β (Aβ) peptide, generated by β-and γ-secretases. The accumulation and deposition of Aβ is hypothesized to cause AD pathogenesis, leading to the debilitating neurological deficits observed in AD patients. However, it is unclear how processing of APP to generate Aβ corresponds with the age-dependent pattern of brain-regional neurodegeneration common in AD. We have previously shown that overexpression of BACE1, the primary β-secretase gene, in mice expressing an AD mutant form of APP leads to significantly elevated regional Aβ levels, which coincide with the regional pattern of Aβ deposition. In the current study, we have used our genomic-based β-secretase transgenic mice to determine how BACE1 regulates the spatial and temporal pattern of Aβ production throughout post-natal development. Specifically, we observed unique differences in the brain-regional expression pattern between neonatal and adult BACE1 transgenic mice. These alterations in the BACE1 expression profile directly corresponds with age-related differences in regional Aβ production and deposition. These studies indicate that modulation of BACE1 expression leads to dramatic alterations in APP processing and AD-like neuropathology. Furthermore, our studies provide further evidence that BACE1 plays a major role in the regulation of the APP processing pathway, influencing the agedependent onset of AD pathogenesis.

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“…12,13 The vector plasmids that expressed green fluorescent protein (L-GFP), wildtype NEP (L-NEP) or enzymatically inactive mutant NEP (L-NEPmu) driven by the human cytomegalovirus promoter were constructed as described. 12,13 The lentiviral vector titers were estimated by flow cytometric analyses (fluorescence-activated cell sorting (FACS)) as described previously. 13 DU145 cells were transduced with L-GFP, L-NEP or L-NEPmu at the indicated multiplicity of infection (MOI) in the presence of 6 mg/ml polybrene (Sigma-Aldrich, St Louis, MO, USA) for 72 h.…”

Section: Lentiviral Vector Production and Transductionmentioning

confidence: 99%

“…Lentiviruses such as human immunodeficiency virus type-1 have been developed for in vitro and in vivo gene delivery. 12,13 Lentiviral vectors have numerous advantages for gene delivery, including the ability to infect non-dividing cells, longterm transgene expression and absence of induction of host inflammatory and immune responses. 12 Recent studies have begun to examine their utility in cancer gene therapy.…”

Section: Introductionmentioning

confidence: 99%

Neutral endopeptidase inhibits prostate cancer tumorigenesis by reducing FGF-2-mediated angiogenesis

Horiguchi

Chen

Goodman

et al. 2007

Prostate Cancer Prostatic Dis

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Neutral endopeptidase (NEP) is a cell surface peptidase that catalytically inactivates a variety of physiologically active peptides including basic fibroblast growth factor (FGF-2). We investigated the effect of using lentivirus to overexpress NEP in NEP-deficient DU145 prostate cancer cells. Third-generation lentiviral vectors encoding wild-type NEP (L-NEP), catalytically inactive mutant NEP (L-NEPmu), and green fluorescent protein (L-GFP) were stably introduced into DU145 cells. FGF-2 levels in cell culture supernatants decreased by 80% in L-NEP-infected DU145 cells compared to cells infected with L-NEPmu or L-GFP (Po0.05) while levels of other angiogenic factors were not altered. In vitro tubulogenesis of human vascular endothelial cells induced by conditioned media from DU145 cells infected with L-NEP was significantly reduced compared with that from DU145 cells infected with L-GFP (Po0.05). Tumor xenografts from L-NEP-infected DU145 cells were significantly smaller compared to control cell xenografts and vascularity within these tumors was decreased (Po0.05). Our data suggest that stable expression of NEP in DU145 cells inhibits prostate cancer tumorigenicity by inhibiting angiogenesis, with a probable mechanism being proteolytic inactivation of FGF-2.

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Neprilysin Gene Transfer Reduces Human Amyloid Pathology in Transgenic Mice

Cited by 361 publications

References 27 publications

An antiaggregation gene therapy strategy for Lewy body disease utilizing β-synuclein lentivirus in a transgenic model

An antiaggregation gene therapy strategy for Lewy body disease utilizing β-synuclein lentivirus in a transgenic model

Spatial and temporal control of age-related APP processing in genomic-based β-secretase transgenic mice

Neutral endopeptidase inhibits prostate cancer tumorigenesis by reducing FGF-2-mediated angiogenesis

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