Enhanced Prostacyclin Synthesis by Adenoviral Gene Transfer Reduced Glial Activation and Ameliorated Dopaminergic Dysfunction in Hemiparkinsonian Rats

Prostacyclin (PGI2), a potent vasodilator and platelet antiaggregatory eicosanoid, is cytoprotective in cerebral circulation. It is synthesized from arachidonic acid (AA) by the sequential action of cyclooxygenase- (COX-) 1 or 2 and prostacyclin synthase (PGIS). Because prostacyclin is unstablein vivo, PGI2analogs have been developed and demonstrated to protect against brain ischemia. This work attempts to selectively augment PGI2synthesis in mixed glial culture or in a model of Parkinson’s disease (PD) by direct adenoviral gene transfer of prostacyclin biosynthetic enzymes and examines whether it confers protection in cultures orin vivo. Confluent mixed glial cultures actively metabolized exogenous AA into PGE2and PGD2. These PGs were largely NS398 sensitive and considered as COX-2 products. Gene transfer of AdPGIS to the cultures effectively shunted the AA catabolism to prostacyclin synthesis and concurrently reduced cell proliferation. Furthermore, PGIS overexpression significantly reduced LPS stimulation in cultures.In vivo, adenoviral gene transfer of bicistronic COX-1/PGIS to substantia nigra protected 6-OHDA- induced dopamine depletion and ameliorated behavioral deficits. Taken together, this study shows that enhanced prostacyclin synthesis reduced glial activation and ameliorated motor dysfunction in hemiparkinsonian rats. Prostacyclin may have a neuroprotective role in modulating the inflammatory response in degenerating nigra-striatal pathway.

Download Full-text

In vivo electrophysiology of nigral and thalamic neurons in alpha-synuclein-overexpressing mice highlights differences from toxin-based models of parkinsonism

Journal of Neurophysiology ◽

10.1152/jn.00441.2013 ◽

2013 ◽

Vol 110 (12) ◽

pp. 2792-2805 ◽

Cited By ~ 10

Author(s):

C. J. Lobb ◽

A. K. Zaheer ◽

Y. Smith ◽

D. Jaeger

Keyword(s):

Primary Motor Cortex ◽

Alpha Synuclein ◽

Motor Dysfunction ◽

Motor Deficits ◽

Dopamine Depletion ◽

Wild Type ◽

Beta Oscillations ◽

Nigrostriatal Dopamine ◽

6 Hydroxydopamine

Numerous studies have suggested that alpha-synuclein plays a prominent role in both familial and idiopathic Parkinson's disease (PD). Mice in which human alpha-synuclein is overexpressed (ASO) display progressive motor deficits and many nonmotor features of PD. However, it is unclear what in vivo pathophysiological mechanisms drive these motor deficits. It is also unknown whether previously proposed pathophysiological features (i.e., increased beta oscillations, bursting, and synchronization) described in toxin-based, nigrostriatal dopamine-depletion models are also present in ASO mice. To address these issues, we first confirmed that 5- to 6-mo-old ASO mice have robust motor dysfunction, despite the absence of significant nigrostriatal dopamine degeneration. In the same animals, we then recorded simultaneous single units and local field potentials (LFPs) in the substantia nigra pars reticulata (SNpr), the main basal ganglia output nucleus, and one of its main thalamic targets, the ventromedial nucleus, as well as LFPs in the primary motor cortex in anesthetized ASO mice and their age-matched, wild-type littermates. Neural activity was examined during slow wave activity and desynchronized cortical states, as previously described in 6-hydroxydopamine-lesioned rats. In contrast to toxin-based models, we found a small decrease, rather than an increase, in beta oscillations in the desynchronized state. Similarly, synchronized burst firing of nigral neurons observed in toxin-based models was not observed in ASO mice. Instead, we found more subtle changes in pauses of SNpr firing compared with wild-type control mice. Our results suggest that the pathophysiology underlying motor dysfunction in ASO mice is distinctly different from striatal dopamine-depletion models of parkinsonism.

Download Full-text

Formulation and in vitro and in vivo evaluation of a cationic emulsion as a vehicle for improving adenoviral gene transfer

International Journal of Pharmaceutics ◽

10.1016/j.ijpharm.2014.08.024 ◽

2014 ◽

Vol 475 (1-2) ◽

pp. 49-59 ◽

Cited By ~ 11

Author(s):

Soo-Yeon Kim ◽

Sang-Jin Lee ◽

Soo-Jeong Lim

Keyword(s):

Gene Transfer ◽

In Vivo Evaluation ◽

Adenoviral Gene Transfer

Download Full-text

An adenoviral gene transfer system to express high levels of soluble allogeneic MHC class I molecules in vitro and in vivo

Transplantation Proceedings ◽

10.1016/s0041-1345(02)03252-9 ◽

2002 ◽

Vol 34 (6) ◽

pp. 2318-2319

Author(s):

C Graeb ◽

M Justl ◽

S Tange ◽

K.-W Jauch ◽

E.K Geissler

Keyword(s):

Gene Transfer ◽

Mhc Class I ◽

Class I ◽

Transfer System ◽

Adenoviral Gene Transfer ◽

Gene Transfer System ◽

Mhc Class I Molecules

Download Full-text

Ultrasound targeted gene therapy in a mouse model of prostate cancer: Evaluation of immune response.

Journal of Clinical Oncology ◽

10.1200/jco.2018.36.6_suppl.120 ◽

2018 ◽

Vol 36 (6_suppl) ◽

pp. 120-120

Author(s):

Flavia De Carlo ◽

Litty Thomas ◽

Rounak Nande ◽

Olivia Boskovic ◽

Gailen Marshall ◽

...

Keyword(s):

Prostate Cancer ◽

Gene Transfer ◽

Cancer Cells ◽

Transduction Efficiency ◽

Acquired Immunity ◽

Prostate Cancer Cells ◽

Adenoviral Gene Transfer ◽

Specific Tissue

120 Background: Gene transfer to malignant sites using human adenoviruses (hAd) has been limited because of their immunogenicity. Murine cells often lack some of the receptors needed for hAd infection; therefore, are generally non-permissive for hAd infection and replication, which limits translational studies of adenoviral gene transfer techniques. We developed a gene transfer method, which uses a combination of lipid-encapsulated perfluorocarbon microbubbles (MBs) and ultrasound (US) to shield and deliver hAds to a specific tissue bypassing the requirement of the coxsackie and adenovirus receptor (CAR). Methods: Transduction efficiency and GFP protein expression of hAd.GFP was assessed by flow cytometry and fluorescence microscopy in murine TRAMP-C2 and human DU145 prostate cancer cells. Innate and acquired immunity response was determined by ELISA and CTL assay in C57BL/6 mice bearing TRAMP-C2 syngeneic tumor grafts following injections of MBs-Ad.GFP complexes in the presence or absence of ultrasound. Results: We observed that the murine prostate cancer cells TRAMP-C2 were transduced less efficiently by hAd.GFP than the human DU145 cells. We showed in vitro that the transduction rate was increased significantly in both TRAMP-C2 and DU145 prostate cancer cells when delivering the Ad particles by a combination of MBs and US. Moreover, we observed expression of the GFP transgene in both cell lines at 48 hours and 72 hours. Lack of activation of the innate and acquired immunity was observed in vivo by quantifying IL-6 and TNF-α cytokines, and by assaying neutralizing IgG antibodies and CTLs activity, following intratumoral or intravenous injections of MBs-Ad.GFP complexes in the presence or absence of ultrasound. Conclusions: This study demonstrates the feasibility of using the TRAMP-C2 murine model of prostate adenocarcinoma to translate our ultrasound-mediated MB-Ad delivery system from the bench to the clinic. Our data provides evidence that the TRAMP-C2 prostate cancer graft model is a suitable system to study in immune competent animals the capacity of lipid-encapsulated perfluorocarbon MBs and US, to shield and deliver hAds to a site-specific tissue bypassing the requirement of specific receptors.

Download Full-text

Enhancement of adenoviral gene transfer to adult rat cardiomyocytes in vivo by immobilization and ultrasound treatment of the heart

Gene Therapy ◽

10.1038/sj.gt.3302476 ◽

2005 ◽

Vol 12 (11) ◽

pp. 936-941 ◽

Cited By ~ 15

Author(s):

M Sato ◽

P O'Gara ◽

S E Harding ◽

S J Fuller

Keyword(s):

Gene Transfer ◽

Ultrasound Treatment ◽

Rat Cardiomyocytes ◽

Adenoviral Gene Transfer ◽

Adult Rat

Download Full-text

Limited functional and metabolic improvements in hypertrophic and healthy rat heart overexpressing the skeletal muscle isoform of SERCA1 by adenoviral gene transfer in vivo

AJP Heart and Circulatory Physiology ◽

10.1152/ajpheart.01023.2008 ◽

2008 ◽

Vol 295 (6) ◽

pp. H2483-H2494 ◽

Cited By ~ 15

Author(s):

J. Michael O'Donnell ◽

Aaron Fields ◽

Xianyao Xu ◽

Shamim A. K. Chowdhury ◽

David L. Geenen ◽

...

Keyword(s):

Skeletal Muscle ◽

Gene Transfer ◽

Ex Vivo ◽

Rate Pressure Product ◽

Substrate Selection ◽

Nmr Analysis ◽

Sprague Dawley ◽

Important Distinction ◽

Adenoviral Gene Transfer

Adenoviral gene transfer of sarco(endo)plasmic reticulum Ca2+-ATPase (SERCA)2a to the hypertrophic heart in vivo has been consistently reported to lead to enhanced myocardial contractility. It is unknown if the faster skeletal muscle isoform, SERCA1, expressed in the whole heart in early failure, leads to similar improvements and whether metabolic requirements are maintained during an adrenergic challenge. In this study, Ad.cmv.SERCA1 was delivered in vivo to aortic banded and sham-operated Sprague-Dawley rat hearts. The total SERCA content increased 34%. At 48–72 h posttransfer, echocardiograms were acquired, hearts were excised and retrograded perfused, and hemodynamics were measured parallel to NMR measures of the phosphocreatine (PCr)-to-ATP ratio (PCr/ATP) and energy substrate selection at basal and high workloads (isoproterenol). In the Langendorff mode, the rate-pressure product was enhanced 27% with SERCA1 in hypertrophic hearts and 10% in shams. The adrenergic response to isoproterenol was significantly potentiated in both groups with SERCA1. 31P NMR analysis of PCr/ATP revealed that the ratio remained low in the hypertrophic group with SERCA1 overexpression and was not further compromised with adrenergic challenge. 13C NMR analysis revealed fat and carbohydrate oxidation were unaffected at basal with SERCA1 expression; however, there was a shift from fats to carbohydrates at higher workloads with SERCA1 in both groups. Transport of NADH-reducing equivalents into the mitochondria via the α-ketoglutamate-malate transporter was not affected by either SERCA1 overexpression or adrenergic challenge in both groups. Echocardiograms revealed an important distinction between in vivo versus ex vivo data. In contrast to previous SERCA2a studies, the echocardiogram data revealed that SERCA1 expression compromised function (fractional shortening) in the hypertrophic group. Shams were unaffected. While our ex vivo findings support much of the earlier cardiomyocyte and transgenic data, the in vivo data challenge previous reports of improved cardiac function in heart failure models after SERCA intervention.

Download Full-text