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Mariana Puntel,
Ghulam Muhammad A K M,
Catherine Farrokhi,
Nathan Vanderveen,
Christopher Paran,
Ashley Appelhans,
Kurt M Kroeger,
Alireza Salem,
Liliana Lacayo,
Robert N Pechnick,
Kyle R Kelson,
Sukhpreet Kaur,
Sean Kennedy,
Donna Palmer, Philip Ng,
Chunyan Liu,
Johnny Krasinkiewicz,
Pedro R Lowenstein,
Maria G Castro
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ABSTRACT: Adenoviral vectors (Ads) are promising gene delivery vehicles due to their high transduction efficiency; however, their clinical usefulness has been hampered by their immunogenicity and the presence of anti-Ad immunity in humans. We reported the efficacy of a gene therapy approach for glioma consisting of intratumoral injection of Ads encoding conditionally cytotoxic herpes simplex type 1 thymidine kinase (Ad-TK) and the immunostimulatory cytokine fms-like tyrosine kinase ligand 3 (Ad-Flt3L). Herein, we report the biodistribution, efficacy, and neurological and systemic effects of a bicistronic high-capacity Ad, i.e., HC-Ad-TK/TetOn-Flt3L. HC-Ads elicit sustained transgene expression, even in the presence of anti-Ad immunity, and can encode large therapeutic cassettes, including regulatory elements to enable turning gene expression "on" or "off" according to clinical need. The inclusion of two therapeutic transgenes within a single vector enables a reduction of the total vector load without adversely impacting efficacy. Because clinically the vectors will be delivered into the surgical cavity, normal regions of the brain parenchyma are likely to be transduced. Thus, we assessed any potential toxicities elicited by escalating doses of HC-Ad-TK/TetOn-Flt3L (1×10(8), 1×10(9), or 1×10(10) viral particles [vp]) delivered into the rat brain parenchyma. We assessed neuropathology, biodistribution, transgene expression, systemic toxicity, and behavioral impact at acute and chronic time points. The results indicate that doses up to 1×10(9) vp of HC-Ad-TK/TetOn-Flt3L can be safely delivered into the normal rat brain and underpin further developments for its implementation in a phase I clinical trial for glioma.
Toxicology and Applied Pharmacology 02/2013; · 4.45 Impact Factor
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Nunzia Pastore,
Keith Blomenkamp,
Fabio Annunziata,
Pasquale Piccolo,
Pratibha Mithbaokar,
Rosa Maria Sepe,
Francesco Vetrini,
Donna Palmer, Philip Ng,
Elena Polishchuk,
Simona Iacobacci,
Roman Polishchuk,
Jeffrey Teckman,
Andrea Ballabio,
Nicola Brunetti-Pierri
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ABSTRACT: Alpha-1-anti-trypsin deficiency is the most common genetic cause of liver disease in children and liver transplantation is currently the only available treatment. Enhancement of liver autophagy increases degradation of mutant, hepatotoxic alpha-1-anti-trypsin (ATZ). We investigated the therapeutic potential of liver-directed gene transfer of transcription factor EB (TFEB), a master gene that regulates lysosomal function and autophagy, in PiZ transgenic mice, recapitulating the human hepatic disease. Hepatocyte TFEB gene transfer resulted in dramatic reduction of hepatic ATZ, liver apoptosis and fibrosis, which are key features of alpha-1-anti-trypsin deficiency. Correction of the liver phenotype resulted from increased ATZ polymer degradation mediated by enhancement of autophagy flux and reduced ATZ monomer by decreased hepatic NFκB activation and IL-6 that drives ATZ gene expression. In conclusion, TFEB gene transfer is a novel strategy for treatment of liver disease of alpha-1-anti-trypsin deficiency. This study may pave the way towards applications of TFEB gene transfer for treatment of a wide spectrum of human disorders due to intracellular accumulation of toxic proteins.
EMBO Molecular Medicine 02/2013; · 10.33 Impact Factor
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ABSTRACT: Helper-dependent adenoviral (HDAd) vectors can mediate long-term, high-level transgene expression from transduced hepatocytes with no chronic toxicity. However, a toxic acute response with potentially lethal consequences has hindered their clinical applications. Liver sinusoidal endothelial cells (LSECs) and Kupffer cells are major barriers to efficient hepatocyte transduction. Understanding the mechanisms of adenoviral vector uptake by non-parenchymal cells may allow the development of strategies aimed at overcoming these important barriers and to achieve preferential hepatocyte gene transfer with reduced toxicity. Scavenger receptors on Kupffer cells bind adenoviral particles and remove them from the circulation, thus preventing hepatocyte transduction. In the present study, we show that HDAd particles interact in vitro and in vivo with scavenger receptor-A (SR-A) and with scavenger receptor expressed on endothelial cells-I (SREC-I) and we exploited this knowledge to increase the efficiency of hepatocyte transduction by HDAd vectors in vivo through blocking of SR-A and SREC-I with specific fragments antigen-binding (Fabs).Molecular Therapy (2013); doi:10.1038/mt.2012.287.
Molecular Therapy 01/2013; · 6.87 Impact Factor
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ABSTRACT: We previously dissected the components of the innate immune response to Helper-dependent adenoviral vectors (HDAds) using genetic models, and demonstrated that multiple pattern recognition receptor signaling pathways contribute to this host response to HDAds in vivo. Based on analysis of cytokine expression profiles, type I interferon (IFN) mRNA is induced in host mouse livers at 1 hour post-injection. This type I IFN signaling amplifies cytokine expression in liver independent of the nature of vector DNA sequences after 3 hours post-injection. This type I IFN signaling in response to HDAds administration contributes to transcriptional silencing of both HDAd prokaryotic and eukaryotic DNA in liver. This silencing occurs early and is mediated by epigenetic modification as shown by in vivo chromatin immunoprecipitation (ChIP) with anti-histone deacetylase (HDAC) and promyelocytic leukemia protein (PML). In contrast, self-complementary adeno-associated viral vectors (scAAVs) showed significantly lower induction of type I IFN mRNA in liver compared to HDAds at both early and late time points. These results show that the type I IFN signaling dependent transgene silencing differs between AAV and HDAd vectors after liver-directed gene transfer.Molecular Therapy (2013); doi:10.1038/mt.2012.277.
Molecular Therapy 01/2013; · 6.87 Impact Factor
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Kilian Guse,
Masataka Suzuki,
Gautam Sule,
Terry K Bertin,
Henna Tyynismaa,
Sofia Ahola-Erkkilä,
Donna Palmer,
Anu Suomalainen, Philip Ng,
Vincenzo Cerullo,
Akseli Hemminki,
Brendan Lee
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Maria Castro,
Weidong Xiong,
Mariana Puntel,
Catherine Farrokhi,
Kurt M Kroeger,
Robert N Pechnick, Philip Ng,
Pedro Lowenstein,
Akm Ghulam Muhammad,
Alireza Salem,
Liliana Lacayo,
Kyle R Kelson,
Donna J Palmer,
Chungyan Liu,
Ashley Appelhans
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ABSTRACT: Adenoviral vectors (Ads) have been evaluated in clinical trials for glioma. However, systemic immunity against the vectors can hamper therapeutic efficacy. We demonstrated that combined immunostimulation and cytotoxic gene therapy provides long-term survival in pre-clinical glioma models. Since helper-dependent high capacity Ads (HC-Ads) elicit sustained transgene expression, in the presence of anti-adenoviral immunity, we engineered HC-Ads encoding conditional cytotoxic herpes simplex type 1 thymidine kinase and immunostimulatory cytokine fms-like tyrosine kinase ligand 3 under the control of TetOn system. Escalating doses of combined HC-Ads (1x108, 1x109, 1x1010 vp) were delivered into the rat brain. We assessed neuropathology, biodistribution, transgene expression, systemic toxicity and behavioral impact at acute and chronic time points post-vector delivery. Histopathological analysis did not reveal any evidence of toxicity or long-term inflammation at the lower doses tested. Vector genomes were restricted to the injection site. Serum chemistry did not uncover adverse systemic side effects at any of the doses tested. Taken together our data indicates that doses of up to 1x109 vp of each HC-Ad can be safely administered into the normal brain. This comprehensive toxicity and biodistribution study will lay the foundations for implementation of a Phase I clinical trial for GBM using HC-Ads.
Human gene therapy methods. 10/2012;
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A K M Ghulam Muhammad,
Weidong Xiong,
Mariana Puntel,
Catherine Farrokhi,
Kurt M Kroeger,
Alireza Salem,
Liliana Lacayo,
Robert N Pechnick,
Kyle R Kelson,
Donna Palmer, Philip Ng,
Chunyan Liu,
Pedro R Lowenstein,
Maria G Castro
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ABSTRACT: Abstract Adenoviral vectors (Ads) have been evaluated in clinical trials for glioma. However, systemic immunity against the vectors can hamper therapeutic efficacy. We demonstrated that combined immunostimulation and cytotoxic gene therapy provides long-term survival in preclinical glioma models. Because helper-dependent high-capacity Ads (HC-Ads) elicit sustained transgene expression, in the presence of antiadenoviral immunity, we engineered HC-Ads encoding conditional cytotoxic herpes simplex type 1 thymidine kinase and immunostimulatory cytokine Fms-like tyrosine kinase ligand-3 under the control of the TetOn system. Escalating doses of combined HC-Ads (1×10(8), 1×10(9), and 1×10(10) viral particles [VP]) were delivered into the rat brain. We assessed neuropathology, biodistribution, transgene expression, systemic toxicity, and behavioral impact at acute and chronic time points after vector delivery. Histopathological analysis did not reveal any evidence of toxicity or long-term inflammation at the lower doses tested. Vector genomes were restricted to the injection site. Serum chemistry did not uncover adverse systemic side effects at any of the doses tested. Taken together, our data indicate that doses of up to 1×10(9) VP of each HC-Ad can be safely administered into the normal brain. This comprehensive toxicity and biodistribution study will lay the foundations for implementation of a phase 1 clinical trial for GBM using HC-Ads.
Human gene therapy methods. 09/2012; 23(4):271-84.
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Kilian Guse,
Masataka Suzuki,
Gautam Sule,
Terry K Bertin,
Henna Tyynismaa,
Sofia Ahola-Erkkilä,
Donna Palmer,
Anu Suomalainen, Philip Ng,
Vincenzo Cerullo,
Akseli Hemminki,
Brendan Lee
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ABSTRACT: Abstract Skeletal muscle represents an attractive target tissue for adenoviral gene therapy to treat muscle disorders and as a production platform for systemic expression of therapeutic proteins. However, adenovirus serotype 5 vectors do not efficiently transduce adult muscle tissue. Here we evaluated whether capsid modifications on adenoviral vectors could improve transduction in mature murine muscle tissue. First-generation and helper-dependent serotype 5 adenoviral vectors featuring the serotype 3 knob (5/3) showed significantly increased transduction of skeletal muscle after intramuscular injection in adult mice. Furthermore, we showed that full-length dystrophin could be more efficiently transferred to muscles of mdx mice using a 5/3-modified helper-dependent adenoviral vector. In contrast to first-generation vectors, helper-dependent adenoviral vectors mediated stable marker gene expression for at least 1 year after intramuscular injection. In conclusion, 5/3 capsid-modified helper-dependent adenoviral vectors show enhanced transduction in adult murine muscle tissue and mediate long-term gene expression, suggesting the suitability of these vectors for muscle-directed gene therapy.
Human gene therapy 08/2012; 23(10):1065-70. · 4.20 Impact Factor
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Nicola Brunetti-Pierri,
Aimee Liou,
Priti Patel,
Donna Palmer,
Nathan Grove,
Milton Finegold,
Pasquale Piccolo,
Elizabeth Donnachie,
Karen Rice,
Arthur Beaudet,
Charles Mullins, Philip Ng
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ABSTRACT: Hemophilia B is an excellent candidate for gene therapy because low levels of factor IX (FIX) (≥1%) result in clinically significant improvement of the bleeding diathesis. Helper-dependent adenoviral (HDAd) vectors can mediate long-term transgene expression without chronic toxicity. To determine the potential for HDAd-mediated liver-directed hemophilia B gene therapy, we administered an HDAd expressing hFIX into rhesus macaques through a novel and minimally invasive balloon occlusion catheter-based method that permits preferential, high-efficiency hepatocyte transduction with low, subtoxic vector doses. Animals given 1 × 10(12) and 1 × 10(11) virus particle (vp)/kg achieved therapeutic hFIX levels for the entire observation period (up to 1,029 days). At 3 × 10(10) and 1 × 10(10) vp/kg, only subtherapeutic hFIX levels were achieved which were not sustained long-term. Balloon occlusion administration of HDAd was well tolerated with negligible toxicity. Five of six animals developed inhibitors to hFIX. These results provide important information in assessing the clinical utility of HDAd for hemophilia B gene therapy.
Molecular Therapy 07/2012; 20(10):1863-70. · 6.87 Impact Factor
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ABSTRACT: Prestin is a membrane protein in the outer hair cell (OHC) that has been shown to be essential for electromotility. OHCs from
prestin-null mice do not express prestin, do not have a nonlinear capacitance (the electrical signature of electromotility),
and are smaller in size than wild-type OHCs. We sought to determine whether prestin-null OHCs can be transduced to incorporate
functional prestin protein in a normal fashion. A recombinant helper-dependent adenovirus expressing prestin and green fluorescent
protein (HDAd–prestin–GFP) was created and tested in human embryonic kidney cells (HEK cells). Transduced HEK cells demonstrated
membrane expression of prestin and nonlinear capacitance. HDAd–prestin–GFP was then applied to cochlear sensory epithelium
explants harvested from wild-type and prestin-null mice at postnatal days2–3, the age at which native prestin is just beginning
to become functional in wild-type mice. At postnatal days4–5, we investigated transduced OHCs for (1) their prestin expression
pattern as revealed by immunofluorescence; (2) their cell surface area as measured by linear capacitance; and (3) their prestin
function as indicated by nonlinear capacitance. HDAd–prestin–GFP efficiently transduced OHCs of both genotypes and prestin
protein localized to the plasma membrane. Whole-cell voltage clamp studies revealed a nonlinear capacitance in transduced
wild-type and prestin-null OHCs, but not in non-transduced cells of either genotype. Prestin transduction did not increase
the linear capacitance (cell surface area) for either genotype. In peak nonlinear capacitance, voltage at peak nonlinear capacitance,
charge density of the nonlinear capacitance, and shape of the voltage-capacitance curves, the transduced cells of the two
genotypes resembled each other and previously reported data from adult wild-type mouse OHCs. Thus, prestin introduced into
prestin-deficient OHCs segregates normally to the cell membrane and generates a normal nonlinear capacitance, indicative of
normal prestin function.
Journal of the Association for Research in Otolaryngology 04/2012; 9(3):307-320. · 2.84 Impact Factor
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ABSTRACT: Successful liver-directed gene therapy has the potential to revolutionize medicine. Helper-dependent adenoviral vectors (HDAds) are devoid of all viral coding sequences and have shown tremendous potential for liver-direct gene therapy. In small and large animals, hepatic transduction with HDAd has resulted in high level, long-term transgene expression without chronic toxicity in a variety of disease models. Recent advancements in the large-scale manufacture of HDAd have permitted contemplation of clinical application. However, dose-dependent activation of the host innate inflammatory response remains an obstacle for clinical translation. Recent advancements in vector capsid modifications, immune modulation regimes, as well as novel routes of vector administration may yet permit clinical liver-directed gene therapy with HDAd.
Current pharmaceutical design 07/2011; 17(24):2488-99. · 4.41 Impact Factor
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ABSTRACT: As much as 90% of an intravenously (i.v.) injected dose of adenovirus serotype 5 (Ad5) is absorbed and destroyed by liver Kupffer cells. Viruses that escape these cells can then transduce hepatocytes after binding factor X (FX). Given that interactions with FX and Kupffer cells are thought to occur on the Ad5 hexon protein, we replaced its exposed hypervariable regions (HVR) with those from Ad6. When tested in vivo in BALB/c mice and in hamsters, the Ad5/6 chimera mediated >10 times higher transduction in the liver. This effect was not due to changes in FX binding. Rather, Ad5/6 appeared to escape Kupffer cell uptake as evidenced by producing no Kupffer cell death in vivo, not requiring predosing in vivo, and being phagocytosed less efficiently by macrophages in vitro compared to Ad5. When tested as a helper-dependent adenovirus (Ad) vector, Ad5/6 mediated higher luciferase and factor IX transgene expression than either helper-dependent adenoviral 5 (HD-Ad5) or HD-Ad6 vectors. These data suggest that the Ad5/6 hexon-chimera evades Kupffer cells and may have utility for systemic and liver-directed therapies.
Molecular Therapy 04/2011; 19(7):1254-62. · 6.87 Impact Factor
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ABSTRACT: Helper-dependent adenoviral (HDAd) vectors devoid of all viral-coding sequences are promising non-integrating vectors for liver-directed gene therapy because they have a large cloning capacity, can efficiently transduce a wide variety of cell types from various species independent of the cell cycle and can result in long-term transgene expression without chronic toxicity. The main obstacle preventing clinical applications of HDAd for liver-directed gene therapy is the host innate inflammatory response against the vector capsid proteins that occurs shortly after intravascular vector administration resulting in acute toxicity, the severity of which is dependent on vector dose. Intense efforts have been focused on elucidating the factors involved in this acute response and various strategies have been investigated to improve the therapeutic index of HDAd vectors. These strategies have yielded encouraging results with the potential for clinical translation.
Human Molecular Genetics 04/2011; 20(R1):R7-13. · 7.64 Impact Factor
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ABSTRACT: INTRODUCTION This protocol describes the characterization of helper-dependent adenoviral vectors (HDAd) with respect to physical titer, helper virus contamination, and genomic structure using standard molecular biology techniques. At a minimum, purified vector should be assessed for its physical titer, degree of helper virus contamination, and analysis of the HDAd genomic structure. If desired and depending on the application, additional characterization (i.e., endotoxin, sterility, adventitious agents, etc.) should be performed according to protocols established for E1-deleted Ad vectors.
Cold Spring Harbor Protocols 01/2011; 2011(7):867-70. · 4.63 Impact Factor
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ABSTRACT: INTRODUCTION This protocol describes in detail the rescue, amplification, and large-scale production of helper-dependent adenoviral vectors (HDAds) for gene transfer and gene therapy. Production of HDAd can be divided into three parts: (1) rescue, which involves converting pHDAd (plasmid form) to HDAd (viral form); (2) amplification, in which the amount of HDAd is increased by serial coinfections (passages) of the producer cells with the HDAd and the helper virus; and (3) large-scale production to generate large quantities of HDAd. HDAds (also referred to as gutless, gutted, mini, fully deleted, high-capacity, Δ, pseudo) are deleted of all virus-coding sequences. HDAds retain the advantages of early-generation Ad vectors including high-efficiency in vivo transduction and high-level transgene expression. However, the absence of viral gene expression in transduced cells permits long-term transgene expression in the absence of chronic toxicity. Moreover, the deletion of the viral sequences permits a cloning capacity of ∼37 kb. This allows for the delivery of whole-genomic loci, multiple transgenes, and large cis-acting elements to enhance, prolong, and regulate transgene expression. In addition, because the vector genome exists episomally in transduced cells, the risks of germ-line transmission and insertional mutagenesis leading to oncogenic transformation are negligible.
Cold Spring Harbor Protocols 01/2011; 2011(7):857-66. · 4.63 Impact Factor
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ABSTRACT: Adenovirus serotype (Ad5) is the most studied Ad. Ad1, 2, and 6 are also members of species C Ad and are presumed to have biologies similar to Ad5. In this work, we have compared the ability of Ad1, 2, 5, and 6 to infect liver and muscle after intravenous and intramuscular injection. We found that Ad6 was surprisingly the most potent at liver gene delivery and that Ad1 and Ad2 were markedly weaker than Ad5 and 6. To understand these differences, we sequenced the Ad6 genome. This revealed that the Ad6 fiber protein is surprisingly three shaft repeats shorter than the others which may explain differences in virus infectivity in vitro, but not in the liver. Comparison of hexon hypervariable regions (HVRs) suggests that the higher transduction by Ad5 and 6 as compared to Ad1 and 2 may be related to differences in charge and length.
Virology 01/2011; 412(1):19-27. · 3.35 Impact Factor
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ABSTRACT: Murine leukemia virus (MLV)-based replication-competent retrovirus (RCR) vectors have been shown to mediate efficient, selective, and persistent tumor transduction, thereby achieving significant therapeutic benefit in a wide variety of cancer models. To further augment the efficiency of this strategy, we have developed a delivery method employing a gutted adenovirus encoding an RCR vector (AdRCR); thus, tumor cells transduced with the adenoviral vector transiently become RCR vector producer cells in situ. As expected, high-titer AdRCR achieved significantly higher initial transduction levels in human cancer cells both in vitro and in vivo, as compared to the original RCR vector itself. Notably, even at equivalent initial transduction levels, more secondary RCR progeny were produced from AdRCR-transduced cells as compared to RCR-transduced cells, resulting in further acceleration of subsequent RCR replication kinetics. In pre-established tumor models in vivo, prodrug activator gene therapy with high-titer AdRCR could achieve enhanced efficacy compared to RCR alone, in a dose-dependent manner. Thus, AdRCR hybrid vectors offer the advantages of high production titers characteristic of adenovirus and secondary production of RCR in situ, which not only accelerates subsequent vector spread and progressive tumor transduction, but can also significantly enhance the therapeutic efficacy of RCR-mediated prodrug activator gene therapy.
Molecular Therapy 01/2011; 19(1):76-82. · 6.87 Impact Factor
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ABSTRACT: Adenoviral vectors (AdV) activate multiple signaling pathways associated with innate immune responses, including mitogen-activated protein kinases (MAPKs). In this study, we investigated how systemically-injected AdVs activate two MAPK pathways (p38 and ERK) and the contribution of these kinases to AdV-induced cytokine and chemokine responses in mice. Mice were injected intravenously either with a helper-dependent Ad2 vector that does not express viral genes or transgenes, or with the Ad2 mutant ts1, which is defective in endosomal escape. We found that AdV induced rapid phosphorylation of p38 and ERK as well as a significant cytokine response, but ts1 failed to activate p38 or ERK and induced only a limited cytokine response. These results demonstrate that endosomal escape of virions is a critical step in the induction of these innate pathways and responses. We then examined the roles of p38 and ERK pathways in the innate cytokine response by administering specific kinase inhibitors to mice prior to AdV. The cytokine and chemokine response to AdV was only modestly suppressed by a p38 inhibitor, while an ERK inhibitor has mixed effects, lowering some cytokines and elevating others. Thus, even though p38 and ERK are rapidly activated after i.v. injection of AdV, cytokine and chemokine responses are mostly independent of these kinases.
PLoS ONE 01/2011; 6(10):e26755. · 4.09 Impact Factor
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ABSTRACT: Although adenovirus vectors (Ads) have been widely utilized for gene delivery, their clinical application has been hampered by host immune responses. It has been shown that macrophages can induce inflammatory response against Ads in vivo, but they are not easily activated by Ads in vitro, suggesting their activation requires interaction with other cells. In this study, we investigated the interaction between macrophages and epithelial cells during Ad infection. Ad infection of the macrophage-epithelial cell co-culture resulted in rapid and drastic changes in the cell culture such as decrease in pH within 24h, indicating macrophage activation. Ad infected co-culture showed several characteristics of inflammation including cytotoxicity, induction of pro-inflammatory cytokines, and generation of nitric oxide and reactive oxygen species. These signs of macrophage activation and inflammation were observed exclusively in the co-culture and were absent or significantly weaker in the macrophage mono-culture suggesting that there was a synergistic response by the interaction between macrophages and epithelial cells. We found that inhibition of NF-κB activation significantly reduced the inflammatory responses in the co-culture. Furthermore, we show that only the macrophages adjacent to epithelial cells were activated during Ad infection demonstrating that the interaction between macrophages and epithelial cells are crucial for Ad-induced inflammatory response.
Immunology letters 11/2010; 134(1):93-102. · 2.91 Impact Factor
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Jan-Michael Prill,
Sigrid Espenlaub,
Ulrike Samen,
Tatjana Engler,
Erika Schmidt,
Francesco Vetrini,
Amanda Rosewell,
Nathan Grove,
Donna Palmer, Philip Ng,
Stefan Kochanek,
Florian Kreppel
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ABSTRACT: In vivo gene transfer with adenovirus vectors would significantly benefit from a tight control of the adenovirus-inherent liver tropism. For efficient hepatocyte transduction, adenovirus vectors need to evade from Kupffer cell scavenging while delivery to peripheral tissues or tumors could be improved if both scavenging by Kupffer cells and uptake by hepatocytes were blocked. Here, we provide evidence that a single point mutation in the hexon capsomere designed to enable defined chemical capsid modifications may permit both detargeting from and targeting to hepatocytes with evasion from Kupffer cell scavenging. Vector particles modified with small polyethylene glycol (PEG) moieties specifically on hexon exhibited decreased transduction of hepatocytes by shielding from blood coagulation factor binding. Vector particles modified with transferrin or, surprisingly, 5,000 Da PEG or dextran increased hepatocyte transduction up to 18-fold independent of the presence of Kupffer cells. We further show that our strategy can be used to target high-capacity adenovirus vectors to hepatocytes emphasizing the potential for therapeutic liver-directed gene transfer. Our approach may lead to a detailed understanding of the interactions between adenovirus vectors and Kupffer cells, one of the most important barriers for adenovirus-mediated gene delivery.
Molecular Therapy 10/2010; 19(1):83-92. · 6.87 Impact Factor
