Sstr2A: a relevant target for the delivery of genes into human glioblastoma cells using fiber-modified adenoviral vectors.

Glioblastomas are the most aggressive of the brain tumors occurring in adults and children. Currently available chemotherapy prolongs the median survival time of patients by only 4 months. The low efficiency of current treatments is partly owing to the blood-brain barrier, which restricts the penetration of most drugs into the central nervous system. Locoregional treatment strategies thus become mandatory. In this context, viral tools are of great interest for the selective delivery of genes into tumoral cells. Gliomas express high levels of type 2 somatostatin receptors (sstr2A), pinpointing them as suitable targets for the improvement of transduction efficiency in these tumors. We designed a new adenoviral vector based on the introduction of the full-length somatostatin (SRIF (somatotropin release-inhibiting factor)) sequence into the HI loop of the HAdV fiber protein. We demonstrate that (i) HAdV-5-SRIF uptake into cells is mediated by sstr2A, (ii) our vector drives high levels of gene expression in cells expressing endogenous sstr2A, with up to 65-fold enhancement and (iii) low doses of HAdV-5-SRIF are sufficient to infect high-grade human primary glioblastoma cells. Adenoviral vectors targeting SRIF receptors might thus represent a promising therapeutic approach to brain tumors.

Modeling nonlinearity in coherent transmissions with dominant intrachannel-four-wave-mixing.

By extending a well-established time-domain perturbation approach to dual-polarization propagation, we provide an analytical framework to predict the nonlinear interference (NLI) variance, i.e., the variance induced by nonlinearity on the sampled field, and the nonlinear threshold (NLT) in coherent transmissions with dominant intrachannel-four-wave-mixing (IFWM). Such a framework applies to non dispersion managed (NDM) very long-haul coherent optical systems at nowadays typical baudrates of tens of Gigabaud, as well as to dispersion-managed (DM) systems at even higher baudrates, whenever IFWM is not removed by nonlinear equalization and is thus the dominant nonlinearity. The NLI variance formula has two fitting parameters which can be calibrated from simulations. From the NLI variance formula, analytical expressions of the NLT for both DM and NDM systems are derived and checked against recent NLT Monte-Carlo simulations.

Influence of chimeric human-bovine fibers on adenoviral uptake by liver cells and the antiviral immune response.

Human adenoviruses (HAdV) are widely used for in vitro and in vivo gene transfer. Viral hepatotropism, inflammatory responses and neutralization by pre-existing antibodies (NAbs) are obstacles for clinical applications of HAdV vectors. Although the multifactorial events leading to innate HAdV toxicity are far from being elucidated, there is a consensus that the majority of intravenously injected-HAdV vectors is sequestered by Kuppfer cells, probably independently of coagulation factors. In this study, we show that the adenoviral-associated humoral and innate cytokine immune responses are significantly reduced when HAdV-5 vector carrying human bovine chimeric fibers (HAdV-5-F2/BAdV-4) is intravenously injected into mice. Fiber pseudotyping modified its interaction with blood coagulation factors, as FIX and FX no longer mediate the infection of liver cells by HAdV-5-F2/BAdV-4. As a consequence, at early time points post-infection, several cytokines and chemokines (IFN-gamma, IL-6, IP-10, MCP-1, RANTES and MP1beta) were found to be present at lower levels in the plasma of mice that had been intravenously injected with HAdV-5-F2/BAdV-4 compared with mice injected with the parental vector HAdV-5. Moreover, genetic modification of the fiber allowed HAdV-5-F2/BAdV-4 to partially escape neutralization by NAbs.

Non-heparan sulfate GAG-dependent infection of cells using an adenoviral vector with a chimeric fiber conserving its KKTK motif.

Recently, the potential involvement of the putative heparan sulfate proteoglycans (HSPG) binding motif, KKTK, in mediating HAdV-5 liver cell infection following intravascular virus delivery has been debated. In the present study, we demonstrated that HSPGs were not involved in the in vitro infection process of an adenoviral vector harboring chimeric fibers without mutation in the KKTK motif, HAdV-5-F2/BAdV-4. The entry of HAdV-5-F2/BAdV-4 into cells occurs by two mechanisms 1) the attachment of HAdV-5-F2/BAdV-4 to the surface of cells requires N-glycosylation, 2) the uptake of the virus is effective after interaction with a co-receptor, putatively the chondroitin sulfate C. Together, these results contribute to improving our understanding of the molecular mechanisms determining HAdV's infectivity in vitro and may aid in designing novel HAdV-based vectors for gene therapy applications.