Characterization and adaptive texture synthesis-based compression scheme

International audienceThis paper presents an adaptive texture synthesis-based compression scheme, where textured regions are detected and removed at encoder side, allowing the decoder to use texture synthesis to fill them. The detection relies on locally adaptive resolution segmentation. According to results shown by synthesis algorithms, they need to be parameterized according to the patterns to be synthesized. In this framework, the synthesizer gets its parameters from DCT feature-based texture descriptors. An adaptive pixel-based algorithm is used, relying on the comparison between current pixel neighborhood and those in an atypically shaped sample. Different neighborhood sizes are considered to better match texture patterns. The framework has been validated within an H.264/AVC video codec. Experimental results show significant bit-rate saving at similar visual quality

Enabling the formation of native mAb, Fab ' and Fc-conjugates using a bis-disulfide bridging reagent to achieve tunable payload-to-antibody ratios (PARs)

Either as full IgGs or as fragments (Fabs, Fc, etc.), antibodies have received tremendous attention in the development of new therapeutics such as antibody-drug conjugates (ADCs). The production of ADCs involves the grafting of active payloads onto an antibody, which is generally enabled by the site-selective modification of native or engineered antibodies via chemical or enzymatic methods. Whatever method is employed, controlling the payload-antibody ratio (PAR) is a challenge in terms of multiple aspects including: (i) obtaining homogeneous protein conjugates; (ii) obtaining unusual PARs (PAR is rarely other than 2, 4 or 8); (iii) using a single method to access a range of different PARs; (iv) applicability to various antibody formats; and (v) flexibility for the production of heterofunctional antibody-conjugates (e.g. attachment of multiple types of payloads). In this article, we report a single pyridazinedione-based trifunctional dual bridging linker that enables, in a two-step procedure (re-bridging/click), the generation of either mAb-, Fab′-, or Fc-conjugates from native mAb, (Fab′)2 or Fc formats, respectively. Fc and (Fab′)2 formats were generated via enzymatic digestion of native mAbs. Whilst the same reduction and re-bridging protocols were applied to all three of the protein formats, the subsequent click reaction(s) employed to graft payload(s) drove the generation of a range of PARs, including heterofunctional PARs. As such, exploiting click reactivity and/or orthogonality afforded mAb-conjugates with PARs of 6, 4, 2 or 4 + 2, and Fab′- and Fc-conjugates with a PAR of 3, 2, 1 or 2 + 1 on-demand. We believe that the homogeneity, novelty and variety in accessible PARs, as well as the applicability to various antibody-conjugate formats enabled by our non-recombinant method could be a suitable tool for antibody-drug conjugates optimisation (optimal PAR value, optimal payloads combination) and boost the development of new antibody therapeutics (Fab′- and Fc-conjugates)

Texture refinement framework for improved video coding

ISBN: 978-0-8194-7936-5 - WOSInternational audienceH.264/AVC standard offers an efficient way of reducing the noticeable artefacts of former video coding schemes, but it can be perfectible for the coding of detailed texture areas. This paper presents a conceptual coding framework, utilizing visual perception redundancy, which aims at improving both bit-rate and quality on textured areas. The approach is generic and can be integrated into usual coding scheme. The proposed scheme is divided into three steps: a first algorithm analyses texture regions, with an eye to build a dictionary of the most representative texture sub-regions (RTS). The encoder preserves then them at a higher quality than the rest of the picture, in order to enable a refinement algorithm to finally spread the preserved information over textured areas. In this paper, we present a first solution to validate the framework, detailing then the encoder side in order to define a simple method for dictionary building. The proposed H.264/AVC compliant scheme creates a dictionary of macroblock

Modular Chemical Construction of IgG-like Mono- and Bispecific Synthetic Antibodies (SynAbs)

In recent years there has been rising interest in the field of protein−protein conjugation, especially related to bispecific antibodies (bsAbs) and their therapeutic applications. These constructs contain two paratopes capable of binding two distinct epitopes on target molecules and are thus able to perform complex biological functions (mechanisms of action) not available to monospecific mAbs. Traditionally these bsAbs have been constructed through protein engineering, but recently chemical methods for their construction have started to (re)emerge. While these have been shown to offer increased modularity, speed, and for some methods even the inherent capacity for further functionalization (e.g., with small molecule cargo), most of these approaches lacked the ability to include a fragment crystallizable (Fc) modality. The Fc component of IgG antibodies offers effector function and increased half-life. Here we report a first-in-class disulfide rebridging and click-chemistry-based method for the generation of Fc-containing, IgG-like mono- and bispecific antibodies. These are in the FcZ-(FabX)-FabY format, i.e., two distinct Fabs and an Fc, potentially all from different antibodies, attached in a homogeneous and covalent manner. We have dubbed these molecules synthetic antibodies (SynAbs). We have constructed a T cellengager (TCE) SynAb, FcCD20-(FabHER2)-FabCD3, and have confirmed that it exhibits the expected biological functions, including the ability to kill HER2+ target cells in a coculture assay with T cells

Conception, synthèse et activité biologique de vecteurs peptidiques pour le ciblage et/ou la thérapie du cancer

This thesis work is about conception, synthesis and biological activities of peptide vectors for diagnostic and/or therapeutic applications against cancer.We used cyclodecapeptidic scaffolds chemoselectively handled with targeting elements and effectors. Those scaffolds presenting four c[RGDfK] ligands have a strong affinity for integrin avB3 receptors, wich are overexpressed in various cancers and by endothelial cells from the tumor surrounding. They are poorly expressed in healthy tissues. The multivalent presentation of -RGD- motifs higly increases the internalisation of the scaffold by tumor cells. Thus we developed molecules composed by four -RGD- motifs for tumor targeting, and different effectors for various applications. Thanks to multiple collaborations, we linked the vector to a highly cytotoxic compound (cryptophycine), a photosensitiser (DHP), a pro-apoptotic peptide (BAX), a DOTA complex (for 68-Ga complexation, for PET applications). We also grafted the cyclodecapeptide bearing four -RGD- motifs to polymers or silica nanoparticles, both fluorescent.The main project of this thesis was the conception of dual targeting vectors. Our objective was to simultaneously target two receptors overexpressed in the tumor periphery. Beside the targeting of avB3, we decided to target the NRP1 receptor, which is also overexpressed during tumor angiogenesis. We exploited various chemoselective reactions (oxime, huisgen cycloaddition, peptide coupling) to synthesise fluorescent vectors targeting one of the two receptors, or both. In vitro and in vivo biological experiments were realised. We discovered that dual targeting compounds allow a really good tumor detection, but inferior to mono targeting ones. Nevertheless, the cellular answer triggered by dual targeting compounds is totally different from those obtained with other compounds, including co-injection. We found different elements that tend to show that a NRP1-vector-integrin could be formed, and would be blocked inside the cellular membrane, resulting in its internalisation's blocking.Ces travaux de thèse portent sur la conception, la synthèse et l'étude biologique de vecteurs peptidiques pour des applications en diagnostic et/ou thérapie du cancer.Nous avons utilisé des châssis cyclodécapeptidiques fonctionnalisés de façon chimiosélective par des éléments de ciblage et des effecteurs. Ces châssis, dotés de quatre ligands c[RGDfK], possèdent une forte affinité pour les récepteurs intégrine avB3 qui sont surexprimés dans de nombreux cancers et par les cellules endothéliales de l'environnement tumoral. Ils sont en revanche peu exprimés par les tissus sain. La présentation multivalente des ligands -RGD- permet également au châssis d'être internalisé par les cellules tumorales.Nous avons donc mis au point des molécules composées du châssis peptidique, de quatre ligands -RGD- pour le ciblage tumoral, et de différents effecteurs pour plusieurs applications. A travers de multiples collaborations, nous avons relié ce vecteur à un agent hautement toxique (cryptophycine), un photosensibilisateur (DHP), un peptide pro-apoptotique (BAX), un complexe de Gallium 68 (pour une étude clinique de phase I pour une application en imagerie TEP). Nous avons également greffé ces châssis présentant quatre motifs -RGD- à des polymères ou à des nanoparticules de silice, tous deux fluorescents.Le projet principal de cette thèse était la conception de vecteurs ciblant deux récepteurs tumoraux de manière simultanée. En plus de cibler l'intégrine avB3, nous avons ciblé le récepteur NRP1 qui est lui aussi surexprimé lors de l'angiogenèse tumorale. Nous avons exploité divers réactions chimiosélectives (oxime, cycloaddition de Huisgen, amidation) pour concevoir des vecteurs fluorescents ciblant l'un des deux récepteurs ou les deux à la fois. Des tests biologiques in vitro et in vivo ont été réalisés. Il s'avère que les composés à double ciblage permettent une très bonne détection de la tumeur, mais non supérieure à des composés à mono ciblage. En revanche, la réponse cellulaire déclenchée par les composés à double ciblage est unique, et totalement différente d'une co-injection. Nous avons plusieurs éléments qui tendent à prouver qu'un complexe NRP1-vecteur-Intégrine se formerait et resterait ancré au niveau de la membrane cellulaire, bloquant son internalisation

Design, synthesis and biological activity of peptidic vectors for the diagnostic and/or therapy of tumours

Ces travaux de thèse portent sur la conception, la synthèse et l'étude biologique de vecteurs peptidiques pour des applications en diagnostic et/ou thérapie du cancer.Nous avons utilisé des châssis cyclodécapeptidiques fonctionnalisés de façon chimiosélective par des éléments de ciblage et des effecteurs. Ces châssis, dotés de quatre ligands c[RGDfK], possèdent une forte affinité pour les récepteurs intégrine avB3 qui sont surexprimés dans de nombreux cancers et par les cellules endothéliales de l'environnement tumoral. Ils sont en revanche peu exprimés par les tissus sain. La présentation multivalente des ligands -RGD- permet également au châssis d'être internalisé par les cellules tumorales.Nous avons donc mis au point des molécules composées du châssis peptidique, de quatre ligands -RGD- pour le ciblage tumoral, et de différents effecteurs pour plusieurs applications. A travers de multiples collaborations, nous avons relié ce vecteur à un agent hautement toxique (cryptophycine), un photosensibilisateur (DHP), un peptide pro-apoptotique (BAX), un complexe de Gallium 68 (pour une étude clinique de phase I pour une application en imagerie TEP). Nous avons également greffé ces châssis présentant quatre motifs -RGD- à des polymères ou à des nanoparticules de silice, tous deux fluorescents.Le projet principal de cette thèse était la conception de vecteurs ciblant deux récepteurs tumoraux de manière simultanée. En plus de cibler l'intégrine avB3, nous avons ciblé le récepteur NRP1 qui est lui aussi surexprimé lors de l'angiogenèse tumorale. Nous avons exploité divers réactions chimiosélectives (oxime, cycloaddition de Huisgen, amidation) pour concevoir des vecteurs fluorescents ciblant l'un des deux récepteurs ou les deux à la fois. Des tests biologiques in vitro et in vivo ont été réalisés. Il s'avère que les composés à double ciblage permettent une très bonne détection de la tumeur, mais non supérieure à des composés à mono ciblage. En revanche, la réponse cellulaire déclenchée par les composés à double ciblage est unique, et totalement différente d'une co-injection. Nous avons plusieurs éléments qui tendent à prouver qu'un complexe NRP1-vecteur-Intégrine se formerait et resterait ancré au niveau de la membrane cellulaire, bloquant son internalisation.This thesis work is about conception, synthesis and biological activities of peptide vectors for diagnostic and/or therapeutic applications against cancer.We used cyclodecapeptidic scaffolds chemoselectively handled with targeting elements and effectors. Those scaffolds presenting four c[RGDfK] ligands have a strong affinity for integrin avB3 receptors, wich are overexpressed in various cancers and by endothelial cells from the tumor surrounding. They are poorly expressed in healthy tissues. The multivalent presentation of -RGD- motifs higly increases the internalisation of the scaffold by tumor cells. Thus we developed molecules composed by four -RGD- motifs for tumor targeting, and different effectors for various applications. Thanks to multiple collaborations, we linked the vector to a highly cytotoxic compound (cryptophycine), a photosensitiser (DHP), a pro-apoptotic peptide (BAX), a DOTA complex (for 68-Ga complexation, for PET applications). We also grafted the cyclodecapeptide bearing four -RGD- motifs to polymers or silica nanoparticles, both fluorescent.The main project of this thesis was the conception of dual targeting vectors. Our objective was to simultaneously target two receptors overexpressed in the tumor periphery. Beside the targeting of avB3, we decided to target the NRP1 receptor, which is also overexpressed during tumor angiogenesis. We exploited various chemoselective reactions (oxime, huisgen cycloaddition, peptide coupling) to synthesise fluorescent vectors targeting one of the two receptors, or both. In vitro and in vivo biological experiments were realised. We discovered that dual targeting compounds allow a really good tumor detection, but inferior to mono targeting ones. Nevertheless, the cellular answer triggered by dual targeting compounds is totally different from those obtained with other compounds, including co-injection. We found different elements that tend to show that a NRP1-vector-integrin could be formed, and would be blocked inside the cellular membrane, resulting in its internalisation's blocking

Multimeric RGD-Based Strategies for Selective Drug Delivery to Tumor Tissues

RGD peptides have received a lot of attention over the two last decades, in particular to improve tumor therapy through the targeting of the αVβ3 integrin receptor. This review focuses on the molecular design of multimeric RGD compounds, as well as the design of suitable linkers for drug delivery. Many examples of RGD–drug conjugates have been developed, and we show the importance of RGD constructs to enhance binding affinity to tumor cells, as well as their drug uptake. Further, we also highlight the use of RGD peptides as theranostic systems, promising tools offering dual modality, such as tumor diagnosis and therapy. In conclusion, we address the challenging issues, as well as ongoing and future development, in comparison with large molecules, such as monoclonal antibodies

Recent, non-classical, approaches to antibody lysine modification

International audienc

Multimeric RGD-Based Strategies for Selective Drug Delivery to Tumor Tissues

RGD peptides have received a lot of attention over the two last decades, in particular to improve tumor therapy through the targeting of the αVβ3 integrin receptor. This review focuses on the molecular design of multimeric RGD compounds, as well as the design of suitable linkers for drug delivery. Many examples of RGD–drug conjugates have been developed, and we show the importance of RGD constructs to enhance binding affinity to tumor cells, as well as their drug uptake. Further, we also highlight the use of RGD peptides as theranostic systems, promising tools offering dual modality, such as tumor diagnosis and therapy. In conclusion, we address the challenging issues, as well as ongoing and future development, in comparison with large molecules, such as monoclonal antibodies

Spatiotemporal texture synthesis and region-based motion compensation for video compression

International audienceIn this paper, a content-based approach for video compression is proposed. The main novelty relies on the complete texture analysis/synthesis framework, which enables the use of multiple algorithms, depending on texture characteristics. The idea comes from the efficient MPEG prediction based on a best mode selection. Existing synthesis algorithms cannot be efficient in synthesizing every kind of texture but a certain range of them. This approach is designed to be jointly used with current and future standard compression schemes. At encoder side, texture analysis includes segmentation and characterization tools, in order to localize candidate regions for synthesis: motion compensation or texture synthesis. The corresponding areas are not encoded. The decoder fills them using texture synthesis. The remaining regions in images are classically encoded. They can potentially serve as input for texture synthesis. The chosen tools are developed and adapted in order to ensure the coherency of the whole scheme. Thus, a texture characterization step provides required parameters to the texture synthesizer. Two texture synthesizers, including a pixel-based and a patch-based approach, are used on different types of texture, complementing each other. The scheme is coupled with a motion estimator in order to segment coherent regions and to interpolate rigid motions using an affine model. Inter frame adapted synthesis is therefore used for non-rigid texture regions. The framework has been validated within an H.264/MPEG4-AVC video codec. Experimental results show significant bit-rate saving at similar visual quality levels, assessed using subjective tests. The method can be coupled with the future HEVC in which blocks can be skipped by the encoder to be synthesized at decoder side