pH-Degradable mannosylated nanogels for dendritic cell targeting

We report on the design of glycosylated nanogels via core-cross linking of amphiphilic non-water-soluble block copolymers composed of an acetylated glycosylated block and a pentafluorophenyl (PFP) activated ester block prepared by reversible addition fragmentation (RAFT) polymerization. Self-assembly, pH-sensitive core-cross-linking, and removal of remaining PFP esters and protecting groups are achieved in one pot and yield fully hydrated sub-100 nm nanogels. Using cell subsets that exhibit high and low expression of the mannose receptor (MR) under conditions that suppress active endocytosis, we show that mannosylated but not galactosylated nanogels can efficiently target the MR that is expressed on the cell surface of primary dendritic cells (DCs). These nanogels hold promise for immunological applications involving DCs and macrophage subsets

Dysregulated innate and adaptive immune responses discriminate disease severity in COVID-19

The clinical spectrum of COVID-19 varies and the differences in host response characterizing this variation have not been fully elucidated. COVID-19 disease severity correlates with an excessive pro-inflammatory immune response and profound lymphopenia. Inflammatory responses according to disease severity were explored by plasma cytokine measurements and proteomics analysis in 147 COVID-19 patients. Furthermore, peripheral blood mononuclear cell cytokine production assays and whole blood flow cytometry were performed. Results confirm a hyperinflammatory innate immune state, while highlighting hepatocyte growth factor and stem cell factor as potential biomarkers for disease severity. Clustering analysis reveals no specific inflammatory endotypes in COVID-19 patients. Functional assays reveal abrogated adaptive cytokine production (interferon-gamma, interleukin-17 and interleukin-22) and prominent T cell exhaustion in critically ill patients, whereas innate immune responses were intact or hyperresponsive. Collectively, this extensive analysis provides a comprehensive insight into the pathobiology of severe to critical COVID-19 and highlight potential biomarkers of disease severity

Gene Therapy for Fibrodysplasia Ossificans Progressiva: Feasibility and Obstacles

Fibrodysplasia ossificans progressiva (FOP) is a rare and devastating genetic disease, in which soft connective tissue is converted into heterotopic bone through an endochondral ossification process. Patients succumb early as they gradually become trapped in a second skeleton of heterotopic bone. Although the underlying genetic defect is long known, the inherent complexity of the disease has hindered the discovery of effective preventions and treatments. New developments in the gene therapy field have motivated its consideration as an attractive therapeutic option for FOP. However, the immune system\u27s role in FOP activation and the as-yet unknown primary causative cell, are crucial issues which must be taken into account in the therapy design. While gene therapy offers a potential therapeutic solution, more knowledge about FOP is needed to enable its optimal and safe application

Synthetic carbohydrate biomaterials for innate immune-engineering

Cancer still remains one of the leading causes of death, responsible for one in six deaths globally according to the WHO. Until recently cancer treatment was founded on three classical pillars: surgical removal of tumors if possible (1), often in combination with high energy X-ray radiotherapy (2) and/or chemotherapeutics (3).1 But faced with serious limitations including unresponsive neoplasms, very moderate results, severe side effects, huge comorbidities, and lack of specificity, scientific research was pushed to devise more effective therapies. By looking into the underlying mechanisms that caused cancer, new treatment options have emerged. Amongst these, hormone therapy and targeted therapies were introduced in the past decades to tackle a select group of specific malignancies. Recent insights in the immuno-oncology field sparked the development of cancer immunotherapy. Here the mechanisms on which tumor cells rely to actively evade immune surveillance and escape cell death have led to a plethora of new therapies that make use of the patient’s own immune system. Cancer immunotherapy has revolutionized the oncology field, creating new and better therapeutic avenues for patients suffering neoplasms that are sometimes unresponsive to more traditional treatments. Although immune-based therapies have been at the origin of spectacular remissions, it still faces significant drawbacks. These include life-threatening side effects, 20-30 % response rates in treated patients and high treatment costs. Hence, there is an urgent need for more effective immunotherapy to either improve upon or complement the current state of the art. In this dissertation, we focused on the design of novel carbohydrate based biomaterials to help engineer an immune response directed against cancer cells. We investigated the potential of mannosylated nanogels as a potential carrier for in vivo cancer vaccination and antibody-recruiting polymers that can act as an in situ cancer vaccine

Replication package for the Docker Inheritance network analysis

<p><strong>Container Image Inheritance on DockerHub: Empirical Analysis and Insights</strong></p> <p>This repository represents a replication package for our SCAM paper on DockerHub inheritance network.</p> <p>This replication package requires Python 3.5+ to be installed.</p> <p>These experiments were executed on a Linux Ubuntu OS.</p> <p>This replication package contains four folders:<br> - data: contains all datasets required.<br> - scripts: contains all scripts needed to collect the data.<br> - notebooks: contains notebooks where we analyze data. <br> - figures: contains figures saved from the notebooks.</p> <p>To obtain the analysis used in the paper, one should execute ``jupyter notebook`` at the root of this replication package, and open the notebook contained in ``notebooks``.</p> <p>The data is under the Creative Commons Attribution Share-Alike 4.0 license. The source code is under the GNU General Public License.</p&gt

Cell surface clicking of antibody-recruiting polymers to metabolically azide-labeled cancer cells

Triggering antibody-mediated innate immune mechanisms to kill cancer cells is an attractive therapeutic avenue. In this context, recruitment of endogenous antibodies to the cancer cell surface could be a viable alternative to the use of monoclonal antibodies. We report on antibody-recruiting polymers containing multiple antibody-binding hapten motifs and cyclooctynes that can covalently conjugate to azides introduced onto the glycocalyx of cancer cells by metabolic labeling with azido sugars

Tyrosine-Triazolinedione Bioconjugation as Site-Selective Protein Modification Starting from RAFT-Derived Polymers

The electrophilic aromatic substitution (S_EAr) reaction of triazolinediones (TADs) with the phenol moiety of tyrosine amino acid residues is a potent method for the site-selective formation of polymer–protein conjugates. Herein, using poly­(<i>N</i>,<i>N</i>-dimethylacrylamide) (pDMA) and bovine serum albumin (BSA) as model reagents, the performance of this tyrosine-TAD bioconjugation in aqueous solutions is explored. At first, reversible addition–fragmentation chain transfer (RAFT) polymerization with a functional urazole, a precursor for TAD, chain transfer agent is used for the synthesis of a TAD end-functionalized pDMA. Eventually, the BSA ligation efficiency and selectivity of this polymer was evaluated in different aqueous solvent mixtures using SDS-PAGE and mass spectroscopy after trypsin digestion

Transiently responsive protein–polymer conjugates via a 'grafting-from' RAFT approach for intracellular co-delivery of proteins and immune-modulators

We report on transiently responsive protein-polymer conjugates that temporarily change their protein conformation from the soluble to the particle-like state. 'Grafting-from' RAFT polymerization of a dioxolane-containing acrylamide with a protein macroCTA is used to design polymer-protein conjugates that self-assemble into nano-particles at physiological temperature and pH. Acid triggered hydrolysis of the dioxolane units into diolmoeities rendered the conjugates fully water soluble irrespective of temperature

Well-defined polymer-paclitaxel prodrugs by a grafting-from-drug approach

We report on the design of a polymeric prodrug of the anticancer agent paclitaxel (PTX) by a grafting-from-drug approach. A chain transfer agent for reversible addition fragmentation chain transfer (RAFT) polymerization was efficiently and regioselectively linked to the C2' position of paclitaxel, which is crucial for its bioactivity. Subsequent RAFT polymerization of a hydrophilic monomer yielded well-defined paclitaxel-polymer conjugates with high drug loading, water solubility, and stability. The versatility of this approach was further demonstrated by w-end post-functionalization with a fluorescent tracer. In vitro experiments showed that these conjugates are readily taken up into endosomes where native PTX is efficiently cleaved off and then reaches its subcellular target. This was confirmed by the cytotoxicity profile of the conjugate, which matches those of commercial PTX formulations based on mere physical encapsulation