Memory NK cell features exploitable in anticancer immunotherapy

Besides their innate ability to rapidly produce effector cytokines and kill virus-infected or transformed cells, natural killer (NK) cells display a strong capability to adapt to environmental modifications and to differentiate into long-lived, hyperfunctional populations, dubbed memory or memory-like NK cells. Despite significant progress in the field of NK cell-based immunotherapies, some factors including their short life span and the occurrence of a tumor-dependent functional exhaustion have limited their clinical efficacy so that strategies aimed at overcoming these limitations represent one of the main current challenges in the field. In this scenario, the exploitation of NK cell memory may have a considerable potential. This article summarizes recent evidence in the literature on the peculiar features that render memory NK cells an attractive tool for antitumor immunotherapy, including their long-term survival and in vivo persistence, the resistance to tumor-dependent immunosuppressive microenvironment, the amplified functional responses to IgG-opsonized tumor cells, and in vitro expansion capability. Along with highlighting these issues, we speculate that memory NK cell-based adoptive immunotherapy settings would greatly take advantage from the combination with tumor-targeting therapeutic antibodies (mAbs), as a strategy to fully unleash their clinical efficacy

Perianal-fistelassoziiertes Karzinom bei Morbus Crohn: eine multizentrische retrospektive Fallkontrollstudie

Perianale fistelassoziierte Karzinome sind eine seltene, aber schwerwiegende Komplikation bei Patienten mit Morbus Crohn. Es fehlt an einheitlichen Leitlinien für die Diagnose, Behandlung und Nachsorge dieser Untergruppe von Patienten. Außerdem ist es nicht klar, welche Patienten gefährdet sind und für welche ein Screening-Programm sinnvoll wäre. Ziel dieser Studie ist, die Population der Patienten mit perianalen fistelassoziierten Karzinomen bei Morbus Crohn herauszuarbeiten sowie die Risikofaktoren, die Diagnose, den Verlauf und die Prognose zu analysieren. Diese multizentrische retrospektive Fall-Kontroll-Studie wurde an vier deutschen Krankenhäusern durchgeführt. Die Analyse umfasste vierzig Patienten mit nachgewiesenem Malignom in Verbindung mit perianalen Fisteln bei Morbus Crohn und vierzig Kontrollen mit fistulierendem Morbus Crohn ohne Nachweis eines anorektalen Karzinoms. Univariate sowie multivariate statistische Analysen wurden durchgeführt, um Risikofaktoren für die Krebsentwicklung zu identifizieren und um das Überleben zu beschreiben. Histologisch ergab sich ein Adenokarzinom bei 33/40 Patienten und ein Plattenepithelkarzinom bei 7/40 Patienten. Die klinischen Risikofaktoren für die Entwicklung eines fistelassoziierten Karzinoms waren: transsphinktäre Fisteltyp (OR=3,37, 95%CI: 1,07- 10,61), postoperativ persistierende Fistel (OR=4,00, 95%CI: 1,48-10,79) komplexe Fisteln (OR=7,21, 95%CI: 1,48-25,07) chronische Fistelaktivität (OR=3,44, 95%CI 1,31-9,06). Signifikante multivariate Hazard Ratios für die Gesamtmortalität und das progressionsfreie Überleben wurden für den histologischen Krebstyp, die metastasierte Erkrankung und die R1- Resektion gezeigt. Das Gesamtüberleben betrug 45.1 ± 28.6 Monate und die 5-Jahres- Überlebensrate lag bei 65%. Zusammenfassend lässt sich sagen, dass eine frühe Diagnose für einen günstigen Krankheitsverlauf entscheidend ist, während die Behandlung der chronischen Fistelaktivität der Schlüssel zur Verhinderung von Malignität sein kann. Eine spezialisierte multimodale Therapie ist für die erfolgreiche Behandlung von perianalen fistelassoziierten Malignomen von größter Bedeutung

Design of hybrid gels based on gellan-cholesterol derivative and P90G liposomes for drug depot applications

Gels are extensively studied in the drug delivery field because of their potential benefits in therapeutics. Depot gel systems fall in this area, and the interest in their development has been focused on long-lasting, biocompatible, and resorbable delivery devices. The present work describes a new class of hybrid gels that stem from the interaction between liposomes based on P90G phospholipid and the cholesterol derivative of the polysaccharide gellan. The mechanical properties of these gels and the delivery profiles of the anti-inflammatory model drug diclofenac embedded in such systems confirmed the suitability of these hybrid gels as a good candidate for drug depot applications

Obinutuzumab-mediated high-affinity ligation of FcγRIIIA/CD16 primes NK cells for IFNγ production

Natural killer (NK) cell-mediated antibody-dependent cellular cytotoxicity (ADCC), based on the recognition of IgG-opsonized targets by the low-affinity receptor for IgG FcγRIIIA/CD16, represents one of the main mechanisms by which therapeutic antibodies (mAbs) mediate their antitumor effects. Besides ADCC, CD16 ligation also results in cytokine production, in particular, NK-derived IFNγ is endowed with a well-recognized role in the shaping of adaptive immune responses. Obinutuzumab is a glycoengineered anti-CD20 mAb with a modified crystallizable fragment (Fc) domain designed to increase the affinity for CD16 and consequently the killing of mAb-opsonized targets. However, the impact of CD16 ligation in optimized affinity conditions on NK functional program is not completely understood. Herein, we demonstrate that the interaction of NK cells with obinutuzumab-opsonized cells results in enhanced IFNγ production as compared with parental non-glycoengineered mAb or the reference molecule rituximab. We observed that affinity ligation conditions strictly correlate with the ability to induce CD16 down-modulation and lysosomal targeting of receptor-associated signaling elements. Indeed, a preferential degradation of FcεRIγ chain and Syk kinase was observed upon obinutuzumab stimulation independently from CD16-V158F polymorphism. Although the downregulation of FcεRIγ/Syk module leads to the impairment of cytotoxic function induced by NKp46 and NKp30 receptors, obinutuzumab-experienced cells exhibit an increased ability to produce IFNγ in response to different stimuli. These data highlight a relationship between CD16 aggregation conditions and the ability to promote a degradative pathway of CD16-coupled signaling elements associated to the shift of NK functional progra

Enhancing the Sustainability of Social Housing Policies through the Social Impact Approach: Innovative Perspectives form a “Paris Affordable Housing Challenge” Project in France

The environmental, economic and social challenges re-launched in the European Union Agendas (e.g., Horizon 2020 and Europe 2020–2030) have recently returned to being highly debated. In particular, policies and interventions in the field of social housing (SH) are still remaining crucial issues for urban regeneration. These interventions are aimed to combine sustainability criteria with architectural, urban and environmental quality. In this context, our goal in this article is to provide an innovative perspective on the topic highlighting the positive returns enabled by the logic of the social impact approach (SIA). A pilot project is proposed to be performed in the VI arrondissement of Paris. Starting from the French regulatory context and the requirements set by the “Paris Affordable Housing Challenge” competition, the levers of social finance for new social demands and the levers of incentives are applied to a real case. The research results show that the application of the emerging principles of social impact investing (SII) in areas difficult to access in the private market had positive returns. The final aim of the article is to outline guidelines that consider the quality, management and generation of the social impact requirements highlighted in the proposal to facilitate the application of the SIA to other interventions and contexts

Acute myocardial infarction and stroke registries. The italian experience

Cardiovascular diseases (CVD) are the leading causes of death and hospitalisation in nearly all European countries and accounted for almost 40% of all deaths in 2013. With the exception of few rigorous but limited studies carried out in some geographical areas, available data on CVD incidence and prevalence are generally limited and of poor quality, despite the magnitude of the CVD phenomenon. The EUROCISS Project, supported by the Health Monitoring Programme of the DG SANCO from 2000 to 2007, provided general guidance and updated methods for the surveillance of Acute Myocardial Infarction (AMI) and Stroke. The Italian population-based registry of major coronary and cerebrovascular events was set up following EUROCISS Project recommendations; it also took into account the experience acquired by Italy in the WHO-MONICA project since the mid-1980s and continued with the coordination of the EUROCISS Project. The project: “A population-based AMI register: assessing the feasibility for a pilot study to implement a surveillance system of AMI in Mediterranean countries according to EUROCISS recommendations”, in the framework of the EuroMed Programme, followed major practical and operative issues for the implementation of a population-based registry for coronary and cerebrovascular events, which are here described. This paper includes the definition of target population, data sources, events, indicators, quality methods, and the description of a software used to implement the registry

Challenges and opportunities in establishing an Health Examination Survey

In Italy, the last 30 years witnessed the implementation of cross-sectional surveys providing baseline data on numerous risk factors collected from random samples of the adult general population. In order to support those groups who would like to implement an health examination survey (HES), according to the experience of  the CUORE Project surveys, the objective of this paper is to describe some information related to the organization of a survey (examination sites and sampling, selection of analytic laboratory, coordination and personnel involved, sample selection, recruitment and appointment scheduling, informative notice and informed consent, participation rate, non-participation bias, quality assurance, survey data, long term storage of the samples, internal quality control, external quality assessment, feedback to participants, error checking, correction and documentation of the data, transfer and storage of the data, statistical analyses and interpretation of results, dissemination of results), usually shortly described  in scientific papers but relevant when an HES is planned

Tumor-Targeting Anti-CD20 Antibodies Mediate In Vitro Expansion of Memory Natural Killer Cells: Impact of CD16 Affinity Ligation Conditions and In Vivo Priming

Natural Killer (NK) cells represent a pivotal player of innate anti-tumor immune responses. The impact of environmental factors in shaping the representativity of different NK cell subsets is increasingly appreciated. Human Cytomegalovirus (HCMV) infection profoundly affects NK cell compartment, as documented by the presence of a CD94/NKG2C+Fc∝RI≥- long-lived “memory” NK cell subset, endowed with enhanced CD16-dependent functional capabilities, in a fraction of HCMV seropositive subjects. However, the requirements for memory NK cell pool establishment/maintenance and activation have not been fully characterised yet. Here we describe the capability of anti-CD20 tumor-targeting therapeutic monoclonal antibodies (mAbs) to drive the selective in vitro expansion of memory NK cells, and we show the impact of donor' HCMV serostatus and CD16 affinity ligation conditions on this event. In vitro expanded memory NK cells maintain the phenotypic and functional signature of their freshly isolated counterpart; furthermore, our data demonstrate that CD16 affinity ligation conditions differently affect memory NK cell proliferation and functional activation, as rituximab-mediated low-affinity ligation represents a superior proliferative stimulus, while high-affinity aggregation mediated by glycoengineered obinutuzumab results in improved multifunctional responses. Our work also expands the molecular and functional characterization of memory NK cells, and investigates the possible impact of CD16 functional allelic variants on their in vivo and in vitro expansion. These results reveal new insights in Ab-driven memory NK cell responses in a therapeutic setting, and may ultimately inspire new NK cell-based intervention strategies against cancer, in which the enhanced responsiveness to mAb-bound target could significantly impact therapeutic efficacy

Caveolin-1 implicated as a pro-invasive gene in high-grade glioma cell models: implementation of a 3d spheroid matrix invasion assay

INTRODUCTION: The poor prognosis associated with Glioblastoma multiforme (GBM) is multifactorial but includes the capacity of residual tumour cells not removed by surgery and resistant to radio-/chemo-therapy undergoing diffuse invasion into the surrounding brain tissue. Caveolin-1 (Cav-1) is the major structural and functional component of caveolae. In a number of tumour types Cav-1 is recognised to participate in cytoskeletal rearrangement, integrin-mediated adhesion and/or matrix remodelling. We proposed Cav-1 serves to promote invasion of GBM cells. To investigate this we have employed in an in-vitro 3D cellular invasion assay. METHOD: The human GBM cell lines, UP007 and UP029 established from primary cultures of biopsy-derived brain tumours (University of Portsmouth), U-87 MG (ECACC) and U-373 MG (ECACC) were genetically modified to stably knock-out Cav-1 using a Lentiviral Cav-1 shRNA approach; corresponding stably transfected non-target (NT) shRNA cell lines were generated as controls. Neuropheres were formed and embedded within an extracellular matrix (Matrigel™). Over a two-/four-day period (depending on cell line) the migration of cells away from the neurophere core (CORE) was quantified by image capture and processing (Image J) using a custom-developed MatLab script for pixel density analysis indicative of the density of migrating cellular material. RESULTS: Cav-1 knockout resulted in significant (P0.05) towards reduced invasion. Depending upon the cell line the Cav-1 knockdown also resulted in reduced size and cellular density of the neurosphere core (UP007 and UP029) indicative of reduced proliferation and/or cell survival capacity. CONCLUSION: Using an in-vitro 3D cellular invasion assay we have found Cav-1 expression in a series of three GBM cell lines to promote cellular invasion capacity. Ongoing studies are addressing signalling mechanisms and the influence of the microenvironment

Discovering Business Processes models expressed as DNF or CNF formulae of Declare constraints

In the field of Business Process Management, the Process Discovery task is one of the most important and researched topics. It aims to automatically learn process models starting from a given set of logged execution traces. The majority of the approaches employ procedural languages for describing the discovered models, but declarative languages have been proposed as well. In the latter category there is the Declare language, based on the notion of constraint, and equipped with a formal semantics on LTLf. Also, quite common in the field is to consider the log as a set of positive examples only, but some recent approaches pointed out that a binary classification task (with positive and negative examples) might provide better outcomes. In this paper, we discuss our preliminary work on the adaptation of some existing algorithms for Inductive Logic Programming, to the specific setting of Process Discovery: in particular, we adopt the Declare language with its formal semantics, and the perspective of a binary classification task (i.e., with positive and negative examples