Stratification of ovarian cancer borderline from high-grade serous carcinoma patients by quantitative serum NMR spectroscopy of metabolites, lipoproteins, and inflammatory markers

Background: Traditional diagnosis is based on histology or clinical-stage classification which provides no information on tumor metabolism and inflammation, which, however, are both hallmarks of cancer and are directly associated with prognosis and severity. This project was an exploratory approach to profile metabolites, lipoproteins, and inflammation parameters (glycoprotein A and glycoprotein B) of borderline ovarian tumor (BOT) and high-grade serous ovarian cancer (HGSOC) for identifying additional useful serum markers and stratifying ovarian cancer patients in the future.Methods: This project included 201 serum samples of which 50 were received from BOT and 151 from high-grade serous ovarian cancer (HGSOC), respectively. All the serum samples were validated and phenotyped by 1H-NMR-based metabolomics with in vitro diagnostics research (IVDr) standard operating procedures generating quantitative data on 38 metabolites, 112 lipoprotein parameters, and 5 inflammation markers. Uni- and multivariate statistics were applied to identify NMR-based alterations. Moreover, biomarker analysis was carried out with all NMR parameters and CA-125.Results: Ketone bodies, glutamate, 2-hydroxybutyrate, glucose, glycerol, and phenylalanine levels were significantly higher in HGSOC, while the same tumors showed significantly lower levels of alanine and histidine. Furthermore, alanine and histidine and formic acid decreased and increased, respectively, over the clinical stages. Inflammatory markers glycoproteins A and B (GlycA and GlycB) increased significantly over the clinical stages and were higher in HGSOC, alongside significant changes in lipoproteins. Lipoprotein subfractions of VLDLs, IDLs, and LDLs increased significantly in HGSOC and over the clinical stages, while total plasma apolipoprotein A1 and A2 and a subfraction of HDLs decreased significantly over the clinical stages. Additionally, LDL triglycerides significantly increased in advanced ovarian cancer. In biomarker analysis, glycoprotein inflammation biomarkers behaved in the same way as the established clinical biomarker CA-125. Moreover, CA-125/GlycA, CA-125/GlycB, and CA-125/Glycs are potential biomarkers for diagnosis, prognosis, and treatment response of epithelial ovarian cancer (EOC). Last, the quantitative inflammatory parameters clearly displayed unique patterns of metabolites, lipoproteins, and CA-125 in BOT and HGSOC with clinical stages I–IV.Conclusion:1H-NMR-based metabolomics with commercial IVDr assays could detect and identify altered metabolites and lipoproteins relevant to EOC development and progression and show that inflammation (based on glycoproteins) increased along with malignancy. As inflammation is a hallmark of cancer, glycoproteins, thereof, are promising future serum biomarkers for the diagnosis, prognosis, and treatment response of EOC. This was supported by the definition and stratification of three different inflammatory serum classes which characterize specific alternations in metabolites, lipoproteins, and CA-125, implicating that future diagnosis could be refined not only by diagnosed histology and/or clinical stages but also by glycoprotein classes

Loss of function of RIMS2 causes a syndromic congenital cone-rod synaptic disease with neurodevelopmental and pancreatic involvement

Congenital cone-rod synaptic disorder (CRSD), also known as incomplete congenital stationary night blindness (iCSNB), is a non-progressive inherited retinal disease (IRD) characterized by night blindness, photophobia, and nystagmus, and distinctive electroretinographic features. Here, we report bi-allelic RIMS2 variants in seven CRSD-affected individuals from four unrelated families. Apart from CRSD, neurodevelopmental disease was observed in all affected individuals, and abnormal glucose homeostasis was observed in the eldest affected individual. RIMS2 regulates synaptic membrane exocytosis. Data mining of human adult bulk and single-cell retinal transcriptional datasets revealed predominant expression in rod photoreceptors, and immunostaining demonstrated RIMS2 localization in the human retinal outer plexiform layer, Purkinje cells, and pancreatic islets. Additionally, nonsense variants were shown to result in truncated RIMS2 and decreased insulin secretion in mammalian cells. The identification of a syndromic stationary congenital IRD has a major impact on the differential diagnosis of syndromic congenital IRD, which has previously been exclusively linked with degenerative IRD

Antibody-guided in vivo imaging of Aspergillus fumigatus lung infections during anti-fungal azole treatment

This is the final version. Available on open access from Nature Research via the DOI in this recordData availability: Due to their large size, the raw imaging data that support the findings of this study are directly available from the corresponding authors upon reasonable request. Derived data have been compiled in the Source Data file provided with this paper. Any remaining data supporting the findings from this study are available from the corresponding author upon reasonable request.Invasive pulmonary aspergillosis (IPA) is a life-threatening lung disease of immunocompromised humans, caused by the opportunistic fungal pathogen Aspergillus fumigatus. Inadequacies in current diagnostic procedures mean that early diagnosis of the disease, critical to patient survival, remains a major clinical challenge, and is leading to the empiric use of antifungal drugs and emergence of azole resistance. A non-invasive procedure that allows both unambiguous detection of IPA, and its response to azole treatment, is therefore needed. Here, we show that a humanised Aspergillus-specific monoclonal antibody, dual labelled with a radionuclide and fluorophore, can be used in immunoPET/MRI in vivo and 3D light sheet fluorescence microscopy ex vivo to quantify early A. fumigatus lung infections and to monitor the efficacy of azole therapy. Our antibody-guided approach reveals that early drug intervention is critical to prevent complete invasion of the lungs by the fungus, and demonstrates the power of molecular imaging as a non-invasive procedure for tracking IPA in vivo.Ministry of Culture and Science of North Rhine-WestphaliaGoverning Mayor of Berlin including Science and ResearchFederal Ministry of Education and ResearchEuropean Union FP7Werner Siemens Foundatio

Optimisation du concept d'inhibition de Cyclooxygénase dans le traitement du cancer de la prostate

Acetylsalicylic acid, a molecule found in willow bark's extracts, has been used for thousands of years as an antiinflammatory and analgesic agent. During the 80's, the discovery of cyclooxygenase as a target for this enzyme inhibitor revived the interest in non steroidal anti-inflammatory drugs (NSAIDs) research. The existence of several isoforms of COX has been proven more recently, and each one has been associated with different effects NSAIDs can have. This led to the emergence of therapeutics targeting a single isoform, in an attempt to lower the side effects of non-specific drugs. Even if still discussed, the usefulness of this new type of molecules in the cancer pathology has recently been showed, particularly on prostate cancer cells. Our study consists in optimizing cyclooxygenase inhibitors to achieve more efficacy in chemotherapy and chemoprevention of this kind of disease. First of all, the use of a peptide vector that can be recognized and cleaved by the prostate specific antigen (a protease active exclusively in the prostate) while carrying a COX-2 inhibitor and capable of, after cleavage followed by intramolecular rearrangement, releasing the active moiety appealed to us in a targeted therapy approach. The conception, synthesis and study of a novel spacer allowing us to link the COX-2 inhibitor to a peptide while permitting a cleaving – rearrangement sequence to occur after PSA recognition were conducted and gave promising results. In parallel, adding to a COX inhibitor an other active moiety under the form of a prodrug was looked into. Nitric oxide (NO), an ubiquitous radical messenger, displays surprising pharmacological properties in a wide array of tissues, be it as a vasodilatator, gastric mucosa protector and even in cancer chemotherapy. The synthesis of a wide panel of innovative compounds, COX-2 selective or not, able to release both NO and the inhibitor seemed interesting, especially when considering the amount of contradictory bibliographic data presenting the mode of action of this type of drugs. Once obtained, the in-depth analysis of the metabolic pathway underwent by our molecules had to be studied in order to shed light on the biological mechanisms underlying their effects. Using various techniques, such as ex vivo vasodilatation experiments on rat aorta strips and electron spin resonance measurements, we were able to access metabolization kinetics of these prodrugs; while classical enzymatic inhibition assays on human whole blood and proliferation assays on human prostatic cancer cell lines revealed the therapeutic interest of our compounds. Finally, combining the peptide vector and the NO donor on a cyclooxygenase inhibitor was attempted.L'acide acétylsalicylique, molécule dérivée d'extraits de saule, est utilisé depuis des milliers d'années comme antiinflammatoire et analgésique. La découverte au cours des années 80 de la cyclooxygénase, cible de cet inhibiteur enzymatique, a relancé la recherche de nouveaux anti-inflammatoires non stéroïdiens (AINS). C'est la mise en évidence de l'existence de plusieurs isoformes de la COX et l'attribution des effets pharmacologiques des inhibiteurs de cyclooxygénase à ces différents isoformes qui a permis l'émergence de nouvelles thérapeutiques minimisant les effets secondaires gastriques des AINS. Bien qu'encore controversé, l'intérêt de ce type de composés dans la pathologie cancéreuse a été récemment montré, en particulier dans le cancer de la prostate. Notre étude consiste en l'optimisation de composés inhibiteurs de cyclooxygénase afin de les rendre plus efficaces dans la chimioprévention et la chimiothérapie de ce type de maladie. Tout d'abord, la vectorisation d'un inhibiteur sélectif COX-2 par un peptide pouvant être reconnu et clivé par l'antigène prostatique spécifique (protéase active uniquement au niveau de la prostate), et capable, après ce clivage, d'amorcer une réaction intramoléculaire conduisant à la libération du principe actif, nous a paru pertinente dans le cadre d'une thérapie ciblée. La conception, la synthèse et l'étude d'un espaceur novateur permettant de lier l'inhibiteur COX au peptide et d'obtenir ce type de réarrangement ont ainsi été menées et ont débouché sur des résultats prometteurs. En parallèle, l'adjonction d'une seconde entité active à un composé inhibiteur COX sous la forme d'une prodrug a été envisagée. Le monoxyde d'azote NO, radical messager ubiquitaire, possède des propriétés pharmacologiques étonnantes dans un grand nombre de tissus, que ce soit en vasodilatation, protection des muqueuses gastriques mais également dans le cadre du traitement d'un cancer. Synthétiser un nombre important de composés, inhibiteurs sélectifs ou non de COX-2, pouvant, après métabolisation libérer à la fois NO et l'inhibiteur nous a semblé intéressant, en particulier compte tenu de la controverse actuelle sur les modes d'actions réels de ces composés. Après leurs synthèses, l'analyse approfondie du devenir métabolique de ces molécules a dû être menée pour tenter de comprendre leur prise en charge par le système enzymatique. C'est par le biais de différentes évaluations pharmacologiques aussi variées que l'étude de la vasodilatation aortique ex-vivo ou la résonance paramagnétique électronique, que les cinétiques de métabolisation ont pu être abordées ; et c'est en réalisant des mesures d'inhibitions enzymatiques (sur sang humain) et de prolifération cellulaire (sur lignées cancéreuses humaines) que le potentiel thérapeutique de nos composés a pu être estimé. Enfin, la combinaison de ces deux approches par la conception et la synthèse de composés inhibiteurs de cyclooxygénase / donneurs de NO vectorisés a été abordée

Advances in the in vivo molecular imaging of invasive pulmonary aspergillosis

This is the final version. Available on open access from MDPI via the DOI in this recordInvasive pulmonary aspergillosis (IPA) is a life-threatening infection of immunocompromised patients with Aspergillus fumigatus, a ubiquitous environmental mould. While there are numerous functioning antifungal therapies, their high cost, substantial side effects and fear of overt resistance development preclude permanent prophylactic medication of risk-patients. Hence, a fast and definitive diagnosis of IPA is desirable, to quickly identify those patients that really require aggressive antimycotic treatment and to follow the course of the therapeutic intervention. Yet, despite decades of research into this issue such a diagnostic procedure is still not available. Here we discuss the array of currently available methods for IPA detection and their limits. We then show that molecular imaging using positron-emission-tomography (PET) combined with morphological computed tomography or magnetic imaging is highly promising to become a future non-invasive approach for IPA diagnosis and therapy monitoring, albeit still requiring thorough validation and relying on further acceptance and dissemination of the approach. Thereby our approach using the A. fumigatus-specific humanized monoclonal antibody hJF5 labelled with 64Cu as PET-tracer has proven highly effective in pre-clinical models and hence bears high potential for human application.European Union FP

Monoclonal antibody-targeted PEGylated liposome-ICG encapsulating doxorubicin as a potential theranostic agent

Indocyanine green (ICG) is an FDA-approved, strongly photo-absorbent/fluorescent probe that has been incorporated into a clinically-relevant PEGylated liposome as a flexible optoacoustic contrast agent platform. This study describes the engineering of targeted PEGylated liposome-ICG using the anti-MUC-1 "humanized" monoclonal antibody (MoAb) hCTM01 as a tumour-specific theranostic system. We aimed to visualise non-invasively the tumour accumulation of these MoAb-targeted liposomes over time in tumour-bearing mice using multispectral optoacoustic tomography (MSOT). Preferential accumulation of targeted PEGylated liposome-ICG was studied after intravenous administration in comparison to non-targeted PEGylated liposome-ICG using both fast growing (4T1) and slow growing (HT-29) MUC-1 positive tumour models. Monitoring liposomal ICG in the tumour showed that both targeted and non-targeted liposome-ICG formulations preferentially accumulated into the tumour models studied. Rapid accumulation was observed for targeted liposomes at early time points mainly in the periphery of the tumour volume suggesting binding to available MUC-1 receptors. In contrast, non-targeted PEGylated liposomes showed accumulation at the centre of the tumour at later time points. In an attempt to take this a step further, we successfully encapsulated the anticancer drug, doxorubicin (DOX) into both targeted and non-targeted PEGylated liposome-ICG. The engineering of DOX-loaded targeted ICG liposome systems present a novel platform for combined tumour-specific therapy and diagnosis. This can open new possibilities in the design of advanced image-guided cancer therapeutics

Metabolic stability of superoxide and hydroxyl radical adducts of a cyclic nitrone toward rat liver microsomes and cytosol: A stopped-flow ESR spectroscopy study

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