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

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

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

International audienc

siRNA liposome-gold nanorod vectors for multispectral optoacoustic tomography theranostics.

Therapeutic applications of gene silencing using siRNA have seen increasing interest over the past decade. The optimization of the delivery and biodistribution of siRNA using liposome-gold nanorod (AuNRs) nanoscale carriers can greatly benefit from adept imaging methods that can visualize the time-resolved delivery performance of such vectors. In this work, we describe the effect of AuNR length incorporated with liposomes and show their complexation with siRNA as a novel gene delivery vehicle. We demonstrate the application of multispectral optoacoustic tomography (MSOT) to longitudinally visualize the localisation of siRNA carrying liposome-AuNR hybrids within tumors. Combination of in vivo MSOT with ex vivo fluorescence cryo-slice imaging offers further insight into the siRNA transport and activity obtained

High-Resolution Multispectral Optoacoustic Tomography of the Vascularization and Constitutive Hypoxemia of Cancerous Tumors

Diversity of the design and alignment of illumination and ultrasonic transducers empower the fine scalability and versatility of optoacoustic imaging. In this study, we implement an innovative high-resolution optoacoustic mesoscopy for imaging the vasculature and tissue oxygenation within subcutaneous and orthotopic cancerous implants of mice in vivo through acquisition of tomographic projections over 180° at a central frequency of 24 MHz. High-resolution volumetric imaging was combined with multispectral functional measurements to resolve the exquisite inner structure and vascularization of the entire tumor mass using endogenous and exogenous optoacoustic contrast. Evidence is presented for constitutive hypoxemia within the carcinogenic tissue through analysis of the hemoglobin absorption spectra and distribution. Morphometric readouts obtained with optoacoustic mesoscopy have been verified with high-resolution ultramicroscopic studies. The findings described herein greatly extend the applications of optoacoustic mesoscopy toward structural and multispectral functional measurements of the vascularization and hemodynamics within solid tumors in vivo and are of major relevance to basic and preclinical oncological studies in small animal models

Lactate production precedes inflammatory cell recruitment in arthritic ankles : an imaging study

Purpose: Inflammation is involved in many disease processes. However, accurate imaging tools permitting diagnosis and characterization of inflammation are still missing. As inflamed tissues exhibit a high rate of glycolysis, pyruvate metabolism may offer a unique approach to follow the inflammatory response and disease progression. Therefore, the aim of the study was to follow metabolic changes and recruitment of inflammatory cells after onset of inflammation in arthritic ankles using hyperpolarized 1-13C-pyruvate magnetic resonance spectroscopy (MRS) and 19F magnetic resonance imaging (MRI), respectively. Procedure: Experimental rheumatoid arthritis (RA) was induced by intraperitoneal injection of glucose-6-phosphate-isomerase-specific antibodies (GPI) containing serum. To monitor pyruvate metabolism, the transformation of hyperpolarized 1-13C-pyruvate into hyperpolarized 1-13C-lactate was followed using MRS. To track phagocytic immune cell homing, we intravenously injected a perfluorocarbon emulsion 48 h before imaging. The animals were scanned at days 1, 3, or 6 after GPI-serum injection to examine the different stages of arthritic inflammation. Finally, to confirm the pyruvate metabolic activity and the link to inflammatory cell recruitment, we conducted hematoxylin-eosin histopathology and monocarboxylase transporter (MCT-1) immune histochemistry (IHC) of inflamed ankles. Results: Hyperpolarized 1-13C-pyruvate MRS revealed a high rate of lactate production immediately at day 1 after GPI-serum transfer, which remained elevated during the progression of the disease, while 19F-MRI exhibited a gradual recruitment of phagocytic immune cells in arthritic ankles, which correlated well with the course of ankle swelling. Histopathology and IHC revealed that MCT-1 was expressed in regions with inflammatory cell recruitment, confirming the metabolic shift identified in arthritic ankles. Conclusions: Our study demonstrated the presence of a very early metabolic shift in arthritic joints independent of phagocytic immune cell recruitment. Thus, hyperpolarized 1-13C-pyruvate represents a promising tracer to monitor acute arthritic joint inflammation, even with minor ankle swelling. Furthermore, translated to the clinics, these methods add a detailed characterization of disease status and could substantially support patient stratification and therapy monitoring