Harnessing tumor angiogenesis to explore ovarian cancer immune suppression and address target-therapies outcomes

In 2020, ovarian cancer still remains the "Biggest Enemy" faced by Gynecology Oncologists, due to the lack of biological and clinical tools for early diagnosis, high recurrence rates despite the recent introduction of targeted-therapies for the management of advanced disease, and 5-year overall survival below 40%. In particular, high grade serous ovarian cancer (HGSOC), recently classified as a Type II ovarian carcinoma, still accounts alone for the 80% of all ovarian cancer deaths. In light of these numbers and to the urgent need to reduce mortality from high grade serous ovarian cancer, in 2015 an international group of opinion leaders summarized a comprehensive list of priority issues which are still considered as “unmet needs” in the understanding of high grade serous ovarian cancer. Among these issues, the Authors identify the following biological landscapes to be explored for HGSOC patients’s future prognosis change: 1) to exploit HGSOC patients’ immune response and interaction between host immune system and tumor microenvironment; 2) to analyse recurrent and end-stage disease samples, in order to shed light on acquired resistance mechanisms; 3) to integrate all –omics data on individual samples with immune and other tumor microenvironment components in primary and recurrent samples. As a consequence, the research work carried out during this international PhD programme, and here discussed, aimed to add answers to these universally recognized priority issues in ovarian cancer understanding. In particular, the leading research line developed and carried out aimed to explore the modulation of tumor-derived neoangiogenesis during ovarian cancer progression and the role of anti-angiogenetic agents in reverting tumor immune suppressive circuit thus triggering host’s anti-tumoral immunological response. The 3 research studies here presented* were both carried out in the context of the “OCTIPS Consortium”, a 7th Framework Program research Project, funded by the European Commission in 2012, which currently holds in its biobank at Charité Medical University (Berlin, Germany) one of the largest European collection of paired primary and recurrent ovarian cancers samples; and in the context of the “Laboratory of Tumor Immunology and Cell therapy Unit“ directed by Prof. Marianna Nuti, at Sapienza University (Rome, Italy).Im Jahr 2020, bleibt das Ovarialkarzinom noch der schlimmste der gyneco onkologische Erkrankungen wegen diesen Gruenden: 1. Fehlen einer wirksamen screeing Methode 2. hohe Rezidivrate, trotz der neuen “Target therapies” fuer die fortgeschrittenen Stadien dieser Krankheit 3. kurzes 5-Jahren gesamt-Ueberleben (<40%) Bei dem fortgeschrittenen seroesen Eierstockkrebs (high grade serous ovarian cancer, HGSOC), kuerzlich genauso bekannt wie “Ovarialkarzinoma Typ II”, liegt die Sterblichkeit bei 80% unter allen Todesfaelle vom Ovarialkarzinom. Im Jahr 2015, ist eine Liste von unbeantworteten Schwerpunkten in Ovarialkarzinoma von einer Internationalen Expertengruppe erstellt worden. Zu diesem Ziel, konzentrieren sich die Autoren auf den biologischen Hintergrund von HGSOC, wie folgt: 1) Beziehung zwischen die Tumor-Mikroumgebung und das Immunsystem 2) Gewebeanalysen zur Bestimmung von neu aufgetretenen Resistenz Mechanism 3) Integration mit omics Daten von Proben mit immunologische und andere Tumor mivroinironment Komponenten. Demensprechend ist die Forschung die waehrend diesem internationalen Phd Programm durchgefuehrt wurde, und hier discutiert wird, zur Vorstellung von Antworten zu diesen prioritaeren Fragestellungen zum Verstaendnis vom Ovarialkarzinom gerichtet. Die folgende Forschungslinien wurden entwickelt: Untersuchung der Modulation von der Tumor gesteuerte Neoangiogenese zur Zeit der Progession von Ovarialkarzinom und von der Rolle die von den antiagiogenteischen Medikamenten in der Aenderung der Tumor assoziierte immunologische Hemmung spielen. Die 3 Studien die hier vorgestellt werden* sind im Rahmen von dem “OCTIPS Consortium” einen Forschungsprojeckt der von der Europaeische Kommission 2012 finanziert wurde, durchgefuehrt worden. Die Biobank vom OPTICS Consortium ist einer der groessten in Europa und enthaelt Proben von Ovarialkarzinome zur Zeit der Diagnose sowie zur Zeit vom Rezidiv und befindet sich an der Charite’ Medical University (Berlin, Germany). Zusaetzlich hat es eine Kollaboration mit dem Laboratory of Tumor Immunology and Cell therapy Unit“ unter der Leitung von Frau Prof. Marianna Nuti, an der Universitaet Sapienza (Rom, Italien) gegeben

Immunobiology of solid cancers: cellular and molecular pathways as potential diagnostic and therapeutic targets

In the last four decades, tumor immunology has shed light on identity and functions of cells and molecules involved in tumor rejection through the involvement of the immune system [1]. Several groups of immune cells have been demonstrated to be able to contrast tumor occurrence and tumor progression by killing immunogenic tumor cells, a phenomenon recognized under the definition of “immunosurveillance” [2]. Unfortunately, cancer may evade immunosurveillance and progress through the modifications of its own antigens, which can reduce tumor immunogenicity and/or increase its immunosuppressive action [3]. After years of investigations, harnessing the immune system to attack cancer has recently led scientists to gather enough clinical data to show what a powerful sword immunotherapy can be

Tumor-derived microvesicles modulate antigen cross-processing via reactive oxygen species-mediated alkalinization of phagosomal compartment in dendritic cells

Dendritic cells (DCs) are the only antigen-presenting cells able to prime naïve T cells and cross-prime antigen-specific CD8+ T cells. Their functionality is a requirement for the induction and maintenance of long-lasting cancer immunity. Albeit intensively investigated, the in vivo mechanisms underlying efficient antigen cross-processing and presentation are not fully understood. Several pieces of evidence indicate that antigen transfer to DCs mediated by microvesicles (MVs) enhances antigen immunogenicity. This mechanism is also relevant for cross-presentation of those tumor-associated glycoproteins such as MUC1 that are blocked in HLA class II compartment when internalized by DCs as soluble molecules. Here, we present pieces of evidence that the internalization of tumor-derived MVs modulates antigen-processing machinery of DCs. Employing MVs derived from ovarian cancer ascites fluid and established tumor cell lines, we show that MV uptake modifies DC phagosomal microenvironment, triggering reactive oxygen species (ROS) accumulation and early alkalinization. Indeed, tumor MVs carry radical species and the MV uptake by DCs counteracts the chemically mediated acidification of the phagosomal compartment. Further pieces of evidence suggest that efficacious antigen cross-priming of the MUC1 antigen carried by the tumor MVs results from the early signaling induced by MV internalization and the function of the antigen-processing machinery of DCs. These results strongly support the hypothesis that tumor-derived MVs impact antigen immunogenicity by tuning the antigen-processing machinery of DCs, besides being carrier of tumor antigens. Furthermore, these findings have important implications for the exploitation of MVs as antigenic cell-free immunogen for DC-based therapeutic strategies

Effects of commercial formulations of glyphosate on marine crustaceans and implications for risk assessment under temperature changes.

Abstract Glyphosate-based formulations are the most commonly used herbicides worldwide with the risk of potential contamination of aquatic bodies. The present study assessed the response of four marine crustaceans to three different brands of herbicides Roundup®Platinum, Efesto® and Taifun® MK CL.T, under two selected temperatures of 20 °C and 30 °C. The harpacticoid copepod Tigriopus fulvus, the anostracan Artemia franciscana, the amphipod Corophium insidiosum and the isopod Sphaeroma serratum were chosen as testing organisms. Effects of herbicides and temperatures were assessed by estimating lethal concentrations. The results showed that the high temperature rises the toxicity of glyphosate with an increase of mortality of all the tested species. This is an important aspect for future risk assessments of pesticides under global climate change scenarios. Efesto® resulted the most toxic brand, showing C. insidiosum the most sensitive with 96 h-LC50 values of 3.25 mg/L acid equivalent (a.e.) at 30 °C and 7.94 mg/L a.e. at 20 °C followed by T. fulvus while A. franciscana and S. serratum were the less sensitive. This study provides important information for assessing the toxic effects of three different brands of glyphosate-based herbicides on non-target marine organisms suggesting that they should be carefully managed to minimize any negative impact on marine organisms

Rast, smrtnost i prinos olige (Atherina boyeri Risso, 1810) iz lagune Lesina (Jadransko more, Italija)

Population structure, growth, age, mortality and exploitation status of Atherina boyeri, caught in the period from June 2013 to May 2014 in the Lesina lagoon (Apulia region, southern Adriatic) was studied. Samples were taken each month by using beach seines . The length–weight relationship of all sand smell specimens was described by the equation W=0.012*TL 3.10; (R2= 0.84). Population parameters including the asymptotic length (L∞) and growth coefficient (K) were assessed to evaluate the stock status. The recruitment pattern was modeled with a FiSAT routine. The asymptotic length (L∞) was 111.0 mm, while the growth coefficient (K) was 0.68 year−1. The growth performance index ((φ’) reached 3.92. The total mortality coefficient “Z”, the natural mortality coefficient “M” and the fishing mortality coefficient “F” were estimated as 2.24, 1.48 and 0.76 year-1, respectively. Exploitation rate for the population of A. boyeri in the Lesina lagoon estimated as 0.34 was resulted still lower than the predicted maximum value of Emax 0.59. The probability of capture indicated that L50 was of 55.57 mm, indicating that the stock of sand smelt in Lesina lagoon is not being over-fished.Istraživana je struktura populacije, rast, starost, smrtnost i status eksploatacije olige Atherinaboyeri, ulovljene u razdoblju od lipnja 2013. do svibnja 2014. u laguni Lesina (regija Apulija, južni Jadran). Uzorci su uzimani svaki mjesec pomoću mreže potegače.Dužinsko-maseni odnos opisan je jednadžbom W = 0,012 * TL 3,10; (R2 = 0,84). Utvrđeni su parametri populacije, uključujući asimptotsku duljinu (L∞) i koeficijent rasta (K), kako bi se procijenilo stanje zaliha. Obrazac novačenja modeliran je FiSAT rutinom. Asimptotska duljina (L∞) iznosila je 111,0 mm, dok je koeficijent rasta (K) iznosio 0,68 godina − 1. Indeks učinka rasta (φ’) dosegnuo je 3,92. Ukupni koeficijent smrtnosti „Z“, koeficijent prirodne smrtnosti „M“ i koeficijent mortaliteta ribolova „F“ procijenjeni su na 2,24, 1,48 i 0,76 godina-1.Stopa eksploatacije populacije olige u Lesinskoj laguni procijenjena na 0,34 rezultirala je i dalje nižom od predviđene maksimalne vrijednosti Emax 0,59

Microgel particles with distinct morphologies and common chemical compositions: a unified descrip-tion of the responsivity to temperature and osmotic stress

Poly(N-isopropylacrylamide) (PNIPAM) hydrogel microparticles with different core-shell morphologies have been designed, while maintaining an unvaried chemical composition: a morphology with (i) an un-crosslinked core with a crosslinked shell of PNIPAM chains and (ii) PNIPAM chains crosslinked to form the core with a shell consisting of tethered un-crosslinked PNIPAM chains to the core. Both morphologies with two different degrees of crosslinking have been assessed by confocal microscopy and tested with respect to their temperature responsivity and deformation by applying an osmotic stress. The thermal and mechanical behavior of these architectures have been framed within a Flory-Rehner modified model in order to describe the microgel volume shrinking occurring as response to a temperature increase or an osmotic perturbation. This study provides a background for assessing to what extent the mechanical features of the microgel particle surface affect the interactions occurring at the interface of a microgel particle with a cell, in addition to the already know ligand/receptor interaction. These results have direct implications in triggering a limited phagocytosis of microdevices designed as injectable drug delivery systems

Tumor derived Microvesicles enhance cross-processing ability of clinical grade Dendritic Cells

Tumor cells release extracellular microvesicles (MVs) in the microenvironment to deliver biological signals to neighbouring cells as well as to cells in distant tissues. Tumor-derived MVs appear to play contradictory role promoting both immunosuppression and tumor growth and both evoking tumor specific immune response. Recent evidences indicate that tumor-derived MVs can positively impact Dendritic Cells (DCs) immunogenicity by reprogramming DC antigen processing machinery and intracellular signaling pathways, thus promoting anti-tumor response. DCs are considered pivot cells of the immune system due to their exclusive ability to coordinate the innate and acquired immune responses, cross-present exogenous antigens and prime naïve T cells. DCs are required for the induction and maintenance of long-lasting anti-tumor immunity and their exploitation has been extensively investigated for the design of anti-tumor vaccines. However, the clinical grade culture conditions that are required to generate DCs for therapeutic use can strongly affect their functions. Here, we investigated the immunomodulatory impact of MVs carrying the MUC1 tumor glycoantigen (MVsMUC1) as immunogen formulation on clinical grade DCs grown in X-VIVO 15 (X-DCs). Results indicated that X-DCs displayed reduced performance of the antigen processing machinery in term of diminished phagocytosis and acidification of the phagosomal compartment suggesting an altered immunogenicity of clinical grade DCs. Pulsing DCs with MVsMUC1 restored phagosomal alkalinization, triggering ROS increase. This was not observed when a soluble MUC1 protein was employed (rMUC1). Concurrently, MVsMUC1 internalization by X-DCs allowed MUC1 cross-processing. Most importantly, MVsMUC1 pulsed DCs activated IFNγ response mediated by MUC1 specific CD8+ T cells. These results strongly support the employment of tumor-derived MVs as immunogen platforms for the implementation of DC-based vaccine

Estimativas da diversidade genética de acessos de Paspalum spp. com ouso de marcadores microssatélites.

Editores técnicos: João de Mendonça Naime, Caue Ribeiro, Maria Alice Martins, Elaine Cristina Paris, Paulino Ribeiro Villas Boas, Ladislau Marcelino Rabello

Triple peptide vaccination as consolidation treatment in women affected by ovarian and breast cancer: clinical and immunological data of a phase I/II clinical trial

Vaccination with priming and expansion of tumour reacting T cells is an important therapeutic option to be used in combination with novel checkpoint inhibitors to increase the specificity of the T cell infiltrate and the efficacy of the treatment. In this phase I/II study, 14 high-risk disease-free ovarian (OC) and breast cancer (BC) patients after completion of standard therapies were vaccinated with MUC1, ErbB2 and carcinoembryonic antigen (CEA) HLA-A2+-restricted peptides and Montanide. Patients were subjected to 6 doses of vaccine every two weeks and a recall dose after 3 months. ECOG grade 2 toxicity was observed at the injection site. Eight out of 14 patients showed specific CD8+ T cells to at least one antigen. None of 4 patients vaccinated for compassionate use showed a CD8 activation. An OC patient who suffered from a lymph nodal recurrence, showed specific anti-ErbB2 CD8+ T cells in the bulky aortic lymph nodes suggesting homingof the activated T cells. Results confirm that peptide vaccination strategy is feasible, safe and well tolerated. In particular OC patients appear to show a higher response rate compared to BC patients. Vaccination generates a long-lasting immune response, which is strongly enhanced by recall administrations. The clinical outcome of patients enrolled in the trial appears favourable, having registered no deceased patients with a minimum follow-up of 8 years. These promising data, in line with the results of similar studies, the high compliance of patients observed and the favourable toxicity profile, support future trials of peptide vaccination in clinically disease-free patients who have completed standard treatments