​Department of Biotechnology and Biosciences

Intestinal microbiota and gut-born T helper-17 (Th17) lymphocytes may act as drivers of smoldering multiple myeloma (SMM) to MM evolution. We demonstrate here that administering the human commensal Prevotella melaninogenica to transgenic Vk*MYC mice affected by Early-MM, mimicking human SMM, significantly delayed evolution to full-blown MM. Mechanistically, P. melaninogenica increased the production of short-chain fatty acids (SCFAs), preventing skew of dendritic cells towards a pro-Th17 phenotype and accumulation of Th17 cells in the bone marrow of treated mice. P. melaninogenica or butyrate synergized with anti-PD-L1 antibodies by restraining Th17 cell expansion while unleashing immune checkpoint blockade (ICB)-induced effector CD8+ T cells. P. melaninogenica also attenuated IL-17-mediated skin lesions that mimicked ICB-induced immune-related adverse events. Thus, modulation of the gut microbiota or SCFAs administration with or without ICB might represent treatment options for patients affected by plasma cell dyscrasias and other hematologic or solid tumors where IL-17 acts as driving force. The present dataset contains: - 1H NMR spectra acquired on control and growth media for PM and PH; - CMP and .exp files created for metabolite quantification in MestreNova software by SMA analysis.

Supplementary data and underlying data supporting the finding described in "Production and carbon footprint of microbial oil from waste lemon peel extract". Abstract Background The agricultural sector is one of the leading producers of agro-industrial solid organic waste. This waste is mainly disposed of by incineration or landfilled, representing a huge loss of potential resources, which could be used for the production of high-value chemicals. In this study, a fermentation process for the production of microbial oil from waste lemon extract (LE), an aqueous side stream deriving from waste lemon peel and pulp processing, was developed and assessed for impacts. Microbial oil can have many and diverse applications, from plasticisers in plastic and rubber compounds to moisturizers in cosmetic formulation. Methods and results Characterization of LE revealed that its autoclaving process is effective for increasing the concentration of readily available glucose and fructose, reaching 28.77 ± 0.08 g L-1 and 25.68 ± 0.27 g L-1. Nitrogen content was measured too, revealing a C/N ratio of 85, optimal for triggering lipid accumulation in the selected microbial cell factory. Therefore, the oleaginous yeast Cutaneotrichosporon oleaginosum was cultivated in an unmodified LE-based medium in 2 L bioreactors, resulting in a lipid accumulation of 0.47 ± 0.08 goil gCDW-1. Finally, a new lipid extraction method using green solvents was developed, which allowed to extract and purify 11.29 g of oils, corresponding to 35% of the cell dry weight. The carbon footprint of this laboratory-scale production was estimated to be 71 - 434 kgCO2eq kg-1 microbial oil, with electricity consumption of the fermentation step as the main factor. Simulation of the process in a 300L fermenter suggests that the electricity consumption, and therefore the overall impact, can be drastically reduced with scale-up. Conclusions The proposed process is promising in terms of production and has the advantage of not being in competition with edible resources and land use. However, the microbial oil yield and the extraction process must be optimized to make the process sustainable.

NMR File of the paper "Cleaner synthesis of preclinically validated vaccine adjuvants" in publication on Frontiers in Chemistry

Metabolite libraries built from NMR spectra for the metabolomics analysis of antarctic soils extracts, using the Simple Mixture Analysis (SMA) tool implemented in MestreNova 14.3.0-30573 software.

Metabolite library built from NMR spectra for metabolomic analysis of cheese whey permeate (CWP) using the Simple Mixture Analysis (SMA) tool implemented in MestreNova 14.3 software. Metabolites included: 2,3-Butandiol, Acetate, Alanine, Betaine, Choline, Citrate, Creatine, Creatinine, Ethanol, Fumarate, Hippurate, Lactate, Lactose, Orotate, Succinate, α-D-Galactose, β-D-Galactose Sample CWP in PB 10 mM (10% D2O) pH 7.4, TSP (1mM), Diluition Factor (10%) File included: SMA library .exp files and optimized .cmp files

Abstract Background The microbial production of succinic acid (SA) from renewable carbon sources via the reverse TCA (rTCA) pathway is a process potentially accompanied by net-fixation of carbon dioxide (CO2). Among reduced carbon sources, glycerol is particularly attractive since it allows a nearly twofold higher CO2-fixation yield compared to sugars. Recently, we described an engineered Saccharomyces cerevisiae strain which allowed SA production in synthetic glycerol medium with a maximum yield of 0.23 Cmol Cmol−1. The results of that previous study suggested that the glyoxylate cycle considerably contributed to SA accumulation in the respective strain. The current study aimed at improving the flux into the rTCA pathway accompanied by a higher CO2-fixation and SA yield. Results By changing the design of the expression cassettes for the rTCA pathway, overexpressing PYC2, and adding CaCO3 to the batch fermentations, an SA yield on glycerol of 0.63 Cmol Cmol−1 was achieved (i.e. 47.1% of the theoretical maximum). The modifications in this 2nd-generation SA producer improved the maximum biomass-specific glycerol consumption rate by a factor of nearly four compared to the isogenic baseline strain solely equipped with the dihydroxyacetone (DHA) pathway for glycerol catabolism. The data also suggest that the glyoxylate cycle did not contribute to the SA production in the new strain. Cultivation conditions which directly or indirectly increased the concentration of bicarbonate, led to an accumulation of malate in addition to the predominant product SA (ca. 0.1 Cmol Cmol−1 at the time point when SA yield was highest). Off-gas analysis in controlled bioreactors with CO2-enriched gas-phase indicated that CO2 was fixed during the SA production phase. Conclusions The data strongly suggest that a major part of dicarboxylic acids in our 2nd-generation SA-producer was formed via the rTCA pathway enabling a net fixation of CO2. The greatly increased capacity of the rTCA pathway obviously allowed successful competition with other pathways for the common precursor pyruvate. The overexpression of PYC2 and the increased availability of bicarbonate, the co-substrate for the PYC reaction, further strengthened this capacity. The achievements are encouraging to invest in future efforts establishing a process for SA production from (crude) glycerol and CO2.

Additional file 4: Figure S4 Intracellular Ca2+ mobilization in CD34+ HSPCs w/wo GLP-1R stimulation. A Spontaneous Ca2+ transients elicited in single FLUO4 loaded cells before and during LIRA treatment. B Mean FLUO4 fluorescence before and after LIRA injection from a population of CD34+ HSPCs (N = 8).

Additional file 5.

Additional file 6.

Abstract Background Glioblastoma multiforme (GBM) is an incurable tumor, with a median survival rate of only 14–15 months. Along with heterogeneity and unregulated growth, a central matter in dealing with GBMs is cell invasiveness. Thus, improving prognosis requires finding new agents to inhibit key multiple pathways, even simultaneously. A subset of GBM stem-like cells (GSCs) may account for tumorigenicity, representing, through their pathways, the proper cellular target in the therapeutics of glioblastomas. GSCs cells are routinely enriched and expanded due to continuous exposure to specific growth factors, which might alter some of their intrinsic characteristic and hide therapeutically relevant traits. Methods By removing exogenous growth factors stimulation, here we isolated and characterized a subset of GSCs with a “mitogen-independent” phenotype (I-GSCs) from patient’s tumor specimens. Differential side-by-side comparative functional and molecular analyses were performed either in vitro or in vivo on these cells versus their classical growth factor (GF)-dependent counterpart (D-GSCs) as well as their tissue of origin. This was performed to pinpoint the inherent GSCs’ critical regulators, with particular emphasis on those involved in spreading and tumorigenic potential. Transcriptomic fingerprints were pointed out by ANOVA with Benjamini-Hochberg False Discovery Rate (FDR) and association of copy number alterations or somatic mutations was determined by comparing each subgroup with a two-tailed Fisher’s exact test. The combined effects of interacting in vitro and in vivo with two emerging GSCs’ key regulators, such as Wnt5a and EphA2, were then predicted under in vivo experimental settings that are conducive to clinical applications. In vivo comparisons were carried out in mouse-human xenografts GBM model by a hierarchical linear model for repeated measurements and Dunnett’s multiple comparison test with the distribution of survival compared by Kaplan–Meier method. Results Here, we assessed that a subset of GSCs from high-grade gliomas is self-sufficient in the activation of regulatory growth signaling. Furthermore, while constitutively present within the same GBM tissue, these GF-independent GSCs cells were endowed with a distinctive functional and molecular repertoire, defined by highly aggressive Wnt5aHigh/EphA2Low profile, as opposed to Wnt5aLow/EphA2High expression in sibling D-GSCs. Regardless of their GBM subtype of origin, I-GSCs, are endowed with a raised in vivo tumorigenic potential than matched D-GSCs, which were fast-growing ex-vivo but less lethal and invasive in vivo. Also, the malignant I-GSCs’ transcriptomic fingerprint faithfully mirrored the original tumor, bringing into evidence key regulators of invasiveness, angiogenesis and immuno-modulators, which became candidates for glioma diagnostic/prognostic markers and therapeutic targets. Particularly, simultaneously counteracting the activity of the tissue invasive mediator Wnt5a and EphA2 tyrosine kinase receptor addictively hindered GSCs’ tumorigenic and invasive ability, thus increasing survival. Conclusion We show how the preservation of a mitogen-independent phenotype in GSCs plays a central role in determining the exacerbated tumorigenic and high mobility features distinctive of GBM. The exploitation of the I-GSCs' peculiar features shown here offers new ways to identify novel, GSCs-specific effectors, whose modulation can be used in order to identify novel, potential molecular therapeutic targets. Furthermore, we show how the combined use of PepA, the anti-Wnt5a drug, and of ephrinA1-Fc to can hinder GSCs’ lethality in a clinically relevant xenogeneic in vivo model thus being conducive to perspective, novel combinatorial clinical application.