The GSK3\u3b2 inhibitor BIS I reverts YAP-dependent EMT signature in PDAC cell lines by decreasing SMADs expression level

The Yes-associated protein, YAP, is a transcriptional co-activator, mediating the Epithelial to Mesenchymal Transition program in pancreatic ductal adenocarcinoma (PDAC). With the aim to identify compounds that can specifically modulate YAP functionality in PDAC cell lines, we performed a small scale, drug-based screening experiment using YAP cell localization as the read-out. We identified erlotinib as an inducer of YAP cytoplasmic localization, an inhibitor of the TEA luciferase reporter system and the expression of the bona fide YAP target gene, Connective Tissue Growth Factor CTGF. On the other hand, BIS I, an inhibitor of PKC\u3b4 and GSK3\u3b2, caused YAP accumulation into the nucleus. Activation of \u3b2-catenin reporter and interfering experiments show that inhibition of the PKC\u3b4/GSK3\u3b2 pathway triggers YAP nuclear accumulation inducing YAP/TEAD transcriptional response. Inhibition of GSK3\u3b2 by BIS I reduced the expression levels of SMADs protein and reduced YAP contribution to EMT. Notably, BIS I reduced proliferation, migration and clonogenicity of PDAC cells in vitro, phenocopying YAP genetic down-regulation. As shown by chromatin immunoprecipitation experiments and YAP over-expressing rescue experiments, BIS I reverted YAP-dependent EMT program by modulating the expression of the YAP target genes E-cadherin, vimentin, CTGF and of the newly identified target, CD133. In conclusion, we identified two different molecules, erlotinib and BIS I, modulating YAP functionality although via different mechanisms of action, with the second one specifically inhibiting the YAP-dependent EMT program in PDAC cell lines

Transcriptional induction of the heat shock protein B8 mediates the clearance of misfolded proteins responsible for motor neuron diseases

Neurodegenerative diseases (NDs) are often associated with the presence of misfolded protein inclusions. The chaperone HSPB8 is upregulated in mice, the human brain and muscle structures affected during NDs progression. HSPB8 exerts a potent pro-degradative activity on several misfolded proteins responsible for familial NDs forms. Here, we demonstrated that HSPB8 also counteracts accumulation of aberrantly localized misfolded forms of TDP-43 and its 25 KDa fragment involved in most sporadic cases of Amyotrophic Lateral Sclerosis (sALS) and of Fronto Lateral Temporal Dementia (FLTD). HSPB8 acts with BAG3 and the HSP70/HSC70-CHIP complex enhancing the autophagic removal of misfolded proteins. We performed a high-through put screening (HTS) to find small molecules capable of inducing HSPB8 in neurons for therapeutic purposes. We identified two compounds, colchicine and doxorubicin, that robustly up-regulated HSPB8 expression. Both colchicine and doxorubicin increased the expression of the master regulator of autophagy TFEB, the autophagy linker p62/SQSTM1 and the autophagosome component LC3. In line, both drugs counteracted the accumulation of TDP-43 and TDP-25 misfolded species responsible for motoneuronal death in sALS. Thus, analogs of colchicine and doxorubicin able to induce HSPB8 and with better safety and tolerability may result beneficial in NDs models

A Screening of Native (Poly)phenols and Gut-Related Metabolites on 3D HCT116 Spheroids Reveals Gut Health Benefits of a Flavan-3-ol Metabolite

Scope Epidemiological evidence suggests that a reduced risk of colorectal cancer (CRC) is correlated with high consumption of fruits and vegetables, which are major sources of fiber and phytochemicals, such as flavan-3-ols. However, it remains unknown how these phytochemicals and their specific gut-related metabolites may alter cancer cell behavior. Methods and results A focused screening using native (poly)phenols and gut microbial metabolites (GMMs) on 3D HCT116 spheroids is carried out using a high-throughput imaging approach. Dose-responses, IC50, and long-term exposure are calculated for the most promising native (poly)phenols and GMMs. As a result, this research shows that (poly)phenol catabolites may play a key role in preventing cancer propagation. Indeed, mu M concentration levels of (4R)-5-(3MODIFIER LETTER PRIME,4MODIFIER LETTER PRIME-dihydroxyphenyl)-gamma-valerolactone significantly decrease spheroid size at early stages of spheroid aggregation and gene expression of matrix metalloproteinases. Conclusion A chronic exposure to (4R)-5-(3MODIFIER LETTER PRIME,4MODIFIER LETTER PRIME-dihydroxyphenyl)-gamma-valerolactone may lead to a reduced CRC risk. Daily intake of monomeric, oligomeric, and polymeric flavan-3-ols may increase the colonic concentrations of this metabolite, and, in turn, this compound may act locally interacting with intestinal epithelial cells, precancerous and cancer cells

sample codes and their origin.

<p>Products, active ingredients and target microorganisms used in IPM and organic production in the area and during the season of the study.</p><p>sample codes and their origin.</p

Networks representing sample/OTU interaction.

<p>In both networks edge visibility (line width and opacity) is enhanced based on eweights, to better highlight the most relevant connections. A: sample nodes are shown according to grapevine cultivar (yellow: Chardonnay; blue: Merlot), OTU nodes are white, with edges indicated according to pest management type (red: IPM; green: organic production). B: sample nodes are indicated according to pest management type (red: IPM; green: organic production), OTU nodes are white, with edges indicated according to taxonomic assignment at phylum level (colour legend as for <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0112763#pone-0112763-g003" target="_blank">Fig. 3</a>). C: zoomed in view of <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0112763#pone-0112763-g004" target="_blank">Figure 4B</a>, with eweight significance for edge visibility emphasised.</p

Microbial community analysis plots based on 16S rDNA pyrosequencing.

<p>A: alpha diversity metrics based on observed OTUs, richness (Chao's richness and Abundance-Based Coverage estimators) and diversity (Shannon's and Simpson's diversity indices) B: histogram representing taxonomic composition and relative abundance (over 2%) at family and <i>genus</i> level for each cultivar in each treatment.</p

Multivariate analysis of beta-diversity: two-dimensional scatter plots of endophytic community composition in vineyards.

<p>A: PCoA of B-ARISA markers; B: CAP of B-ARISA markers; C: PCoA of 16S rDNA data; D: CAP of 16S rDNA markers. Ellipses and triangles represent samples from IPM and organic vineyards respectively; samples taken from Merlot and Chardonnay cvs are shown in red and green respectively.</p

A Screening of native (poly)phenols and gut-related metabolites on 3D HCT116 spheroids reveals gut health benefits of a flavan-3-ol metabolite

Scope: Epidemiological evidence suggests that a reduced risk of colorectal cancer (CRC) is correlated with high consumption of fruits and vegetables, which are major sources of fiber and phytochemicals, such as flavan-3-ols. However, it remains unknown how these phytochemicals and their specific gut-related metabolites may alter cancer cell behavior. Methods and results: A focused screening using native (poly)phenols and gut microbial metabolites (GMMs) on 3D HCT116 spheroids is carried out using a high-throughput imaging approach. Dose–responses, IC50, and long-term exposure are calculated for the most promising native (poly)phenols and GMMs. As a result, this research shows that (poly)phenol catabolites may play a key role in preventing cancer propagation. Indeed, μM concentration levels of (4R)-5-(3ʹ,4ʹ-dihydroxyphenyl)--valerolactone significantly decrease spheroid size at early stages of spheroid aggregation and gene expression of matrix metalloproteinases. Conclusion: A chronic exposure to (4R)-5-(3ʹ,4ʹ-dihydroxyphenyl)-- valerolactone may lead to a reduced CRC risk. Daily intake of monomeric, oligomeric, and polymeric flavan-3-ols may increase the colonic concentrations of this metabolite, and, in turn, this compound may act locally interacting with intestinal epithelial cells, precancerous and cancer cells

Interkingdom transfer of the acne causing agent, Propionibacterium acnes, from human to grapevine

Here we report the surprising and, to our knowledge, unique example of horizontal interkingdom transfer of a human opportunistic pathogen (Propionibacterium acnes) to a crop plant (the domesticated grapevine Vitis vinifera L.). Humans, like most organisms, have established a long-lasting cohabitation with a variety of microbes, including pathogens and gut-associated bacteria. Studies which have investigated the dynamics of such associations revealed numerous cases of bacterial host switches from domestic animals to humans. Much less is, however, known about the exchange of microbial symbionts between humans and plants. Fluorescent in-situ hybridization localized P. acnes in the bark, in xylem fibers, and, more interestingly, inside pith tissues. Phylogenetic and population genetic analyses suggest that the establishment of the grapevine-associated P. acnes as obligate endophyte is compatible with a recent transfer event, likely during the Neolithic, when grapevine was domesticated