Background High-grade serous tubo-ovarian cancer (HGSTOC) is characterised by extensive inter- and intratumour heterogeneity, resulting in persistent therapeutic resistance and poor disease outcome. Molecular subtype classification based on bulk RNA sequencing facilitates a more accurate characterisation of this heterogeneity, but the lack of strong prognostic or predictive correlations with these subtypes currently hinders their clinical implementation. Stromal admixture profoundly affects the prognostic impact of the molecular subtypes, but the contribution of stromal cells to each subtype has poorly been characterised. Increasing the transcriptomic resolution of the molecular subtypes based on single-cell RNA sequencing (scRNA-seq) may provide insights in the prognostic and predictive relevance of these subtypes. Methods We performed scRNA-seq of 18,403 cells unbiasedly collected from 7 treatment-naive HGSTOC tumours. For each phenotypic cluster of tumour or stromal cells, we identified specific transcriptomic markers. We explored which phenotypic clusters correlated with overall survival based on expression of these transcriptomic markers in microarray data of 1467 tumours. By evaluating molecular subtype signatures in single cells, we assessed to what extent a phenotypic cluster of tumour or stromal cells contributes to each molecular subtype. Results We identified 11 cancer and 32 stromal cell phenotypes in HGSTOC tumours. Of these, the relative frequency of myofibroblasts, TGF-β-driven cancer-associated fibroblasts, mesothelial cells and lymphatic endothelial cells predicted poor outcome, while plasma cells correlated with more favourable outcome. Moreover, we identified a clear cell-like transcriptomic signature in cancer cells, which correlated with worse overall survival in HGSTOC patients. Stromal cell phenotypes differed substantially between molecular subtypes. For instance, the mesenchymal, immunoreactive and differentiated signatures were characterised by specific fibroblast, immune cell and myofibroblast/mesothelial cell phenotypes, respectively. Cell phenotypes correlating with poor outcome were enriched in molecular subtypes associated with poor outcome. Conclusions We used scRNA-seq to identify stromal cell phenotypes predicting overall survival in HGSTOC patients. These stromal features explain the association of the molecular subtypes with outcome but also the latter’s weakness of clinical implementation. Stratifying patients based on marker genes specific for these phenotypes represents a promising approach to predict prognosis or response to therapy.

Electrochemical micromachining (micro-ECM) is an unconventional micromachining technology that has capability to fabricate high aspect ratio micro-holes, micro-cavities, microchannels and grooves on conductive and difficult-to-cut materials. Both academia and industry have the consensus that it offers promising machining performance especially in terms of high surface finish, no tool wear and absence of thermally induced defects. Furthermore in order to machine novel materials with extreme properties, novel hybrid electrochemical micromachining technologies are under development. With these hybrid micro-ECM technologies, capabilities of micro-ECM can be expanded by combining it with other processes. To fully exploit the potential as well as improve micro-ECM technology and related hybrid processes, a wide spectrum of multidisciplinary knowledge is needed. The present review systematically discusses process capabilities and research developments of electrochemical micromachining and its hybrid variants considering knowledge of both basic and applied research fields. After few introductory review articles in prior state of the art, this review fills an important gap in research literature by presenting first time an extended literature source with a wide coverage of recent research developments in electrochemical micromachining technology and its hybrid variants. This paper outlines the research and engineering developments in electrochemical micromachining technology and its hybrid variants, review of the related concepts, aspects of tooling, advanced process capabilities and process energy sources. It also provides new sights into technological understanding of micro-ECM technology which will be helpful in future engineering developments of this technology.

No abstract

Synopsis: IL1β is implicated in cancer progression. The authors show that IL1β promotes neutrophil accumulation in tumors and suppresses antitumor immunity independently of the inflammasome, suggesting that therapeutic inflammasome inhibition will not limit IL1β production in certain cancer types.

High fluxes and low fouling are practically highly desired in all membrane processes. One generic way to realize this is by introducing surface patterns on the membrane to increase the active area and the surface roughness. A new method to prepare flat-sheet patterned membranes using a patterned knife combined with a modified phase inversion process is now presented. After casting the polymer solution, the patterned knife shapes the top surface into the desired pattern. Non-solvent is sprayed immediately after the passage of the knife to rapidly solidify the polymer and preserve the pattern. To construct the patterned knife, different materials and techniques were evaluated: (1) an aluminium knife shaped by electrical discharge micromachining, (2) a gypsum knife shaped by powder-based 3D-printing, and (3) an acrylate resin, shaped by photopolymerization 3D-printing (stereo-lithography). As such, replacing the conventional nonsolvent contact in the phase inversion via immersion by spraying already increased membrane permeance, but a significant further increase of permeance resulted from the increased surface area realized through the patterning. A minimum viscosity of the casting solution of around 7.5-10 Pa.s was required to realize well-pronounced patterns on the membrane, as screened for polyacrylonitrile and cellulose acetate, both dissolved in dimethyl sulfoxide and using water as non-solvent.

No abstract

To date, the overall response rate of PD-1 blockade remains unsatisfactory, partially due to limited understanding of tumor immune microenvironment (TIME). B-cell lymphoma 9 (BCL9), a key transcription co-activator of the Wnt pathway, is highly expressed in cancers. By genetic depletion and pharmacological inhibition of BCL9 in tumors, we found that BCL9 suppression reduced tumor growth, promoted CD8+ T cell tumor infiltration, and enhanced response to anti-PD-1 treatment in mouse colon cancer models. To determine the underlying mechanism of BCL9’s role in TIME regulation, single-cell RNA-seq was applied to reveal cellular landscape and transcription differences in the tumor immune microenvironment upon BCL9 inhibition. CD155-CD226 and CD155-CD96 checkpoints play key roles in cancer cell/CD8+ T cell interaction. BCL9 suppression induces phosphorylation of VAV1 in CD8+ T cells and increases GLI1 and PATCH expression to promote CD155 expression in cancer cells. In The Cancer Genome Atlas database analysis, we found that BCL9 expression is positively associated with CD155 and negatively associated with CD226 expression. BCL9 is also linked to adenomatous polyposis coli (APC) mutation involved in patient survival following anti-PD-1 treatment. This study points to cellular diversity within the tumor immune microenvironment affected by BCL9 inhibition and provides new insights into the role of BCL9 in regulating CD226 and CD96 checkpoints

No abstract