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A subset of colorectal cancer (CRC) with a mesenchymal phenotype (CMS4) displays an aggressive disease, with an increased risk of recurrence after surgery, reduced survival, and resistance to standard treatments. It has been shown that the AXL and TGFβ signaling pathways are involved in epithelial-to-mesenchymal transition, migration, metastatic spread, and unresponsiveness to targeted therapies. However, the prognostic role of the combination of these biomarkers and the anti-tumor effect of AXL and TGFβ inhibition in CRC still has to be assessed. To evaluate the role of AXL and TGFβ as negative biomarker in CRC, we conducted an in-depth in silico analysis of CRC samples derived from the Gene Expression Omnibus. We found that AXL and TGFβ receptors are upregulated in CMS4 tumors and are correlated with an increased risk of recurrence after surgery in stage II/III CRC and a reduced overall survival. Moreover, we showed that AXL receptor is differently expressed in human CRC cell lines. Dual treatment with the TGFβ galunisertib and the AXL inhibitor, bemcentinib, significantly reduced colony formation and migration capabilities of tumor cells and displayed a strong anti-tumor activity in 3D spheroid cultures derived from patients with advanced CRC. Our work shows that AXL and TGFβ receptors identify a subgroup of CRC with a mesenchymal phenotype and correlate with poor prognosis. Dual inhibition of AXL and TGFβ could represent a novel therapeutic strategy for patients with this aggressive disease.

Background: RAS mutations are the only validated biomarkers in metastatic colorectal cancer (mCRC) for anti-epidermal growth factor receptor (EGFR) therapy. Limited clinical information is available on AXL expression, marker of epithelial to mesenchymal transition, in mCRC., Methods: AXL was retrospectively assessed by immunohistochemistry in 307 patients. RAS wild-type (WT) patients (N = 136) received first-line anti-EGFR-based therapy; RAS mutant patients (N = 171) received anti-angiogenic-based regimens. Preclinical experiments were performed using human RAS WT CRC cell lines and xenograft models. AXL RNA levels were assessed in a cohort of patients with available samples at baseline and at progression to anti-EGFR treatment and in the GSE5851 dataset., Results: AXL was expressed in 55/307 tumour tissues, correlating with worse survival in the overall population (AXL-positive, 23.7 months; AXL-negative, 30.8 months; HR, 1.455, P = 0.032) and in RAS WT patients (AXL-positive, 23.0 months; AXL-negative, 35.8 months; HR,1.780, P = 0.032). Progression-free survival (PFS) in the RAS WT cohort was shorter in the AXL-positive cohort (6.2 months versus 12.1 months; HR, 1.796, P = 0.013). Three-dimensional cultures obtained from a patient following anti-EGFR therapy resulted AXL-positive, showing resistance to anti-EGFR drugs and sensitivity to AXL inhibition. AXL transfection in CRC cell lines induced AXL overexpression and resistance to the EGFR blockade. At progression to cetuximab, 2/10 SW48-tumour xenograft mice showed AXL expression. Consistently, AXL RNA levels increased in 5/7 patients following anti-EGFR therapy. Moreover, in the GSE5851 dataset higher AXL RNA levels correlated with worse PFS with cetuximab in KRAS-exon2 WT chemorefractory patients., Conclusions: AXL is a marker of poor prognosis in mCRC with consistent clinical and preclinical evidences of involvement in primary and acquired resistance to anti-EGFR drugs in RAS WT patients., Competing Interests: Conflict of interest statement TT reports advisory board for Amgen, Bayer, Merck, Novartis, Roche and Sanofi. FM reports advisory board for Lilly and MSD. ChCr reports personal fees from Roche, Amgen, Bayer and Servier, research funding from Merck Serono and consulting or advisory role for Roche, Bayer and Amgen. FP reports honoraria/speaker bureau from Amgen, Roche, Merck Serono, Lilly, Sanofi, Bayer and Servier and research grants from BMS. GWP reports personal fees from advisory boards and/or speaker's honorarium: Amgen, Bayer, BMS, Boston Biomedical, Celgene, Halozyme, Lilly, Merck Serono, Roche, Sanofi, Servier, Shire, MSD, Taiho and CECOG. NN reports personal financial interests as speaker's fee and/or advisory boards from MSD, Qiagen, Bayer, Biocartis, Incyte, Roche, BMS, Merck, Thermo Fisher, Boehringer Ingelheim, AstraZeneca, Sanofi and Eli Lilly; institutional financial interests (financial support to research projets) from Merck, Sysmex, Thermo Fisher, Qiagen, Roche, AstraZeneca and Biocartis; non-financial interests: President, International Quality Network for Pathology (IQN Path), President Elect, Italian Cancer Society (SIC). EvMa reports personal financial interests honoraria for advisory role for advisory board from: Astra Zeneca, Sanofi Genzyme, Servier, Celgene, Merck, Lilly and Roche; research grants from MSD and BSD; travel grants from Merck and Roche; institutional financial interests as financial support for clinical trials from Roche, Sanofi Genzyme, Teva and Merck. GA reports personal financial interests from Hoffman-La Roche, Bristol Myers Squibb, Bayer, Servier, Amgen, Merck Serono and Menarini; institutional financial interests from Bayer, Servier, Novartis, Boehringer Ingelheim, Boston Pharmaceuticals, Hoffman-La Roche and Genentech. EE reports personal financial interests, honoraria for advisory role, travel grants, research grants from Hoffman-La Roche, Sanofi Aventis, Amgen, Merck Serono and Servier, MSD; institutional financial interests from Hoffman-LaRoche, Sanofi Aventis, Amgen, Merck Serono, MSD, Boehringer Ingelheim, AbbVie, Array Pharmaceuticals, Pierre Fabre and Novartis. AF reports personal financial interests (advisory role/honoraria): Bayer, Bristol, Lilly, Merck, Pierre Fabre, Roche, Servier; institutional financial interests as financial support for clinical trials from AstraZeneca, Bayer, Bristol, Lilly, Merck, MSD, Novartis, Roche, Sanofi and Servier. JT reports scientific consultancy roles for Bayer, Boehringer Ingelheim, Chugai, Genentech, Inc., Ipsen, Lilly, MSD, Merck Serono, Merrimack, Merus, Novartis, Peptomyc, Pfizer, Rafael Pharmaceuticals, F. Hoffmann-La Roche Ltd, Sanofi, Seattle Genetics, Symphogen and Taiho. FC reports being in the advisory boards for Merck, Roche, Amgen, Bayer, Servier, Symphogen and Pfizer and research funding from Roche, Merck, Amgen, Bayer and Ipsen. ErMa reports being in the advisory boards for Amgen, Bayer, Merck, Roche, Sanofi and Servier and giving expert opinion for ESMO (European Society of Medical Oncology). ClCa, BB, VMV, GM, VS, PPV, DC, VB, NM, FZ, MD, AN, AB, CA, GS, RF and MS report no competing conflicts of interest., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

Background: Glioblastoma is the most aggressive type of brain cancer. Dendritic cell (DC)-based immunotherapy against glioblastoma depends on the effectiveness of loaded antigens. Sphere-inducing culture conditions are being studied by many as a potential antigen source. Here, we investigated two different in vitro conditions (spheroid culture versus adherent culture) in relation to DC immunotherapy: (1) We studied the specific spheroid-culture proteome and assessed the clinical importance of spheroid proteins. (2) We evaluated the immunogenicity of spheroid lysate - both compared to adherent conditions., Methods: We used seven spheroid culture systems, three of them patient-derived. Stemness-related markers were studied in those three via immunofluorescence. Spheroid-specific protein expression was measured via quantitative proteomics. The Cancer Genome Atlas (TCGA) survival data was used to investigate the clinical impact of spheroid proteins. Immunogenicity of spheroid versus adherent cell lysate was explored in autologous ELISPOT systems (DCs and T cells from the three patients)., Results: (1) The differential proteome of spheroid versus adherent glioblastoma culture conditions could successfully be established. The top 10 identified spheroid-specific proteins were associated with significantly decreased overall survival (TCGA MIT/Harvard cohort; n = 350, P = 0.014). (2) In exploratory experiments, immunogenicity of spheroid lysate vis-á-vis interferon (IFN)γ production was lower than that of adherent cell lysate (IFNγ ELISPOT; P = 0.034)., Conclusions: Spheroid culture proteins seem to represent survival-relevant targets, supporting the use of spheroid culture conditions as an antigen source for DC immunotherapy. However, immunogenicity enhancement should be considered for future research. Transferability of our findings in terms of clinical impact and regarding different spheroid-generation techniques needs further validation., (Copyright © 2019 International Society for Cell and Gene Therapy. Published by Elsevier Inc. All rights reserved.)

Glioblastoma is the most dangerous brain cancer. One reason for glioblastoma's aggressiveness are glioblastoma stem-like cells. To target them, a number of markers have been proposed (CD133, CD44, CD15, A2B5, CD36, CXCR4, IL6R, L1CAM, and ITGA6). A comprehensive study of co-expression patterns of them has, however, not been performed so far. Here, we mapped the multidimensional co-expression profile of these stemness-associated molecules. Gliomaspheres - an established model of glioblastoma stem-like cells - were used. Seven different gliomasphere systems were subjected to multicolor flow cytometry measuring the nine markers CD133, CD44, CD15, A2B5, CD36, CXCR4, IL6R, L1CAM, and ITGA6 all simultaneously based on a novel 9-marker multicolor panel developed for this study. The viSNE dimensionality reduction algorithm was applied for analysis. All gliomaspheres were found to express at least five different glioblastoma stem-like cell markers. Multi-dimensional analysis showed that all studied gliomaspheres consistently harbored a cell population positive for the molecular signature CD44+/CD133+/ITGA6+/CD36+. Glioblastoma patients with an enrichment of this combination had a significantly worse survival outcome when analyzing the two largest available The Cancer Genome Atlas datasets (MIT/Harvard Affymetrix: P = 0.0015, University of North Carolina Agilent: P = 0.0322). In sum, we detected a previously unknown marker combination - demonstrating feasibility, usefulness, and importance of high-dimensional gliomasphere marker combinatorics., (© 2018 The Authors. Journal of Cellular and Molecular Medicine published by John Wiley & Sons Ltd and Foundation for Cellular and Molecular Medicine.)

Maintaining dendritic cells (DC) in a state of dysfunction represents a key mechanism by which tumour cells evade recognition and elimination by the immune system. Limited knowledge about the intracellular mediators of DC dysfunction restricts success of therapies aimed at reactivating a DC-driven anti-tumour immune response. Using a cell type-specific murine knock-out model, we have identified MAPK-activated protein kinase 2 (MK2) as a major guardian of a suppressive DC phenotype in the melanoma tumour microenvironment. MK2 deletion in CD11c + cells led to an expansion of stimulatory CD103 + DCs, mounting a potent CD8 + T cell response that resulted in elimination of highly aggressive B16-F10 tumours upon toll-like receptor (TLR) activation in the presence of tumour antigen. Moreover, tumour infiltration by suppressive myeloid cells was strongly diminished. These insights into the regulation of DC functionality reveal MK2 as a targetable pathway for DC-centred immunomodulatory cancer therapies.

We demonstrate non-degenerate down-conversion at 810 and 1550 nm for long-distance fiber based quantum communication using polarization entangled photon pairs. Measurements of the two-photon visibility, without dark count subtraction, have shown that the quantum correlations (raw visibility 89%) allow secure quantum cryptography after 100 km of non-zero dispersion shifted fiber using commercially available single photon detectors. In addition, quantum state tomography has revealed little degradation of state negativity, decreasing from 0.99 at the source to 0.93 after 100 km, indicating minimal loss in fidelity during the transmission.

Interference of photons emerging from independent sources is essential for modern quantum-information processing schemes, above all quantum repeaters and linear-optics quantum computers. We report an observation of nonclassical interference of two single photons originating from two independent, separated sources, which were actively synchronized with a rms timing jitter of 260 fs. The resulting (two-photon) interference visibility was (83+/-4)%.