Epithelial Cell Line Derived from Endometriotic Lesion Mimics Macrophage Nervous Mechanism of Pain Generation on Proteome and Metabolome Levels

Endometriosis is a benign disease affecting one in ten women of reproductive age worldwide. Although the pain level is not correlated to the extent of the disease, it is still one of the cardinal symptoms strongly affecting the patients’ quality of life. Yet, a molecular mechanism of this pathology, including the formation of pain, remains to be defined. Recent studies have indicated a close interaction between newly generated nerve cells and macrophages, leading to neurogenic inflammation in the pelvic area. In this context, the responsiveness of an endometriotic cell culture model was characterized upon inflammatory stimulation by employing a multi-omics approach, including proteomics, metabolomics and eicosanoid analysis. Differential proteomic profiling of the 12-Z endometriotic cell line treated with TNFα and IL1β unexpectedly showed that the inflammatory stimulation was able to induce a protein signature associated with neuroangiogenesis, specifically including neuropilins (NRP1/2). Untargeted metabolomic profiling in the same setup further revealed that the endometriotic cells were capable of the autonomous production of 7,8-dihydrobiopterin (BH2), 7,8-dihydroneopterin, normetanephrine and epinephrine. These metabolites are related to the development of neuropathic pain and the former three were found up-regulated upon inflammatory stimulation. Additionally, 12-Z cells were found to secrete the mono-oxygenated oxylipin 16-HETE, a known inhibitor of neutrophil aggregation and adhesion. Thus, inflammatory stimulation of endometriotic 12-Z cells led to specific protein and metabolite expression changes suggesting a direct involvement of these epithelial-like cells in endometriosis pain development

Selective Arylation of Selenocysteine of Thioredoxin Reductase 1 by an Organogold Compound: Expanding the Tool-Box of Metal-Templated Reactions in Cancer Cells

Cyclometalated gold(III) complexes have been reported to template C-S cross-coupling reactions in a biological environment and acting as modifiers of cysteine residues. To broaden the scope of organogold complexes for covalent protein post-translational modification in cancer cells, an oxime-containing C^N-cyclometalated gold(III) compound was synthesised featuring a carboxylic acid group for either immobilisation on amine-bearing solid support (Au), or functionalisation with a fluorescent tag (Au-Fluo). Live-cell imaging revealed that Au-Fluo distributed evenly into SW480 colon carcinoma cells, with a slight preference for the nuclear and nucleolar compartments. Thioredoxin reductase 1 (TXNRD1) was observed as the major interactor of Au from SW480 cell lysates in chemoproteomic approaches and a 2 : 1 binding stoichiometry resulted from titration-dependent pull-downs. Direct interactions confirmed a high reactivity of Au towards the catalytic CysSec-dyad at the C-terminus of TXNRD1 and revealed double arylation events at this motif. Therefore, the observed Au-templated arylation of selenocysteine likely contributes to the compound’s biological effects. Proteome profiling of SW480 cancer cells treated with sub-cytotoxic concentrations of Au revealed an apparent reduction of the available selenium pool by down-regulating the detected selenoproteins, except TXNRD1. Additionally, Au treatment induced the NRF2-KEAP1 stress response, pointing towards a disturbance of the intracellular redox balance by Au-mediated covalent targeting of TXNRD1. Inhibition of heme oxygenase-1 (HMOX1), the most strongly induced NRF2-target, showed pronounced synergism with Au treatment. Overall, organogold compounds, templating the formation of C–S(Se) bonds in cells as a novel mode of action, hold promise for the targeted modification of (onco)proteins

A multi-omics based anti-inflammatory immune signature characterizes long COVID-19 syndrome

Summary: To investigate long COVID-19 syndrome (LCS) pathophysiology, we performed an exploratory study with blood plasma derived from three groups: 1) healthy vaccinated individuals without SARS-CoV-2 exposure; 2) asymptomatic recovered patients at least three months after SARS-CoV-2 infection and; 3) symptomatic patients at least 3 months after SARS-CoV-2 infection with chronic fatigue syndrome or similar symptoms, here designated as patients with long COVID-19 syndrome (LCS). Multiplex cytokine profiling indicated slightly elevated pro-inflammatory cytokine levels in recovered individuals in contrast to patients with LCS. Plasma proteomics demonstrated low levels of acute phase proteins and macrophage-derived secreted proteins in LCS. High levels of anti-inflammatory oxylipins including omega-3 fatty acids in LCS were detected by eicosadomics, whereas targeted metabolic profiling indicated high levels of anti-inflammatory osmolytes taurine and hypaphorine, but low amino acid and triglyceride levels and deregulated acylcarnitines. A model considering alternatively polarized macrophages as a major contributor to these molecular alterations is presented

Single-cell transcriptomics and epigenomics unravel the role of monocytes in neuroblastoma bone marrow metastasis

Metastasis is the major cause of cancer-related deaths. Neuroblastoma (NB), a childhood tumor has been molecularly defined at the primary cancer site, however, the bone marrow (BM) as the metastatic niche of NB is poorly characterized. Here we perform single-cell transcriptomic and epigenomic profiling of BM aspirates from 11 subjects spanning three major NB subtypes and compare these to five age-matched and metastasis-free BM, followed by in-depth single cell analyses of tissue diversity and cell-cell interactions, as well as functional validation. We show that cellular plasticity of NB tumor cells is conserved upon metastasis and tumor cell type composition is NB subtype-dependent. NB cells signal to the BM microenvironment, rewiring via macrophage mgration inhibitory factor and midkine signaling specifically monocytes, which exhibit M1 and M2 features, are marked by activation of pro- and anti-inflammatory programs, and express tumor-promoting factors, reminiscent of tumor-associated macrophages. The interactions and pathways characterized in our study provide the basis for therapeutic approaches that target tumor-to-microenvironment interactions