Glutathionylation is an important posttranslational modification that protects proteins from further oxidative damage as well as influencing protein structure and activity. In the present study, we demonstrate that the cysteine-42 residue in protein arginine N-methyltransferase 5 (PRMT5) is glutathionylated in aged mice or in cells that have been exposed to oxidative stress. Deglutathionylation of this protein is catalyzed by glutaredoxin-1 (Grx1). Using mutagenesis and subsequent biochemical analyses, we show that glutathionylation decreased the binding affinity of PRMT5 with methylosome protein-50 (MEP50) and reduced the methyltransferase activity of PRMT5. Furthermore, overexpression of PRMT5-C42A mutant caused a significant increase in histone methylation in HEK293T and A549 cells and promoted cell growth, whereas overexpression of the PRMT5-C42D mutant, a mimic of glutathionylated PRMT5, inhibited cell proliferation. Taken together, our results demonstrate a new mechanism of regulation of PRMT5 methyltransferases activity and suggest that PRMT5 glutathionylation is partly responsible for reactive oxygen species-mediated cell growth inhibition.

Studies in the yeast Saccharomyces cerevisiae have helped define mechanisms underlying the activity of the ubiquitin–proteasome system (UPS), uncover the proteasome assembly pathway, and link the UPS to the maintenance of cellular homeostasis. However, the spectrum of UPS substrates is incompletely defined, even though multiple techniques—including MS—have been used. Therefore, we developed a substrate trapping proteomics workflow to identify previously unknown UPS substrates. We first generated a yeast strain with an epitope tagged proteasome subunit to which a proteasome inhibitor could be applied. Parallel experiments utilized inhibitor insensitive strains or strains lacking the tagged subunit. After affinity isolation, enriched proteins were resolved, in-gel digested, and analyzed by high resolution liquid chromatography-tandem MS. A total of 149 proteasome partners were identified, including all 33 proteasome subunits. When we next compared data between inhibitor sensitive and resistant cells, 27 proteasome partners were significantly enriched. Among these proteins were known UPS substrates and proteins that escort ubiquitinated substrates to the proteasome. We also detected Erg25 as a high-confidence partner. Erg25 is a methyl oxidase that converts dimethylzymosterol to zymosterol, a precursor of the plasma membrane sterol, ergosterol. Because Erg25 is a resident of the endoplasmic reticulum (ER) and had not previously been directly characterized as a UPS substrate, we asked whether Erg25 is a target of the ER associated degradation (ERAD) pathway, which most commonly mediates proteasome-dependent destruction of aberrant proteins. As anticipated, Erg25 was ubiquitinated and associated with stalled proteasomes. Further, Erg25 degradation depended on ERAD-associated ubiquitin ligases and was regulated by sterol synthesis. These data expand the cohort of lipid biosynthetic enzymes targeted for ERAD, highlight the role of the UPS in maintaining ER function, and provide a novel tool to uncover other UPS substrates via manipulations of our engineered strain.

Co-fractionation MS (CF-MS) is a technique with potential to characterize endogenous and unmanipulated protein complexes on an unprecedented scale. However this potential has been offset by a lack of guidelines for best-practice CF-MS data collection and analysis. To obtain such guidelines, this study thoroughly evaluates novel and published Saccharomyces cerevisiae CF-MS data sets using very high proteome coverage libraries of yeast gold standard complexes. A new method for identifying gold standard complexes in CF-MS data, Reference Complex Profiling, and the Extending 'Guilt-by-Association' by Degree (EGAD) R package are used for these evaluations, which are verified with concurrent analyses of published human data. By evaluating data collection designs, which involve fractionation of cell lysates, it is found that near-maximum recall of complexes can be achieved with fewer samples than published studies. Distributing sample collection across orthogonal fractionation methods, rather than a single high resolution data set, leads to particularly efficient recall. By evaluating 17 different similarity scoring metrics, which are central to CF-MS data analysis, it is found that two metrics rarely used in past CF-MS studies – Spearman and Kendall correlations – and the recently introduced Co-apex metric frequently maximize recall, whereas a popular metric—Euclidean distance—delivers poor recall. The common practice of integrating external genomic data into CF-MS data analysis is also evaluated, revealing that this practice may improve the precision and recall of known complexes but is generally unsuitable for predicting novel complexes in model organisms. If studying nonmodel organisms using orthologous genomic data, it is found that particular subsets of fractionation profiles (e.g. the lowest abundance quartile) should be excluded to minimize false discovery. These assessments are summarized in a series of universally applicable guidelines for precise, sensitive and efficient CF-MS studies of known complexes, and effective predictions of novel complexes for orthogonal experimental validation.

After ejaculation, mammalian spermatozoa must undergo a process known as capacitation in order to successfully fertilize the oocyte. Several post-translational modifications occur during capacitation, including sialylation, which despite being limited to a few proteins, seems to be essential for proper sperm-oocyte interaction. Regardless of its importance, to date, no single study has ever identified nor quantified which glycoproteins bearing terminal sialic acid (Sia) are altered during capacitation. Here we characterize sialylation during mouse sperm capacitation. Using tandem MS coupled with liquid chromatography (LC–MS/MS), we found 142 nonreductant peptides, with 9 of them showing potential modifications on their sialylated oligosaccharides during capacitation. As such, N-linked sialoglycopeptides from C4b-binding protein, endothelial lipase (EL), serine proteases 39 and 52, testis-expressed protein 101 and zonadhesin were reduced following capacitation. In contrast, mitochondrial aconitate hydratase (aconitase; ACO2), a TCA cycle enzyme, was the only protein to show an increase in Sia content during capacitation. Interestingly, although the loss of Sia within EL (N62) was accompanied by a reduction in its phospholipase A₁ activity, a decrease in the activity of ACO2 (i.e. stereospecific isomerization of citrate to isocitrate) occurred when sialylation increased (N612). The latter was confirmed by N612D recombinant protein tagged with both His and GFP. The replacement of Sia for the negatively charged Aspartic acid in the N612D mutant caused complete loss of aconitase activity compared with the WT. Computer modeling show that N612 sits atop the catalytic site of ACO2. The introduction of Sia causes a large conformational change in the alpha helix, essentially, distorting the active site, leading to complete loss of function. These findings suggest that the switch from oxidative phosphorylation, over to glycolysis that occurs during capacitation may come about through sialylation of ACO2.

Renal Cell Carcinoma (RCC) is one of the most commonly diagnosed cancers worldwide with research efforts dramatically improving understanding of the biology of the disease. To investigate the role of the immune system in treatment-naïve clear cell Renal Cell Carcinoma (ccRCC), we interrogated the immune infiltrate in patient-matched ccRCC tumor samples, benign normal adjacent tissue (NAT) and peripheral blood mononuclear cells (PBMCs isolated from whole blood, focusing our attention on the myeloid cell infiltrate. Using flow cytometric, MS, and ExCYT analysis, we discovered unique myeloid populations in PBMCs across patient samples. Furthermore, normal adjacent tissues and ccRCC tissues contained numerous myeloid populations with a unique signature for both tissues. Enrichment of the immune cell (CD45⁺) fraction and subsequent gene expression analysis revealed a number of myeloid-related genes that were differentially expressed. These data provide evidence, for the first time, of an immunosuppressive and pro-tumorigenic role of myeloid cells in early, clinically localized ccRCC. The identification of a number of immune proteins for therapeutic targeting provides a rationale for investigation into the potential efficacy of earlier intervention with single-agent or combination immunotherapy for ccRCC.

Protein synthesis on the endoplasmic reticulum (ER) requires the dynamic coordination of numerous cellular components. Together, resident ER membrane proteins, cytoplasmic translation factors, and both integral membrane and cytosolic RNA-binding proteins operate in concert with membrane-associated ribosomes to facilitate ER-localized translation. Little is known, however, regarding the spatial organization of ER-localized translation. This question is of growing significance as it is now known that ER-bound ribosomes contribute to secretory, integral membrane, and cytosolic protein synthesis alike. To explore this question, we utilized quantitative proximity proteomics to identify neighboring protein networks for the candidate ribosome interactors SEC61β (subunit of the protein translocase), RPN1 (oligosaccharyltransferase subunit), SEC62 (translocation integral membrane protein), and LRRC59 (ribosome binding integral membrane protein). Biotin labeling time course studies of the four BioID reporters revealed distinct labeling patterns that intensified but only modestly diversified as a function of labeling time, suggesting that the ER membrane is organized into discrete protein interaction domains. Whereas SEC61β and RPN1 reporters identified translocon-associated networks, SEC62 and LRRC59 reporters revealed divergent protein interactomes. Notably, the SEC62 interactome is enriched in redox-linked proteins and ER luminal chaperones, with the latter likely representing proximity to an ER luminal chaperone reflux pathway. In contrast, the LRRC59 interactome is highly enriched in SRP pathway components, translation factors, and ER-localized RNA-binding proteins, uncovering a functional link between LRRC59 and mRNA translation regulation. Importantly, analysis of the LRRC59 interactome by native immunoprecipitation identified similar protein and functional enrichments. Moreover, [³⁵S]-methionine incorporation assays revealed that siRNA silencing of LRRC59 expression reduced steady state translation levels on the ER by ca. 50%, and also impacted steady state translation levels in the cytosol compartment. Collectively, these data reveal a functional domain organization for the ER and identify a key role for LRRC59 in the organization and regulation of local translation.

The EGFR tyrosine kinase inhibitor gefitinib is commonly used for lung cancer patients. However, some patients eventually become resistant to gefitinib and develop progressive disease. Here, we indicate that ecto-ATP synthase, which ectopically translocated from mitochondrial inner membrane to plasma membrane, is considered as a potential therapeutic target for drug-resistant cells. Quantitative multi-omics profiling reveals that ecto-ATP synthase inhibitor mediates CK2-dependent phosphorylation of DNA topoisomerase IIα (topo IIα) at serine 1106 and subsequently increases the expression of long noncoding RNA, GAS5. Additionally, we also determine that downstream of GAS5, p53 pathway, is activated by ecto-ATP synthase inhibitor for regulation of programed cell death. Interestingly, GAS5-proteins interactomic profiling elucidates that GAS5 associates with topo IIα and subsequently enhancing the phosphorylation level of topo IIα. Taken together, our findings suggest that ecto-ATP synthase blockade is an effective therapeutic strategy via regulation of CK2/phospho-topo IIα/GAS5 network in gefitinib-resistant lung cancer cells.

Small admixtures in water, e.g. of metal ions, often act as cell growth regulators. Here we report that enrichment of deuterium content in water, normally found at 8 mm concentration, two-three folds increases cell proliferation and lowers the oxidative stress level as well. Acting as an anti-oxidant, deuterium-enriched water prevents the toxic effect of such oxidative agents as hydrogen peroxide and auranofin. This action is opposite to that of deuterium depletion that is known to suppress cell growth and induce oxidative stress in mitochondria. We thus hypothesize that deuterium may be a natural cell growth regulator that controls mitochondrial oxidation-reduction balance. Because growth acceleration is reduced approximately by half by addition to water a minute amount (0.15%) of ¹⁸O isotope, at least part of the deuterium effect on cell growth can be explained by the isotopic resonance phenomenon. A slight (2-fold) enrichment of deuterium in water accelerates human cell growth. Quantitative MS based proteomics determined changes in protein abundances and redox states and found that deuterium-enriched water acts mainly through decreasing ROS production in mitochondria. This action is opposite to that of deuterium depletion that suppresses cell growth by inducing oxidative stress. Thus deuterium may be a natural cell growth regulator that controls mitochondrial oxidation-reduction balance. The role of isotopic resonance in this effect was validated by further experiments on bacteria.

Amino acid hydroxylation is a common post-translational modification, which generally regulates protein interactions or adds a functional group that can be further modified. Such hydroxylation is currently considered irreversible, necessitating the degradation and re-synthesis of the entire protein to reset the modification. Here we present evidence that the cellular machinery can reverse FIH-mediated asparagine hydroxylation on intact proteins. These data suggest that asparagine hydroxylation is a flexible and dynamic post-translational modification akin to modifications involved in regulating signaling networks, such as phosphorylation, methylation and ubiquitylation.

We performed an in-depth characterization and comparison of the pediatric and adult urinary glycomes using a nanoLC-MS/MS based glycomics method, which included normal healthy pediatric (1–10 years, n = 21) and adult (21–50 years, n = 22) individuals. A total of 116 N-glycan compositions were identified, and 46 of them could be reproducibly quantified. We performed quantitative comparisons of the 46 glycan compositions between different age and sex groups. The results showed significant quantitative changes between the pediatric and adult cohorts. The pediatric urinary N-glycome was found to contain a higher level of high-mannose (HM), asialylated/afucosylated glycans (excluding HM), neutral fucosylated and agalactosylated glycans, and a lower level of trisialylated glycans compared with the adult. We further analyzed gender-associated glycan changes in the pediatric and adult group, respectively. In the pediatric group, there was almost no difference of glycan levels between males and females. In adult, the majority of glycans were more abundant in males than females, except the high-mannose and tetrasialylated glycans. These findings highlight the importance to consider age-matching and adult sex-matching for urinary glycan studies. The identified normal pediatric and adult urinary glycomes can serve as a baseline reference for comparisons to other disease states affected by glycosylation.

High-speed analysis of large (prote)omics sample sets at the rate of thousands or millions of samples per day on a single platform has been a challenge since the beginning of proteomics. For many years, ESI-based MS methods have dominated proteomics because of their high sensitivity and great depth in analyzing complex proteomes. However, despite improvements in speed, ESI-based MS methods are fundamentally limited by their sample introduction, which excludes off-line sample preparation/fractionation because of the time required to switch between individual samples/sample fractions, and therefore being dependent on the speed of on-line sample preparation methods such as liquid chromatography. Laser-based ionization methods have the advantage of moving from one sample to the next without these limitations, being mainly restricted by the speed of modern sample stages, i.e. 10 ms or less between samples. This speed matches the data acquisition speed of modern high-performing mass spectrometers whereas the pulse repetition rate of the lasers (>1 kHz) provides a sufficient number of desorption/ionization events for successful ion signal detection from each sample at the above speed of the sample stages. Other advantages of laser-based ionization methods include the generally higher tolerance to sample additives and contamination compared with ESI MS, and the contact-less and pulsed nature of the laser used for desorption, reducing the risk of cross-contamination. Furthermore, new developments in MALDI have expanded its analytical capabilities, now being able to fully exploit high-performing hybrid mass analyzers and their strengths in sensitivity and MS/MS analysis by generating an ESI-like stable yield of multiply charged analyte ions. Thus, these new developments and the intrinsically high speed of laser-based methods now provide a good basis for tackling extreme sample analysis speed in the omics.

Coronavirus disease 2019 (COVID-19) is a highly contagious infection and threating the human lives in the world. The elevation of cytokines in blood is crucial to induce cytokine storm and immunosuppression in the transition of severity in COVID-19 patients. However, the comprehensive changes of serum proteins in COVID-19 patients throughout the SARS-CoV-2 infection is unknown. In this work, we developed a high-density antibody microarray and performed an in-depth proteomics analysis of serum samples collected from early COVID-19 (n = 15) and influenza (n = 13) patients. We identified a large set of differentially expressed proteins (n = 132) that participate in a landscape of inflammation and immune signaling related to the SARS-CoV-2 infection. Furthermore, the significant correlations of neutrophil and lymphocyte with the CCL2 and CXCL10 mediated cytokine signaling pathways was identified. These information are valuable for the understanding of COVID-19 pathogenesis, identification of biomarkers and development of the optimal anti-inflammation therapy.

MS-based proteome profiling has become increasingly comprehensive and quantitative, yet a persistent shortcoming has been the relatively large samples required to achieve an in-depth measurement. Such bulk samples, typically comprising thousands of cells or more, provide a population average and obscure important cellular heterogeneity. Single-cell proteomics capabilities have the potential to transform biomedical research and enable understanding of biological systems with a new level of granularity. Recent advances in sample processing, separations and MS instrumentation now make it possible to quantify >1000 proteins from individual mammalian cells, a level of coverage that required an input of thousands of cells just a few years ago. This review discusses important factors and parameters that should be optimized across the workflow for single-cell and other low-input measurements. It also highlights recent developments that have advanced the field and opportunities for further development.

Cross-linking MS (XL-MS) has been recognized as an effective source of information about protein structures and interactions. In contrast to regular peptide identification, XL-MS has to deal with a quadratic search space, where peptides from every protein could potentially be cross-linked to any other protein. To cope with this search space, most tools apply different heuristics for search space reduction. We introduce a new open-source XL-MS database search algorithm, OpenPepXL, which offers increased sensitivity compared with other tools. OpenPepXL searches the full search space of an XL-MS experiment without using heuristics to reduce it. Because of efficient data structures and built-in parallelization OpenPepXL achieves excellent runtimes and can also be deployed on large compute clusters and cloud services while maintaining a slim memory footprint. We compared OpenPepXL to several other commonly used tools for identification of noncleavable labeled and label-free cross-linkers on a diverse set of XL-MS experiments. In our first comparison, we used a data set from a fraction of a cell lysate with a protein database of 128 targets and 128 decoys. At 5% FDR, OpenPepXL finds from 7% to over 50% more unique residue pairs (URPs) than other tools. On data sets with available high-resolution structures for cross-link validation OpenPepXL reports from 7% to over 40% more structurally validated URPs than other tools. Additionally, we used a synthetic peptide data set that allows objective validation of cross-links without relying on structural information and found that OpenPepXL reports at least 12% more validated URPs than other tools. It has been built as part of the OpenMS suite of tools and supports Windows, macOS, and Linux operating systems. OpenPepXL also supports the MzIdentML 1.2 format for XL-MS identification results. It is freely available under a three-clause BSD license at https://openms.org/openpepxl.

Trypsin is the protease of choice in bottom-up proteomics. However, its application can be limited by the amino acid composition of target proteins and the pH of the digestion solution. In this study we characterize ProAlanase, a protease from the fungus Aspergillus niger that cleaves primarily on the C-terminal side of proline and alanine residues. ProAlanase achieves high proteolytic activity and specificity when digestion is carried out at acidic pH (1.5) for relatively short (2 h) time periods. To elucidate the potential of ProAlanase in proteomics applications, we conducted a series of investigations comprising comparative multi-enzymatic profiling of a human cell line proteome, histone PTM analysis, ancient bone protein identification, phosphosite mapping and de novo sequencing of a proline-rich protein and disulfide bond mapping in mAb. The results demonstrate that ProAlanase is highly suitable for proteomics analysis of the arginine- and lysine-rich histones, enabling high sequence coverage of multiple histone family members. It also facilitates an efficient digestion of bone collagen thanks to the cleavage at the C terminus of hydroxyproline which is highly prevalent in collagen. This allows to identify complementary proteins in ProAlanase- and trypsin-digested ancient bone samples, as well as to increase sequence coverage of noncollagenous proteins. Moreover, digestion with ProAlanase improves protein sequence coverage and phosphosite localization for the proline-rich protein Notch3 intracellular domain (N3ICD). Furthermore, we achieve a nearly complete coverage of N3ICD protein by de novo sequencing using the combination of ProAlanase and tryptic peptides. Finally, we demonstrate that ProAlanase is efficient in disulfide bond mapping, showing high coverage of disulfide-containing regions in a nonreduced mAb.

Over the past decade, modern methods of MS (MS) have emerged that allow reliable, fast and cost-effective identification of pathogenic microorganisms. Although MALDI-TOF MS has already revolutionized the way microorganisms are identified, recent years have witnessed also substantial progress in the development of liquid chromatography (LC)-MS based proteomics for microbiological applications. For example, LC-tandem MS (LC-MS²) has been proposed for microbial characterization by means of multiple discriminative peptides that enable identification at the species, or sometimes at the strain level. However, such investigations can be laborious and time-consuming, especially if the experimental LC-MS² data are tested against sequence databases covering a broad panel of different microbiological taxa. In this proof of concept study, we present an alternative bottom-up proteomics method for microbial identification. The proposed approach involves efficient extraction of proteins from cultivated microbial cells, digestion by trypsin and LC–MS measurements. Peptide masses are then extracted from MS¹ data and systematically tested against an in silico library of all possible peptide mass data compiled in-house. The library has been computed from the UniProt Knowledgebase covering Swiss-Prot and TrEMBL databases and comprises more than 12,000 strain-specific in silico profiles, each containing tens of thousands of peptide mass entries. Identification analysis involves computation of score values derived from correlation coefficients between experimental and strain-specific in silico peptide mass profiles and compilation of score ranking lists. The taxonomic positions of the microbial samples are then determined by using the best-matching database entries. The suggested method is computationally efficient – less than 2 mins per sample - and has been successfully tested by a test set of 39 LC-MS¹ peak lists obtained from 19 different microbial pathogens. The proposed method is rapid, simple and automatable and we foresee wide application potential for future microbiological applications.

Pathway analyses are key methods to analyze 'omics experiments. Nevertheless, integrating data from different 'omics technologies and different species still requires considerable bioinformatics knowledge.

Here we present the novel ReactomeGSA resource for comparative pathway analyses of multi-omics datasets. ReactomeGSA can be used through Reactome's existing web interface and the novel ReactomeGSA R Bioconductor package with explicit support for scRNA-seq data. Data from different species is automatically mapped to a common pathway space. Public data from ExpressionAtlas and Single Cell ExpressionAtlas can be directly integrated in the analysis. ReactomeGSA greatly reduces the technical barrier for multi-omics, cross-species, comparative pathway analyses.

We used ReactomeGSA to characterize the role of B cells in anti-tumor immunity. We compared B cell rich and poor human cancer samples from five of the Cancer Genome Atlas (TCGA) transcriptomics and two of the Clinical Proteomic Tumor Analysis Consortium (CPTAC) proteomics studies. B cell-rich lung adenocarcinoma samples lacked the otherwise present activation through NFkappaB. This may be linked to the presence of a specific subset of tumor associated IgG+ plasma cells that lack NFkappaB activation in scRNA-seq data from human melanoma. This showcases how ReactomeGSA can derive novel biomedical insights by integrating large multi-omics datasets.

Despite the crucial function of the small intestine in nutrient uptake our understanding of the molecular events underlying the digestive function is still rudimentary. Recent studies demonstrated that enterocytes do not direct the entire dietary triacylglycerol toward immediate chylomicron synthesis. Especially after high-fat challenges, parts of the resynthesized triacylglycerol are packaged into cytosolic lipid droplets for transient storage in the endothelial layer of the small intestine. The reason for this temporary storage of triacylglycerol is not completely understood. To utilize lipids from cytosolic lipid droplets for chylomicron synthesis in the endoplasmic reticulum, stored triacylglycerol has to be hydrolyzed either by cytosolic lipolysis or lipophagy. Interestingly, triacylglycerol storage and chylomicron secretion rates are unevenly distributed along the small intestine, with the proximal jejunum exhibiting the highest intermittent storage capacity. We hypothesize that correlating hydrolytic enzyme activities with the reported distribution of triacylglycerol storage and chylomicron secretion in different sections of the small intestine is a promising strategy to determine key enzymes in triacylglycerol remobilization. We employed a serine hydrolase specific activity-based labeling approach in combination with quantitative proteomics to identify and rank hydrolases based on their relative activity in 11 sections of the small intestine. Moreover, we identified several clusters of enzymes showing similar activity distribution along the small intestine. Merging our activity-based results with substrate specificity and subcellular localization known from previous studies, carboxylesterase 2e and arylacetamide deacetylase emerge as promising candidates for triacylglycerol mobilization from cytosolic lipid droplets in enterocytes.

Extracellular vesicles (EVs) secreted by the epididymal epithelium transfer to spermatozoa key proteins that are essential in promoting motility and subsequent fertilization success. Using the domestic cat model, the objectives were to (1) characterize and compare protein content of EVs between segments of the epididymis, and (2) compare EV protein compositions between normo- and teratospermic individuals (producing >60% of abnormal spermatozoa). Epididymal EVs from adult cats were isolated and assessed via liquid chromatography tandem MS. Both male types shared 3008 proteins in total, with 98 and 20 EV proteins unique to normospermic and teratospermic males, respectively. Expression levels of several proteins changed between epididymal segments in both male types. Several proteins in both groups were related to sperm motility (e.g. hexokinase 1, adenylate kinase isoenzyme) and zona pellucida or oolemma binding (e.g. disintegrin and metalloproteinase domain proteins, zona binding proteins 1 and 2). Interestingly, seven cauda-derived EV proteins trended downward in teratospermic compared with normospermic males, which may relate to poor sperm quality. Collective results revealed, for the first time, EV proteins related to sequential sperm maturation with differences observed between normospermic and teratospermic individuals.

Endometrial carcinoma (EC) is the most common gynecologic malignancy in the United States, with limited effective targeted therapies. Endometrial tumors exhibit frequent alterations in protein kinases, yet only a small fraction of the kinome has been therapeutically explored. To identify kinase therapeutic avenues for EC, we profiled the kinome of endometrial tumors and normal endometrial tissues using Multiplexed Inhibitor Beads and Mass Spectrometry (MIB-MS). Our proteomics analysis identified a network of kinases overexpressed in tumors, including Serine/Arginine-Rich Splicing Factor Kinase 1 (SRPK1). Immunohistochemical (IHC) analysis of endometrial tumors confirmed MIB-MS findings and showed SRPK1 protein levels were highly expressed in endometrioid and uterine serous cancer (USC) histological subtypes. Moreover, querying large-scale genomics studies of EC tumors revealed high expression of SRPK1 correlated with poor survival. Loss-of-function studies targeting SRPK1 in an established USC cell line demonstrated SRPK1 was integral for RNA splicing, as well as cell cycle progression and survival under nutrient deficient conditions. Profiling of USC cells identified a compensatory response to SRPK1 inhibition that involved EGFR and the up-regulation of IGF1R and downstream AKT signaling. Co-targeting SRPK1 and EGFR or IGF1R synergistically enhanced growth inhibition in serous and endometrioid cell lines, representing a promising combination therapy for EC.

The absence of the dystrophin protein in Duchenne muscular dystrophy (DMD) results in myofiber fragility and a plethora of downstream secondary pathologies. Although a variety of experimental therapies are in development, achieving effective treatments for DMD remains exceptionally challenging, not least because the pathological consequences of dystrophin loss are incompletely understood. Here we have performed proteome profiling in tibialis anterior muscles from two murine DMD models (mdx and mdx52) at three ages (8, 16, and 80 weeks of age), all n = 3. High-resolution isoelectric focusing liquid chromatography-tandem MS (HiRIEF-LC–MS/MS) was used to quantify the expression of 4974 proteins across all 27 samples. The two dystrophic models were found to be highly similar, whereas multiple proteins were differentially expressed relative to WT (C57BL/6) controls at each age. Furthermore, 1795 proteins were differentially expressed when samples were pooled across ages and dystrophic strains. These included numerous proteins associated with the extracellular matrix and muscle function that have not been reported previously. Pathway analysis revealed multiple perturbed pathways and predicted upstream regulators, which together are indicative of cross-talk between inflammatory, metabolic, and muscle growth pathways (e.g. TNF, INF, NF-B, SIRT1, AMPK, PGC-1α, PPARs, ILK, and AKT/PI3K). Upregulation of CAV3, MVP and PAK1 protein expression was validated in dystrophic muscle by Western blot. Furthermore, MVP was upregulated during, but not required for, the differentiation of C2C12 myoblasts suggesting that this protein may affect muscle regeneration. This study provides novel insights into mutation-independent proteomic signatures characteristic of the dystrophic phenotype and its progression with aging.

Sepsis-induced acute kidney injury (S-AKI) is the most common complication in hospitalized and critically ill patients, highlighted by a rapid decline of kidney function occurring a few hours or days after sepsis onset. Systemic inflammation elicited by microbial infections is believed to lead to kidney damage under immunocompromised conditions. However, although AKI has been recognized as a disease with long-term sequelae, partly because of the associated higher risk of chronic kidney disease (CKD), the understanding of kidney pathophysiology at the molecular level and the global view of dynamic regulations in situ after S-AKI, including the transition to CKD, remains limited. Existing studies of S-AKI mainly focus on deriving sepsis biomarkers from body fluids. In the present study, we constructed a mid-severity septic murine model using cecal ligation and puncture (CLP), and examined the temporal changes to the kidney proteome and phosphoproteome at day 2 and day 7 after CLP surgery, corresponding to S-AKI and the transition to CKD, respectively, by employing an ultrafast and economical filter-based sample processing method combined with the label-free quantitation approach. Collectively, we identified 2,119 proteins and 2950 phosphosites through multi-proteomics analyses. Among them, we identified an array of highly promising candidate marker proteins indicative of disease onset and progression accompanied by immunoblot validations, and further denoted the pathways that are specifically responsive to S-AKI and its transition to CKD, which include regulation of cell metabolism regulation, oxidative stress, and energy consumption in the diseased kidneys. Our data can serve as an enriched resource for the identification of mechanisms and biomarkers for sepsis-induced kidney diseases.

Specific E3 ligases target tumor suppressors for degradation. Inhibition of such E3 ligases may be an important approach to cancer treatment. RNF146 is a RING domain and PARylation-dependent E3 ligase that functions as an activator of the β-catenin/Wnt and YAP/Hippo pathways by targeting the degradation of several tumor suppressors. Tankyrases 1 and 2 (TNKS1/2) are the only known poly-ADP-ribosyltransferases that require RNF146 to degrade their substrates. However, systematic identification of RNF146 substrates have not yet been performed. To uncover substrates of RNF146 that are targeted for degradation, we generated RNF146 knockout cells and TNKS1/2-double knockout cells and performed proteome profiling with label-free quantification as well as transcriptome analysis. We identified 160 potential substrates of RNF146, which included many known substrates of RNF146 and TNKS1/2 and 122 potential TNKS-independent substrates of RNF146. In addition, we validated OTU domain-containing protein 5 and Protein mono-ADP-ribosyltransferase PARP10 as TNKS1/2-independent substrates of RNF146 and SARDH as a novel substrate of TNKS1/2 and RNF146. Our study is the first proteome-wide analysis of potential RNF146 substrates. Together, these findings not only demonstrate that proteome profiling can be a useful general approach for the systemic identification of substrates of E3 ligases but also reveal new substrates of RNF146, which provides a resource for further functional studies.

AAA+ ATPases constitute a large family of proteins that are involved in a plethora of cellular processes including DNA disassembly, protein degradation and protein complex disassembly. They typically form a hexametric ring-shaped structure with six subunits in a (pseudo) 6-fold symmetry. In a subset of AAA+ ATPases that facilitate protein unfolding and degradation, six subunits cooperate to translocate protein substrates through a central pore in the ring. The number and type of nucleotides in an AAA+ ATPase hexamer is inherently linked to the mechanism that underlies cooperation among subunits and couples ATP hydrolysis with substrate translocation. We conducted a native MS study of a monodispersed form of PAN, an archaeal proteasome AAA+ ATPase, to determine the number of nucleotides bound to each hexamer of the WT protein. We utilized ADP and its analogs (TNP-ADP and mant-ADP), and a nonhydrolyzable ATP analog (AMP-PNP) to study nucleotide site occupancy within the PAN hexamer in ADP- and ATP-binding states, respectively. Throughout all experiments we used a Walker A mutant (PAN^K217A) that is impaired in nucleotide binding as an internal standard to mitigate the effects of residual solvation on mass measurement accuracy and to serve as a reference protein to control for nonspecific nucleotide binding. This approach led to the unambiguous finding that a WT PAN hexamer carried – from expression host – six tightly bound ADP molecules that could be exchanged for ADP and ATP analogs. Although the Walker A mutant did not bind ADP analogs, it did bind AMP-PNP, albeit at multiple stoichiometries. We observed variable levels of hexamer dissociation and an appearance of multimeric species with the over-charged molecular ion distributions across repeated experiments. We posit that these phenomena originated during ESI process at the final stages of ESI droplet evolution.

Plasmodium, the malaria parasite, undergoes a complex life cycle alternating between a vertebrate host and a mosquito vector of the genus Anopheles. In red blood cells of the vertebrate host, Plasmodium multiplies asexually or differentiates into gamete precursors, the male and female gametocytes, responsible for parasite transmission. Sexual stage maturation occurs in the midgut of the mosquito vector, where male and female gametes egress from the host erythrocytes to fuse and form a zygote. Gamete egress entails the successive rupture of two membranes surrounding the parasite, the parasitophorous vacuole membrane and the erythrocyte plasma membrane. In this study, we used the rodent model parasite Plasmodium berghei to design a label-free quantitative proteomic approach aimed at identifying gender-related proteins differentially released/secreted by purified mature gametocytes when activated to form gametes. We compared the abundance of molecules secreted by wild type gametocytes of both genders with that of a transgenic line defective in male gamete maturation and egress. This enabled us to provide a comprehensive data set of egress-related molecules and their gender specificity. Using specific antibodies, we validated eleven candidate molecules, predicted as either gender-specific or common to both male and female gametocytes. All of them localize to punctuate, vesicle-like structures that relocate to cell periphery upon activation, but only three of them localize to the gametocyte-specific secretory vesicles named osmiophilic bodies. Our results confirm that the egress process involves a tightly coordinated secretory apparatus that includes different types of vesicles and may put the basis for functional studies aimed at designing novel transmission-blocking molecules.

Mallory-Denk-bodies (MDBs) are hepatic protein aggregates associated with inflammation both clinically and in MDB-inducing models. Similar protein aggregation in neurodegenerative diseases also triggers inflammation and NF-B activation. However, the precise mechanism that links protein aggregation to NF-B-activation and inflammatory response remains unclear. Herein we find that treating primary hepatocytes with MDB-inducing agents (N-methylprotoporphyrin (NMPP), protoporphyrin IX (PPIX), or Zinc-protoporphyrin IX (ZnPP)) elicited an IBα-loss with consequent NF-B activation. Four known mechanisms of IBα-loss i.e. the canonical ubiquitin-dependent proteasomal degradation (UPD), autophagic-lysosomal degradation, calpain degradation and translational inhibition, were all probed and excluded. Immunofluorescence analyses of ZnPP-treated cells coupled with 8 M urea/CHAPS-extraction revealed that this IBα-loss was due to its sequestration along with IBβ into insoluble aggregates, thereby releasing NF-B. Through affinity pulldown, proximity biotinylation by antibody recognition, and other proteomic analyses, we verified that NF-B subunit p65, which stably interacts with IBα under normal conditions, no longer binds to it upon ZnPP-treatment. Additionally, we identified 10 proteins that interact with IBα under baseline conditions, aggregate upon ZnPP-treatment, and maintain the interaction with IBα after ZnPP-treatment, either by cosequestering into insoluble aggregates or through a different mechanism. Of these 10 proteins, the nucleoporins Nup153 and Nup358/RanBP2 were identified through RNA-interference, as mediators of IBα-nuclear import. The concurrent aggregation of IBα, NUP153, and RanBP2 upon ZnPP-treatment, synergistically precluded the nuclear entry of IBα and its consequent binding and termination of NF-B activation. This novel mechanism may account for the protein aggregate-induced inflammation observed in liver diseases, thus identifying novel targets for therapeutic intervention. Because of inherent commonalities this MDB cell model is a bona fide protoporphyric model, making these findings equally relevant to the liver inflammation associated with clinical protoporphyria.

At neuronal synapses, activation of group I metabotropic glutamate receptors (mGluR1/5) triggers a form of long-term depression (mGluR-LTD) that relies on new protein synthesis and the internalization of AMPA-type glutamate receptors. Dysregulation of these processes has been implicated in the development of mental disorders such as autism spectrum disorders and therefore merit a better understanding on a molecular level. Here, to study mGluR-induced signaling pathways, we integrated quantitative phosphoproteomics with the analyses of newly synthesized proteins via bio-orthogonal amino acids (azidohomoalanine) in a pulsed labeling strategy in cultured hippocampal neurons stimulated with DHPG, a specific agonist for group I mGluRs. We identified several kinases with important roles in DHPG-induced mGluR activation, which we confirmed using small molecule kinase inhibitors. Furthermore, changes in the AMPA receptor endocytosis pathway in both protein synthesis and protein phosphorylation were identified, whereby Intersectin-1 was validated as a novel player in this pathway. This study revealed several new insights into the molecular pathways downstream of group I mGluR activation in hippocampal neurons, and provides a rich resource for further analyses.

Huanglongbing (HLB) is the most devastating and widespread citrus disease. All commercial citrus varieties are susceptible to the HLB-associated bacterium, Candidatus Liberibacter asiaticus (CLas), which resides in the phloem. The phloem is part of the plant vascular system and is involved in sugar transport. To investigate the plant response to CLas, we enriched for proteins surrounding the phloem in an HLB susceptible sweet orange variety, Washington navel (Citrus sinensis (L) Osbeck). Quantitative proteomics revealed global changes in the citrus proteome after CLas inoculation. Plant metabolism and translation were suppressed, whereas defense-related proteins such as peroxidases, proteases and protease inhibitors were induced in the vasculature. Transcript accumulation and enzymatic activity of plant peroxidases in CLas infected sweet orange varieties under greenhouse and field conditions were assessed. Although peroxidase transcript accumulation was induced in CLas infected sweet orange varieties, peroxidase enzymatic activity varied. Specific serine proteases were up-regulated in Washington navel in the presence of CLas based on quantitative proteomics. Subsequent activity-based protein profiling revealed increased activity of two serine proteases, and reduced activity of one protease in two C. sinensis sweet orange varieties under greenhouse and field conditions. The observations in the current study highlight global reprogramming of the citrus vascular proteome and differential regulation of enzyme classes in response to CLas infection. These results open an avenue for further investigation of diverse responses to HLB across different environmental conditions and citrus genotypes.

Stroke remains a leading cause of death and disability worldwide. Despite continuous advances, the identification of key molecular signatures in the hyper-acute phase of ischemic stroke is still a primary interest for translational research on stroke diagnosis, prognosis, and treatment. Data integration from high-throughput -omics techniques has become crucial to unraveling key interactions among different molecular elements in complex biological contexts, such as ischemic stroke. Thus, we used advanced data integration methods for a multi-level joint analysis of transcriptomics and proteomics data sets obtained from mouse brains at 2 h after cerebral ischemia. By modeling net-like correlation structures, we identified an integrated network of genes and proteins that are differentially expressed at a very early stage after stroke. We validated 10 of these deregulated elements in acute stroke, and changes in their expression pattern over time after cerebral ischemia were described. Of these, CLDN20, GADD45G, RGS2, BAG5, and CTNND2 were next evaluated as blood biomarkers of cerebral ischemia in mice and human blood samples, which were obtained from stroke patients and patients presenting stroke-mimicking conditions. Our findings indicate that CTNND2 levels in blood might potentially be useful for distinguishing ischemic strokes from stroke-mimicking conditions in the hyper-acute phase of the disease. Furthermore, circulating GADD45G content within the first 6 h after stroke could also play a key role in predicting poor outcomes in stroke patients. For the first time, we have used an integrative biostatistical approach to elucidate key molecules in the initial stages of stroke pathophysiology and highlight new notable molecules that might be further considered as blood biomarkers of ischemic stroke.

A new transatlantic relationship? 4 October 2022 — 6:30PM TO 7:30PM Anonymous (not verified) 22 September 2022 Chatham House and Online

US senator Jeanne Shaheen examines the implications of new UK leadership, the war in Ukraine, and NATO expansion for the US–UK relationship.

In recent weeks, the UK has ushered in a new prime minister and a new monarch. The US will hold potentially power-shifting mid-term elections in November after nearly two years of the Biden presidency that promised to bring the US ‘back’ as a global leader in international affairs.

These leadership changes come at a time when Europe is at war, NATO is expanding and US–China competition is re-ordering long-held alliances. Old assumptions about foreign policy are in flux in the midst of huge international challenges.

Democratic senator Shaheen, a senior member of the Senate Foreign Relations Committee, explores how these changes might influence the US–UK ‘special’ relationship.

• How will the trajectory of Russia’s war on Ukraine influence the bilateral relationship? What leadership is needed now?

• What does Russia’s war on Ukraine mean for NATO in responding to other pressing security challenges?

• What domestic constraints might limit the US’s power to reinsert itself as a global leader?

As with all Chatham House member events, questions from the members drive the conversation.

Read the transcript. 

War on Ukraine: The state of the global response 17 October 2022 — 6:30PM TO 7:30PM Anonymous (not verified) 3 October 2022 Chatham House and Online

Implications of the war for the future of multilateralism.

Russia’s war on Ukraine has tested the capacity for a unified global response to grave violations of the UN Charter. The world is in unchartered territory as a nuclear member of the United Nations Security Council attacks a non-nuclear country. 

Multilateral institutions that were born out of an effort to prevent war are struggling to prove their relevance in the face of growing existential threats to humanity. The war is exacerbating divisions within the global community, disrupting food and energy supplies worldwide and contributing to a profound crisis of multilateralism.

The longer these divisions last, the longer the war in the middle of Europe and the harder it will be to respond to the interconnected global crises that threaten everyone.

This discussion offers a unique insight into the macro-geopolitical questions in relation to the war in Ukraine with members of the Elders and other experts:

The panel considers:

• How can a more united global response to Russian aggression be built?

• What stands in the way of an effective multilateral response based on international norms?

• In what ways are divisions between UN member states influencing the trajectory of the war or prospects for peace?

• How is the conflict changing geopolitics and the ability of the multilateral system to address global challenges?

As with all members events, questions from the audience drive the conversation. This event is organised in partnership between Chatham House and The Elders, the group of independent global leaders founded by Nelson Mandela who work for peace, justice and human rights.

Read the transcript.

Black perspectives on international relations 27 October 2022 — 5:00PM TO 6:00PM Anonymous (not verified) 4 October 2022 Chatham House and Online

How can black perspectives help the world tackle global challenges and expand our understanding of international relations?

As social boundaries change, the viewpoints of black academics, thought leaders and policymakers have grown in both influence and strength worldwide, challenging western and institutional norms. 

However, many institutions and organizations, long established with the exclusion of black voices, have to adapt if there is to be greater inclusion and diversity of thought when tackling major global issues.

Growing reflection on the legacy of colonialism and the importance of the power of diversity may be needed for today’s problems.

The search for global racial equality has seen a growing commitment to ensuring the black experience is at the heart of geopolitical discussions.

This panel discussion looks at what changes are occurring now and how is the conversation shifting. It also examines the challenges posed by the increasing politicization of race and culture issues in the current political environment.

Key questions discussed by the panel include:

• What is the importance of black voices in international relations and where are the main challenges to greater incorporation?

• What are some of the leading perspectives, approaches and beliefs within Africa and across the black diaspora regarding international relations?

• To what extent are governments, businesses and leading global institutions making efforts to include more black voices in decision-making?

• How are black academics changing wider geopolitical conversations and to what extent can deeper conversations lead to change?

• Will an ‘African Century’ bring black perspectives on international relations to the fore of the geopolitical agenda?

As with all members events, questions from the audience drive the conversation.

Read the transcript. 

War on Ukraine: The energy crisis and Europe’s impending long winter 2 November 2022 — 5:00PM TO 6:00PM Anonymous (not verified) 4 October 2022 Online

Can Europe remain unified over the long winter?

Since Russia’s invasion of Ukraine, the global community has been responding to significant price shocks, especially energy. As Europe heads into a particularly difficult winter, policymakers are grappling with the costs, both political and economic, required to make sure Russian energy blackmail does not succeed.

Retaining a unified front against Russia and providing continued support to the Ukrainian government will be great challenges. As the cold begins to bite, war fatigue may accelerate among the populations of Europe. Providing their people with adequate heat will not come cheaply for governments across the continent at a time of economic uncertainty.

At this critical moment of Russia’s invasion, experts discuss:

• Have European preparations been sufficient to stave off an energy crisis this winter?
• What will be Russia’s reaction during and after the winter period, particularly if Europe avoids energy market failures?
• How will this ‘energy crisis’ ensure future dependencies on single state actors of goods and services do not occur in the future?

Read the transcript. 

Chatham House Primer: Inside China’s government 30 November 2022 — 6:00PM TO 6:45PM Anonymous (not verified) 4 October 2022 Chatham House

How are decisions made in Beijing, across China and where does the CCP fit in?

Still little is known in Western circles about the inner workings of China’s government. In power since 1949, the ruling Chinese Communist Party has evolved over decades to its current embodiment under President Xi Jinping.

The need for a better understanding of China’s government has been heightened, particularly as the country navigates post-COVID troubles, global economic downturns, wars in Europe, climate change and heightened tension with the United States.

This Primer has been prepared to pull back the veil on the Chinese government. Key issues to be tackled include:

• What is the decision-making process in China’s government?

• How is the party–government relationship best explained?

• How has the party evolved in recent years with new forms of governance and leadership?

• How has China’s government evolved in recent years, particularly in a globalized environment?

• A description of the central government–province dynamic?

• How are citizens engaged in the political process?

• What are the major centres of power in the Chinese political system?

• Has the COVID-19 pandemic altered attitudes towards and the operation of government?

As with all Chatham House member events, questions from members drive the conversation.

Expanding and enhancing the global cyber workforce 17 November 2022 — 5:00PM TO 6:00PM Anonymous (not verified) 5 October 2022 Chatham House and Online

How can we address the cybersecurity workforce shortage and skills gap?

Accelerated digital transformation and heightened geopolitical tensions on the international stage have increased the need for effective cybersecurity practices and policies as well as a skilled workforce. Despite this, the demand for cybersecurity professionals continues to outpace the supply for societies and businesses globally, resulting in a cybersecurity workforce gap.

To ensure that digital transformation is available, safe and beneficial to all, significant efforts are needed to encourage cyber workforce capacity-building and knowledge-sharing at both national and international levels.

This discussion, supported by (ISC)² and the UK Cyber Security Council, will explore how to effectively address the twin challenges of the global cyber workforce shortage and skills gap.

• What are the implications of the global cyber workforce and skills gaps for businesses and societies?

• What shape do these gaps take within society? Where are they most prevalent and how do they vary?

• What is the role of education and private-public partnerships in effectively addressing these gaps?

• How does the UK National Cyber Strategy seek to address these challenges? What are the key lessons from this strategy?

• What other efforts are being made internationally to bridge this gap? What opportunities are there for knowledge-sharing and capacity-building?

• What is the role of diversity, equity and inclusion in tackling these gaps?

As with all members events, questions from the audience drive the conversation. If you are not a member of Chatham House but would still like to attend the event please email Eleanor Macmillan-Fox to enquire about registration.

Read the transcript. 

Advanced technologies in the face of war 24 October 2022 — 1:00PM TO 2:00PM Anonymous (not verified) 5 October 2022 Online

How is NATO strengthening its technological edge?

Russia’s invasion of Ukraine has brought with it a heavy focus on technology and weaponry, particularly as casualties mount and large numbers of equipment are lost on both sides. The conflict has highlighted how states and their militaries seek technological superiority and how access to advanced capabilities can help shape the course of the war.

Aiming to sharpen the Alliance’s technological edge, NATO is working to support the development of emerging and potentially disruptive technologies such as artificial intelligence (AI), autonomous systems, biotechnologies and quantum technologies that are seen as presenting both risks and opportunities for the Alliance.

As part of this work, NATO’s newly formed Defence Innovation Accelerator for the North Atlantic (DIANA), hosted by both the UK and Estonia, brings together academia, industry and government to support the development of critical technologies to deter and defend against existing and future threats.

Key questions to be considered by the panel include:

• How will the technologies that form the focus of DIANA’s efforts strengthen the Alliance and prepare it to better deal with threats to peace and security across the region?

• How will these technologies be applied and used in war?

• To what extent can a war be won by technology?

• Is Ukraine, and other future conflict zones, in danger of becoming a testing ground for emerging technologies?

• What has the war in Ukraine taught NATO about modern warfare and how should the Alliance respond to this?

• After the commotion of AUKUS, how will the Alliance manage the sharing of technologies and IP among member states?

As with all members events, questions from the audience drive the conversation.

Read the transcript. 

American democracy in 2022: Trump, insurrection, and midterm elections 31 October 2022 — 2:00PM TO 3:00PM Anonymous (not verified) 7 October 2022 Online

How much has Donald Trump changed US politics and democracy, and will Trump and ‘Trumpism’ be more or less significant in the years ahead?

America’s democracy is divided, polarized and fragmenting. Inequality and internal division have a long history. But Trump’s lasting influence on the Republican party, and politics more broadly, continues to leave a mark. Repeated denials of President Joe Biden’s 2020 election win, wrapped in claims of electoral fraud, have eroded faith in the democratic institutions.

The memories of 6 January are still fresh, reminding all of the dangers posed by such actions. All told, America’s democracy has taken a beating in recent years.

To help make sense of the events over recent years and consequences for the coming mid-terms, Peter Baker and Susan Glasser (authors of The Divider: Trump in the White House 2017–2021) walk through in detail how the American politics of today has been arrived at.

Key questions discussed include:

• What has been learned from the January 6 Committee?

• Is there a likelihood of a similar event in the future?

• When and how will Trump lose his influence over the Republican party? 

• What are the broader ramifications of the Trump era?

• What did the events of 6 January mean for America’s relationships globally?

 As with all Chatham House member events, questions from members drive the conversation.

Read the transcript. 

Members' drinks and exhibition 27 October 2022 — 6:00PM TO 8:00PM Anonymous (not verified) 7 October 2022 Chatham House

A showcase of a selection of the Black Cultural Archives photo exhibition of up-and-coming Black British leaders.

Chatham House is pleased to showcase a selection of the Black Cultural Archives photo exhibition of up-and-coming Black British leaders. These portraits illuminate the talent and pipeline of ‘next gen’ leaders in the Black British community – some of whom will be in attendance at the reception. The chair of the Black British Cultural Archives, Dr Yvonne Thompson, will make short remarks at around 18:20 BST.

We hope you will join your fellow members and Chatham House staff for a chance to connect and celebrate.

You are welcome to attend the event Black perspectives on International Relations preceding the reception.

Please note as space is limited, this event is operating a ballot for registrations. Your place will be confirmed by Tuesday 25 October if you are successful.

The battle for truth: The BBC's role at 100 22 November 2022 — 6:30PM TO 7:30PM Anonymous (not verified) 13 October 2022 Chatham House and Online

Tim Davie, director general of the BBC, and others discuss how the BBC shapes, and is shaped by, the world today.

This year, the BBC turns 100 and the World Service 90 in a world facing crises of increasing scale and frequency and in an age of disinformation, democratic disruption and a growing assault on truth and free reporting worldwide. What does this mean for Britain’s foremost news provider at home and across the globe?

This conversation considers:

• How does the BBC navigate a drastically changing media landscape?

• What does the BBC represent in the UK and to the world?

• What ‘soft power’ does the BBC have and how does it use it best?

As with all members events, questions from the audience drive the conversation.

Read the transcript. 

Russia's war on everybody 6 December 2022 — 5:00PM TO 6:00PM Anonymous (not verified) 14 October 2022 Chatham House and Online

Experts discuss the methods Moscow has employed to exert influence around the world over recent decades.

Russia’s assault on Ukraine has reminded the world about the threat it faces from Moscow. But that’s not the only war that Russia has been fighting and Ukraine is not the only target.

Long before February 2022, Russia was already engaged in semi-covert campaigns across Europe and around the world, using any means possible to expand its power and influence and leaving a trail of destruction along the way.

In his new book Russia’s War on Everybody, Chatham House associate fellow Keir Giles examines what this longer war means for us all. Instead of talking only to diplomats, politicians and generals, Giles has looked instead at the effect of Russia’s ambition on ordinary people.

Interviewing 40 eyewitnesses from four continents, he has tried to tell the stories the world doesn’t hear about the impact of Russia’s hostility on individuals and societies that may not even realize they are a target.

At this event, Giles introduces the book at Chatham House. He is joined by experts to talk about the human impact of Russia’s campaigns waged through leveraging corruption and cyber offensives respectively.

As with all members events, questions from the audience drive the conversation.

Read the transcript. 

The future of global trade: Beyond ‘peak globalization’? 23 November 2022 — 11:00AM TO 12:00PM Anonymous (not verified) 17 October 2022 Online

Is globalization in retreat?

The COVID-19 pandemic and Russia’s war on Ukraine have highlighted how vulnerable international trade is. But, even before these recent shocks, rising protectionism in major economies around the world and concerns about the environment have weighed heavily on trade.

According to some key measures, the globalization trend appears to have slowed. But is ‘peak globalization’ a reality or a myth? What are the major phases of globaliszation and what might come next? The answer differs between trade in goods, services, capital, technology, data and people. And whether the future is a more integrated or fragmented world economy also depends on politics and the stability of the international order.

Key questions to be tackled at this event includes:

• How do recent shocks, such as the COVID-19 pandemic and Russia’s war on Ukraine, change globalization? 

• What are the key indicators for the global integration of major economies?

• Will there be a split between a US and China-dominated ’trading sphere of influence’?

• Could trade in services offer ‘globalization’ a new phase of rapid growth?

• What impact will technology continue to have on global trade and the future of globalization?

As with all members events, questions from the audience drive the conversation.

The discussion is part of the Chatham House Global Trade Policy Forum. The Global Trade Policy Forum is supported by founding partner AIG, associate partner Boston Consulting Group and supporting partners Clifford Chance LLP, Diageo PLC and UPS.

Read the transcript. 

The Commonwealth reimagined 8 November 2022 — 6:00PM TO 7:00PM Anonymous (not verified) 18 October 2022 Chatham House and Online

Ghana’s minister of foreign Affairs, the Hon. Shirley Ayorkor Botchwey, discusses her vision for a modern Commonwealth and how it can evolve and match demands from its members.

The death of HM Queen Elizabeth II has focused attention on the future of the Commonwealth. The Commonwealth is an expanding voluntary organization of 56 independent countries in Africa, Asia, the Americas, Europe, and the Pacific.

Its appeal is increasingly beyond the circle of former British colonies – ex-French colonies Togo and Gabon officially joined in October 2022 and the ex-Portuguese colony, Angola, has applied. The Commonwealth Secretariat, established in 1965, is its main intergovernmental agency, which coordinates and carries out much of the Commonwealth’s work, supported by a network of more than 80 organizations.

King Charles III now heads the Commonwealth, which is focused on shared goals of prosperity, democracy and peace. However, the future of the Commonwealth and its purpose are unclear, and the organization needs to develop a sharper agenda on what its international contribution can be across its 56 state members and their peoples.

The minister discusses key questions including:

• What should a modern Commonwealth look like and how can it best operate?

• How can the organization impact policies and actions at a country level?

• What role will young people play in the future of the Commonwealth?

• How can the organization harness collective resources and technology to tackle major global issues such as climate change?

• Can the issue of mobility and immigration among member states be managed?

Iran: Protests, politics and power 16 November 2022 — 6:00PM TO 7:00PM Anonymous (not verified) 18 October 2022 Online

Join Robert Macaire, UK ambassador to Iran (2018-21), and others to discuss what the protests mean for Iran’s domestic, regional and global power.

Protests in Iran, spurred after Masha Amini died in police custody, have drawn focus on how Iranians feel about state repression, a struggling economy and global isolation. Iran is facing the most adamant challenge to its power structure since the ‘green movement’ in 2009 with protests taking place in more than 50 cities and towns across the country. There is no sign that the government will back down but what will that decision mean for the power it can wield at home and abroad?

This conversation examines how the protests impact Iran’s domestic power, its regional relationships and its relations with the US.

• What do the protests demonstrate about Iran’s power domestically and regionally?

• How do the protests influence the JCPOA?

• What will the government gain if they hold a hard line on protesters?

• How do the protests impact Iran’s regional activities?

As with all members events, questions from the audience drive the conversation.

Read the transcript. 

What’s next in UK monetary policy? 4 November 2022 — 4:00PM TO 5:30PM Anonymous (not verified) 19 October 2022 Chatham House and Online

A panel of leading experts discuss the future direction of UK monetary policy.

The UK’s so-called ‘mini-budget’ on 23 September led to a severe market reaction and a wave of criticism at home and abroad that ultimately forced the sacking of UK chancellor Kwasi Kwarteng and contributed to the downfall of Liz Truss’s government.

The new chancellor Jeremy Hunt is due to deliver what will essentially be an entirely new budget in mid-November, with a full assessment from the Office of Budget Responsibility. This will follow the meeting of the Bank of England’s interest rate setting Monetary Policy Committee (MPC) on Thursday 3 November.

Given rising inflationary pressures worldwide, it seems highly likely that the MPC will increase interest rates once again, but by how much and how far there will have to be an additional premium linked to the government’s fiscal strategy is far from clear.

Chatham House’s Global Economy and Finance programme is pleased to partner with Fathom Consulting to host a special session of Fathom’s Monetary Policy Forum.

A presentation of Fathom’s latest economic outlook, fully updated to take account of the previous day’s MPC decision, will be followed by a discussion among four of the MPC’s original former external members. Key questions will include:

• How far has the government been able to restore its fiscal credibility?
• Did the MPC make the right decision on 3 November?
• What is the likely pace and extent of monetary tightening in the UK going forward?
• What will be the long-term consequences for the UK economy of the past month’s policy experiment?
• What are the international implications?

As with all members events, questions from the audience drive the conversation. 

This event is in partnership with Fathom Consulting.

The road to COP27: In conversation with US Special Presidential Envoy for Climate John Kerry 27 October 2022 — 3:00PM TO 4:00PM Anonymous (not verified) 20 October 2022 Chatham House and Online

What will progress on climate change look like at COP27?

With global attention zeroing in on COP27, policymakers and world leaders will meet in Egypt to take the next step in the fight against the climate crisis. The planet is on course to warm well beyond 1.5°C and climate hazards are increasing our exposure to climate risk. Violent and unpredictable weather events increasingly leave devastation among communities, particularly in vulnerable countries.

At the same time, the ripple effects of the conflict in Ukraine will have wide-ranging economic, social and geopolitical consequences for years to come. Whilst some finance is being made available, more is needed to properly address the damage caused by climate change and fund the transition to net zero worldwide. These challenges have become more acute as the world grapples with a growing energy crisis, the war in Ukraine and a troubling economic outlook.

Joined by US Special Presidential Envoy for Climate John Kerry, the following questions are considered:

• Is ‘1.5 degrees’ still on track?

• How can countries better collaborate to move to net zero faster?

• How can we achieve progress on adaptation, climate finance, and loss and damage?

As with all members events, questions from the audience drive the conversation.

Read the transcript. 

Middle East and great power competition 28 November 2022 — 12:00PM TO 1:00PM Anonymous (not verified) 25 October 2022 Chatham House and Online

Experts discuss how the Middle East is changing in a fast-moving geopolitical environment.

The war in Ukraine and great power competition define not only global politics but also regional ones. The Middle East is a microcosm for observing how the great power rivalry informs regional affairs.

OPEC+’s decision to reduce oil supply to international markets and many regional states’ balancing act between the West and Russia, for that matter China as well, are only a few recent policy choices that clearly illustrate how the global and regional levels interact with each other.

Plus this is now a region in which the US has downsized its security commitments, whereas Russia has increased its footprint in regional security and China in economy.

This event tries to unpack how the great power rivalry and the war in Ukraine affect regional politics and how the Middle East adjusts itself to this new phase in global politics.

Thinking out loud: Is disinformation here to stay? 10 November 2022 — 6:00PM TO 6:45PM Anonymous (not verified) 25 October 2022 Chatham House

This event is postponed.

Have you ever wondered how Chatham House researchers approach the big deals that become research? Do you enjoy meeting other Chatham House members and engaging with questions that open your mind? ‘Thinking Out Loud’ invites a small group of members to a live, unscripted discussion with a Chatham House researcher. This in-person event is a way for researchers and members to think out loud to help shape ideas for future research.

Kate Jones, Associate Fellow, International Law Programme at Chatham House will pose some key questions facing how speech is governed in an online world:

• How has big tech influenced the way we think about speech and its limitations?

• Can disinformation be eliminated or even greatly reduced?

• Where should the responsibilities fall between government and business when it comes to speech regulation?

• What might the information landscape look like in 10 years’ time? Should that affect how we tackle disinformation today?

As with all members events, questions from the audience drive the conversation.

Weathering the storm: The UK’s role in the world today 29 November 2022 — 12:00PM TO 1:00PM Anonymous (not verified) 7 November 2022 Chatham House and Online

In conversation with David Miliband, examining the risks and opportunities for the UK in a critical year ahead. 

With a new government in the midst of a global order in flux, the UK’s position in the world needs re-examining.

Just 20 months since the UK’s Integrated Review on international policy and security, Britain’s global blueprint is being reviewed and updated in light of major global developments.

Today, Brexit and the Russia’s invasion of Ukraine require adjustments to the UK’s strategic thinking and positioning in the world.

As the economic and political turmoil of previous weeks begins to abate, this is an important moment to once again determine Britain’s role in Europe and beyond. 
 
Realigning British foreign policy in a rapidly shifting international order will be a major challenge for the new administration.  

International Rescue Committee’s CEO and President, and former UK Foreign Secretary, David Miliband, examines the risks and opportunities for a critical year ahead. 
 
Key questions include:

• What are the crucial decisions the UK needs to make in the coming 12 months?
• What should the UK’s priorities be for its role in the world? How should it project itself amidst geopolitical fracturing?
• How can Britain best respond to humanitarian crises around the world?
• Does the UK have the strategic and economic clout to keep up with its foreign policy and development commitments?

As with all Chatham House member events, questions from members drive the conversation.

Read the transcript. 

Members' Christmas drinks 6 December 2022 — 6:00PM TO 8:00PM Anonymous (not verified) 9 November 2022 Chatham House

Join us at 10 St James’s Square for a chance to raise a glass with fellow Chatham House members and staff.

This evening is a special opportunity to meet fellow Chatham House members and staff around the Christmas tree.

Please note this reception is open to members of Chatham House only. Regrettably, we are unable to register non-member guests.

The Director’s Annual Lecture 2023 10 January 2023 — 6:00PM TO 7:00PM Anonymous (not verified) 17 November 2022 Chatham House and Online

Bronwen Maddox looks ahead to the challenges of the year and sets out Chatham House’s recommendations for change.

Read a transcript of the event

A lecture and discussion on the year ahead in international relations with Bronwen Maddox, director and chief executive of Chatham House.

Although Russia’s war on Ukraine rightly dominated headlines in 2022, other challenges also grew, the climate became warmer, US/China competition intensified, deglobalization became a much-analysed theme, and the global economy suffered significant blows.

The UK has its third prime minister in less than one year as it grapples with its changing place in the world. And the world is still living in the shadow of COVID-19 and what the pandemic revealed about strengths and vulnerabilities, global inequity, and North/South divides.

This event examines how the forces that shaped 2022 may manifest in 2023, and what that means for progress in international relations:

• What will progress look like on the climate agenda?
• How will the new US Congress reposition America’s role in the world?
• What does the North/South divergence on Russia’s war in Ukraine tell us about shared values and prospects for working together?
• After a turbulent year, how will the UK recover its standing in Europe and beyond?