New types of foldamer scaffolds are formidably challenging to design and synthesize, yet highly desirable as structural mimics of peptides/proteins with a wide repertoire of functions. In particular, the development of peptidomimetic helical foldamers holds promise for new biomaterials, catalysts, and drug molecules. Unnatural l-sulfono-γ-AApeptides were recently developed and shown to have potential applications in both biomedical and material sciences. However, d-sulfono-γ-AApeptides, the enantiomers of l-sulfono-γ-AApeptides, have never been studied due to the lack of high-resolution three-dimensional structures to guide structure-based design. Herein, we report the first synthesis and X-ray crystal structures of a series of 2:1 l-amino acid/d-sulfono-γ-AApeptide hybrid foldamers, and elucidate their folded conformation at the atomic level. Single-crystal X-ray crystallography indicates that this class of oligomers folds into well-defined right-handed helices with unique helical parameters. The helical structures were consistent with data obtained from solution 2D NMR, CD studies, and molecular dynamics simulations. Our findings are expected to inspire the structure-based design of this type of unique folding biopolymers for biomaterials and biomedical applications.
In order to address the issue of excessive intake of aluminium (Al) from Al-containing food additives in the Chinese diet, this study conducted a dietary exposure assessment of Al in the general population based on the national surveillance data of Al content in foods and national food consumption data. It was found that the mean dietary exposure of the whole Chinese population to Al from Al-containing food additives was 1.795 mg kg‒1 bw week‒1, not exceeding the PTWI, while high dietary exposures (e.g., 97.5th percentile) to Al were 7.660 and 2.103-2.903 mg kg‒1 bw week‒1 for children, respectively, both exceeding the PTWI. It was found that the dietary exposure to Al for 32.5% of the total Chinese population and 42.6% of children aged 4-6 years exceeded the PTWI. Wheat flour and wheat-based products are the main source of dietary A l exposure (85% of the total intake); and puffed foods are the major source of Al intake for children. These findings suggested that consumption of Al-containing food additives could be a health concern for consumers with high food consumption (97.5th percentile) and children under the age of 14 years.
Initiation of B-cell receptor (BCR) 1 signaling, and subsequent antigen-encounter in germinal centers 2,3 represent milestones of B-lymphocyte development that are both marked by sharp increases of CD25 surface-expression. Oncogenic signaling in B-cell leukemia (B-ALL) 4 and lymphoma 5 also induced CD25-surface expression. While CD25 is known as an IL2-receptor chain on T- and NK-cells 6-9 , the significance of its expression on B-cells was unclear. Our experiments based on genetic mouse models and engineered patient-derived xenografts revealed that, rather than functioning as an IL2-receptor chain, CD25 expressed on B-cells assembled an inhibitory complex including PKCδ and SHIP1 and SHP1 phosphatases for feedback control of BCR-signaling or its oncogenic mimics. Recapitulating phenotypes of genetic ablation of PKCδ 10 - 12 , SHIP1 13,14 and SHP1 14, 15,16 , conditional CD25-deletion decimated early B-cell subsets but expanded mature B-cell populations and induced autoimmunity. In B-cell malignancies arising from early (B-ALL) and late (lymphoma) stages of B-cell development, CD25-loss induced cell death in the former and accelerated proliferation in the latter. Clinical outcome annotations mirrored opposite effects of CD25-deletion: high CD25 expression levels predicted poor clinical outcomes for patients with B-ALL, in contrast to favorable outcomes for lymphoma-patients. Biochemical and interactome studies revealed a critical role of CD25 in BCR-feedback regulation: BCR-signaling induced PKCδ-mediated phosphorylation of CD25 on its cytoplasmic tail (S 268 ). Genetic rescue experiments identified CD25-S 268 tail-phosphorylation as central structural requirement to recruit SHIP1 and SHP1 phosphatases to curb BCR-signaling. A single point mutation CD25 S268A abolished recruitment and activation of SHIP1 and SHP1 to limit duration and strength of BCR-signaling. Loss of phosphatase-function, autonomous BCR-signaling and Ca 2+ -oscillations induced anergy and negative selection during early B-cell development, as opposed to excessive proliferation and autoantibody production in mature B-cells. These findings highlight the previously unrecognized role of CD25 in assembling inhibitory phosphatases to control oncogenic signaling in B-cell malignancies and negative selection to prevent autoimmune disease.
Abstract The structurally disordered intracellular loops (ICLs) of G protein-coupled receptors (GPCRs) play a critical role in G protein coupling. In our previous work, we used a combination of FRET-based and computational methodologies to show that the third intracellular loop (ICL3) modulates the activity and G protein coupling selectivity in GPCRs. In the current study, we have uncovered the role of several lipid components in modulating the conformational ensemble of ICL3 of the β2-adrenergic receptor (β2AR). Our findings indicate that phosphatidylinositol 4,5-bisphosphate (PIP2) in the inner leaflet of the membrane bilayer acts as a stabilizing anchor for ICL3, opening the intracellular cavity to facilitate G protein coupling. This interaction between PIP2 and ICL3 causes tilting of β2AR within the cellular membrane. Notably, this tilting of the receptor is supported by ganglioside GM3 stabilizing the extracellular loops on the outer leaflet of the bilayer, thereby exerting an allosteric effect on the orthosteric ligand binding pocket. Our results underscore the significance of lipids in modulating GPCR activity, proposing an allosteric mechanism that occurs through the receptor’s orientation within the membrane.
Abstract Immune checkpoint inhibitor (ICI) therapy has drastically improved the treatment strategies for mucosal head and neck squamous cell cancer (HNSCC). To increase treatment response rates for highly targeted ICI therapies, predictive and prognostic biomarkers are being actively explored. Specifically, the cytokine expression patterns in the tumor microenvironment (TME) are now recognized as key to understanding immune responsive and resistant phenotypes. Cytokines play an essential role in the regulation of the TME, specifically in modulating the proliferation and differentiation of immune cells. Here, we have studied spatial signatures of various cytokines within the TME of metastatic/recurrent HNSCC tumors treated with Pembrolizumab/Nivolumab. We utilized the PhenoCycler-Fusion to perform whole-slide, single cell resolution spatial phenotyping of the TME of HNSCC tumors from a cohort of n=40 patients. The discovery cohort consisted of patients who had complete vs. partial vs. stable vs. progressive responses to ICI therapy. Transcriptomic profiling of more than 60 RNA targets for various subfamily of chemokines, interleukins, and immune cell lineages was achieved using Akoya’s novel high plex RNA detection technology. Our study identified distinct spatial signatures that implicate certain cytokines in either tumor progression or regression. Specifically, we have identified areas of high and low CXCL9 and CXCL10 expression in several tumor regions that reflect immune-cell landscapes associated with resistance and sensitivity to immunotherapy. Our study demonstrates the power of unbiased spatial phenotyping with whole-slide imaging to identify biomarkers associated with response to ICI therapy in HNSCC.
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