Bridging the gap between organelles: Membrane contact sites as regulator of endocytic dynamics.

Chimeric antigen receptor (CAR) T-cell therapy has shown remarkable efficacy in hematologic malignancies but remains limited in solid tumors because of the immunosuppressive microenvironment, tumor heterogeneity, poor immune-cell infiltration, and progressive T-cell dysfunction. Because cytokine costimulation is critical for maintaining T-cell fitness, we developed a modular engineering strategy, distinct from previous approaches based on direct insertion of large cytokine receptor fragments, in which the intracellular CAR signaling domain was reconstructed to incorporate compact IL-2/IL-15 receptor-derived activation motifs, thereby enabling antigen-dependent coactivation while preserving the overall architecture of the parental CAR. Through systematic screening, we identified S71 as the optimal construct, with significantly greater antitumor activity than other mutants across multiple solid and hematologic tumor targets. Mechanistically, S71 rewired CAR signaling and reprogrammed tumor-induced metabolic responses through a self-sustaining mechanism, improving mitochondrial function and supporting durable T-cell activity. Functionally, S71 promoted enhanced persistence and robust immune memory responses against solid tumors. These findings demonstrate that modular integration of cytokine signaling motifs into CAR intracellular domains can improve CAR T-cell fitness and antitumor efficacy, and they establish S71 as a promising strategy for overcoming barriers to CAR T-cell therapy in solid tumors.

Acrodysostosis type 1 (ACRDYS1) is a rare multisystem developmental disorder affecting skeletal growth, endocrine function, neurodevelopment, metabolism, and tooth formation. It is caused by heterozygous mutations in PRKAR1A, which encodes the type Iα regulatory subunit (RIα) of protein kinase A (PKA), a central mediator of cyclic AMP (cAMP)-dependent signalling. Although ACRDYS1 belongs to the broader family of Gsα-cAMP-PKA-related disorders, its underlying mechanism is distinct. Disease-associated PRKAR1A mutations cluster within regions of RIα that bind cAMP and undergo conformational rearrangements required for PKA activation. These variants impair cAMP binding and disrupt the structural transitions needed to disinhibit catalytic subunits. Importantly, mutant RIα is expressed at near-normal levels and assembles efficiently into PKA holoenzymes, but these complexes respond weakly and sluggishly to physiological cAMP signals. Drawing on structural, biochemical, cellular, and in vivo studies, we define a dual pathogenic mechanism underlying ACRDYS1. First, defective cAMP-driven conformational changes reduce the sensitivity and amplitude of type I PKA activation, producing a hypomorphic signalling state despite intact upstream receptor coupling and cAMP production. Second, activation-resistant RIα holoenzymes impose a dominant-negative constraint by retaining catalytic subunits, further limiting the pool available for productive signalling in heterozygous cells. We relate this core defect in signal responsiveness to tissue-specific vulnerability. Impaired RIα-dependent decoding of cAMP signals disrupts the Ihh-PTHrP feedback loop in the growth plate, blunts hormone-responsive transcriptional programmes in endocrine epithelia, and alters spatially restricted PKA signalling domains in neurons and metabolically active tissues. Despite the diversity of affected organs, the unifying defect is an inability to generate appropriately timed and scaled PKA responses. This framework establishes ACRDYS1 as a disorder of signal decoding rather than signal generation, clarifies its mechanistic distinction from PRKAR1A-related Carney Complex, and highlights therapeutic strategies aimed at restoring local cAMP-PKA signalling dynamics rather than globally amplifying pathway activity.

Birds and mammals exhibit extraordinary facial diversity, reflecting adaptations to distinct ecological niches and feeding strategies. While core face-building developmental programs are conserved and orchestrated by interactions between ectodermal organizers and the underlying mesenchyme, mechanisms driving facial shape variation remain poorly understood. Here, we integrate single-cell transcriptomic and chromatin accessibility profiling of mouse and chicken developing face to construct a comparative regulatory map. Although both ectodermal and mesenchymal populations display distinct regulatory features in each species, the mesenchyme exhibits markedly greater divergence, pointing to its central role in shaping facial morphology. We further reveal unexpected molecular complexity in the main face-shaping organizer, including a mouse-specific Shh/Wnt5a expression domain. At key morphogen loci (Bmp4, Fgf8, and Wnt5a), conserved and lineage-specific enhancers exhibit spatially restricted activity patterns that mirror divergent signaling domains. These findings demonstrate how cis-regulatory evolution modulates conserved developmental programs to generate morphological novelty, providing a valuable resource for studying vertebrate facial evolution.

The red/far-red sensing photoreceptor phytochrome B (phyB) governs multifaceted plant development and responses to light and temperature stimuli. PhyB photoconversion between red-absorbing, inactive Pr and far red-absorbing, active Pfr states, imparted by its covalently bound bilin chromophore, enables rapid switching and plasticity of phyB signaling activities. The phyB Y276H variant (YHB) is photochemically inert but adopts a constitutively active Pfr-like structure regardless of light conditions, which becomes a versatile model to dissect phyB signaling mechanisms. Here, we conducted a large-scale EMS mutagenesis screen on YHB -expressing transgenic lines, mining intragenic suppressor mutations that would unveil critical residues for phyB structure-function relationships. Comparative analyses of 26 nonsense variants suggested modular organization of phyB overall structure and dispensability of the C-terminal HKRD domain for phyB signaling. Amongst fourteen novel and nine known loss-of-function missense variants identified herein, G284E was of particular interest for its fully suppressed constitutive activity in darkness and its restored photochemistry and light responsiveness. The G284E mutation was further tested to also nullify another constitutively active phyB Y303V allele by eliminating chromophore attachment. P309L was the sole variant identified which fully suppressed YHB in both dark and light conditions. C402Y profoundly elicited YHB protein instability. Three variants G118R, C402Y and G538D markedly reduced chromophorylation levels of YHB. Although the chromophore binding site variant C357Y was a strong loss-of-function allele, it retained residual signaling activity with respect to PIF3 protein turnover in dark-grown seedlings, presumably due to its ability to noncovalently bind chromophore. Two tandem prolines (P799, P800) proved critical to YHB structural integrity/stability as well as signaling activity. In summary, these diverse variants shed new insights into multiple levels by which the YHB (and thereby phyB) signaling is initiated, tuned, and disseminated.

Chimeric antigen receptor (CAR) T-cell treatment has developed among major substantial improvements for modern cancer treatment, providing sustained responses in patients with otherwise resistant blood cancers. This method comprises of patients reprogramming or donor's T lymphocytes to interpret tumor associated antigens self-sufficiently of major histocompatibility multifaceted presentation, thereby circumventing a key limitation of natural immune surveillance. The approval of CD19- and BCMA-targeted therapies demonstrated remarkable clinical impact and validated the approach. Over successive generations, CAR constructs have been refined with additional costimulatory elements, cytokine support, and multifunctional signaling domains, improving both their persistence and therapeutic activity. Despite such progress, important challenges remain, including risks of relapse, toxicity including neurotoxicity and cytokine release syndrome with limited efficacy in solid tumors. Current research is focused on strategies, such as armored CARs, gene editing, and combination therapies to expand clinical benefit. A comprehensive literature search was conducted using PubMed, Scopus, and Web of Science databases, covering publications from 2000 to 2026 till date. Relevant peer-reviewed articles were selected based on their relevance to CAR T-cell therapy, including preclinical and clinical studies. Detailed search strategy, inclusion criteria, and screening methods are described in the main manuscript. This review explores the evolution, applications, and future outlook of CAR T-cell rehabilitation.

Chimeric antigen receptor (CAR) T cells have emerged as an effective immunotherapy for hematologic malignancies. The non-signaling domain of CARs, comprising the spacer and transmembrane regions, is a key structural component that can be engineered to influence CAR expression and function. In this study, we evaluated three non-signaling domain configurations-IgG2.CH3/CD28, IgG2/CD28, and CD8/CD8-within a CD19 CAR construct incorporating 4-1BB and interleukin-7 receptor alpha (IL-7Rα) signaling domains. CARs incorporating the IgG2/CD28 domain exhibited reduced surface expression and diminished functional responses compared with IgG2.CH3/CD28 and CD8/CD8 constructs. The CD8/CD8 configuration supported the highest CAR expression and sustained surface density. In contrast, IgG2.CH3/CD28 CAR T cells displayed increased IL-2 and TNF-α secretion and enhanced CD107α upregulation following antigen stimulation. In a serial tumor cell rechallenge assay, IgG2.CH3/CD28 CAR T cells maintained cytotoxic activity and persistence compared with CD8/CD8 CAR T cells. In a NALM-6 xenograft model, IgG2.CH3/CD28 CAR T cells achieved durable tumor control and were associated with improved survival relative to CD8/CD8 CAR T cells. Collectively, these findings support the IgG2.CH3/CD28 non-signaling domain as a suitable structural component for CD19 CARs incorporating IL-7Rα signaling and provide insight into CAR design strategies aimed at improving T cell persistence and anti-leukemic activity.

We propose new scattering networks for signals measured on simplicial complexes, which we call Multiscale Hodge Scattering Networks (MHSNs). Our construction builds on multiscale basis dictionaries on simplicial complexes—namely, the κ -GHWT and κ -HGLET—which we recently developed for simplices of dimension κ ∈ N in a given simplicial complex by generalizing the node-based Generalized Haar–Walsh Transform (GHWT) and Hierarchical Graph Laplacian Eigen Transform (HGLET). Both the κ -GHWT and the κ -HGLET form redundant sets (i.e., dictionaries) of multiscale basis vectors and the corresponding expansion coefficients of a given signal. Our MHSNs adopt a layered structure analogous to a convolutional neural network (CNN), cascading the moments of the modulus of the dictionary coefficients. The resulting features are invariant to reordering of the simplices (i.e., node permutation of the underlying graphs). Importantly, the use of multiscale basis dictionaries in our MHSNs admits a natural pooling operation—akin to local pooling in CNNs—that can be performed either locally or per scale. Such pooling operations are more difficult to define in traditional scattering networks based on Morlet wavelets and in geometric scattering networks based on Diffusion Wavelets. As a result, our approach extracts a rich set of descriptive yet robust features that can be combined with simple machine learning models (e.g., logistic regression or support vector machines) to achieve high-accuracy classification with far fewer trainable parameters than most modern graph neural networks require. Finally, we demonstrate the effectiveness of MHSNs on three distinct problem types: signal classification, domain (i.e., graph/simplex) classification, and molecular dynamics prediction.

Increased NMDA receptor GluN2A-type ionotropic signaling is sufficient to improve spatial memory in immature mice.

The Critical Appraisal Tool for Homeopathic Intervention Studies (CATHIS) core is a streamlined appraisal tool for homeopathic intervention studies focusing on credibility, coherence, and clinical relevance. The aim of the research project was to evaluate its inter-rater reliability, feasibility, and face validity. In a preregistered cross-sectional study, four raters independently applied CATHIS core to 28 trials (21 randomised controlled trials, 7 non-randomised studies on interventions) drawn from reviews on insomnia and hypertension; two external reviewers provided consensus ratings. Inter-rater reliability (IRR) was estimated using percent agreement, Fleiss' κ, and Gwet's AC2 (95% CIs). Feasibility was quantified as rating time and consensus time. Associations among the three domains were explored with correlation analyses and sensitivity checks. IRR varied markedly by domain. Credibility showed good agreement (Fleiss' κ=0.66, 95% CI 0.57-0.74; AC2 =0.76, 0.71-0.82). Coherence yielded only poor-to-fair agreement (κ=0.28, 0.16-0.40; AC2 =0.41, 0.30-0.51). Clinical relevance was similarly limited (κ=0.32, 0.23-0.41; AC2 =0.36, 0.28-0.44). Individual ratings required on average 65.8 min, while consensus discussions averaged 17.7 min. Correlation analyses indicated heterogeneous and partly overlapping domain signals with limited interpretability. Face-validity responses reflected moderate-to-high acceptance but difficulties in consistent application. CATHIS core yielded reproducible credibility ratings but only fair and operationally fragile agreement for coherence and clinical relevance, alongside non-trivial rating burden. Taken together, the current reliability profile is insufficient for confident use in systematic reviews. Targeted refinement appears warranted before broader implementation.

Positive and balancing selection on pattern recognition receptors (PRRs) is widely thought to target ligand-binding domains and affect the specificity of recognition of different pathogens. Alternatively, positive/balancing selection on PRRs could affect general responsiveness by targeting for example signaling domains or cis-regulatory variation. Studies of a wild rodent (the bank vole, Clethrionomys glareolus) have shown that Tlr2-a lipoprotein-binding PRR-is highly polymorphic with divergent haplotypes and signatures of balancing selection, and that Tlr2 genotype is associated with susceptibility to Borrelia afzelii infection in the wild. To investigate what aspect of Tlr2 function has been under selection, we here perform integrated population genetic and functional analyses. Ex vivo infection experiments show that the protective Tlr2 haplotype produces a stronger proinflammatory response to B. afzelii compared to the haplotype associated with susceptibility. Tlr2 genotype has a similar, albeit not statistically significant, effect on responsiveness to the phylogenetically distant pathogen Streptococcus pyogenes. We find that the strongest signature of balancing selection is 4.6 kb upstream of the Tlr2 coding sequence, near a putative enhancer, and that Tlr2 exhibits allele-specific expression such that the protective haplotype is more expressed. Collectively these results indicate that balancing selection has primarily acted on cis-regulatory variation affecting the general responsiveness via Tlr2-signaling rather than on polymorphisms affecting Tlr2 ligand-binding specificity.

Vectorcardiogram (VCG) signal compression is very much in demand in the present-day scenario due to the increasing number of cardiac patients. Hence, in this paper, a new technique is proposed that compresses VCG signal by optimizing the tunable quality wavelet transform (TQWT) parameters. The noise in VCG signal is firstly removed by applying a Savitzky-Golay filter, and then passing noise-free signal to an optimization algorithm that optimizes the TQWT parameters, and obtains the frequency domain signal. This signal is then quantized through dead-zone quantization and processed by a lossless compression mechanism: run-length encoding (RLE) to improve the compression ratio & encode the signal. This compressed signal is reconstructed by Inverse RLE to obtain the decoded signal. Inverse of TQWT is applied to get the reconstructed signal back from the transformed frequency domain to time domain. The parameters of TQWT, especially theQandR, are optimized to get the highestCRat lowest percent root-mean-square-difference(PRD)with best reconstruction quality and least distortions, along with acceptable values of signal-to-noise-ratio(SNR), quality score(QS), andSimilaritywith lowest mean-square-error(MSE). The comparative analysis of different optimization methods indicates that the sparse-particle swarm optimization is best among all the approaches for the tuning of parameters in TQWT for VCG signal compression and reconstruction achieving aCRof 48.18 at aPRDof 3.68,SNRof 29.39,QSof 15.71, similarity of 0.99845,MSEof 0.00016, withQvalue of 2.04307 andRvalue of 1.20568 withcomputational timeof 4.48508 s.

Effectively utilization of disaster warning signals is crucial for mitigating impacts and enhancing operational resilience in disaster management. Although an emerging area of scholarly interest, the literature remains fragmented and lacking a unifying framework to guide research and practice. To consolidate knowledge and direct future research, this study conducts a systematic review of the literature on disaster warning signal research published in leading journals in operations management, management science, operations research, and related supply chain and logistics. Building upon the classic communication model proposed by Shannon, this study develops a conceptual framework for systematically classifying the relevant literature according to three dimensions of a warning signal: (1) signal type (natural, engineering, behavioral, informational, composite), (2) signal transmission (source, channel, receiver) and (3) signal purpose (directing the response of the authority, guiding the protection of the public). We then cross-tabulate this framework with the disaster management domain (e.g., disaster phase, type, and function) and the data domain (e.g., data type, analytics techniques). By synthesizing academic contributions with practical challenges, we articulate the specific value that operations management research on warning signals offers to disaster management practice. Finally, we propose a structured agenda for future research focused on the intersections of signal, disaster, and data domains.

The unique ability of chimeric antigen receptor (CAR) T cells to infiltrate tissues is revolutionizing our perspectives for tackling severe, refractory and otherwise untreatable diseases. In HIV, CAR-T cells have been designed to target viral biomarkers, with limited success so far. Here, we investigated the possibility of redirecting CAR-T cells against a cellular biomarker of the HIV reservoir, the programmed cell death protein 1 (PD-1). We designed two second-generation 4-1BB-CARs using the scFv of either a blocking (bPD1-CAR) or a nonblocking (nbPD1-CAR) anti-PD-1 monoclonal antibody. The CAR avidity modulated T cell sensitivity, trogocytosis, and effector functions, independently of the PD-1 signaling domain. Both anti-PD-1 CAR-T cells could persist for 70 days in HIV-infected humanized mice, correlating with viral protection and a disruption of the lymphoid architecture in the white pulp of the spleen. Together, our results open strategic avenues for reducing the HIV reservoir by demonstrating the feasibility of depleting specific T cell subpopulations.

Actin-rich protrusions densely cover the surface of T cells and are well characterised for their role in migration. Recent studies have uncovered their contribution to antigen surveillance and immune signalling. To further explore how protrusions initiate signalling pathways mediating T-cell activation, we performed live-cell imaging of endogenously tagged proteins in HER2-specific chimeric antigen receptor (CAR) T cells targeting HER2⁺ breast-cancer cells. Quantitative STED microscopy allowed us to monitor protein rearrangement and to correlate it with membrane topology over time. Before activation, key signalling proteins (including Lck, CD45, LAT, and the CAR) were not enriched in protrusions. Upon contact with target cells, rapid protein reorganisation occurred preferentially within protrusions, initiating signalling. HER2-CAR clustering, accompanied by ZAP-70 and LAT recruitment, was enhanced in protrusions. While Lck distribution remained unchanged, exclusion of the phosphatase CD45 was enhanced at protrusion-cell contacts, independently of the CAR signalling domain. Overall, signalling machinery rearranged faster and more effectively at protrusive contacts than at main plasma membrane regions. Together, our data re-frame protrusions as sites of enhanced receptor activation by exclusion and clustering dynamics rather than by pre-enrichment of the signalling machinery.

Gastric anastomotic leakage is a life-threatening postoperative complication that necessitates early detection and timely intervention. Implantable electronics offer direct access to deep tissue and enable accurate recognition of homeostatic imbalances yet are constrained by miniaturization, biodegradability, and power sustainability. Here, we propose an implantable-wearable integrated strategy for early detection of gastric anastomotic leakage. Specifically, a bioresorbable hydrogel patch serves as the sole implant, responding to gastric acid to modulate ultrasound echo signals in both time and frequency domains. This enables early leakage detection (within 1 hour) and millimeter-level spatial localization via a wearable ultrasound array. Meanwhile, the hydrogel patch exhibits acid-enhanced leak sealing capability and degrades naturally after the healing period, thereby eliminating the need for surgical removal. We confirmed its capabilities in spatiotemporal monitoring, anastomotic repair, and bioabsorbability in a rat gastric perforation model. Our system operates without complex electronic implants, highlighting its clinical potential for postoperative monitoring and management of gastric anastomotic leaks.

Co-activating the intrinsic FcRγ/TLR4 signaling axis enhances the antitumor activity of NKG2D CAR-macrophages against prostate cancer.

Proteins normally localized in the intracellular compartments of healthy cells have been observed at the surface of cancer cells, despite lacking a transmembrane domain or secretion signals. This unexpected localization likely reflects yet-unknown functions and presents a unique opportunity to develop cancer cell-specific antibody- or peptide-based therapeutic strategies. While ribosomal proteins (RPs) are primarily involved in translation, several display moonlighting functions in the cytoplasm and nucleus. In this study, we uncover an extracellular form of the ribosomal protein L21 (eL21) in triple-negative breast cancer (TNBC) cells. Using complementary approaches and a broad set of antibodies, we demonstrate that eL21 localizes to the surface of cancer cells. Remarkably, we show that anti-eL21 antibodies trigger a potent, rapid and dose-dependent anti-proliferative effect, including TNBC cell cycle arrest and apoptosis. These findings identify eL21 as a novel ribosomal protein with extra-ribosomal functions at the cancer cell surface and highlight its potential as a therapeutic target in TNBC.

Restoring immune tolerance by engineered regulatory T cell (Treg) therapy is a promising strategy to treat patients suffering from autoimmune and inflammatory diseases. However, in many of these conditions, relevant disease-driving antigens are unknown. Therefore, suitable target (auto-)antigens for antigen-specific Treg cell therapy are rarely available. We present a novel artificial immune biosensor for Treg cells that circumvents this limitation by targeting the immune costimulatory protein CD40-ligand (CD40L), transiently expressed by activated T cells. The artificial immune receptor (AIR) comprises a CD40-derived extracellular binding domain, an intracellular costimulatory signaling domain, and a T cell receptor signaling domain of the CD3-ζ-chain. After interaction with its membrane-bound ligand, this synthetic receptor triggers a TCR-like activation program in Treg cells including induction of Treg effector molecules and cell proliferation. In a mouse model of graft-versus-host disease, transfer of CD40-AIR Treg cells significantly improved survival and demonstrated immune control of the alloantigen-reactive T cell compartment. Expression and signaling of the corresponding human CD40-AIR illustrate the potential for translating this concept. Engineering Treg cells with a CD40L-targeting sensor that detects activated T cells presents a promising therapeutic approach for a broad range of T cell-mediated inflammatory diseases.

Gene-engineered T-cell products have been developed for immunotherapy to treat cancers, with great success observed in haematological malignancies but limited efficacy in treating solid cancers. TCR-engineered T cells utilize transferred TCRs targeting tumour-associated and cancer-specific peptides presented by MHC molecules. The CD3ζ chains are part of the TCR-CD3 complex expressed by T cells and mediate signal transduction when the TCR binds to MHC-presented peptides. In this study, we explored whether co-stimulation domains, that were effective in improving the function of T cells engineered with chimeric antigen receptors (CARs), can be exploited to improve the functionality of TCR-engineered T cells. We inserted the signalling domains of CD28 or 4–1BB at the membrane proximal or the membrane distal position of the intracellular tail of CD3ζ and engineered human T cells to express a specific TCR in combination with either modified CD3ζ or unmodified control. Antigen-specific in vitro stimulation assays revealed that T cells expressing CD3ζ constructs with CD28 signal domains displayed enhanced peptide-specific IL-2 production and, following repeated antigen stimulation, expanded to substantially greater numbers than T cells expressing unmodified CD3ζ. Importantly, greater expansion seen with the CD28-containing ζ did not result in any reduction of effector function as assessed by peptide-specific cytotoxicity and cytokine production. The data indicate that modification of the CD3ζ chain with a CD28 signal motif provides an opportunity to improve antigen-specific expansion and effector function of TCR-engineered T cells by combining signal 1 and co-stimulatory signal 2 in one molecular TCR-CD3 complex.