Disseminated intravascular coagulation (DIC) occurs in a variety of diseases and syndromes. Platelet count, prothrombin time, and levels of fibrinogen and D-dimer are used to determine if a patient has DIC. The tissue factor (TF)/factor (F)VIIa complex plays a central role in the activation of coagulation in sepsis. In Gram-negative sepsis, lipopolysaccharide induces TF expression in monocytes and release TF-positive extracellular vesicles (EVs). This study determined whether EV-TF activity is associated with DIC in patients with sepsis. We studied 2 cohorts of patients with sepsis: a cohort of sequential patients that included 143 patients without DIC and 45 patients with DIC, and a cohort of selected patients that included 39 patients without DIC and 36 patients with DIC. The first cohort had longitudinal samples taken, and the second cohort had samples taken from pre-DIC patients (who developed DIC during the study). Healthy subjects were used as controls. Plasma EV-TF activity was measured using an in-house assay. We used the updated 2025 International Society on Thrombosis and Haemostasis (ISTH) DIC score. EV-TF activity was significantly higher in patients with sepsis than in healthy controls. There was no significant difference in the level of EV-TF activity between pre-DIC patients and patients without DIC. Importantly, EV-TF activity was associated with DIC in both cohorts of patients with sepsis. We observed a time-dependent decrease in EV-TF activity in patients with DIC. Our study suggests that increased levels of circulating EV-TF activity contribute to the activation of coagulation in patients with sepsis.
Messenger RNA vaccines and gene therapies enable rapid and programmable biological intervention, but their therapeutic durability remains variable. Existing nanomedicine research primarily evaluates delivery efficiency, targeting, RNA protection and early protein expression, which do not fully explain why some biological effects persist while others rapidly decline. This Hypothesis and Theory article presents a narrative conceptual synthesis of peer-reviewed literature on lipid nanoparticle delivery, mRNA vaccinology, RNA therapeutics, genome editing, trained innate immunity, epigenetic regulation and regenerative medicine. Evidence was integrated to develop an operational and falsifiable framework for evaluating the contribution of nanocarriers to therapeutic durability. We propose the Biological Memory Buffer Hypothesis, according to which programmable nanocarriers may influence biological systems that encode, maintain or recall therapeutic information after RNA delivery. The framework distinguishes payload persistence from biological-outcome persistence and introduces three measurable constructs: therapeutic memory engineering, nanocarrier memory capacity and the therapeutic persistence window. It further proposes matched-cargo and matched-early-exposure experiments, candidate monophasic and biphasic decay models, a minimum durability reporting set and safety monitoring for maladaptive innate immune imprinting. The framework does not assume that adaptive immune memory, trained immunity, epigenetic regulation and regenerative repair share a single molecular mechanism. Rather, it treats them as distinct biological substrates with a common functional relevance to durability. The hypothesis is falsifiable: failure to detect reproducible carrier-attributable differences under matched conditions would argue against nanocarrier memory capacity as an independent determinant. Incorporating longitudinal, tissue-specific and host-stratified durability measurements may support the rational development of longer-lasting RNA vaccines and gene therapies.
Stress fractures are overuse injuries that develops in response to repetitive loads applied to bone with normal structural integrity and is highly prevalent among physically active populations, but their underlying mechanisms remain incompletely understood. This article reviews the complex pathogenesis and healing mechanisms of stress fractures. Stress fractures develop when repetitive mechanical loading on the bone exceeds its threshold for adaptive repair, leading to the progressive accumulation of microdamage and ultimately disrupting the physiological equilibrium between bone resorption and formation. The healing of stress fractures is characterized by intramembranous ossification, a process that begins with periosteal woven bone formation to stabilize the fracture and proceeds through subsequent bone remodeling to repair the cracks. The development and repair processes of stress fractures involve dynamic alterations in cell types and tissue constituents, along with active signaling activities within and among the involved cells involved. Future research should prioritize the use of larger animal models such as rabbits and minipigs, and the development of stress fracture models that more accurately replicate the clinical pathogenesis of stress fractures. Although the efficacy of anti-osteoporotic agents, non-steroidal anti-inflammatory drugs (NSAIDs), and low-intensity pulsed ultrasound (LIPUS) have been reported, future research should explore additional physical therapy modalities to elucidate their specific role in the management of stress fractures and underlying mechanisms. Overall, by reviewing the latest research advances in the pathogenesis and treatment of stress fractures and exploring targeted therapeutic strategies, this article holds the promise to offer novel insights into their prevention and management, thereby driving improvements and innovations in clinical treatment approaches and demonstrating significant translational potential.
Arginine, also known as L-arginine residue, is a dibasic, cationic, conditionally essential amino acid that serves as an integral part of polypeptide chains. Along with its conventional role in protein synthesis, numerous biological processes depend on L-arginine, such as the synthesis of nitric oxide (NO) and regulation of vascular tone, the urea cycle and acid/base homeostasis, nuclear localization, post-transcriptional and post-translational modifications, signal transduction, the transport system, and viral genome assembly. In the physiological context, it plays diverse roles; for example, it correlates with airflow abnormalities in severe asthma, reduces inflammation, promotes muscle regeneration, and improves blood flow through vasodilation. Keeping in view the above, this review is an effort to collate the current knowledge and discuss and appraise the biological and physiological roles of L-arginine as an integral entity within a biological system.
Antibody-based therapeutics has revolutionized disease treatment, and recent advances in messenger RNA (mRNA) technologies have opened new opportunities for their intracellular production. In particular, in vitro-transcribed mRNA encapsulated in lipid nanoparticles (LNPs) enables targeted delivery to specific cells, where it can enable the synthesis of therapeutic antibodies with prolonged half-lives in a cost-effective manner. Despite rapidly growing experimental data, a modeling framework that integrates mRNA delivery, intracellular expression kinetics, and whole-body antibody disposition remains unavailable. To address this gap, we extended a physiologically based pharmacokinetic model with a novel multiscale layer describing mRNA trafficking, cellular uptake, translation, and degradation. The integrated model was calibrated and validated using five datasets of mRNA-based cancer therapeutics, demonstrating strong predictive performance for the biodistribution of mRNA-encoded antibodies. The newly introduced mRNA layer, while minimally parameterized, effectively represents complex intracellular and systemic processes, enabling quantitative investigation of antibody biodistribution, optimization of dose scheduling, and providing an initial framework for future exploration of how LNP-mRNA formulation influences delivery and pharmacokinetics.
Lathyrol, a bioactive natural compound derived from plants of the Euphorbiaceae family, exhibits antitumor activity, and its molecular targets and underlying mechanisms remain incompletely understood. In this study, thermal proteome profiling (TPP) was applied to systematically identify lathyrol-binding proteins in non-small cell lung cancer (NSCLC) cells. TPP analysis identified glucose-6-phosphate dehydrogenase (G6PD) as a candidate target of lathyrol. A cellular thermal shift assay (CETSA) confirmed increased thermal stability of G6PD upon treatment. Molecular docking indicated a potential interaction between lathyrol and G6PD. The peptide-centric local stability assay (PELSA) revealed localized conformational changes in the C-terminal region of G6PD consistent with the predicted interaction interface. Enzymatic assays showed reduced G6PD activity accompanied by decreased intracellular NADPH levels. Quantitative proteomics indicated alterations in the pathways associated with glucose metabolism and redox regulation. These findings identify G6PD as a functional target of lathyrol in NSCLC cells and link its inhibition to disruption of cellular redox balance and metabolic homeostasis.
Rheumatoid arthritis (RA) is driven in part by hyperactivated fibroblast-like synoviocytes (FLS) that invade articular structures. Iguratimod (IGU), a conventional synthetic DMARD, is clinically effective, but its direct molecular target and impact on synovial cell-cell crosstalk remain unclear. We aimed to elucidate how IGU regulates FLS invasiveness and inflammatory signaling, identify its upstream target within the JAK-STAT pathway, and develop a prodrug with improved pharmacokinetics while preserving disease-modifying activity. We combined in vitro assays in MH7A cells and rat RA-FLS with RNA sequencing and conditioned-medium fast-astral DIA proteomics to characterize IGU's effects on TNF-α-induced migration, invasion, and signaling. STAT1 dependence was interrogated by siRNA knockdown, phosphorylation-deficient mutant reconstitution and IFN-γ rescue. Integrated single-cell RNA-seq of RA and healthy synovium, together with CellChat analysis and complement component 3a (C3a) stimulation of THP-1-derived macrophages, was used to define FLS-macrophage crosstalk. Reverse virtual screening, molecular docking, thermal shift assays, cellular thermal shift assays, kinase assays, and molecular dynamics simulations were applied to characterize IGU-TYK2 interactions. A sulfonamide N-acyl IGU prodrug (AD811) was rationally designed and evaluated for pH-dependent stability, plasma and microsomal metabolism, pharmacokinetics, efficacy, and short-term safety in collagen-induced arthritis rats. IGU suppressed TNF-α-induced FLS migration and invasion without cytotoxicity by selectively inhibiting STAT1 Y701 phosphorylation and nuclear translocation, while sparing STAT1 Y727 phosphorylation and STAT2 Y690 phosphorylation. Bulk and single-cell transcriptomic analyses revealed STAT1 hyperactivation in RA lining-layer FLS and uncovered a STAT1-C3-TNFα feedback loop in which FLS-derived C3/C3a enhances macrophage TNF-α production, thereby reinforcing FLS activation; IGU disrupted this loop by reducing STAT1 activity, C3 transcription, and C3a-driven macrophage TNF-α induction. Biochemical and biophysical studies showed that IGU directly engages the TYK2 JH2 pseudokinase domain, alters its thermal behavior, and inhibits kinase activity of a TYK2 construct containing JH2 and JH1, while not measurably inhibiting the isolated JH1 catalytic domain, consistent with JH2-dependent allosteric modulation of TYK2 output. The prodrug AD811 exhibited pH-sensitive stability, rapid plasma conversion to IGU, favorable oral bioavailability, and therapeutic efficacy, joint protection, and preliminary hepatic and gastric safety comparable to equimolar IGU in vivo. In FLS-centered experimental systems, IGU reduces RA-FLS invasiveness by targeting TYK2 JH2 and disrupting a STAT1-C3-TNFα feedback loop between lining-layer FLS and macrophages, thereby attenuating both intrinsic fibroblast aggressiveness and inflammatory crosstalk. The prodrug AD811 maintains these disease-modifying actions while improving pharmacokinetic properties, nominating AD811 as a promising candidate for further translational development in RA. This study mechanistically links IGU, a clinically used csDMARD, to selective modulation of TYK2 JH2 and downstream STAT1 signaling in synovial lining-layer FLS. By showing that, in FLS-centered models, IGU disrupts a STAT1-C3-TNFα feedback loop between FLS and macrophages and thereby reduces FLS invasiveness and inflammatory crosstalk, our data provide a concrete cellular and molecular basis for its disease-modifying effects in rheumatoid arthritis. Furthermore, the rationally designed prodrug AD811 exhibits improved pharmacokinetic properties and a favorable short-term safety profile in vivo, supporting its further evaluation as a potential oral small-molecule candidate.
Members of the Vps10p receptor family regulate protein trafficking and cellular differentiation in the nervous system. Previous structural studies of the dimeric Vps10p family member SorCS2 have focused on isolated ectodomains, revealing substantial structural plasticity but overlooking the influence of the membrane association on receptor organization. Here we establish two complementary tools for reconstituting the SorCS2 ectodomain on proteoliposomes in its native orientation: non-covalent coupling via a C-terminal His-tag and nickel affinity, and covalent attachment via strain-promoted alkyne-azide cycloaddition using a C-terminal azide. We visualize the SorCS2 membrane-associated protein organization using electron cryo-tomography and obtain a nanometer resolution subtomogram average of the His-tag coupled SorCS2 ectodomain dimer. Four distinct, previously unreported, SorCS2 dimer-of-dimer arrangements are observed. The two most prominent interactions form through "head-to-side" docking of a Vps10p domain to the Vps10p and PKD core of another dimer, and "head-to-head" symmetric interactions between the Vps10p and SoMP domains of two dimers. Two less frequent assemblies comprise "side-by-side" interactions between the beta-propeller and 10CC domains and symmetrical "face-to-face" beta-propeller top face interactions. Together these interactions organize SorCS2 into two distinct helical arrangements and small receptor clusters on liposome surfaces. The promiscuity of membrane-stabilized SorCS2 cis interactions supports a more general mechanism in which the organization of receptor systems is influenced by membrane association. The tools presented here provide a versatile platform for visualizing ectodomain-mediated receptor assemblies in a membrane context.
Vitamin B2 (riboflavin), a precursor of the essential coenzymes flavin mononucleotide and flavin adenine dinucleotide, plays a critical role in numerous cellular metabolic pathways, including redox reactions, energy production, and biosynthetic processes. Due to its fundamental importance in human and animal nutrition, riboflavin has found wide application across the food, pharmaceutical, nutraceutical, and cosmetic industries. In recent decades, the development of metabolic engineering techniques has revolutionized riboflavin production, transitioning from traditional chemical synthesis to environmentally sustainable microbial fermentation. Microorganisms such as A. gossypii, Candida famata, and Bacillus subtilis have been extensively engineered to enhance riboflavin yield at an industrial scale. Key metabolic engineering strategies include reprogramming the purine metabolic network, mutating ribulose-5-phosphate 3-epimerase, and relieving regulatory constraints on purine biosynthesis. These advances have significantly improved the efficiency, cost-effectiveness, and sustainability of industrial riboflavin production. This review article presents an in-depth analysis of biotechnological methods aimed at increasing riboflavin production, ensuring a consistent supply of this crucial micronutrient for both human and animal nutrition. It stands as a significant reference for researchers and industry stakeholders.
CD19-directed chimeric antigen receptor (CAR) T-cell therapy has transformed the management of relapsed or refractory large B-cell lymphoma (LBCL), producing durable remissions in a subset of patients whose disease previously had few curative options. Axicabtagene ciloleucel, tisagenlecleucel, and lisocabtagene maraleucel established CAR T-cell therapy in the third-line setting, and randomized studies subsequently moved axicabtagene ciloleucel and lisocabtagene maraleucel into second-line treatment for primary refractory or early relapsed disease. This review provides a clinically anchored, mechanism-focused synthesis of CAR T-cell therapy in LBCL. We critically compare pivotal trials, long-term follow-up, patient-selection principles, and real-world evidence, emphasizing that apparent differences across products must be interpreted in light of eligibility criteria, analytic denominators, bridging therapy, manufacturing intervals, toxicity grading, and treatment crossover. We then examine resistance and relapse as systems-level phenomena arising from antigen modulation, tumor-intrinsic evolution, impaired CAR T-cell fitness, suppressive myeloid and stromal networks, systemic inflammation, metabolic stress, and incomplete immune recovery. The biological basis and clinical implications of cytokine release syndrome, immune effector cell-associated neurotoxicity syndrome, prolonged cytopenias, infections, and late nonrelapse mortality are also reviewed. Finally, we discuss circulating tumor DNA, metabolic imaging, single-cell and multi-omic profiling, artificial intelligence, dual-target and armored constructs, allogeneic platforms, and in vivo CAR programming as components of precision cellular therapy. The central clinical challenge is no longer whether CAR T-cell therapy can work, but how to select patients, deliver treatment rapidly, anticipate failure, and preserve long-term immune and functional health.
The pathology of diabetic foot ulcer (DFU) is characterized by keratinocyte dysfunction, non-resolving inflammation, and oxidative stress. We aim to investigate the effects and mechanisms of piroxicam on DFU healing through regulating mitochondrial function and suppressing inflammation. DFU was established in male C57BL/6 J mice and ovariectomized female mice. Piroxicam (1% or 0.33%) solution or saline was then applied for 9 days. HaCaT cells were induced with high glucose (HG) and subsequently incubated with piroxicam (0, 1.2, 3.7, 11, 33, 100 nM). Piroxicam significantly promoted DFU healing and inhibited the fibrosis in male diabetic mice at a low dose. Consistently, piroxicam enhanced proliferation and migration, and inhibited inflammation, fibrosis, and cellular senescence in HG-induced HaCaT cells. Mechanistically, piroxicam alleviated HG-induced mitochondrial dysfunction by stabilizing the mitochondrial respiratory chain, increasing biogenesis, and enhancing mitophagy. These effects further attenuated oxidative stress and inhibited the cGAS-STING-NF-κB inflammatory pathway, thereby reducing the release of pro-inflammatory factors. Furthermore, molecular docking revealed that piroxicam bound to ERα, a finding further confirmed by a cellular thermal shift assay. HG induced a significant decrease in nuclear ERα protein levels, which was reversed by piroxicam, especially at 11 and 33 nM. Additionally, piroxicam's pro-healing and anti-inflammation effects were attenuated in ovariectomized female DFU mice. Piroxicam's protection of mitochondrial function and suppression of oxidative stress was also abolished upon blocking ERα by tamoxifen. In conclusion, piroxicam alleviates mitochondrial dysfunction and suppresses inflammatory responses by binding to ERα, which ultimately promotes DFU healing at low doses.
Cold stress (CS) is a significant obstacle in tobacco (Nicotiana tabacum L.) farming, significantly affecting plant development, photosynthetic activity, and cellular redox balance. In recent decades, bio-stimulants have created environmentally friendly substances that make the plant resistant to abiotic stresses, such as cold stress. The recent developments highlight bio-stimulants as a sustainable solution to improve cold stress tolerance in tobacco production. These substances promote plant growth, thereby increasing plant resilience to unfavorable temperature conditions. This review assesses the role of bio-stimulants in improving cold stress tolerance in tobacco, focusing on physiological, biochemical, and molecular responses. It summarizes the effects of various bio-stimulants on plant growth, antioxidant defense systems, and photosynthetic performance under low-temperature conditions. The enhancement of enzymatic antioxidants and non-enzymatic antioxidants by bio-stimulants helps overcome oxidative damage. Evidence in molecular biology studies to understand bio-stimulant-mediated regulation of stress responsive genes is also critically discussed in order to understand the role bio-stimulants play in enhancing the genetic potential of tobacco to cold stress. This review presents an integrated scheme of the multifarious functions of bio-stimulants in improving cold stress tolerance of tobacco. It also highlights existing knowledge gaps and provides research directions on how to explore efficient, sustainable, and climate resilient tobacco production systems in the future.
This work aimed to develop and characterize a three-dimensional culture platform that recreates the tumor microenvironment of multiple myeloma cells (MMCs), focusing on how atypical plasma cells (aPCs) interact with other bone marrow cells and extracellular matrix (ECM) components to develop drug resistance. The biomimetic platform consists of a static system based on a conical agarose geometry that facilitates the agglomeration and proliferation of aPCs. Magnetic alginate microgels, manufactured by microfluidics, configure the ECM through a layer-by-layer functionalization with collagen, hyaluronic acid, chondroitin sulfate, and heparin. In addition, bone marrow mesenchymal stem cell (BMMSC) pellets were integrated to simulate direct and indirect interactions between BMMSCs and MMCs. The platform has been validated with three lines of MMCs (RPMI8226, MM1S, U266) and the patients' mononuclear cell fraction collected from bone marrow aspirates extracted in two Spanish Hospitals. The microgel analysis aimed to verify stability and to characterize the presence of the different coatings. Biological studies have investigated the proliferation and resistance to drug treatments in MM cell lines. In addition, patient-derived cells were cultured for up to 14 days to study the survival and possible immunophenotypic changes of the atypical plasma cell (aPC) fraction.
Claudin-3 is a tight junction protein expressed in the distal nephron, notably in the thick ascending limb of Henle's loop and the distal convoluted tubule, where it acts as an aldosterone-regulated paracellular barrier to sodium and chloride transport. Here, we demonstrate that claudin-3 knockout (KO) mice exhibit an unexpected diet-dependent alteration in glomerular filtration rate (GFR): under a normal sodium (NS) diet, GFR decreases significantly, whereas under a low sodium (LS) diet, GFR increases paradoxically compared with wild-type controls. To identify the underlying mechanism, we examined claudin-3 expression in the macula densa, the specialized epithelial structure located at the junction between the thick ascending limb and the distal convoluted tubule that governs tubuloglomerular feedback (TGF). Immunofluorescence analysis revealed that claudin-3 colocalizes with neuronal nitric oxide synthase (nNOS) and the Na+-K+-2Cl- cotransporter (NKCC2) in a distinct cluster of cells characterized by densely packed nuclei, positioned in direct contact with the glomerulus and immediately adjacent to an NCC-positive distal convoluted tubule, while remaining itself NCC-negative. Notably, within the tubule containing the macula densa, nNOS expression was strictly confined to macula densa cells, clearly distinguishing them from the surrounding tubular epithelium. These results identify claudin-3 as a previously unrecognized component of macula densa biology and suggest a role for claudin-3 in the control of GFR possibly via the tubuloglomerular feedback.
Systemic lupus erythematosus (SLE) is a multisystem autoimmune disease; hematologic involvement is common and strongly associated with prognosis. Platelets-the second most abundant cellular component of peripheral blood-are anucleate cytoplasmic fragments. Their canonical roles include hemostasis, thrombosis, and vascular repair. More recently, platelets have emerged as regulators of innate and adaptive immunity that amplify inflammatory signaling and may facilitate tumor dissemination, suggesting underappreciated pathogenic roles in the initiation and progression of SLE. This review synthesizes platelet pathobiology in SLE and structures it into four modules: (i) pro-inflammatory actions of activated platelets; (ii) pathogenic effects of platelet-derived microparticles (PMPs); (iii) mechanisms by which platelets exacerbate lupus nephritis; and (iv) pathways linking platelets to SLE-associated cardiovascular disease. We also evaluate platelet-targeted therapeutic strategies and their translational prospects. Our aim is to provide a coherent framework for the platelet-SLE interface that informs mechanistic studies, guides biomarker development, and supports the design of more precise diagnostics and therapies.
This commemorative article reflects on a research journey spanning neural development, stem cell biology, regenerative medicine, and iPSC-based drug discovery. My early work focused on RNA-mediated regulation in the nervous system, including studies on myelin basic protein gene regulation and the identification and functional characterization of the RNA-binding protein Musashi. These studies contributed to the conceptual foundation of neural stem cell biology and helped establish methods for identifying and isolating neural stem/progenitor cells, including those present in the adult human brain. Building on this foundation, my colleagues and I pursued translational research in spinal cord injury, ranging from analyses of injury pathophysiology and molecular interventions to preclinical studies using rodent and non-human primate models. These efforts ultimately led to the first-in-human clinical study of induced pluripotent stem cell-derived neural stem/progenitor cell transplantation for subacute spinal cord injury. In parallel, we developed patient-derived iPSC platforms for neurological disease modeling and drug discovery, particularly for amyotrophic lateral sclerosis, where iPSC-based screening identified Ropinirole as a therapeutic candidate and enabled reverse translational research linking cellular phenotypes with clinical responses. Looking ahead, I argue that the future of regenerative therapy will depend on the continued integration of developmental biology, stem cell science, disease modeling, rehabilitation, and clinical translation to address unmet medical needs in disorders of the central nervous system.
Benign prostatic hyperplasia (BPH) represents a highly prevalent age-related disorder, traditionally managed through androgen-driven pathways. However, the limited efficacy of conventional hormonal therapies in a substantial subset of patients necessitates the investigation of alternative pathogenic mechanisms. The emergence of single-cell RNA sequencing (scRNA-seq) has provided the necessary resolution to map cellular heterogeneity and microenvironmental dynamics within the prostate. This review synthesizes recent advancements in applying scRNA-seq to BPH research, highlighting a transition from a homogeneous, hormone-centric view toward recognizing BPH as a complex, heterogeneous process driven by multifaceted cell-immune interactions. We detail how scRNA-seq has identified distinct cellular subsets within the hyperplastic transition zone, including novel basal epithelial subtypes and activated fibroblast populations that contribute directly to nodule formation and disease progression. Furthermore, we examine the central role of chronic inflammation, mediated by immune cell infiltration and senescence-associated secretory phenotypes (SASP), in perpetuating a proliferative microenvironment. The technology also clarifies mechanisms underlying treatment resistance and identifies potential biomarkers and novel therapeutic targets beyond the androgen axis, such as the CXCL13/CD4+T cell axis and granzyme K pathways. Looking forward, integrating scRNA-seq with spatial multi-omics aims to construct a comprehensive spatiotemporal atlas of BPH, facilitating molecular subtyping and the development of precision, microenvironment-targeted therapies. In conclusion, scRNA-seq is redefining the pathophysiological landscape of BPH, offering a path toward innovative, non-androgenic therapeutic strategies aimed at achieving true disease modification.
Interleukin-17 receptor A (IL17RA) acts as a core mediator of pro-inflammatory signaling and contributes crucially to the pathogenesis of autoimmune disorders including psoriasis and rheumatoid arthritis. Monoclonal antibodies against IL17RA have achieved clinical application, yet their clinical utility is restricted by multiple drawbacks: high production cost, complicated manufacturing procedures, mandatory injection delivery, and absent oral bioavailability. To overcome these bottlenecks, this study develops a novel inhibitory peptide WMX-8 intended to specifically disrupt the binding between IL17A and IL17RA. The polypeptide WMX-8 was rationally designed and chemically synthesized, followed by purification to obtain high-purity samples. A series of systematic in vitro biological assessments were conducted using IL17RA-expressing cell models (keratinocytes and monocyte-macrophages), alongside IL17RA-deficient cells as negative control groups. Biochemical and cellular assays verified that WMX-8 binds IL17RA with strong affinity and markedly suppresses the secretion of pro-inflammatory cytokines in IL17RA-positive keratinocytes and monocyte-macrophages. Its anti-inflammatory efficacy is equivalent to that of the reference anti-IL17RA monoclonal antibody. The inhibitory activity fully relies on IL17RA expression, since the anti-inflammatory effect disappears entirely in IL17RA-knockout cells. In addition, WMX-8 displays desirable pharmaceutical features, including low immunogenicity and convenient large-scale synthesis.This study validates the promising druggable properties of WMX-8 targeting the IL17A-IL17RA axis. Our findings lay a solid foundation for further preclinical investigation of peptide therapeutics against IL17A/IL17RA for the treatment of autoimmune diseases.
Fibroblast activation protein (FAP) was a prominent target overexpressed in the stroma of various tumors. However, most current FAP-targeted radiotracers suffer from rapid systemic clearance and limited tumor retention, which limits their imaging contrast and therapeutic efficacy. In this study, we exploited diethylamino coumarin as a novel small-molecule albumin binder to optimize the pharmacokinetic profile of FAP-targeted radiopharmaceuticals. Two probes with different linker lengths, [68Ga]-Ga-PEG2-COU and [68Ga]-Ga-PEG6-COU, were synthesized with high purity (>95%) and stability. The PEG2 variant demonstrated superior cellular uptake (6.72 ± 1.94 IA%/106 cells at 120 min) compared to PEG6 (4.73 ± 0.51 IA%/106 cells). It achieved high tumor accumulation with a tumor-to-heart (T/H) ratio of 14.64 ± 7.24 at 60 min. Crucially, unlike [68Ga]-Ga-FAPI-04, whose tumor-to-muscle ratio declined from 4.18 ± 0.14 to 2.44 ± 0.15 over 120 min, the PEG2 probe's T/M ratio continuously increased from 3.65 ± 0.18 at 30 min to 5.17 ± 0.54 at 120 min. Collectively, these findings highlight the potential of the coumarin-based albumin-binding platform to enhance the pharmacokinetics of FAP-targeted radiopharmaceuticals.
Chronic inflammatory demyelinating polyneuropathy (CIDP) is an immune-mediated peripheral neuropathy with heterogeneous and often incomplete responses to current immunotherapies, but the underlying immune basis remains poorly defined. Although CIDP shares features of immune-mediated demyelination with multiple sclerosis (MS), the two diseases affect distinct anatomical compartments and exhibit divergent therapeutic responses, suggesting fundamentally different underlying immune programs. Here, we address this gap by defining the peripheral immune architecture of CIDP using an integrated, multi-modal approach. Peripheral blood was obtained from 20 patients with CIDP and 20 age- and sex-matched healthy controls. Single-cell RNA sequencing was performed in a discovery subset and integrated with publicly available MS peripheral blood datasets to provide a cross-disease reference framework. The single-cell analysis was designed as an exploratory discovery step to identify candidate immune signatures. Transcriptomic, pathway, and ligand-receptor analyses were complemented by cytokine profiling and flow-cytometric validation in the full cohort. CIDP exhibited broad inflammatory activation with preferential enrichment of type I interferon and inflammasome-related programs compared with MS. Despite reduced B-cell frequencies, CIDP showed transcriptional enrichment of germinal center-associated programs, indicating a dissociation between cell number and activation state. In parallel, CD8 effector T cells demonstrated enhanced cytotoxicity and cytoskeletal remodeling programs, supported by increased expression of actin-regulatory genes and strengthened intercellular signaling interactions. In contrast, MS showed greater enrichment of integrin-talin-vinculin signaling pathways in B cells and CD4 T-cell subsets, consistent with trafficking-related immune mechanisms. Together, these findings indicate a coordinated immune axis linking B-cell dysregulation and cytotoxic CD8 T-cell activation in CIDP. Integrated peripheral immune profiling identified candidate CIDP-associated immune signatures including dysregulated B-cell activation despite numerical reduction and a prominent cytotoxic CD8 T-cell program within a type I interferon- and inflammasome-skewed inflammatory milieu. These findings provide an exploratory framework for understanding peripheral immune dysregulation in CIDP and warrant further translational studies in larger, treatment-stratified cohorts.