ID: 4054211
Title: Stereospecific binding sites for [3H]nicotinamide adenine dinucleotide in the rat brain.
Abstract: NAD is a potent inhibitor of electrical activity in the dentate gyrus of the guinea pig hippocampus. NAD is rapidly degraded by an NADase enzyme present on synaptosomal membranes that we have recently found to be inhibited by nicotinamide mononucleotide. In this report we have characterised the binding sites present on brain membranes for [3H]NAD in the presence of this inhibitor. We have demonstrated two binding sites of KdS 49 nM and 4.26 microM that are modulated by GTP. From structure-activity studies we have shown the binding to be stereospecific for the beta-isomer of NAD requiring the whole of the molecule for full receptor affinity. The binding sites are distinct from those reported for adenosine and their presence has significance for the physiological role of NAD in the mammalian brain.

ID: 7359526
Title: Trapping of metabolically generated electrophilic species with cyanide ion: metabolism of 1-benzylpyrrolidine.
Abstract: Incubations of 1-benzylpyrrolidine (4) and specifically deuterium-labeled analogues of 4 with rabbit liver microsomal preparations in the presence of cyanide ion have led to the characterization of 1-benzyl-2-cyanopyrrolidine (13), cis- and trans-1-benzyl-2,5-dicyanopyrrolidine (14a and 14b, respectively), and 1-benzyl-5-cyano-2-pyrrolidinone (15). The cyano adducts of the amine are thought to result from nucleophilic attack by cyanide ion on metabolically generated iminium species. The cyanolactam may be produced by mixed function oxidation of the dicyano compounds. Incubations of tritium-labeled 1-benzylpyrrolidine with rabbit liver microsomal preparations led to the reduced nicotinamide adenine dinucleotide phosphate dependent incorporation of the label into the macromolecular fraction isolated from the postincubates. Although the level of incorporation was low compared to the amount of cyano adducts formed, it is comparable to that reported for other metabolically activated cytotoxic agents. Attempts to identify the possible arene oxide rearrangement product 1-(4-hydroxybenzyl)pyrrolidine (24) as a metabolite of 4 were unsuccessful. The results have prompted us to postulate that metabolically generated iminium ions are capable of alkylating nucleophilic functionalities present on microsomal macromolecules.

ID: 25268725
Title: Design, synthesis and SAR studies of NAD analogues as potent inhibitors towards CD38 NADase.
Abstract: Nicotinamide adenine dinucleotide (NAD), one of the most important coenzymes in the cells, is a substrate of the signaling enzyme CD38, by which NAD is converted to a second messenger, cyclic ADP-ribose, which releases calcium from intracellular calcium stores. Starting with 2'-deoxy-2'-fluoroarabinosyl-β-nicotinamide adenine dinucleotide (ara-F NAD), a series of NAD analogues were synthesized and their activities to inhibit CD38 NAD glycohydrolase (NADase) were evaluated. The adenosine-modified analogues showed potent inhibitory activities, among which 2'-deoxy-2'-fluoroarabinosyl-β-nicotinamide guanine dinucleotide (ara-F NGD) was the most effective one. The structure-activity relationship of NAD analogues was also discussed.

ID: 31439792
Title: NAD+ cleavage activity by animal and plant TIR domains in cell death pathways.
Abstract: SARM1 (sterile alpha and TIR motif containing 1) is responsible for depletion of nicotinamide adenine dinucleotide in its oxidized form (NAD+) during Wallerian degeneration associated with neuropathies. Plant nucleotide-binding leucine-rich repeat (NLR) immune receptors recognize pathogen effector proteins and trigger localized cell death to restrict pathogen infection. Both processes depend on closely related Toll/interleukin-1 receptor (TIR) domains in these proteins, which, as we show, feature self-association-dependent NAD+ cleavage activity associated with cell death signaling. We further show that SARM1 SAM (sterile alpha motif) domains form an octamer essential for axon degeneration that contributes to TIR domain enzymatic activity. The crystal structures of ribose and NADP+ (the oxidized form of nicotinamide adenine dinucleotide phosphate) complexes of SARM1 and plant NLR RUN1 TIR domains, respectively, reveal a conserved substrate binding site. NAD+ cleavage by TIR domains is therefore a conserved feature of animal and plant cell death signaling pathways.

ID: 33053563
Title: The NAD+-mediated self-inhibition mechanism of pro-neurodegenerative SARM1.
Abstract: Pathological degeneration of axons disrupts neural circuits and represents one of the hallmarks of neurodegeneration1-4. Sterile alpha and Toll/interleukin-1 receptor motif-containing protein 1 (SARM1) is a central regulator of this neurodegenerative process5-8, and its Toll/interleukin-1 receptor (TIR) domain exerts its pro-neurodegenerative action through NADase activity9,10. However, the mechanisms by which the activation of SARM1 is stringently controlled are unclear. Here we report the cryo-electron microscopy structures of full-length SARM1 proteins. We show that NAD+ is an unexpected ligand of the armadillo/heat repeat motifs (ARM) domain of SARM1. This binding of NAD+ to the ARM domain facilitated the inhibition of the TIR-domain NADase through the domain interface. Disruption of the NAD+-binding site or the ARM-TIR interaction caused constitutive activation of SARM1 and thereby led to axonal degeneration. These findings suggest that NAD+ mediates self-inhibition of this central pro-neurodegenerative protein.

ID: 35476981
Title: Products of gut microbial Toll/interleukin-1 receptor domain NADase activities in gnotobiotic mice and Bangladeshi children with malnutrition.
Abstract: Perturbed gut microbiome development has been linked to childhood malnutrition. Here, we characterize bacterial Toll/interleukin-1 receptor (TIR) protein domains that metabolize nicotinamide adenine dinucleotide (NAD), a co-enzyme with far-reaching effects on human physiology. A consortium of 26 human gut bacterial strains, representing the diversity of TIRs observed in the microbiome and the NAD hydrolase (NADase) activities of a subset of 152 bacterial TIRs assayed in vitro, was introduced into germ-free mice. Integrating mass spectrometry and microbial RNA sequencing (RNA-seq) with consortium membership manipulation disclosed that a variant of cyclic-ADPR (v-cADPR-x) is a specific product of TIR NADase activity and a prominent, colonization-discriminatory, taxon-specific metabolite. Guided by bioinformatic analyses of biochemically validated TIRs, we find that acute malnutrition is associated with decreased fecal levels of genes encoding TIRs known or predicted to generate v-cADPR-x, as well as decreased levels of the metabolite itself. These results underscore the need to consider microbiome TIR NADases when evaluating NAD metabolism in the human holobiont.

ID: 36087583
Title: Uncompetitive, adduct-forming SARM1 inhibitors are neuroprotective in preclinical models of nerve injury and disease.
Abstract: Axon degeneration is an early pathological event in many neurological diseases. The identification of the nicotinamide adenine dinucleotide (NAD) hydrolase SARM1 as a central metabolic sensor and axon executioner presents an exciting opportunity to develop novel neuroprotective therapies that can prevent or halt the degenerative process, yet limited progress has been made on advancing efficacious inhibitors. We describe a class of NAD-dependent active-site SARM1 inhibitors that function by intercepting NAD hydrolysis and undergoing covalent conjugation with the reaction product adenosine diphosphate ribose (ADPR). The resulting small-molecule ADPR adducts are highly potent and confer compelling neuroprotection in preclinical models of neurological injury and disease, validating this mode of inhibition as a viable therapeutic strategy. Additionally, we show that the most potent inhibitor of CD38, a related NAD hydrolase, also functions by the same mechanism, further underscoring the broader applicability of this mechanism in developing therapies against this class of enzymes.

ID: 42341897
Title: Programmed axon degeneration gene variants in human disease.
Abstract: Programmed axon degeneration (PAD; also known as Wallerian degeneration) is a conserved pathway controlling axon breakdown following injury or metabolic stress. PAD is driven by the depletion of nicotinamide adenine dinucleotide (NAD) through loss of the pro-survival enzyme NMNAT2 and activation of the pro-degenerative NADase SARM1. Recent genetic studies have identified pathogenic variants in PAD pathway enzymes associated with severe neurodegenerative phenotypes. Pathogenic variants in NAMPT, NMNAT1, NMNAT2, and SARM1 have been identified and will be discussed in this review. NAMPT variants cause sensory and motor neuropathy with neurodevelopmental symptoms. NMNAT1 variants are well-characterized causes of Leber Congenital Amaurosis type 9, while NMNAT2 variants result in peripheral neuropathies with childhood onset. SARM1 gain-of-function variants with constitutively active NADase activity are enriched in amyotrophic lateral sclerosis patients. These findings demonstrate that maintaining proper NAD homeostasis is crucial for axon survival, and disruption through genetic variants leads to distinct neurodegenerative outcomes. Understanding these rare variants provides insight into PAD mechanisms and supports development of broad-spectrum neuroprotective therapies targeting this pathway. Current therapeutic approaches include SARM1 inhibitors in clinical trials, gene therapy, and NAD precursor supplementation, offering hope for treating multiple neurodegenerative diseases.

ID: 42385602
Title: CD38-driven NAD⁺ depletion governs crypt structure and barrier dysfunction in fluorouracil-induced colonic mucositis: From metabolic collapse to architectural remodeling.
Abstract: 5-fluorouracil (5-FU)-induced colonic mucositis is characterized by epithelial injury, crypt architectural disruption, inflammatory infiltration, oxidative stress, and loss of barrier integrity. The metabolic determinants that regulate epithelial resilience and architectural recovery remain incompletely defined, although 5-FU exposure reproducibly induces these structural lesions. Nicotinamide adenine dinucleotide (NAD+) supports epithelial energy balance and redox homeostasis, while accelerated NAD+ consumption may amplify tissue injury during toxic stress. CD38 is a principal NAD+ hydrolase; yet, its contribution to mucosal structural injury and barrier failure has not been fully elucidated. Herein, we tested whether pharmacological inhibition of CD38 using the small-molecule CD38 inhibitor 78c preserves colonic tissue architecture in a mouse 5-FU-induced mucositis model. We also evaluated the requirement for NAD+ biosynthesis. 5-FU produced prominent histopathological injury marked by epithelial damage/apoptosis, crypt disorganization, inflammatory-cell infiltration, oxidative-injury, and disrupted tight junctions with barrier dysfunction. CD38 inhibition mitigated these abnormalities to preserve crypt-architecture and epithelial continuity. As a result, inflammatory infiltration, oxidative stress, and apoptotic signaling were attenuated, with concurrent restoration of tight junction integrity and intestinal barrier function. Protection was associated with significant improvement in colonic NAD+ levels and the NAD+/NADH balance, increased SIRT1 activity, reduced PARP activation, and activation of Nrf2-linked stress response. Markedly, inhibition of NAD⁺ biosynthesis with a NAMPT inhibitor abrogated the structural and barrier-protective effects of CD38 inhibition across histological, biochemical, and molecular readouts, indicating dependence on NAD⁺ availability. These findings identify the CD38-NAD⁺ axis as a metabolic regulator of colonic-crypt architecture and epithelial barrier organization during 5-FU-induced mucositis.

ID: 42442093
Title: Ergothioneine attenuates age-related declines in circadian rhythmicity.
Abstract: Aging is accompanied by the progressive deterioration of circadian clock function, characterized by reduced amplitude, increased period variability, and impaired metabolic coupling. Declining intracellular nicotinamide adenine dinucleotide (NAD+) levels and SIRT1 activity have been implicated as key mediators of age-associated circadian disruption. Ergothioneine (EGT), a diet-derived antioxidant with longevity-associated effects, has recently been reported to improve healthspan and modulate redox metabolism. However, its effects on the circadian clock remain unclear. Here, we examined whether EGT mitigates age-related decline in circadian rhythmicity using PER2::LUC mouse embryonic fibroblasts (MEFs). Chronic EGT treatment enhanced the amplitude of the PER2::LUC rhythm in a dose-dependent manner without markedly altering the baseline period length. Aging-associated NAD+ decline was modeled using FK866, a NAMPT inhibitor that depletes intracellular NAD+, which reduced rhythm amplitude, lengthened the period, and increased cycle-to-cycle variability. Notably, co-treatment with EGT significantly restored rhythm amplitude, attenuated FK866-induced period lengthening, and reduced period variability. Biochemical analyses revealed that EGT increased the NAD+/NADH ratio under basal conditions and significantly elevated both NAD+ levels and the NAD+/NADH ratio under FK866-induced NAD+ depletion. This effect was not attributable solely to cytoprotection. This study demonstrates that EGT enhances circadian rhythm robustness and counteracts NAD+-depletion-induced clock dysfunction. EGT may ameliorate age-related circadian decline by improving intracellular redox balance and NAD+ metabolism. Given that EGT crosses the blood-brain barrier, it may represent a novel nutritional strategy to preserve circadian function during aging.

ID: 42445026
Title: Novel Pyrrolidinone Derivatives as SARM1 Inhibitors for Treating Axonal Degeneration.
Abstract: Provided herein are novel pyrrolidinone derivatives as SARM1 inhibitors, pharmaceutical compositions, use of such compounds in treating axonal degeneration, and processes for preparing such compounds.

ID: 42449637
Title: Targeting Sirtuins in Thyroid Cancer: Mechanisms, Drug Development, and Emerging Roles in Tumor Immunity and Ferroptosis.
Abstract: Thyroid cancer (TC) is the most common endocrine malignancy, with incidence increasing worldwide. Although most differentiated TCs have a favorable prognosis, radioiodine (RAI)-refractory differentiated thyroid cancer (DTC), BRAF inhibitor-resistant papillary thyroid cancer, and anaplastic thyroid cancer (ATC) remain major areas of unmet clinical need. The sirtuin (SIRT) family of NAD+-dependent enzymes has emerged as a multifaceted regulator of TC biology, with isoform-specific dichotomous roles: SIRT1, SIRT6, and SIRT7 act as tumor promoters through engagement of BRAF/MAPK, PI3K/AKT, epithelial-mesenchymal transition (EMT), and Hippo pathways, while SIRT3 and SIRT4 function as tumor suppressors via mitochondrial metabolic regulation. This review synthesizes recent developments that expand the therapeutic landscape: (i) the recognition that SIRT7 functions as a desuccinylase with preclinically identified oncogenic substrates, modifying KIF23 in ATC and LATS1 in PTC; (ii) the emerging roles of isoform-specific SIRT axes, including the NAMPT-SIRT1-PD-L1 axis, SIRT6-associated regulatory T-cell biology, and SIRT2 as a T-cell metabolic checkpoint, as determinants of immune microenvironment state and potential modulators of immune checkpoint inhibitor response; and (iii) the SIRT6-nuclear receptor coactivator 4 (NCOA4) ferritinophagy axis as a supported ferroptosis vulnerability in ATC, with potential but still hypothesis-generating relevance to dedifferentiated and RAI-refractory DTC. Importantly, the therapeutic logic for SIRT6 is disease-state-specific rather than contradictory: SIRT6 inhibition is rationalized in BRAF-driven aggressive PTC and DTC contexts where SIRT6 supports MAPK signaling, EMT, and ferroptosis resistance, whereas in SIRT6-high ATC, the same enzyme's NCOA4-dependent ferritinophagy activity may instead be exploited to enhance ferroptosis sensitivity. We review the current SIRT modulator pharmacological toolkit-including EX-527, OSS_128167, and emerging SIRT7-selective inhibitors-and identify the substantial clinical translation gap, with no SIRT-targeted clinical trial yet conducted in TC, despite strong preclinical rationale. We outline biomarker-stratified combination strategies with BRAF/MEK inhibitors, multikinase inhibitors, immune checkpoint inhibitors, and ferroptosis inducers, prioritizing biomarker-driven preclinical validation and, where supported by efficacy and safety data, subsequent early-phase evaluation in BRAF V600E-mutant and SIRT6-high thyroid cancer. Sirtuins thus represent a mechanistically promising and potentially biomarker-stratifiable therapeutic hypothesis for difficult-to-treat thyroid cancer; however, clinical translation remains at an early stage and requires validated biomarkers, isoform-selective compounds, and disease-specific in vivo evidence.

ID: 42453345
Title: Recent Advances in Organofluoroprobes for Brain Imaging: Progress, Challenges, and Future Directions.
Abstract: The precise and early diagnosis of neurodegenerative diseases remains a major challenge because of their highly complex and multifactorial pathophysiology. However, in recent years, the potential of organofluoroprobesorganic molecules designed to function as fluorescent probes in bioimaginghas become increasingly prominent. These probes enable visualization of essential biomarkers, including amyloid-beta (Aβ), tau, reactive oxygen and nitrogen species (ROS/RNS), neurotransmitters, and abnormal metal ions, facilitating early detection and monitoring of diseases such as Alzheimer's and Parkinson's. This review focuses on small-molecule organofluoroprobes, which consist of curcumin-, boron-dipyrromethane (BODIPY)-, cyanine-, coumarin-, benzothiazole-, thiophene-, naphthalene-, oxazine-, and organometallic-based systems. We also address biomarker-activated probes, nanoengineered aggregation-induced emission (AIE) luminogens, and photoacoustic probes that provide greater tissue penetration and multimodal imaging. Nevertheless, these imaging probes still have shortcomings, including poor specificity, limited penetration through the blood-brain barrier (BBB), and inefficient near-infrared II (NIR-II) emission. Notably, the design and optimization of fluoroprobes are being revolutionized by the integration of artificial intelligence (AI) and computational methods, such as deep learning, generative models, and virtual screening. These approaches provide new avenues for predictive modeling of physicochemical properties, target affinity and in vivo performance, thereby significantly reducing the time and cost of development. The review concludes by discussing current challenges and future perspectives, including the convergence of AI-assisted molecular design and synthetic chemistry in bioimaging, ultimately leading to the clinical translation of next-generation brain imaging probes.

ID: 42453383
Title: In silico drug discovery and molecular dynamics simulation for targeting neonatal pneumonia and bronchopulmonary dysplasia.
Abstract: Neonatal pneumonia and bronchopulmonary dysplasia (BPD) are major causes of morbidity and mortality in preterm infants, driven by excessive inflammation involving the NOD-like receptor family pyrin domain-containing 3 (NLRP3) inflammasome. This study employed in silico drug discovery, including virtual screening, molecular docking, ADMET profiling, molecular dynamics (MD) simulations, and MM/PBSA calculations, followed by preliminary in vitro validation to identify novel NLRP3 inhibitors from Traditional Chinese Medicine (TCM) compounds for these conditions. The NLRP3 NACHT domain (PDB ID: 7ALV) served as the target. A library of FDA-approved drugs and TCM-derived compounds underwent molecular docking with AutoDock Vina. Top hits were evaluated for ADMET properties using SwissADME, pkCSM, and admetSAR. Selected complexes (Hinokiflavone, Theaflavin, Sciadopitysin, Liquiritin apioside, Tigogenin) were subjected to 200 ns all-atom MD simulations in GROMACS and MM/PBSA binding free energy analysis. In vitro cytoprotective effects were assessed via MTT assay in LPS-stimulated BEAS-2B and MLE-12 lung epithelial cells, with glyburide as positive control. Hinokiflavone and Theaflavin exhibited the strongest docking scores (-10.8 and -10.4 kcal/mol), superior MD stability (lowest RMSD: 0.21 ± 0.02 nm and 0.23 ± 0.03 nm; high hydrogen bond occupancy: 78% and 82%), and most favorable MM/PBSA binding energies (-55.8 and -55.2 kcal/mol), driven by van der Waals and electrostatic interactions. They showed acceptable ADMET profiles with high intestinal absorption and low BBB penetration. In vitro, Hinokiflavone restored cell viability to 89.6% ± 3.2% at 50 µM (comparable to glyburide at 91.2% ± 2.8%), while Theaflavin reached 82.4% ± 3.9%, demonstrating dose-dependent protection against LPS-induced cytotoxicity. Hinokiflavone and Theaflavin emerge as promising NLRP3 inhibitors with stable binding to the NACHT domain and cytoprotective effects in lung epithelial cells. These TCM-derived compounds warrant further preclinical investigation as potential targeted therapies to mitigate inflammation in neonatal pneumonia and BPD.

ID: 42453397
Title: Artificial intelligence in biologic drug discovery: A review of methodological evolution and therapeutic applications.
Abstract: Biologic drugs, primarily comprising proteins and nucleic acids, have emerged as powerful therapeutic modalities; however, their discovery and optimization are often hindered by their inherent complexity. The advent of artificial intelligence (AI), particularly deep learning, is catalyzing a paradigm shift in this field, transitioning it from a process reliant on serendipity and laborious experimentation to a data-driven engineering discipline. This review systematically charts the co-evolution of AI methodologies and their transformative applications across the modern biologic drug development pipeline. We first outline AI's methodological progression, from language models deciphering biological sequence grammar to structure prediction models like AlphaFold making macromolecular folds computationally accessible, and finally to generative models enabling de novo molecular creation. We then explore the practical impact of these technologies in two core phases: the de novo design of novel biologics with bespoke functions and the subsequent multi-parameter engineering and optimization of these candidates for clinical viability. While the potential is immense, significant strategic challenges remain, including the need to build a new AI-native experimental ecosystem and bridge the profound complexity gap between molecular-level predictions and systemic in vivo outcomes. Overcoming these obstacles will usher in a new era of AI-driven, automated closed-loop drug discovery.

ID: 42453411
Title: Molecular glue degraders: Rational design, specificity engineering, and advanced delivery.
Abstract: Molecular glue degraders (MGDs) have emerged as a transformative modality in the field of targeted protein degradation (TPD), enabling the selective elimination of disease-relevant proteins, including those traditionally considered undruggable. Unlike bifunctional proteolysis-targeting chimeras (PROTACs), MGDs operate through monovalent architectures that induce protein-protein interactions (PPIs) between E3 ligases and neosubstrates, offering advantages in chemical simplicity, cell permeability, and target scope. However, MGD discovery remains serendipitously, and a translational framework that links rational design to predictable selectivity and tissue exposure is still lacking. In this review, we present an integrated framework for advancing next-generation MGDs through three critical dimensions: rational design, specificity optimization, and delivery systems. First, we examined cutting-edge strategies in MGD design, including covalent handle-based reprogramming, PPI-driven stabilization, and multi-site, multi-functional constructs. Second, we explored structure-guided engineering and chemoinformatic models, such as cereblon degron motifs, zone-based design and multiparameter optimization, to improve neosubstrate selectivity while minimizing off-target liabilities. Third, we summarized delivery platforms, including antibody‒drug conjugates, nanoparticle-enabled systems, and folate-mediated targeting, which are primarily intended to improve tissue selectivity and targeted distribution, thereby promoting local tissue accumulation. Finally, we discussed emerging opportunities at the intersection of artificial intelligence, structural biology, and systems pharmacology for accelerating MGD discovery and clinical translation. Collectively, these interdisciplinary insights underscore the therapeutic promise of MGDs and lay the groundwork for their next-generation evolution in precision medicine.

ID: 42453426
Title: Revolutionizing drug discovery from natural products: The roles of artificial intelligence and multi-omics in accelerating innovation.
Abstract: Natural products and their derivatives have long been crucial in drug therapy, especially in traditional medicine. However, challenges in screening, isolation, characterization, and optimization have slowed their development in the pharmaceutical industry. Recent advancements in artificial intelligence (AI) and multi-omics technologies are revitalizing this field. AI offers powerful tools for understanding natural compounds, enhancing molecular representations, and supporting tasks such as binding prediction, drug repurposing, and retrosynthesis. Moreover, generative models are aiding in natural product optimization and the creation of pseudo-natural compounds. At the same time, multi-omics technologies, including genomics, transcriptomics, proteomics, and metabolomics, have enabled high-throughput studies of plant traits, synthesis, regulatory mechanisms, and quality control, providing valuable data for AI model development. These advancements help accelerate the discovery of new compounds with medicinal potential. Furthermore, in the field of traditional Chinese medicine research, which is largely based on natural plant sources, AI systems exemplified by UNIQ system, combining AI and multi-omics, have been instrumental in mechanistic studies and new drug development. This study comprehensively discusses the algorithms and applications of AI and multi-omics technologies in the drug development of natural compounds and plants, as well as summarizing relevant databases which might provide high-quality data for the future development of AI algorithms targeting natural products.

ID: 42453484
Title: Targeting hexokinase 2 for cancer therapy: advances, limitations, and emerging opportunities in inhibitors and degraders.
Abstract: Hexokinase 2 (HK2) is an important target in cancer metabolism because it supports both glycolytic flux and mitochondria-associated survival signaling. In recent years, multiple HK2-directed strategies have emerged, including active-site inhibitors, repurposed scaffolds, electrophilic glycolysis blockers, and targeted protein degraders. Although several compounds have shown promising antitumor activity in preclinical models, their translation has been constrained by high structural similarity to HK1, the polarity and exposure liabilities of glucose-mimetic chemotypes, and, in some cases, limited mechanistic specificity. This review summarizes recent progress in HK2 inhibitor and degrader discovery, highlighting representative chemotypes, structure-activity relationships, pharmacological properties, and translational considerations. We also discuss opportunities for future development, including isoform-selective design, disruption of HK2-mitochondrial interactions, improved delivery strategies, and rational combination approaches. Overall, HK2 remains a promising but pharmacologically challenging target for cancer therapy.

ID: 42453940
Title: Perithyroidal Adipose Tissue Drives Thyroid Tumorigenesis through Adipokine Signaling and Immune Suppression.
Abstract: The perithyroidal adipose tissue (PAT), given its direct anatomical proximity to the thyroid gland, has long been postulated as a modulator of the thyroid tumor microenvironment. However, its cellular composition, functional heterogeneity, and specific roles in thyroid cancer progression remain unknown. To address these knowledge gap, we performed single-nucleus RNA sequencing of PAT from patients with papillary thyroid carcinoma (PTC) and multinodular goiter (MNG), combined with machine learning, proteomics, immunofluorescence, ex vivo assays, and human serum analysis. We constructed the first high-resolution atlas of human PAT, revealing an immune-rich niche and previously unrecognized adipocyte heterogeneity, including thermogenic subpopulations (BL-Ad1, BL-Ad2, OXPHOS-Ad) from distinct progenitors. Functionally, the PAT secretome from PTC substantially enhanced thyroid cancer cell proliferation compared to MNG. Integrated analyses identified a pathogenic adipokine triad characterized by loss of ADIPOQ and gain of NAMPT and IGF1. Restoring ADIPOQ signaling or inhibiting NAMPT/IGF1 suppressed tumor growth in vitro and in vivo. Additionally, we identified CCL14, down-regulated in PTC-derived OXPHOS-Ad, as a key immune regulator. Reduced CCL14-CCR1 signaling impaired CD80 expression in M1-like macrophages, disrupting CD80-CD28 costimulation and consequently diminishing T cell proliferation and recruitment. Consistently, circulating CCL14 levels were reduced in PTC patients. In conclusion, PAT acts as a dynamic endocrine and immunomodulatory component of the tumor microenvironment that promotes thyroid tumor growth through adipokine-mediated and immune-dependent mechanisms.

ID: 42454109
Title: Establishment of a machine learning prediction model for Wallerian degeneration after ischemic stroke.
Abstract: Wallerian degeneration (WD) is a common and clinically significant complication of ischemic stroke (IS). Due to the multifactorial and nonlinear characteristics of its underlying mechanisms, accurately identifying high-risk patients early remains challenging. This study aimed to develop and validate an interpretable machine learning (ML) model to predict WD after IS. We retrospectively analyzed clinical data from 269 patients with IS, all admitted to the Xinhua Hospital of Dalian University. The patients were randomly divided into a training set (70%) and an internal validation set (30%). Thirty demographic, imaging, and laboratory variables were assessed, and predictive features were selected through Least Absolute Shrinkage and Selection Operator (LASSO) regression, followed by confirmation using multivariate logistic regression. Nine ML algorithms were constructed and compared. The best-performing model was interpreted using Shapley Additive Explanations (SHAP). Among the 269 patients, 35.32% (95/269) of IS patients developed WD. LASSO regression selected eight candidate predictors (six reaching statistical significance in multivariate logistic regression; MCA and PCA retained based on LASSO selection and biological relevance): smoking history, hyperlipidemia, standard antiplatelet therapy, achieving LDL target with oral statins, maximum cross-sectional area of the stroke, middle cerebral artery (MCA), posterior cerebral artery (PCA), and the number of stroke-affected layers. Within the study population, the Random Forest model showed internally favorable predictive performance (training AUC = 0.946; validation AUC = 0.856) and reasonable internal consistency, surpassing AdaBoost, Logistic Regression, Lasso, Decision Tree, KNN, GaussianNB, XGBoost, and LightGBM. Through SHAP analysis, this study quantified and visualized the contribution of each predictive variable to the Random Forest model's prediction of the occurrence of WD, identifying key factors such as smoking history, MCA, and PCA, and revealing their interactions, thereby enhancing the model's interpretability for research purposes. We developed and validated an interpretable Random Forest model with potential for predicting the occurrence of WD. By integrating demographic, imaging, and laboratory features, this model provides an internally validated framework that shows promise for early risk assessment, with potential to support personalized management pending external validation.

ID: 42454649
Title: Can Cavity Prediction Algorithms Help in Docking Experiments?
Abstract: Blind docking is a method for predicting a binding mode of a ligand with a protein without any prior information about a binding site. Some tools allow this type of docking experiment directly, others, including some established tools, require binding site information being passed as an input. In this latter case, one can use cavity prediction tools and use the results of their prediction as an input in these docking calculations. However, it is still unclear if the results of these predictions can be reliably used in protein-ligand docking and what is the best technical way to pass this information to the docking algorithm. In this study we estimated the applicability of the binding pocket prediction tools in docking experiments to address this gap in knowledge. We use four different computational tools for cavity prediction and use the best predicted cavities represented in different ways to run GOLD docking calculations. Analysis of subsequent use in docking highlights that Fpocket and CAVIAR are the best performing cavity prediction tools in this context. Further analysis shows that accurate binding site input does not guarantee accurate binding pose predictions and, even with the predicted cavities, the more restrained the input is, the more reliable the docking results are.

ID: 42454651
Title: Constructing a NIR AIEgen Based on the Novel Molecular Framework for Imaging Lipid Droplet Accumulation in Parkinson's Disease Mouse Brain.
Abstract: Parkinson's disease (PD) pathogenesis has been linked to the aberrant accumulation of lipid droplets (LDs). Understanding the situation of LD accumulation and its dynamic variations in the neural tissues of PD patients is therefore crucial for deciphering the underlying pathophysiological mechanisms of this neurodegenerative disorder. Developing a specific fluorescent probe for LDs that can also function within the brain presents a significant challenge. This study introduces a NIR (near-infrared) emitting fluorescent probe, QCN-TP, through step-by-step molecular design strategy, based on a novel framework of dye, characterized by multiple advantageous properties, including polarity sensitivity, a substantial Stokes shift (150-280 nm), and aggregation-induced emission (AIE). The probe, QCN-TP, demonstrates remarkable sensitivity and selectivity in the recognition of LD accumulation, with enhanced deep tissue penetration capabilities. Notably, QCN-TP effectively monitors both the presence of intracellular LDs and their dynamic content variations in MPP+-induced PD cell models. Furthermore, the probe enables in vivo imaging of abnormal LD accumulation in the brain tissue of PD model mice. This probe offers a useful tool for real-time monitoring of lipid dynamics, with potential applications in PD research and drug discovery.