Hepatocellular carcinoma (HCC) is a major cause of cancer mortality worldwide. Emerging evidence in cancer neuroscience suggests that tumor–nerve crosstalk can modulate tumor growth, immune remodeling, dissemination, and therapy resistance. Perineural invasion (PNI) is a hallmark of aggressive tumors and correlates with recurrence, metastasis, disease-specific death, and neuropathic pain, yet its prevalence and prognostic relevance in primary HCC remain inconsistently defined. Given the limited and heterogeneous clinical data, we synthesize available observations and propose mechanistic hypotheses for neural–tumor interactions in the hepatic niche. Potential drivers are organized into five domains—neurotrophic factors, axon guidance molecules, neurotransmitter signaling, cell adhesion molecules, and the tumor microenvironment—highlighting neuro–immune–stromal circuits that may create permissive perineural niches. We also outline experimental platforms to test these hypotheses, including co-culture systems, organoids, and microfluidic nerve-on-chip models. Targeting neural–tumor interactions may offer translational opportunities in HCC, but causality and actionable targets cannot be established without improved clinical and experimental rigor. Standardized pathological assessment and dedicated HCC-focused PNI models are needed to validate mechanisms, harmonize reporting, and enable reproducible, clinically meaningful advances.
Liver metastasis relies on the establishment of a pre-metastatic niche (PMN) before tumor cell seeding. Although low-dose irradiation can modulate local immune responses, whether prophylactic low-dose liver irradiation influences hepatic PMN formation and metastatic colonization remains unclear. We used a prophylactic low-dose liver irradiation strategy in mouse models of non-small cell lung cancer (NSCLC) liver metastasis to evaluate its effects on metastatic colonization and hepatic immune remodelling. Bulk RNA sequencing, single-cell RNA sequencing, and in vitro functional assays were performed to characterize irradiation-induced hepatocyte-monocyte signaling, and in vivo CXCL10 neutralization was used to assess its functional relevance. Prophylactic low-dose liver irradiation suppressed NSCLC liver metastasis and reshaped the hepatic immune microenvironment during PMN formation. A single 2 Gy dose showed the strongest antimetastatic effect. Mechanistically, 2 Gy irradiation induced hepatocyte-derived SAA1. SAA1 activated a TLR2–NF-κB/RELA-associated transcriptional program in inflammatory monocytes, leading to SLC7A11 upregulation, improved redox homeostasis, and increased CXCL10 secretion. SLC7A11 activity was required to sustain this chemokine-secretory phenotype. CXCL10 neutralization in vivo reduced CD8 + T-cell recruitment and attenuated the antimetastatic effect of low-dose irradiation. Prophylactic low-dose liver irradiation limits NSCLC liver metastasis and is associated with reprogramming of the PMN toward an anti-metastatic immune state. These findings link irradiation to metastatic niche immune remodeling and highlight the translational relevance of low-dose immune radiobiology in metastasis control.
Diabetic retinopathy (DR) remains a leading cause of visual impairment worldwide and is characterized by retinal microvascular dysfunction and chronic neuroinflammation. The pathogenesis of DR involves the activation of retinal microglia, which release a variety of pro-inflammatory cytokines, thereby amplifying inflammatory signaling and compromising the structural and functional integrity of the retinal vasculature. Piceatannol (PIC), a bioactive polyphenolic compound, has demonstrated potent anti-inflammatory properties and beneficial effects on vascular homeostasis. However, its role and underlying mechanisms in DR remain largely unexplored. An integrated approach combining network pharmacology and transcriptomic analyses was employed to systematically identify the potential molecular targets of PIC in DR. Molecular docking was first performed to predict the interaction between PIC and C-C motif chemokine receptor 5 (CCR5), followed by experimental validation of the binding affinity. In vitro and in vivo DR models were established using high glucose (HG)-stimulated microglia and streptozotocin-induced diabetic mice, respectively. The effects of PIC on microglial polarization, retinal vascular permeability, and inflammatory responses were subsequently evaluated. A total of 159 overlapping targets associated with both PIC and DR were identified, among which CCR5 emerged as a key target. In diabetic mice, PIC treatment restored retinal thickness, reduced the formation of acellular capillaries, and attenuated retinal vascular leakage. CCR5 was predominantly expressed in retinal microglia, whereas its ligand, CCL5, was significantly upregulated in DR. PIC inhibited microglial activation and M1 polarization, reduced the secretion of pro-inflammatory cytokines, and promoted the expression of M2-associated markers through modulation of the CCL5/CCR5/NF-κB signalling pathway. In vitro, PIC suppressed HG-induced M1 polarization of microglia and preserved the expression of endothelial tight junction proteins, including ZO-1 and Claudin-5. Furthermore, CCR5 knockdown and pharmacological activation experiments confirmed the central role of CCR5 in mediating the protective effects of PIC. PIC exerts protective effects on retinal microvascular function in DR by regulating microglial polarization. Specifically, it suppresses the pro-inflammatory M1 phenotype while promoting the anti-inflammatory M2 phenotype through inhibition of the CCL5/CCR5/NF-κB signaling pathway. Collectively, these findings suggest that PIC may represent a promising therapeutic candidate for the prevention and treatment of DR.
Conventional chimeric antigen receptor T-cell (CAR-T) manufacturing requires prolonged ex vivo processing and substantial viral input. We investigated whether brief T-cell activation combined with closed-loop microfluidic recirculation could improve low-multiplicity-of-infection (MOI) lentiviral transduction while generating functional CD19 CAR-T cells within a 24-h core process. Primary human T cells were activated with CD3/CD28 beads for 4 h and transduced with CD19 CAR/green fluorescent protein reporter (CAR/GFP) lentivirus at MOI 0.5 or 1.0 in donor-matched microfluidic-chip and static-plate comparisons. Unless otherwise stated, microfluidic rapid-manufactured CAR-T (MF-rmCAR-T) products were generated at MOI 1.0 by 20 h closed-loop recirculation. Conventionally manufactured CAR-T (cmCAR-T) products served as the product-level comparator. Day 7 CAR/GFP positivity, bulk-product vector copy number (VCN), viability, expansion, and phenotype were assessed, followed by in vitro functional testing and exploratory evaluation in a systemic Raji-Luc xenograft model. Donor-matched data were analyzed using paired t-tests or two-way repeated-measures ANOVA with Šídák correction, as appropriate. Microfluidic processing increased Day 7 CAR/GFP positivity versus matched static transduction at MOI 0.5 (14.6
Lung cancer (LC) remains one of the leading causes of cancer-related morbidity and mortality worldwide. Increasing evidence suggests that both the lung cancer-associated microbiome and the intratumoral microbiome are important components of the tumor ecosystem and contribute to cancer development, progression, and therapeutic response. Distinct microbial alterations have been identified in the lower airway, bronchoalveolar lavage fluid, airway epithelium, and tumor tissues, indicating complex host-microbe interactions across multiple microbial niches. Mechanistically, these microbial communities may influence tumor biology through microbial metabolites, chronic inflammation, immune modulation, and remodeling of the tumor microenvironment. In addition, specific microbial signatures have been associated with disease diagnosis, prognosis, recurrence, and treatment outcomes, highlighting their potential clinical value. This review integrates current advances in lung cancer-associated and intratumoral microbiota, summarizes their biological and clinical significance, and discusses key methodological challenges and future directions for clinical translation.
Parenteral nutrition-associated cholestasis (PNAC) is a major complication in very preterm infants receiving prolonged parenteral nutrition (PN) and is typically diagnosed only after conjugated hyperbilirubinemia develops. Noninvasive biomarkers reflecting disturbances in the gut-liver axis remain limited. This study aimed to identify biomarkers associated with PNAC using an integrated multi-omics approach. In this prospective cohort study, 81 very preterm infants (< 30 weeks’ gestation) receiving prolonged PN were enrolled, including 20 who developed cholestasis and 61 age- and time-matched controls. Urine and stool samples were collected during PN exposure, predominantly when PNAC occurred. Multi-omics profiling included bile acid quantification in urine and feces using ultra-performance liquid chromatography-tandem mass spectrometry, urinary metabolomics using proton nuclear magnetic resonance spectroscopy, and stool microbiome profiling using 16 S ribosomal RNA sequencing. Logistic regression and receiver operating characteristic analyses evaluated associations and discriminatory performance. Infants who developed cholestasis showed alterations across microbiome composition, fecal bile acids, urinary bile acids, and urinary metabolites. Urinary bile acids demonstrated the strongest discriminatory performance among all omics layers. Several urinary sulfated bile acids, including taurolithocholic acid-3-sulfate and taurochenodeoxycholic acid-3-sulfate, were strongly associated with disease risk. A composite score based on the top three urinary bile acids achieved an area under the receiver operating characteristic curve of 0.950, outperforming the corresponding urinary metabolites (0.894), fecal bile acids (0.734), and microbiome features (0.641). Cross-omics analysis revealed coordinated associations among gut microbial features, fecal bile acids, urinary bile acids, and urinary metabolites, supporting an integrated gut-liver-urine framework. Urinary sulfated bile acids represent robust biomarkers associated with PNAC in very preterm infants and may serve as a noninvasive test for PNAC during PN exposure.
Patients with disorders of consciousness (DoC) lack effective non-invasive neuromodulation therapies capable of targeting deep-brain arousal circuits. Temporal interference stimulation (TIS) offers potential for non-invasive deep brain targeting. However, its efficacy, optimal stimulation parameters, and underlying neural mechanisms remain to be systematically investigated. We constructed a rodent TIS electrode configuration with cathode affixed to the masseter muscle to achieve free-moving stimulation in mice, and optimized stimulation parameters in isoflurane-anesthetized C57 mice, validated using righting reflex assays. Target specificity was confirmed by comparing Central Medial Thalamic Nucleus (CM) stimulation with that of the caudate putamen. Neuronal activation was assessed via calcium imaging, c-Fos staining, and electrocorticography. Biosafety was evaluated through acute and 7-day repeated histopathological analyses and open-field testing. Preliminary feasibility was assessed in two patients with chronic DoC using Coma Recovery Scale–Revised and functional near-infrared spectroscopy. Optimal TIS parameters (60 s, 0.8 mA, 10 Hz) significantly promoted arousal, accelerated righting reflex recovery, and increased respiratory rate. CM-targeted TIS preferentially activated CM neurons, enhanced cortical beta/gamma oscillations, and suppressed delta activity. No tissue damage, neuronal loss, microglial activation, or motor deficits were observed under acute or repeated stimulation. In two patients, 10 Hz TIS transiently improved consciousness ratings and enhanced functional connectivity and right Broca’s area activation. CM-targeted TIS is a promising non-invasive deep neuromodulation strategy, supported by a translational evidence chain from preclinical optimization, mechanistic validation, and biosafety assessment to preliminary clinical feasibility. Chinese Clinical Trial Registry, ChiCTR2500114570. Registered 15 December 2025.
Abnormal lipid metabolism is widely recognized as a hallmark of cancer cells. In particular, non-small cell lung cancer (NSCLC) cells exhibit heightened lipid metabolic processes to meet their biosynthetic and energetic demands, while concurrently reprogramming lipid metabolism to remodel the tumour microenvironment (TME). This review details the pivotal roles of lipid metabolic reprogramming in NSCLC. First, we delineate the distinct lipid metabolic characteristics of NSCLC, exploring their regulatory mechanisms and contributions to tumour progression. Given the significant impact of lipids on the TME, we subsequently describe the intricate crosstalk between NSCLC cells and other cellular components within the TME. Furthermore, we investigate the relationship between lipid metabolism and NSCLC treatment, encompassing its role in therapeutic resistance and the potential of targeting lipid metabolism as a therapeutic strategy. Finally, we discuss the prospects and current limitations of targeting lipid metabolism in the clinical management of NSCLC. A substantial body of evidence suggests that lipid metabolic reprogramming plays a critical role in NSCLC. Median time to progression data from a phase I clinical study demonstrated that monotherapy with the FASN inhibitor showed a preliminary signal of clinical activity in KRAS-mutant NSCLC (64
Bone regeneration in individuals with type 2 diabetes mellitus (T2DM) is significantly impaired, even when autologous bone grafts are used. However, previous studies have failed to elucidate the physiological mechanism, and effective treatments are lacking. This study investigated the underlying mechanisms and developed a targeted therapeutic strategy. We established a femoral condyle defect model in diabetic rats and performed autologous bone grafting. Multiomics sequencing analyses revealed core events that occurred in diabetic bone tissue. Seahorse XF analysis was used to detect cellular metabolic reprogramming. Calcium ion flow cytometry and YAP nuclear localization experiments confirmed the changes in mechanosensitivity in BMSCs from diabetic rats. Targeted scaffolds to repair diabetic bone defects were developed through Si ion surface modification and structural design and validated in the diabetic rat bone defect model. Proteomic and targeted energy metabolomics analyses revealed an imbalance in osteogenic/adipogenic differentiation and glycolytic dysfunction. RNA sequencing of BMSCs from diabetic rats revealed that reduced mechanosensitivity may be the core mechanism responsible for the alterations in cell fate. Flow cytometry with a calcium ion probe and YAP nuclear localization experiments confirmed the decreased mechanosensitivity of BMSCs from diabetic rats. Si-CaP was developed to increase glycolysis and the TPMS microstructure was designed to promote YAP nuclear translocation, thereby redirecting BMSCs toward osteogenic differentiation. In vivo experiments in a diabetic rat femoral condyle defect model demonstrated that the structured Si-CaP TPMS scaffold effectively promoted bone regeneration. T2DM-associated bone defect healing impairment is caused by metabolic reprogramming and reduced mechanosensitivity in BMSCs. Targeted modulation of metabolism and mechanosensitivity is an effective treatment for bone defects in T2DM rats.
Currently, numerous studies have clearly established the tumor-suppressive role of Tight Junction Protein 2 (TJP2). Although our previous research confirmed the down-regulation of TJP2 in ccRCC, its biological functions and underlying regulatory mechanisms remain poorly understood. In this study, we characterized the biological functions of TJP2 using cell phenotypic assays and mouse orthotopic renal tumor growth model. Subsequently, we identified mitochondrial import inner membrane translocase subunit Tim21 (TIMM21) as the downstream target protein of TJP2 through TMT (Tandem Mass Tags)-based proteomic profiling and co-immunoprecipitation (co-IP) experiment. Furthermore, we elucidated the downstream signaling mechanisms by which the TJP2/TIMM21 axis inhibits ccRCC progression using a combination of IP, Immunofluorescence, Western blotting, qRT-PCR, and RNA-seq analyses. Our experimental results confirmed that TJP2 over-expression significantly inhibits the growth, migration, and invasion of ccRCC cells and promotes cell apoptosis both in vitro and in vivo. TIMM21 is under-expressed in ccRCC, and its decreased expression correlates with unfavorable pathological characteristics and poor patient prognosis. Mechanistically, TJP2 directly binds to TIMM21 protein, promoting its expression and inhibiting its ubiquitination levels. TJP2 over-expression up-regulates SPRY4 expression and inhibits the ERK1/2 signaling pathway. Notably, knockdown of TIMM21 in TJP2-overexpressing cells significantly reverses TJP2-induced cellular phenotypic inhibition, while suppressing SPRY4 expression and promoting pERK1/2 expression. Consistently, knockdown of SPRY4 similarly rescues the inhibited cellular phenotypes caused by TIMM21 over-expression and restores pERK1/2 levels. Our study demonstrates the mechanism by which the TJP2/TIMM21 axis accelerates SPRY4 expression and restrains the ERK1/2 signaling pathway, thereby suppressing ccRCC progression.
POU4F3 (DFNA15) is an established causative gene for autosomal dominant non-syndromic hearing loss (ADNSHL); however, its mutational spectrum in the Chinese population remains incompletely characterized. This study aimed to expand the mutational spectrum of POU4F3 in the Chinese population, to characterize the clinical phenotypes of affected individuals, to describe the proportion of POU4F3-related families in this referral cohort, and to summarize the global reporting distribution of pathogenic POU4F3 variants in ADNSHL. Among 83 unrelated ADNSHL families evaluated at two tertiary referral centers, nine families harboring POU4F3 variants were identified through whole-exome sequencing and validated by Sanger sequencing with co-segregation analysis. Comprehensive audiological assessments, including pure-tone audiometry, extended high-frequency audiometry, and tinnitus evaluation, were performed on all available family members. Variants were classified according to ACMG/AMP guidelines. Genotype–phenotype correlations were analyzed by integrating data from our cohort with 43 previously published variants. Nine distinct POU4F3 variants were identified, including four novel variants (c.149_152dup, c.687_688delCA, c.704 C > T, and c.709_710delTC) and five previously reported variants (c.371 C > A, c.592 C > A, c.706 C > T, c.952G > A, and EX1-EX2E Del). POU4F3 variants accounted for 10.8
While amyloid-β (Aβ) has historically dominated the research landscape of Alzheimer’s disease (AD), the limited clinical success of Aβ-centric therapies has redirected focus toward tau pathology, which correlates more robustly with cognitive deterioration and synaptic dysfunction. Transcending the traditional linear pathological model, this review reframes tau phosphorylation as a dynamic hub within a multi-scale regulatory network. We first synthesize recent breakthroughs in molecular mechanisms, detailing how the kinase-phosphatase equilibrium, cross-regulation of diverse post-translational modifications (PTMs), nuclear envelope damage, iron metabolism and ferroptosis collectively drive the transition from soluble tau species to neurofibrillary tangles (NFTs). Beyond neuronal boundaries, we elucidate how pathological tau orchestrates systemic neurotoxicity by synergizing with Aβ deposition to trigger neuroinflammation, blood-brain barrier breakdown, and gut-brain axis dysregulation. Finally, we bridge these mechanistic insights with translational advancements, evaluating next-generation biofluid biomarkers and innovative therapeutic modalities—ranging from small-molecule inhibitors to Dephosphorylation-Targeting Chimeras (DEPTACs)—currently under clinical investigation. This integrated perspective offers a holistic framework for understanding AD pathogenesis and provides a roadmap for the development of precision medicine strategies targeting the tau interactome.
High-altitude hypoxia and reoxygenation impose systemic challenges that extend beyond isolated organ injury. However, the host-wide physiological responses, redox-linked metabolic alterations, and host-microbiome interactions underlying these processes remain poorly understood, particularly during the reoxygenation phase. Here, we aimed to characterize systemic and organ-level functional changes across hypoxia and reoxygenation, and identify potential microbial biomarkers associated with physiological adaptation. We analyzed 72 hematological markers in 458 participants across multiple stages of hypoxia and reoxygenation. In a subset of 226 individuals, we profiled the gut microbiome, fecal metabolome, and plasma metabolome of high-altitude returnees. Multi-omics factor analysis was then applied to dissect host-microbiome-metabolism interactions related to redox stress and metabolic homeostasis. Physiological adjustments during hypoxia and reoxygenation were prolonged, with markers indicative of myocardial, hepatic, pancreatic, and inflammatory stress persisting for at least one month after descent. High-altitude returnees exhibited reduced microbial diversity, altered community composition, and pronounced metabolic rewiring that closely correlated with systemic and organ-level stress. Multi-omics factor analysis resolved two major biological axes. Within the reoxygenation-associated stress axis, Rhodococcus emerged as a sensitive indicator of oxidative stress-related metabolic disorder that may serve as a biomarker for evaluating post-descent recovery status. The hypoxic adaptation axis centered on Butyrivibrio, which was linked to lipid metabolism and host energy homeostasis. Altitude-related hypoxia and reoxygenation constitute a full-cycle physiological challenge characterized by sustained redox and metabolic stress. Our findings provide a systems-level framework for understanding host-microbiome responses during oxygen fluctuation and identify candidate biomarkers for future monitoring and intervention.
Following the discontinuation of routine smallpox vaccination and the emergence of mpox outbreaks, immunity against orthopoxviruses has received renewed attention. This study characterized immune responses among individuals with historical smallpox vaccination. Separately, an exploratory mathematical model was used to assess hypothetical transmission scenarios among immunocompromised populations. Cellular and humoral immune responses following historical smallpox vaccination were assessed using enzyme-linked immunosorbent assay and enzyme-linked immunospot assay. Associations between clinical characteristics, laboratory markers, and immune responses were evaluated using regression analyses. An exploratory susceptible-infected-recovered-dead model based on published epidemiological parameters was used to assess hypothetical transmission scenarios among immunocompromised populations. Analysis included 112 peripheral blood mononuclear cell and 3,202 serum samples. Cellular immune responses were limited among individuals with historical smallpox vaccination, with no significant differences observed between healthy and disease groups. Vaccinated individuals with comorbidities had lower vaccinia virus Tianan strain-specific immunoglobulin G titers than the healthy group (P < 0.001), with variations observed across chronic disease subgroups. The proportion of individuals with negative neutralizing antibody responses decreased with age but was higher among disease groups than in the healthy group. Chronic disease (odds ratio [OR], 0.67; 95
Environmental factors play an important role in the natural history of Inflammatory bowel disease (IBD). As a key modifiable environmental factor, diet profoundly influences gut microbiota, mucosal barrier integrity, and host immunity, making dietary strategies an attractive target for IBD prevention and management. Nevertheless, the potential of microbiota-supportive dietary strategies for reducing disease incidence remains unclear. Furthermore, the proteomic signatures linking such a dietary pattern to IBD risk require further exploration. We examined the association between adherence to a microbiota-supportive dietary pattern, quantified using the Dietary Index for Gut Microbiota (DI-GM), and incident IBD in a prospective cohort (n = 208,143). High-throughput plasma proteomic data from a subset (n = 21,919) were further integrated to identify diet-associated circulating proteins, followed by exploratory mediation analysis to investigate candidate proteomic signatures underlying the observed association. Over a mean follow-up of 10.77 years, higher adherence to the microbiota-supportive dietary pattern was associated with a 16
Peripheral artery disease (PAD) affects more than 14 million Americans, and its most advanced form, critical limb-threatening ischemia (CLTI), is associated with chronic inflammation, poor tissue perfusion, and impaired skeletal muscle repair. Although tetraspanins are known to organize immune receptor signaling, their contribution to ischemic tissue injury and recovery in CLTI remains incompletely understood. We investigated the leukocyte tetraspanin CD37 after identifying increased CD37 expression in human PAD gastrocnemius muscle. We analyzed human gastrocnemius muscle transcriptomic data from patients with PAD using deconvolution-based immune profiling to define associations between CD37 expression and immune cell programs. To examine the functional role of CD37 in ischemic tissue repair, we used a murine hindlimb ischemia model in wild-type and Cd37-/- mice. Outcomes included perfusion recovery, skeletal muscle histology, transcriptomic profiling of post-ischemic muscle, tissue immune phenotyping, and ex vivo macrophage stimulation studies. In human PAD muscle, CD37 expression was associated with greater immune infiltration (R = 0.51, p = 0.0002) and showed its strongest relationship with inflammatory macrophage programs (R = 0.48, p = 0.0003). Cd37-/- mice exhibited accelerated revascularization (87.6±9.8
Obesity-induced oxidative stress triggers the cGAS-STING pathway via mitochondrial DNA (mtDNA) leakage, driving chronic white adipose tissue (WAT) inflammation. Nuclear factor erythroid 2-related factor 2 (NRF2) is a master transcriptional regulator of antioxidant responses, but whether it can serve as an endogenous negative regulator of the STING-driven inflammatory cascade in adipocytes remains poorly defined. This study investigated the role of NRF2 in modulating the STING pathway to alleviate adipose inflammation and explored how fat depot heterogeneity influences this regulatory axis. NRF2 activity was assessed in the WAT of high-fat diet-induced obese mice and in differentiated adipocytes. In vitro, palmitic acid-stimulated adipocytes were used to evaluate the anti-inflammatory effects of NRF2 activation. Mechanistic studies involved genetic knockdown/overexpression, transcriptomic profiling, and assessments of cytosolic mtDNA levels and STING translocation. In vivo, obese mice were treated with the NRF2 activator CDDO-Im, alongside in situ AAV-pAdipoq-mediated NRF2 knockdown or STING overexpression in WAT, to assess metabolic phenotypes and inflammatory responses. NRF2 activity was reduced in obese WAT and mature adipocytes. In vitro, NRF2 activation attenuated oxidative stress and inflammation by suppressing the STING cascade, characterized by diminished cytosolic mtDNA leakage and subsequent inhibition of STING-dependent translocation and effector signaling. In vivo, pharmacological NRF2 activation significantly improved obesity, insulin resistance, and systemic inflammation. These effects were largely abolished by local adipocyte-specific NRF2 knockdown or STING overexpression. Transcriptomic profiling revealed depot-specific effects: NRF2 robustly suppressed inflammatory pathways in visceral epididymal WAT (eWAT) but had minimal impact in subcutaneous inguinal WAT (iWAT). Consistently, NRF2 downregulated STING in eWAT while upregulating it in iWAT. Multiplex imaging and cell-fraction analysis demonstrated that active STING signaling operates predominantly in mature adipocytes. Adipocyte differentiation status was identified as a key determinant of this divergence. Due to higher adipogenic potential, NRF2 preferentially inhibited adipogenesis in iWAT, leading to an enrichment of immature adipocytes expressing high STING levels. NRF2 is a critical negative regulator of STING signaling that mitigates obesity-associated inflammation. This study identifies the NRF2–STING axis as a therapeutic target and demonstrates that inherent differences in adipogenicity dictate depot-specific pharmacological responses, providing a framework for precision translational medicine in metabolic diseases.
Neuropathic pain remains a major unmet clinical challenge, as current therapies provide limited efficacy and poor tolerability. A major obstacle to developing effective analgesics is the pronounced cellular and molecular heterogeneity of the dorsal root ganglion (DRG), which integrates neuronal, glial, immune, and stromal responses to injury. Recent advances in single-cell RNA sequencing (scRNA-seq) and single-nucleus RNA sequencing (snRNA-seq) have enabled cell-type-resolved analyses of these responses and revealed disease-associated cellular states that are obscured by bulk tissue profiling, thereby offering new opportunities to redefine disease mechanisms and therapeutic priorities. This review synthesizes evidence from single-cell studies of peripheral nerve injury, diabetic painful neuropathy, and chemotherapy-induced neuropathy. Although these conditions share common biological processes, including neuronal state remodeling, disrupted glial homeostatic support, context-dependent immune responses, and extracellular matrix reorganization, they do not converge on a single conserved molecular program. Peripheral nerve injury is characterized by neuronal injury and repair states, reactive and metabolic glial remodeling, and neuroimmune recruitment and crosstalk. Diabetic neuropathy is associated with altered sensory coding, impaired satellite glial lipid support, and neurodegenerative remodeling. Chemotherapy-induced neuropathy exhibits substantial agent-specific heterogeneity, including subtype-selective neuronal vulnerability and metalloproteinase-dysregulated satellite glial states following paclitaxel treatment, as well as sex-associated fibrotic remodeling following bortezomib treatment. These cellular states may serve adaptive, maladaptive, or degenerative functions. However, because most available studies rely on single-time-point or repeated cross-sectional sampling, they cannot directly establish temporal progression or causal relationships. We further examine how human DRG atlases bridge experimental models and human disease by determining whether candidate molecular targets and cell states are conserved and by identifying their cellular localization. These datasets also highlight species-specific differences in neuronal organization and non-neuronal transcriptional programs that may limit the direct translation of findings from rodent models. Consequently, the primary translational value of DRG single-cell studies lies in elucidating disease mechanisms and prioritizing candidate therapeutic targets for further investigation. Advancing these discoveries toward clinical application will require complementary evidence from human genetics, human DRG transcriptomic datasets, functional validation studies, and clinical pharmacology. Collectively, these complementary approaches may facilitate the development of mechanism-based and cell-type-informed analgesic strategies.
Resistance to therapy is a frequent occurrence in patients with epidermal growth factor receptor (EGFR)-mutant non-small-cell lung cancer (NSCLC) who are treated with EGFR-tyrosine kinase inhibitors (EGFR-TKIs). However, spatial information on how the tumor microenvironment (TME) changes within the same patient from baseline to resistance to first- and then third-generation EGFR-TKIs is scarce. Here, we used rare consecutive re-biopsy samples to build an exploratory, longitudinal, and spatially resolved atlas, aiming to describe within-patient TME remodeling patterns as a resource for hypothesis generation. A patient-matched cohort with serial samples obtained at baseline (T0), after first-generation EGFR-TKI resistance (T1), and after third-generation EGFR- TKI resistance (T2) was established. Using GeoMx Digital Spatial Profiling (DSP), tumor-enriched and stroma-enriched areas of interest (AOIs) were segmented, and paired RNA and protein profiles were quantified. We tracked temporal changes in compartment-specific heterogeneity and immune remodeling and explored associations between early remodeling and subsequent T790M acquisition. We analyzed the data from 15 samples of 6 patients. Tumor- and stroma-enriched compartments were not only transcriptionally but also protein-wise consistently distinct. Spatial heterogeneity was relatively stable from T0 to T1 but increased from T1 to T2, with changes in the stroma-enriched compartments accounting for most of the increase. Early remodeling featured loss of T cell activation programs, reduced neutrophil signatures, increased myeloid remodeling, and impairment of antigen presentation. Longitudinal analyses suggested a biphasic immune trajectory, with early myeloid remodeling and late stromal checkpoint reprogramming. Early spatial remodeling patterns showed potential differences between samples that later acquired T790M and those that did not. This study provides preliminary evidence for dynamic TME changes during sequential EGFR-TKI therapy in EGFR-mutant NSCLC. It suggests a late, stroma-associated expansion of spatial heterogeneity and stage-dependent immune remodeling. Additionally, it offers an initial spatial atlas and generates testable, time-aware hypotheses for future validation.
Colorectal cancer (CRC) exhibits marked cellular heterogeneity, and the cellular context of malignancy-associated epithelial programs remains incompletely defined. We integrated 2,993 CRC samples spanning bulk RNA-seq (n = 2,568; two OS/RFS cohorts), scRNA-seq (281,961 cells/152 specimens), spatial transcriptomics (n = 6), and proteomics (n = 267). Analyses included single-cell integration/annotation, GSVA/HALLMARK, interactome, pseudotime, and ligand–receptor mapping; functional CRISPR assays, EMT immunoblotting, and xenografts; TF profiling (SCENIC/JASPAR/ChIP-qPCR); and exploratory drug-response prediction (OncoPredict), cell-sensitivity assays, and docking/MD modeling. We constructed a stage-stratified single-cell atlas and resolved eleven malignant epithelial subsets, characterizing Epi_4 as late-stage–enriched with EMT, hypoxia, and inflammatory programs and adverse OS/RFS. GPRC5A marked this subset, which we define as GPRC5A+Epi; its expression rose from stage I→IV and was associated with poor outcomes across cohorts, with concordant spatial/proteomic observations. GPRC5A perturbation affected CRC proliferation, migration/invasion, EMT, and xenograft tumorigenicity, supporting a functionally important role in the tested models. SCENIC and ChIP-qPCR supported FOSL1 as an upstream regulator that occupies the GPRC5A promoter. Spatial and ligand–receptor analyses predicted close association and potentially reciprocal signaling between GPRC5A+Epi and POSTN+fibroblasts (COL1A1–SDC4, COL1A1/1A2–ITGA2/ITGB1, PPIA–BSG); concurrent high GPRC5A+Epi/POSTN+Fib signatures were associated with inferior OS/RFS. Drug-response analyses identified an association between GPRC5A status and trametinib sensitivity. Docking/MD produced a computational model of a possible trametinib–GPRC5A interaction, which remains experimentally unvalidated. GPRC5A⁺Epi is a malignancy-associated epithelial state in CRC, and GPRC5A is functionally important for malignant phenotypes in the tested models. Its inferred relationships with POSTN⁺ fibroblasts and the trametinib findings should be regarded as hypothesis-generating pending functional crosstalk, direct-binding, and therapeutic validation.
