Antibody–drug conjugates (ADCs) represent a transformative therapeutic class in non-small cell lung cancer (NSCLC), moving precision oncology beyond traditional targeted therapies. These agents, composed of a monoclonal antibody, linker, and potent cytotoxic payload, enable targeted drug delivery to tumor cells expressing specific antigens like TROP2, HER2, and c-MET. Recent clinical trials have demonstrated remarkable efficacy of ADCs, both as monotherapy in molecularly defined populations and in combination with immunotherapy, offering new hope for patients with advanced disease. This review summarizes the latest clinical progress of ADCs in NSCLC, highlighting key agents and their impact, while also addressing the associated challenges and future directions for optimizing this promising treatment modality.

Renal cell carcinoma (RCC) is a challenging urologic malignancy characterized by its aggressive nature, including invasion, metastasis, and treatment resistance. To explore multi-targeted therapies, we established an advanced clear cell renal cell carcinoma (ccRCC) model via orthotopic tumor transplantation in mice, and established another model simulating post-surgical recurrence by performing radical nephrectomy. We engineered a genetic circuit to reprogram the host liver as a bioreactor, enabling the production and delivery of in vivo self-assembled siRNAs (IVSA-siRNAs) for co-targeting VEGFR2 and mTOR. The efficacy and toxicity of this IVSA-siRNA system were evaluated and compared with the combination therapy of sunitinib and everolimus. In the established models, the combination therapy of sunitinib and everolimus showed efficacy but induced severe adverse effects. In contrast, IVSA-siRNAs potently silenced VEGFR2 and mTOR expression, achieving therapeutic effects in both advanced and radical nephrectomy ccRCC models without discernible toxicity.

Serine hydroxymethyltransferase 2 (SHMT2) is a key mitochondrial enzyme involved in one-carbon metabolism, but its regulation and functional significance in acute kidney injury (AKI) remain unclear. To determine whether SHMT2 downregulation contributes to tubular injury and to elucidate the underlying molecular mechanism, we established murine models of AKI induced by ischemia-reperfusion injury and unilateral ureteral obstruction. In parallel, human proximal tubular HK-2 cells were exposed to hypoxia/reoxygenation or transforming growth factor-β1 stimulation in vitro. We found that SHMT2 expression was markedly reduced in renal tubular epithelial cells across both murine AKI models, showing a significant inverse association with the severity of tubular injury. To elucidate the underlying mechanisms, we subsequently utilized genetic (shRNA) and pharmacological (SHIN1) inhibition of SHMT2, alongside overexpression of the E3 ubiquitin ligase synoviolin (SYVN1). Functional inhibition of SHMT2 aggravated tubular epithelial damage by inducing mitochondrial dysfunction, increasing oxidative stress, promoting the accumulation of nephrotoxic uremic toxins, and impairing both glycolysis and oxidative phosphorylation. Mechanistically, under AKI conditions, elevated SYVN1 directly interacted with SHMT2 via its conserved RING domain, promoting K48-linked polyubiquitination and subsequent proteasomal degradation of SHMT2. Ultimately, this ubiquitin-dependent degradation of SHMT2, mediated by SYVN1, drives AKI progression by inducing metabolic reprogramming and bioenergetic failure in renal tubular cells. These findings highlight the SYVN1/SHMT2 axis as a novel pathogenic mechanism and a promising therapeutic target for preserving tubular metabolic homeostasis and alleviating kidney injury.

Oral squamous cell carcinoma (OSCC), a highly prevalent and poor-prognosis malignancy, is closely associated with tumor metabolic reprogramming, particularly the glutamine-dependent metabolic phenotype. This study systematically investigates the role of N6-methyladenosine (m6A) modification in OSCC through integrated bioinformatics analysis and functional experiments, focusing on the tumor-suppressive function of the m6A reader YTHDC2 and its regulation of glutaminolysis. Analysis based on The Cancer Genome Atlas (TCGA) datasets revealed that YTHDC2 expression was significantly inversely correlated with OSCC malignancy and patient survival. Functional validation showed that YTHDC2 depletion promoted OSCC cell proliferation and stem-like properties, whereas YTHDC2 overexpression markedly suppressed these malignant phenotypes. Mechanistic studies demonstrated that YTHDC2 stabilized VHL mRNA by recognizing m6A modification sites, enhancing VHL protein expression. This promoted VHL-mediated ubiquitin-dependent degradation of HIF-1α, leading to transcriptional repression of its downstream target GLS1. Consequently, this blocked glutaminolysis, tricarboxylic acid (TCA) cycle-driven energy production, and glutathione (GSH)-mediated antioxidant pathways. Additionally, low YTHDC2 expression in OSCC tissues was closely associated with DNA hypermethylation at CpG islands in its promoter, an epigenetic silencing mechanism that sustains the glutamine-addicted phenotype. This study first uncovers the core role of the YTHDC2/m6A/VHL/HIF-1α/GLS1 signaling axis in metabolic regulation of OSCC, providing new insights into the molecular basis of glutamine addiction. YTHDC2 not only serves as a prognostic biomarker for OSCC but also highlights its-mediated metabolic pathway as a theoretical basis for developing targeted therapies against glutaminolysis.

Employing neutrophils (NEs) as endogenous moving gears offers a promising strategy for crossing the blood-brain barrier (BBB) in the treatment of central nervous system (CNS) malignancies such as glioblastoma (GBM). However, in vivo hitchhiking efficiency of NEs is severely limited by spontaneous formation of a nonspecific protein corona. Here, we present pathogen-mimetic liposomes (PM-Lipo) that enable efficient and precise NEs hitchhiking by programming the in vivo protein corona. PM-Lipo is rationally engineered to preferentially enrich complement 3b (C3b) and inactivated C3b (iC3b), thereby recapitulating endogenous complement opsonization and promoting complement receptor 3 (CR3)-mediated recognition by activated NEs. This effect arises from the synergistic integration of two functional lipids that activate distinct complement pathways, triggering a proteolytic cascade with positive feedback to amplify C3b/iC3b deposition. Our results demonstrate that PM-Lipo achieved >85% targeting efficiency toward activated NEs, enabling hitchhiking-mediated transport across the BBB, with 8.94% of the administered PM-Lipo successfully penetrating the brain, leading to marked tumor suppression and more than a twofold extension in median survival. Our research establishes a protein corona programming strategy to harness endogenous immune cells for targeted drug delivery, offering a broadly applicable paradigm for precision nanomedicine in CNS malignancies.

Postnatal depression (PND) increases the risk of neurodevelopmental impairments in offspring, yet the underlying mechanisms remain elusive. Here, we identify an intergenerational signaling pathway through which maternal psychological state is encoded in breastmilk microRNAs (miRNAs) and transmitted to offspring via breastmilk small extracellular vesicles (sEVs). Using cross-fostering models, we demonstrate that maternal depression disrupts the maturation of newborn neurons in the offspring hippocampal dentate gyrus (DG), leading to cognitive deficits and depressive-like behaviors. Strikingly, gastric administration of PND-derived breastmilk sEVs to healthy pups recapitulates these neurodevelopmental phenotypes. Mechanistically, PND-induced miRNAs are packaged into breastmilk sEVs and delivered to infant brain, where they suppress mGluR8 expression in supramammillary nucleus (SuM) neurons projecting to the DG. This presynaptic mGluR8 deficiency dysregulates glutamate release, triggers aberrant synaptic transmission and calcium overload, and ultimately drives a pathological “high-frequency, low amplitude” neuronal activation pattern that compromises the functional integration of newborn neurons into hippocampal circuits. Critically, inhibiting sEV secretion from the mammary gland or sequestering the culprit miRNAs within offspring SuM neurons rescues these neurodevelopmental deficits. Together, these findings establish breastmilk sEV miRNAs as an epigenetic vector that translates maternal psychological state into maladaptive programming of offspring brain circuitry and behavior.

Objective This study investigated the roles of CREB and NMNATs in sevoflurane-induced cognitive deficits and synaptic alterations in developing hippocampal neurons. Methods 160 postnatal day-7 male SD rats were randomized into four groups: Control; Sevo (3% sevoflurane for 6 h); Sevo+D-cycloserine (DCS, an NMDA receptor agonist); and AP-5 (an NMDA receptor antagonist). Cognitive function was assessed at 8 weeks using the Morris water maze. Dendritic spine density/morphology in hippocampal CA1 was analyzed via Golgi-Cox Staining. Protein levels of NMNAT1, NMNAT2, CREB, and p-CREB were measured by western blot and immunofluorescence. Results Compared to controls, sevo-exposed rats exhibited significant spatial memory impairment, demonstrated by increased escape latency, longer path length, and fewer platform crossings. This group also showed reduced dendritic spine density and altered morphology in CA1, alongside decreased p-CREB and NMNAT1/2 expression. In contrast, DCS pretreatment before sevo exposure reversed these deficits, restoring cognitive performance, spine density/morphology, and p-CREB/NMNAT1/2 levels. Conclusion Sevoflurane induces learning/memory deficits and dendritic spine pathology in neonatal rats, likely via NMDA receptor-mediated downregulation of p-CREB and NMNAT1/2. DCS pretreatment effectively mitigates these effects, highlighting that enhancing NMDA receptor activity can counteract sevoflurane-induced neurotoxicity. NMDA receptor modulation represents a promising therapeutic strategy for postoperative cognitive dysfunction.

Postnatal depression (PND) increases the risk of neurodevelopmental impairments in offspring, yet the underlying mechanisms remain elusive. Here, we identify an intergenerational signaling pathway whereby maternal psychological state is encoded in breastmilk microRNAs (miRNAs) and transmitted to offspring via breastmilk small extracellular vesicles (sEVs). Using cross-fostering models, we demonstrate that maternal depression disrupts offspring hippocampal dentate gyrus (DG) newborn neuron maturation, causing cognitive deficits and depressive-like behaviors. Strikingly, gastric administration of PND-derived breastmilk sEVs to healthy pups recapitulates these neurodevelopmental phenotypes. Mechanistically, PND-induced miRNAs are packaged into breastmilk sEVs and delivered to infant brain, where they suppress mGluR8 expression in supramammillary nucleus (SuM) neurons projecting to the DG. This presynaptic mGluR8 deficiency dysregulates glutamate release, triggers aberrant synaptic transmission and calcium overload, and ultimately drives a pathological “high-frequency, low amplitude” neuronal activation pattern that compromises the functional integration of newborn neurons into hippocampal circuits. Critically, inhibiting sEV secretion from the mammary gland or sequestering the culprit miRNAs within offspring SuM neurons rescues these neurodevelopmental deficits. These findings not only uncover a novel mechanism underlying PND-related offspring neurodevelopmental impairments, but also provide a promising therapeutic target for clinical intervention of intergenerational transmission of maternal mental disorders.

Maternal obesity is linked to heightened metabolic disease risk in offspring, but the mediators of this intergenerational effect remain unclear. Using a diet-induced obesity (DIO) mouse model, we showed that maternal circulating small extracellular vesicles (sEVs) crossed the placenta and delivered obesity-associated miRNAs to the fetal liver, with lasting consequences for insulin sensitivity in male offspring. Among these miRNAs, miR-29a-3p was pathologically elevated and targeted both DNA methyltransferases and demethylases, thereby reshaping the DNA methylation landscape. This included hypomethylation of the Pgc-1α locus, a key regulator of gluconeogenesis, which resulted in premature activation of hepatic gluconeogenesis that contributed to the persistent metabolic dysfunction in adulthood in male offspring. These findings identify a transplacental sEV-miRNA-epigenetic axis that perturbs fetal metabolic programming and may represent a conserved mechanism underlying the developmental origins of metabolic disease. The study identifies a transplacental sEV–miRNA–epigenetic axis as a mediator of maternal obesity. Maternal plasma sEVs transfer miR-29a-3p to the fetal liver, epigenetically reprogramming glucose metabolism and driving adult insulin resistance.

Colorectal cancer (CRC) is one of the leading causes of cancer-related mortality worldwide. Ferroptosis, an iron-dependent form of programmed cell death, has emerged as a potential therapeutic target. However, the regulatory mechanisms that allow CRC cells to evade ferroptosis are not fully understood. This study focuses on OTUB2, a deubiquitinating enzyme, and its role in stabilizing U2AF2, which allows CRC cells to resist ferroptosis and autophagy. We analyzed CRC cells and clinical samples to evaluate the effects of OTUB2 on U2AF2 deubiquitination. OTUB2 knockdown and overexpression models were established in CRC cell lines (LoVo, RKO, SW480, HT115) to assess ferroptosis and autophagy activity. Various assays, including western blotting, immunoprecipitation, colony formation, and transwell migration assays, were used to evaluate cell proliferation, migration, and iron metabolism markers. In vivo xenograft models were also employed to assess tumor growth under OTUB2-U2AF2 axis disruption. OTUB2 was highly expressed in CRC tissues compared to normal controls. Knockdown of OTUB2 significantly increased ferroptosis, while enhancing autophagy. Conversely, OTUB2 overexpression reduced ferroptosis and autophagy, maintaining CRC cell survival and proliferation. In vivo studies confirmed that disrupting the OTUB2-U2AF2 axis impaired tumor growth by activating both ferroptosis and autophagy. Importantly, a reciprocal activation relationship between ferroptosis and autophagy was observed under OTUB2-U2AF2 axis deficiency. OTUB2 stabilizes U2AF2 in CRC cells, enabling them to evade ferroptosis and autophagy. Disruption of the OTUB2-U2AF2 axis activates both processes, suppressing tumor growth. Targeting this axis presents a promising therapeutic strategy for CRC treatment.

Drought stress severely limits barley productivity, yet the regulatory roles of long non-coding RNAs (lncRNAs) remain unrevealed. This study aimed to characterize a drought-responsive lncRNA, designated lncDR1, and elucidate its functional mechanism in drought tolerance. Through bioinformatic analysis and biological experiments, we investigated the expression pattern, structural variation, and regulatory mechanism of lncDR1. lncDR1 was specifically induced under drought stress in tolerant barley genotypes and predominantly expressed in leaves. Sequence comparison between tolerant (EC_S1) and sensitive (EC_N1) accessions revealed 23 SNPs and two insertions, with 16 variations clustered in the 33-61 nt region, leading to distinct secondary structures. Overexpression of lncDR1 in both tobacco and barley significantly enhanced drought tolerance, as evidenced by higher fresh weight, root length, Fv/Fm, and survival rate, alongside lower MDA content and reduced water loss. Mechanistically, lncDR1 acted as a competing endogenous RNA (ceRNA) by sponging miR4339, thereby derepressing HvGTG2, a positive regulator of ABA-mediated stomatal closure. Consequently, transgenic lines exhibited enhanced stomatal closure and reduced stomatal conductance under drought stress. This study uncovers a novel lncDR1-miR4339-HvGTG2 regulatory axis that improves drought tolerance by promoting stomatal closure, providing a valuable genetic resource for breeding.

Background:Intratumor heterogeneity (ITH), a critical driver of tumor evolution and immune evasion, remains inadequately characterized at the transcriptomic level in colorectal cancer (CRC), and its clinical implications are not yet fully understood. Methods:We integrated transcriptomic datasets from TCGA-COAD/READ and two independent GEO cohorts (GSE40967 and GSE87211) to develop an RNA-seq-based ITH score using the DEPTH2 algorithm and to con0struct an ITH-related gene (ITRG) prognostic model. A unified cutoff value of 0.64 was established to stratify patients into high- and low-ITH groups. Using 52 survival-associated ITRGs, we generated a nine-gene prognostic signature selected from 101 distinct combinations of feature selection techniques and modeling algorithms and validated its performance in two external datasets. The tumor microenvironment and potential responsiveness to immune checkpoint inhibitors were evaluated using ssGSEA, ESTIMATE, and TIDE algorithms. SHAP analysis, together with in vitro and in vivo experiments, was employed to identify and functionally validate key regulatory genes. Results:Patients in the high-ITH group had markedly poorer overall survival (OS) than those in the low-ITH group. The ITH score correlated strongly with aggressive clinical features, including T3/4 invasion depth, N1/2 nodal status, and AJCC stage III. The nine-gene prognostic signature demonstrated consistent predictive capability across the TCGA training cohort and both GEO validation cohorts. In TCGA, the model yielded time-dependent AUCs of 0.669, 0.664, and 0.645 for 1-, 3-, and 5-year OS, respectively, and retained its status as an independent prognostic indicator in multivariate Cox regression analysis; in GSE40967 and GSE87211, the corresponding AUCs ranged from 0.544-0.573 and 0.648-0.744. The high-risk subgroup was characterized by a stromal immune phenotype enriched with cancer-associated fibroblasts and macrophages, elevated expression of multiple immune checkpoints, increased TIDE scores, and higher tumor mutational burden. SHAP analysis identified IL20RB as the top risk-associated gene, whose knockdown significantly suppressed CRC cell proliferation, migration, invasion, and tumorigenicity in vitro and in vivo. Conclusion:This study introduces and validates a transcriptomic ITH score and a nine-gene ITRG-based prognostic model that delineate the immune landscape and enables effective survival stratification in CRC, complementing the limitations of current staging systems. Additionally, IL20RB is highlighted as a promising therapeutic target, supporting the development of personalized immuno-targeted combination therapies in CRC.

Severe fever with thrombocytopenia syndrome (SFTS) is a highly fatal infectious disease caused by the SFTS virus (SFTSV). Reliable prognostic biomarkers are essential for early intervention, yet specific markers for SFTSV infection remain unidentified. In this study, we identified SFTSV-encoded microRNA-like small RNAs (milRNAs) in patients' sera and evaluated their potential as prognosis biomarker. A multi-phase study involving 170 laboratory-confirmed SFTS patients, 40 patients with other infection and 80 healthy controls was conducted. Small RNA deep sequencing was performed, followed by quantitative reverse transcription polymerase chain reaction (qRT-PCR) and TA cloning for individual validation. The expression dynamics of identified milRNAs were analyzed across different disease stages, and their prognostic potential was assessed using Kaplan-Meier survival analysis and Cox proportional hazards regression. Three SFTSV-encoded milRNAs - SFTSV-S-1480 (S-1480), SFTSV-M-692 (M-692), and SFTSV-L-4706 (L-4706) - were significantly elevated in the sera of severe patients but were nearly undetectable in mild cases and healthy controls. Their expression levels increased notably during the multiple organ dysfunction (MOD) stage, correlating with disease progression. Patients with higher milRNA expression had significantly shorter survival compared to those with lower expression. Receiver operating characteristic (ROC) curve analysis demonstrated that the three-milRNA panel outperformed traditional blood immune cell indicators in predicting disease severity. Our study identifies a panel of three SFTSV-encoded milRNAs as novel prognostic biomarkers for SFTS. Their strong correlation with disease progression and clinical outcomes suggests their potential utility for early risk stratification and targeted intervention.

Achieving precise antitumour drug delivery to tumour sites and selectively inhibiting oncogene function remain core challenges in cancer treatment. While small interfering RNAs (siRNAs) are a powerful means of achieving these goals, their clinical application is limited by delivery barriers, particularly in extrahepatic tissues. Based on the in vivo self-assembled (IVSA) siRNA delivery system, we developed a targeted therapeutic approach for EGFR-positive tumours in this study. We designed an IVSA genetic circuit that can reprogram the liver to produce and self-assemble EGFR siRNAs into small extracellular vesicles (sEVs) tagged with an EGFR-targeting peptide (GE11). The siRNA-encapsulating sEVs can be transported via the blood circulation and guided to EGFR-positive tumour cells by a targeting peptide for tumour therapy. In EGFR-driven NSCLC models, the IVSA siRNA dramatically reduced tumour size and suppressed EGFR expression more effectively than traditional treatments such as gefitinib or osimertinib. We evaluated the efficacy of this system in orthotopic gastric and breast cancer models to further show its therapeutic value for other EGFR-positive tumours. In these models, the IVSA siRNA resulted in significant tumour suppression and enhanced survival outcomes. These findings underscore the versatility and potency of the IVSA platform as a universal therapeutic approach for EGFR-targeted siRNA delivery, providing a promising new avenue for treating a range of EGFR-positive cancers.

KMT2C (lysine methyltransferase 2C), also known as mixed-lineage leukemia 3 (MLL3), is a member of the KMT2 family of histone methyltransferases that catalyzes histone H3 lysine 4 monomethylation (H3K4me1), a hallmark of active enhancer elements. Operating within COMPASS-like complexes (Complex of Proteins Associated with Set1) and in association with the ASCOM coactivator complex (ASC-2–containing complex), KMT2C plays a central role in maintaining enhancer and super-enhancer integrity, thereby sustaining lineage-specific transcriptional programs. Across gastrointestinal malignancies, KMT2C is recurrently altered, predominantly through truncating loss-of-function variants, splice-disrupting events, and structural alterations that impair protein function. Importantly, the biological impact of KMT2C alteration is highly context dependent, shaped by mutation class, co-occurring genomic lesions, and tissue-specific transcriptional circuitry. Rather than inducing linear dysregulation of individual signaling pathways, KMT2C deficiency preferentially destabilizes enhancer and super-enhancer networks, leading to large-scale transcriptional rewiring. Disruption of enhancer modules that enforce cellular identity and homeostasis is frequently accompanied by activation of stress-adaptive and metabolic programs. Concurrently, defects in homologous recombination and replication-stress responses promote genomic instability, while attenuation of cell-cycle checkpoints and senescence barriers facilitates epithelial–mesenchymal transition, stem-like plasticity, and invasive or metastatic behavior. Beyond tumor-intrinsic effects, KMT2C dysfunction can reshape the tumor immune microenvironment through altered antigenic burden, inflammatory signaling, senescence-associated secretory programs, and dynamic stromal interactions, ultimately giving rise to heterogeneous therapeutic vulnerabilities. Clinically, KMT2C alteration has been linked to tumor mutational burden (TMB), microsatellite instability (MSI), immune infiltration patterns, and outcomes following immune checkpoint blockade (ICB). In parallel, KMT2C-associated DNA repair deficiencies provide a mechanistic basis for synthetic-lethal strategies involving poly(ADP-ribose) polymerase (PARP) inhibitors and inhibitors of ataxia telangiectasia and Rad3-related protein (ATR) or checkpoint kinase 1 (CHK1), including rational combinations with epigenetic therapies. In this review, we integrate evidence from hepatocellular carcinoma, pancreatic ductal adenocarcinoma, cholangiocarcinoma, colorectal cancer, gastric cancer, esophageal cancer, and gallbladder cancer within a unified framework that links KMT2C domain architecture to enhancer-network destabilization, phenotypic state transitions, and clinical manifestations. We further propose a functional evaluation paradigm that reframes discrete KMT2C variants as graded states of epigenetic deficiency, coupled with a closed-loop validation strategy integrating tissue-based profiling, liquid biopsy monitoring, and spatial multi-omics analyses.

Osteosarcoma is the most common primary malignant bone tumor in children and adolescents. The purpose of this study is to explore the regulatory mechanism of LINC01116 in osteosarcoma metastasis and its potential association with the urea cycle and chemoresistance. Transcriptomic profiling was performed, and the results were validated by qRT-PCR in 6 paired osteosarcoma and adjacent non-tumor tissues. Functional assays including in vivo xenograft models were used to verify the effect of LINC01116 on osteosarcoma cell proliferation. RNA-protein interaction studies and ChIRP-MS assays were conducted to confirm the binding between LINC01116 and CPS1.LINC01116 was significantly upregulated in metastatic osteosarcoma lesions. Silencing LINC01116 inhibited osteosarcoma cell proliferation both in vitro and in vivo. Mechanistically, LINC01116 directly binds to CPS1 -the rate-limiting enzyme of the urea cycle, which was confirmed by ChIRP-MS. Perturbing the LINC01116/CPS1 axis reduced citrulline levels, indicating impaired urea cycle function. Additionally, CPS1 silencing enhanced osteosarcoma cell sensitivity to cisplatin. This study identifies a novel LINC01116/CPS1 axis that drives osteosarcoma metastasis by regulating the urea cycle. Targeting this axis can sensitize osteosarcoma to cisplatin treatment, which highlights its potential as a therapeutic target for osteosarcoma.

Abnormal accumulation of TAR DNA-binding protein-43 (TDP-43) is a hallmark of amyotrophic lateral sclerosis and frontotemporal lobar degeneration. Small interfering RNAs (siRNAs) targeting TDP-43 offer potential therapeutic strategies for these diseases. However, efficient and safe delivery of siRNAs to the CNS remains a challenge. Here, we present a synthetic biology-based approach that leverages endogenous small RNA processing machinery to self-assemble siRNA-encapsulating small extracellular vesicles and uses the natural circulatory system of the host to transport siRNAs. Specifically, we engineered liver cells to express and package TDP-43-targeting siRNAs into rabies virus glycoprotein-tagged small extracellular vesicles, which are released into the circulation and cross the blood-brain barrier to deliver siRNAs to the CNS. In a mouse model of TDP-43 pathology induced by stereotactic injection of mutant TDP-43 (M337V) virus, treatment with in vivo self-assembled TDP-43 siRNAs (IVSA-siR-TDP43) effectively reduced TDP-43 accumulation, leading to significant improvements in motor function and neuropathology. Additionally, an adeno-associated virus-based delivery system was used to produce IVSA-siR-TDP43, demonstrating sustained therapeutic effects in TDP-43-associated neurodegeneration. These findings highlight a novel, effective and minimally invasive gene therapy platform for addressing TDP-43 pathology in amyotrophic lateral sclerosis and frontotemporal lobar degeneration, offering a promising avenue for future clinical applications.