Search Results (1,595)

Metastasis, chemoresistance, and tumour recurrence are facilitated by cancer stem cells (CSCs), a small subpopulation of cells capable of regenerating a primary tumour while maintaining the tumour’s genetic and phenotypic features. CSCs can be identified by the expression of specific markers; however, the CSC population in ovarian clear cell carcinoma (OCCC), a rare histotype of ovarian cancer, remains poorly defined. Given the well-established role that CSCs play in cancer progression and metastasis, it is critical to identify reliable markers of CSCs in OCCC. Here, we endeavoured to determine whether ALDH1 expression could be used to define OCCC stem cells in OCCC cell lines using a variety of methods including assessing ALDH1A1 expression in spheroids generated under distinct conditions. We also generated and used chemo-resistant cell lines to assess the enrichment of cancer stem cells. Human OCCC cell lines were enriched for CSCs using selective culture conditions and drug resistance methods. CSC-enriched spheroids demonstrated increased expression of stemness markers NANOG and SOX2, while ALDH1A1 expression was enriched only in drug-resistant cell lines, relative to parental cell lines. RNA-seq analyses of CSC-media-derived spheroids versus standard media spheroids provided novel data supporting CSC enrichment and identified transcription factors induced by CSC media. These findings highlight the ambiguous role of ALDH1A1 as a CSC marker in OCCC and demonstrates the utility of CSC enrichment methods for identifying CSC populations in OCCC cell lines. Full article

Objectives: The aim of this study was to evaluate the reliability, accuracy, and morbidity of dynamic sentinel lymph node biopsy (DSNB) in patients with intermediate- and high-risk penile cancer who had clinically negative lymph nodes, with a focus on long-term follow-up outcomes. Methods: Between January 2005 and January 2024, 287 patients with histologically confirmed penile cancer underwent radioisotope-guided DSNB at the Department of Urology, University of Pécs. Patients lost to follow-up within 24 months were excluded. Of the remaining patients, 211 underwent primary DSNB and 37 secondary DSNB. Thirty-nine patients in the primary DSNB group with Tis, T1a, or G1 tumours were additionally excluded, leaving 209 patients for analysis. Variables included tumour stage and grade, sentinel lymph node (SLN) yield and histology, complications, inguinal progression, false-negative rate, and radical lymphadenectomy findings. Results: A total of 573 SLNs were removed from 209 patients, with a median of 2.7 nodes per patient. SLN metastases were identified in 31 patients, involving 37 groins. Among 178 patients with negative SLNs, three developed inguinal metastases. Sensitivity was 91% per patient and 92.5% per groin, with false-negative rates of 8% and 7.5%, respectively. All false-negative cases occurred within two years, with no later inguinal recurrences. DSNB-related complications occurred in 24 patients (11.4%); most were mild or moderate and managed on an outpatient basis. Two patients required surgery and one required hospital readmission. Conclusions: In a high-volume centre, DSNB is a safe and reliable staging procedure with low morbidity in clinically node-negative intermediate- and high-risk penile cancer. Further studies should assess radiotracer injection sites and compare one-day versus two-day protocols. Full article

Background/Objectives: Isocitrate Dehydrogenase (IDH)1/2-mutant diffuse gliomas represent a biologically distinct subgroup of adult brain tumors in which eary metabolic reprogramming and accumulation of the oncometabolite D-2-hydroxyglutarate (D-2HG) drive epigenetic, immunologic, and clinical characteristics, including a high burden of glioma-associated epilepsy. This review summarizes the molecular and metabolic consequences of IDH mutations, their role in glioma-associated epilepsy, and the evolving impact of mutant IDH-targeted therapies in contemporary neuro-oncology. Methods: We conducted a narrative review of key molecular, translational, imaging, and clinical studies on IDH-mutant diffuse gliomas. The literature included the 2021 (World Health Organization) WHO Classification of Tumours of the Central Nervous System, studies investigating D-2HG biology and glioma-associated epilepsy, and prospective clinical trials and real-world evidence evaluating IDH-targeted therapies and contemporary antiseizure management. Particular emphasis was placed on vorasidenib, advanced metabolic imaging, and emerging liquid biopsy approaches. Results: IDH mutations are early driver events that promote D-2HG accumulation, resulting in widespread epigenetic reprogramming, metabolic dysregulation, and an immunosuppressive tumor microenvironment. D-2HG has also been implicated in the development of glioma-associated epilepsy, although the underlying mechanisms remain incompletely understood. Advances in integrated histomolecular diagnostics, magnetic resonance spectroscopy, amino acid positron emission tomography, and cerebrospinal fluid liquid biopsy have improved disease classification and treatment monitoring. Mutant IDH inhibitors, particularly vorasidenib, prolong progression-free survival, delay the need for subsequent treatment, and reduce intratumoral D-2HG concentrations, and have shown encouraging early signals of improved seizure control and preserved health-related quality of life in patients with grade 2 IDH-mutant gliomas, although this evidence remains preliminary and requires confirmation in larger prospective studies. Conclusions: IDH-mutant diffuse gliomas exemplify precision neuro-oncology, in which a single metabolic alteration informs diagnosis, disease monitoring, and targeted therapeutic approach. Additionally, ongoing studies are expected to further define the role of IDH inhibition across different disease stages and in combination with immunotherapy and standard treatments. Lastly, future clinical trials should systematically incorporate seizure outcomes, neurocognitive function, patient-reported outcomes, and immunologic endpoints to optimize both tumor control and quality of life. Full article

Ovarian cancer remains a leading cause of gynaecological cancer mortality, and ultrasound-based deep learning systems for its diagnosis are typically built as separate post hoc processes for classification, segmentation, and interpretability, which introduces workflow inefficiencies and may produce inconsistent predictions. This work addresses that limitation. We propose UM-TOTA (Unified Multi-Task Ovarian Tumour Architecture), a Vision Transformer (ViT)-based architecture that performs eight-class tumour classification, three-class malignancy detection, tumour segmentation, and clinical concept interpretability within a single unified framework. We integrate a concept bottleneck guided by the IOTA and O-RADS clinical guidelines to enable transparent decision-making through medical concepts that clinicians can understand, and we employ combined adaptive t-vMF Dice and boundary-enhanced segmentation losses with progressive task weighting to stabilise multi-task optimisation. We evaluated the model on the Multi-Modality Ovarian Tumor Ultrasound (MMOTU) 2D dataset under two protocols: image-level 5-fold stratified cross-validation, and the patient-disjoint partition released with the dataset. Under cross-validation, UM-TOTA achieved 80.26% ± 1.10% accuracy (97.06% one-vs-rest macro specificity) for eight-class classification, 90.88% ± 1.14% accuracy (90.41% specificity) for malignancy detection, and 77.29% ± 1.29% Dice for segmentation. Under the patient-disjoint partition, which excludes any overlap of patients between training and testing, the corresponding values were 78.46%, 89.13%, and 75.41%, a reduction of under 2.2 percentage points on every metric. The UM-TOTA reduced the computational parameter load by approximately 65.1% relative to sequential single-task pipelines. The learned concepts aligned with established malignancy criteria, identifying vascularisation, solid components, and papillary projections as key predictors. This unified approach offers an efficient and interpretable framework for clinical ovarian ultrasound workflows. Full article

Lemur tail kinase 3 (LMTK3) is an oncogenic Ser/Thr kinase implicated in breast cancer (BC) progression, therapy resistance, and poor clinical outcomes, yet the molecular mechanisms governing its regulation remain poorly understood, particularly the role of its C-terminal intrinsically disordered region (IDR). Given that IDRs frequently harbour hidden structural motifs that control protein dynamics, we combined computational, biophysical, and biochemical approaches to systematically map regulatory elements within the LMTK3 C-terminus, identifying two regions (residues 688–1095 and 1181–1486) that interact with the LMTK3 kinase domain (LMTK3-KD). Characterisation of these interactions revealed that LMTK3^1181–1486 displays preferential binding to inactive wild-type LMTK3-KD over a constitutively active mutant (LMTK3-KD^L313R), a behaviour consistent with a potential autoinhibitory interaction. Guided by AlphaFold3 modelling, we localised this interaction primarily to a short α-helical motif (α-helix 2; residues 1247–1258) within the C-terminal IDR and subsequently identified Ser1258 within this motif as a candidate regulatory phosphorylation site, using [γ-³²P]-ATP kinase assays and mass spectrometry. Phosphorylation at Ser1258 altered interactions between α-helix 2 and the kinase domain, reducing binding to wild-type LMTK3-KD while increasing affinity for LMTK3-KD^L313R. Functionally, phospho-null mutation of Ser1258 impaired oestrogen receptor alpha (ERα) upregulation, proliferation, migration, and clonogenicity in ER-positive BC cell lines, and reduced tumour growth in female BALB/c nude mice bearing orthotopic MCF7 xenografts. Together, these findings identify a previously uncharacterised regulatory element within the LMTK3 C-terminus and support a model in which Ser1258 phosphorylation modulates kinase domain interactions and LMTK3-driven oncogenic functions in BC. Full article

Artificial intelligence (AI) has emerged as a transformative analytical paradigm for modelling cancer progression using medical imaging and complementary clinical data. This systematic review collates and analyses AI-powered systems and techniques for cancer progression risk analytics and prediction. It provides an empirical examination of algorithmic approaches, imaging-derived feature domains, and clinical metrics that underpin progression prediction. Following the PRISMA 2020 reporting guidance, empirical English-language studies published between 2020 and the final search cutoff date of 15 May 2026 were identified from PubMed, EBSCOhost, Google Scholar and Web of Science. Forty (40) eligible studies spanning diverse cancer types were included, appraised using the MMAT and synthesised narratively. The evidence shows that both classical machine learning models and deep learning architectures are widely used to extract predictive information from radiological data. Radiomic descriptors of intratumoural heterogeneity, tumour morphology, functional imaging biomarkers and multiscale transform-based features consistently demonstrate strong associations with disease progression. Imaging-derived features were linked to clinically meaningful progression endpoints, including progression-free survival, disease-free survival, recurrence and metastasis, while clinical and molecular covariates supported risk stratification. This study further develops the AI-driven cancer progression risk analytics framework (AI CanPRAF), which integrates AI taxonomies, imaging phenotypes, clinical context, progression analytics and decision support into one clinically oriented model. The results demonstrate the growing role of multimodal AI systems in the development of clinically grounded cancer progression analytics and prediction solutions. Full article

A 13-year-old German Riding Pony mare was referred for evaluation of progressive respiratory noise and mixed dyspnoea, which rapidly deteriorated to respiratory distress. Clinical examination revealed marked rattling inspiratory and expiratory tracheal sounds and severe, rattling, vesicular lung sounds. Arterial blood gas analysis demonstrated moderate hypoxaemia (partial pressure of arterial oxygen 76 mmHg at rest). Upper airway endoscopy identified a pink, cauliflower-like pedunculated mass originating from the dorsal tracheal wall approximately 15 cm cranial to the tracheal bifurcation, causing an estimated 60% luminal obstruction. The mass was successfully removed via transendoscopic electrosurgical excision. However, complete tumour-free margins were not achieved. Histopathological examination revealed the diagnosis of leiomyosarcoma. Following surgery, the mare exhibited rapid clinical improvement. Peri-operative treatment included antimicrobial, anti-inflammatory, and bronchodilatory therapy. A follow-up endoscopy one month after surgery showed adequate healing of the surgical site. A small, vascularized tissue structure within the scar tissue, considered suspicious for residual tumour growth, was identified. However, no significant progression was observed during a follow-up examination seven months postoperatively. This case highlights tracheal leiomyosarcoma as a rare differential diagnosis in horses with respiratory distress and describes transendoscopic electrosurgical excision as a feasible therapeutic option. Full article

Background/Objectives: Evaluation of renal tumour masses relies on conventional imaging, which cannot reliably characterise the histopathological type, and no validated blood-based diagnostic marker is available for renal cell carcinoma. The cfDNA integrity index, the ratio of long to short circulating cell-free DNA (cfDNA) fragments, may reflect the necrotic origin of tumour-derived DNA and is a candidate qualitative marker of malignancy. This exploratory, single-centre case–control study assessed the preoperative plasma cfDNA integrity index in patients with clear cell renal cell carcinoma (ccRCC). Methods: Plasma concentrations of 90 bp and 222 bp cfDNA fragments were measured by quantitative real-time PCR (qPCR) in 46 patients with histopathologically confirmed ccRCC (before surgery) and 17 healthy volunteers; the two groups were a convenience sample, were not matched, and differed in age and body-mass index. The cfDNA integrity index was defined as the ratio of the 222 bp to the 90 bp fragment concentration. Results: The cfDNA integrity index was higher in patients with ccRCC than in controls (median 0.28 vs. 0.15; p < 0.001, Mann–Whitney test) and rose progressively across healthy, non-metastatic (M0) and metastatic (M1) groups (p < 0.001, Kruskal–Wallis test; M0 vs. M1 p = 0.004). In unadjusted ROC analysis, both the integrity index (AUC 0.83, 95% CI 0.72–0.94) and its long 222 bp fragment component (AUC 0.93) discriminated patients with ccRCC from healthy volunteers, and the index separated metastatic from non-metastatic disease with an AUC of 0.79 (95% CI 0.64–0.88). The index was higher in high-grade (Fuhrman G3 + G4) tumours (p = 0.04) and in tumours with lymphovascular invasion (p < 0.001), and correlated with primary-tumour diameter. Conclusions: In this exploratory cohort, the preoperative plasma cfDNA integrity index was higher in patients with ccRCC than in healthy volunteers and was associated with several adverse pathological features; these findings require confirmation in larger, matched studies that include benign renal masses before any diagnostic role can be claimed. Full article

Resistance to apoptosis, metabolic plasticity, and redox adaptation are major contributors to cancer progression and treatment failure. Ferroptosis and cuproptosis have therefore emerged as metal-dependent forms of regulated cell-death programs with potential relevance for tumours that survive conventional therapy. Ferroptosis is driven by iron-dependent phospholipid peroxidation when glutathione peroxidase 4 (GPX4)-dependent and parallel antioxidant systems fail, whereas cuproptosis depends on mitochondrial copper engagement of lipoylated tricarboxylic-acid-cycle proteins, lipoylated-protein aggregation, iron–sulfur protein destabilization, and proteotoxic stress. This review integrates the molecular basis, genetic architecture, long non-coding RNA (lncRNA)-mediated regulation, mechanistic crosstalk, and therapeutic implications of ferroptosis and cuproptosis in cancer. It emphasizes a critical evidence hierarchy: expression association, computational signature construction, metal accumulation, reactive oxygen species (ROS) generation, or reduced viability should not be interpreted as pathway dependency without pathway-defining biochemical endpoints and rescue experiments. The most credible translational opportunities will depend on functional stratification, tumour-selective delivery, and pharmacodynamic confirmation that distinguishes pathway-defined ferroptosis or cuproptosis from nonspecific metal-induced and oxidative cytotoxicity. Full article

Rheumatoid arthritis (RA) is a chronic, systemic autoimmune disease defined by persistent synovial inflammation, progressive cartilage and bone erosion, and extra-articular manifestations. Current treatments, such as disease-modifying anti-rheumatic drugs (DMARDs), glucocorticoids, and non-steroidal anti-inflammatory drugs (NSAIDs), control disease activity but are limited by toxicity, reduced response over time, and are expensive, demonstrating the need for safer and more effective adjuncts. The marine environment is a rich, largely untapped source of structurally diverse bioactive molecules that could be used to develop new therapeutics. This review compiles current evidence on anti-inflammatory and immunomodulatory compounds from marine organisms relevant to RA, including macroalgae, true marine microalgae, marine microorganisms (bacteria and fungi, including deep-sea taxa), sea cucumbers, sponges, mussels, corals, and jellyfish. Recurring mechanisms of action include inhibition of the NF-κB, MAPK, and JAK/STAT signalling cascades; suppression of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2); reduced production of tumour necrosis factor-α (TNF-α), interleukin (IL)-1β, and IL-6; and activation of the Nrf2 antioxidant response. Particular attention is given to functional lipids (eicosapentaenoic and docosahexaenoic acids, prostaglandin-like oxylipins) and pigments (astaxanthin, fucoxanthin, β-carotene) from marine microalgae and heterotrophic protists, which recent literature identifies as the most clinically advanced marine leads. To clarify translational status, compounds are grouped by their development stage (marketed nutraceutical, clinical trial, or preclinical) and summarised in a dedicated table. Some compounds, such as green-lipped mussel extract, microalgal omega-3 oils, and astaxanthin, have reached the stage of randomised controlled trials for arthritis. However, most other potential treatments are still in the early, preclinical phase. It is worth noting that ocean-derived compounds appear generally safe, but more thorough studies, especially in living organisms and in clinical settings, are needed to confirm their effectiveness for rheumatoid arthritis before any claims can be made about their therapeutic benefits. Full article

Background and Clinical Significance: Steroid cell tumours of the ovary, not otherwise specified (SCT-NOSs), are rare sex cord–stromal neoplasms with a poorly characterised molecular landscape, in which only exceptional CTNNB1 mutations have so far been reported and no recurrent driver alteration is firmly established. A better characterisation of their molecular spectrum has clinical significance for accurate diagnostic categorisation of ovarian sex cord–stromal tumours and for the identification of potentially targetable pathway alterations in this rare entity. Case Presentation: We report two consecutive SCT-NOSs of the right ovary, retrieved from the archives of the Department of Pathology of the Hôpital Universitaire de Bruxelles and of Curepath. Both underwent comprehensive sex cord–stromal and differential immunohistochemistry and targeted next-generation sequencing on a 168-gene panel with a mean coverage of 2690× (Case 1) and a 17-gene panel (Case 2) (MGI DNBSEQ-T7 for Case 1; Ion GeneStudio S5 for Case 2). A 56-year-old post-menopausal woman (Case 1) and a 50-year-old immunosuppressed woman with a history of renal transplantation and lymphoma (Case 2) both presented with rapidly progressive virilisation. The two right ovarian tumours (20 to 25 mm, no Reinke crystals) displayed an unambiguous sex cord–stromal immunophenotype (α-inhibin, calretinin, SF-1 and Melan-A positive; CD10, WT1, EMA, AE1/AE3 and PAX8 negative), with aberrant nuclear and cytoplasmic β-catenin staining. Sequencing identified a pathogenic CTNNB1 c.133T>C p.(Ser45Pro) variant in Case 1 and a pathogenic CTNNB1 c.95A>T p.(Asp32Val) variant in Case 2, with wild-type FOXL2 in both. Conclusions: Three of the four molecularly characterised CTNNB1-mutated SCT-NOSs converge on the two principal GSK-3β phosphorylation residues of β-catenin, identifying Wnt/β-catenin-pathway dysregulation as a potentially recurrent event and providing additional evidence for the involvement of the Wnt/β-catenin pathway in an emerging molecular subset of SCT-NOS. In a tumour with the canonical sex cord–stromal immunophenotype, an exon 3 CTNNB1 hotspot mutation should not be regarded as evidence against the diagnosis of SCT-NOS and may help define a distinct molecular subset. Full article

Background/Objectives: Glioblastoma (GBM) is widely recognised as a highly aggressive form of malignant tumour arising within the central nervous system. The proneural-to-mesenchymal (PN–MES) state transition is a key process underlying malignant progression and therapeutic resistance. The present study was designed to elucidate the association between the PN–MES transition and arachidonic acid (AA) metabolic reprogramming in GBM. Methods: We integrated four public single-cell RNA-sequencing cohorts, two spatial transcriptomic cohorts, and three bulk RNA-sequencing cohorts for GBM. By combining single-cell transcriptomics, spatial transcriptomics, pseudotime trajectory inference, and cell–cell communication analysis, we systematically evaluated the cell-state dependence and spatial heterogeneity of AA metabolism in GBM. We further incorporated machine learning-based survival modelling, molecular docking, molecular dynamics simulations, and in vitro functional assays to identify and validate potential prognostic biomarkers and therapeutic targets. Results: Mesenchymal-like (MES-like) tumour cells showed the highest transcriptionally inferred AA metabolism-related score, and AA metabolism-related gene expression was closely coupled with PN–MES state transition. The peptidylprolyl isomerase A (PPIA)–basigin (BSG) signalling axis was selectively enriched in tumour cells with high expression of arachidonic acid metabolism-related genes (AAMGs), whereas virtual knockout of BSG perturbed MES- and invasion-related gene modules. Spatial transcriptomic analysis confirmed that PPIA–BSG-associated communication was enhanced within MES-like tumour niches and was coupled with inflammatory and hypoxic programmes. AA metabolism-related genes enabled robust prognostic stratification. Molecular simulation and in vitro experiments suggested that Venetoclax could bind BSG and suppress GBM cell viability. Conclusions: AA metabolism-related transcriptional reprogramming defines mesenchymal-associated malignant niches in GBM and may contribute to PN–MES-related state evolution through PPIA–BSG-mediated microenvironmental communication. AA metabolism-related features and the BSG-associated signalling axis may provide candidate directions for prognostic stratification and targeted intervention. Full article

Background/Objectives: Accurate delineation of brain tumours on contrast-enhanced MRI remains difficult because lesions can be small, irregular, and weakly separated from adjacent tissue. This study developed BG-YOLO11s, a boundary-guided single-stage instance-segmentation model for T1 contrast-enhanced MRI. Methods: The public Figshare/Cheng dataset, comprising 3064 slices from 233 patients, was converted to YOLO polygon annotations and evaluated using a fixed 70/15/15 image-level split (2144/459/461 slices). Because patient identifiers were not retained in the exported image-and-polygon data, the split was not guaranteed to be patient-disjoint. Bézier Contour Augmentation generated two contour-perturbed training samples per original slice while leaving validation and test data unchanged. BG-YOLO11s extended YOLO11s-seg with dilated context aggregation in the backbone, boundary-enhanced feature fusion in the neck, and a prototype refinement module with differentiable boundary-aware supervision in the segmentation head. Results: In a single run on the held-out image-level test split, BG-YOLO11s achieved 92.4% precision, 88.7% recall, 94.6% mask mAP@50, 68.9% mAP@50–95, and 86.5% IoU. Relative to YOLO11s-seg, the corresponding gains were 3.8 points in mAP@50, 5.8 points in mAP@50–95, and 4.1 points in IoU. A progressive ablation produced incremental gains along the fixed module-addition sequence, but it did not isolate all component interactions or quantify run-to-run uncertainty. Conclusions: BG-YOLO11s improved single-run mask-overlap estimates under the present image-level benchmark. Patient-disjoint retraining, repeated-seed statistics, boundary-specific metrics, complete failure pattern auditing, and external multi-sequence validation are required before broader clinical or deployment claims can be made. Full article

Breast cancer (BC) progression is strongly influenced by the extracellular matrix (ECM), whose remodelling regulates tumour growth, invasion, metastasis, immune modulation, and therapeutic response. Among ECM components, collagens have emerged as both structural proteins and active mediators of mechanotransduction, stromal interactions, and tumour cell behaviour. This narrative review analyses collagen families and collagen-associated proteins implicated in BC, integrating evidence on their expression patterns, biological functions, clinical significance, and translational potential. We examine fibrillar and non-fibrillar collagens, including fibril-associated collagens with interrupted triple helices (FACITs), membrane-associated collagens with interrupted triple helices (MACITs), basement membrane (BM) collagens, and multiplexins, together with their interactions with cancer-associated fibroblasts (CAFs), immune cells, and signalling pathways involved in tumour progression. Alterations in collagen composition, organization, crosslinking, and degradation regulate ECM stiffness, epithelial–mesenchymal transition (EMT), invasion, metastatic dissemination, and therapy resistance. Several collagen types and collagen-derived fragments also show promise as prognostic biomarkers and therapeutic targets, particularly in aggressive BC subtypes such as human epidermal growth factor receptor 2 (HER2)-positive and triple-negative breast cancer (TNBC). Overall, this review highlights the collagenome as a dynamic component of the breast tumour microenvironment (TME) and supports collagen-informed strategies for improved patient stratification and targeted therapies. Full article

Background/Objectives: PTEN Hamartoma Tumour Syndrome (PHTS) is a rare inherited disorder caused by germline PTEN mutations, presenting with cancer predisposition and neurodevelopmental abnormalities, including macrocephaly. PTEN functions as a tumour suppressor by regulating the PI3K/AKT/mTOR pathway and contributes to cellular adhesion, migration, and genomic stability. As no curative therapy exists, gene therapy represents a promising avenue to correct the underlying genetic defect. Methods: In this study we evaluated the therapeutic potential of AAV9-mediated PTEN gene delivery in a Pten^Δ5/+ mouse model, which is predominantly characterized by progressive lymphoid hyperplasia leading to lymph node tumour development, as well as hepatic focal lesions and macrocephaly. Results: Systemic vector delivery did not improve lymph node enlargement which represents the predominant disease phenotype in the Pten^Δ5/+ mice at the doses tested, due to restricted vector biodistribution, but it did effectively rescue the hepatic lesions, including steatosis and steatohepatitis-like changes. Mechanistically, AAV-mediated PTEN expression significantly reduced elevated p-AKT levels, indicating attenuated hyperactivation of the PI3K/AKT/mTOR pathway. Additionally, intracranial delivery of the same vector in Pten^Δ5/+ mouse neonates ameliorated macrocephaly without apparent adverse effects. Conclusions: These findings highlight the therapeutic potential of AAV9-mediated PTEN gene therapy in PHTS, particularly for hepatic and neuroanatomical manifestations. Optimization of vector biodistribution and transduction efficiency will be critical for clinical translation. Full article