Citations

Abstract Background Genetic testing is increasingly used in presurgical evaluation, but the yield of resection across germline genetic epilepsies remains uncertain. Methods We conducted a systematic review of MEDLINE (PubMed) and Scopus and added cases from three institutional cohorts and the Pediatric Epilepsy Research Consortium (PERC) databases, including individuals with pathogenic/likely pathogenic germline variants besides tuberous sclerosis and neurofibromatosis who underwent resection or laser ablation. Etiologies were grouped into biologically informed categories (GATORopathies, vascular, overgrowth, CNVs, channelopathies, synaptopathies, other). Primary outcome was seizure freedom (Engel I) at last follow-up. Group comparisons used Fisher’s exact and Kruskal–Wallis tests (α=0.05; Bonferroni when applicable). Prespecified sensitivity analyses stratified by lesional status, excluded GATORopathies, and restricted to literature-only cases. Results We included 223 literature cases (64 studies), 35 institutional cases, and 11 PERC cases (n=269). Median follow-up was 24 months (IQR 12–48.0). Seizure freedom was achieved in: vascular disorders 14/19 (73.6%), GATORopathies 79/120 (67.5%), CNVs 18/31 (66.7%), overgrowth 7/13 (53.8%), other 16/25 (41.7%), channelopathies 13/43 (33.3%), and synaptopathies 4/18 (22.2%) (overall p<0.001). Among cases with known imaging, 208/253 (82.2%) had epileptogenic MRI lesions, including 64% of channelopathies and 80% of the synaptopathies. In univariate contrasts (each category vs all others), odds of seizure freedom were higher for vascular disorders (2.66-fold, 95% CI 0.87–9.72) and GATORopathies (2.46-fold, 95% CI 1.46–4.18), and lower for synaptopathies (∼4.2-fold lower, OR 0.24, 95% CI 0.05– 0.78) and channelopathies (∼4.5-fold lower, OR 0.22, 95% CI 0.09–0.48). Direction and magnitude were consistent across prespecified sensitivity analyses (lesional-only, literature-only, exclusion of GATORopathies). Conclusions Resective surgeries can be effective in germline genetic epilepsies, but outcomes vary by pathway. Disorders with discrete, lesional substrates (GATORopathies, vascular) show the highest likelihood of seizure freedom, whereas channelopathies and synaptopathies, despite the presence of MRI lesions, have substantially lower yields even in lesional cases. Prospective, genotype-aware surgical registries with standardized reporting (EEG, imaging, pathology) and time-to-event outcomes are needed to refine the selection of surgical candidates and quantify seizure and non-seizure-related outcomes.

The intensive care unit (ICU) is a highly disease-focused environment where patients are often perceived primarily through their diagnoses rather than as individuals. The Get-To-Know-Me Board (GTKMB) was introduced as a humanizing tool to highlight patients’ personal attributes and foster person-centered care. Despite being found valuable in supporting humanized caring and fostering communication, completion rates at our institution remained low. Through a stakeholder-driven quality improvement initiative, we tested sequential interventions including reminders and checklists, visual magnets, and educational pamphlets to improve and increase the use of the GTKMB. These measures produced modest gains, with reminders and checklists yielding the most notable improvement. Barriers to using the GKTMB included unclear ownership, timing, and inconsistent integration into workflow. Our experience underscores both the promise and limitations of humanizing interventions in critical care. Sustainable implementation requires multidisciplinary buy-in and integration into daily routines. Whereas humanizing tools such as the GTKMB should be evaluated for their impact on meaningful patient outcomes, such outcomes are not clearly established; therefore, we embark on the necessary first step of a method of implementation.

Abstract Neuroblastoma (NB) accounts for approximately 10% of pediatric cancer-related deaths. High-risk NB is often metastatic at diagnosis with a poor 5-year survival rate (&lt;50%) despite intensive multimodal therapy. GD2, a disialoganglioside glycolipid expressed in nearly all NBs, is a well-validated immunotherapy target. Although GD2-CAR-T cells have been clinically evaluated, their efficacy remains limited due to poor tumor infiltration and the immunosuppressive tumor microenvironment (TME), underscoring the need for alternative immune-based approaches. Given the inherent capacity of macrophages to infiltrate solid tumors, phagocytose cancer cells, and modulate the TME, we engineered the GD2-CAR macrophage (GD2-CAR-M) as a novel therapeutic platform for NB. GD2-specific CAR constructs containing the Hu3F8 single-chain variable fragment (scFv) were designed, validated, and introduced into multiple human and murine monocyte/macrophage cell lines via lentiviral transduction. Fluorescence microscopy and flow cytometry demonstrated a significant enhancement in the phagocytosis of GD2-high NB cells and the phagocytic efficiency correlating positively with GD2 expression levels, suggesting antigen-specific phagocytosis by GD2-CAR-M. Our further in vivo studies using an immunocompetent MYCN-driven NB non-genetically engineered mouse model (MYCN-NGEMM) showed that treatment with GD2-CAR-M markedly reduced tumor burden and prolonged survival compared with controls. Notably, mice bearing GD2-high-tumors exhibited significantly improved survival, including one complete remission among ten treated animals. Histological analyses revealed increased connective tissue deposition and decreased tumor cell density, suggesting TME remodeling. Single-cell RNA sequencing and immunohistochemistry further demonstrated enhanced infiltration of macrophages, neutrophils, and T cells, indicating a shift from a “cold” to a “hot” immune TME following GD2-CAR-M treatment. Consistent antitumor effects were also observed in our dbh-MYCN transgenic zebrafish with spontaneous development of NB following GD2-CAR-M administration. Taken together, GD2-CAR-M exhibits potent antigen-specific antitumor activity and promotes favorable TME remodeling in NB models. These findings highlight GD2-CAR-M as a promising translational immunotherapy strategy for high-risk NB. Citation Format: Shizhen (Jane) Zhu, Ke-En Tan, Kok Siong Yeo, Cheng Zhang, Caleb T. Baker, Alexis M. Sosic, Eddie Lim . Yat Yuen, Choong Yong Ung, Stephanie F. Polites, Yi Lin, William A. Weiss, Hu Li. Harnessing GD2-CAR macrophages to target high-risk neuroblastoma [abstract]. In: Proceedings of the AACR Immuno-Oncology Conference (AACR IO): Discovery and Innovation in Cancer Immunology: Revolutionizing Treatment through Immunotherapy; 2026 Feb 18-21; Los Angeles, CA. Philadelphia (PA): AACR; Cancer Immunol Res 2026;14(2 Suppl):Abstract nr B005.

Peroxisome-proliferator–activated receptor delta (PPARδ) regulates metabolic, mitochondrial, and inflammatory pathways implicated in neurodegeneration, making it an attractive therapeutic target for amyotrophic lateral sclerosis (ALS). In this study, we evaluated two PPARδ agonists, KD3010 and T3D-959, in two established ALS/FTD mouse models: an AAV-mediated C9orf72 G4C2-repeat expansion model (C9-149R) and the TDP-43Q331K transgenic model. Drug treatment was initiated prior to the emergence of key disease features and continued for 9–10 months. Comprehensive behavioral, neuropathological, and biomarker analyses revealed marked differences between the two models. C9-149R mice exhibited reduced body weight and subtle behavioral alterations without robust motor deficits, whereas TDP-43Q331K mice developed pronounced, progressive motor and cognitive impairments accompanied by a ~7-fold elevation in plasma neurofilament light chain (NfL). Despite effective target engagement—particularly for T3D-959—neither PPARδ agonist improved motor performance, cognitive behavior, neuroanatomical measures, plasma NfL levels, or disease-associated molecular phenotypes in either model. Prolonged KD3010 treatment resulted in loss of target engagement, consistent with drug tolerance, while T3D-959 sustained PPARδ activation without therapeutic benefit. Together, these findings demonstrate that PPARδ agonism is insufficient to modify disease progression in these ALS/FTD mouse models and underscore the importance of publishing well-powered negative preclinical studies to refine therapeutic strategies for ALS.

Abstract Motivation Illumina DNA methylation arrays have evolved rapidly, expanding genomic coverage while introducing backward incompatibilities by removing many CpG sites present in earlier versions. These changes result in systematic missing values when integrating data across array generations and substantially limiting the reuse of legacy datasets. Results We developed a two-stage framework for imputing missing DNA methylation values. The procedure first imputes randomly missing values using standard imputation techniques and then addresses systematic missingness using multi-output machine learning models, including support vector regression, nearest-neighbor methods, random forest models, and deep neural networks. When evaluated on real datasets with up to fifty percent induced missingness, the proposed framework consistently outperformed conventional imputation approaches. It also accurately imputes the missing CpG sites between methylation arrays and reduced representation bisulfite sequencing data, enabling robust cross-platform data integration. Analyses of large brain tumor methylation datasets demonstrate that the method restores array-specific methylation patterns while preserving biological complexity. Importantly, imputing missing methylation sites significantly improves the performance of epigenetic age prediction models. Availability and implementation This tool is implemented in the Python package “ultra-impute,” freely available at https://github.com/liguowang/ultra-impute. A code snippet demonstrating the usage of the ultra-impute package is provided in a Jupyter Notebook (https://github.com/liguowang/ultra-impute/blob/master/doc/Tutorial.ipynb).

Neutrophils and neutrophil extracellular traps (NETs) contribute to early neuromyelitis optica (NMO) histopathology initiated by IgG targeting astrocytic aquaporin-4 (AQP4) water channels. Yet, the mechanisms underlying neutrophil recruitment and their pathogenic roles in disease progression remain unclear. To investigate molecular-cellular events preceding classical complement cascade activation in a mouse NMO model, we continuously infused, via spinal subarachnoid route, a non-complement-activating mouse monoclonal AQP4-IgG. Parenchymal infiltration of netting neutrophils containing C5a ensued with microglial activation and motor impairment but no blood-brain barrier leakage. Motor impairment and neuronal dysfunction both reversed when AQP4-IgG infusion stopped. Two-photon microscopy and electron microscopy–based reconstructions revealed physical interaction of infiltrating neutrophils with microglia. Ablation of either peripheral neutrophils or microglia attenuated the motor deficit, highlighting their synergistic pathogenic roles. Of note, mice lacking complement receptor C5aR1 exhibited reduction in neutrophil infiltration, microglial lysosomal activation, neuronal lipid droplet burden, and motor impairment. Pharmacological inhibition of C5aR1 recapitulated this protection. Immunohistochemical analysis of an NMO patient’s spinal cord revealed disease-associated microglia surrounding motor neurons in nondestructive lesions. Our study identifies neutrophil-derived C5a signaling through microglial C5aR1 as a key early driver of reversible motor neuron dysfunction in the precytolytic phase of NMO.

GGGGCC (G 4 C 2 ) repeat expansion in C9ORF72 is the most common genetic cause of amyotrophic lateral sclerosis (ALS) and frontotemporal dementia (FTD). Toxicity is thought to result from the accumulation of either repeat RNAs and/or dipeptide repeat proteins (DPRs) translated from repeat-containing transcripts through repeat-associated non-AUG (RAN) translation. To disentangle RNA from DPR toxicity, we mutated a CUG codon predominantly used to initiate DPR translation from all three reading frames. This mutation disrupted DPR synthesis while preserving the expression of repeat-containing RNAs. Despite the accumulation of RNA foci, behavioral deficits and pathological abnormalities, including p-TDP-43 inclusions, STING activation, motor neuron loss, neuroinflammation, and increased plasma neurofilament concentration, were alleviated in C9ORF72 mice. Base editing of the CUG codon also improved molecular phenotypes and survival in patient induced pluripotent stem cell–derived neurons, which highlights the potential of therapeutically targeting DPR production rather than repeat RNAs.