Albert Wang’s research while affiliated with University of California System and other places

OBJECTIVE Cognitive impairment and pain catastrophizing are both associated with worse surgical outcomes. The aim of this study was to define the prevalence of cognitive impairment in patients with adult spinal deformity (ASD) and the relationships between cognitive impairment, pain catastrophizing, patient-reported outcome measures (PROMs), and frailty in the preoperative setting. METHODS This cross-sectional study included patients undergoing evaluation for ASD correction at a single tertiary care center from January 2017 to October 2024. Patients were administered the Montreal Cognitive Assessment (MoCA), Pain Catastrophizing Scale (PCS), Scoliosis Research Society 22-item revised (SRS-22r) questionnaire, the Oswestry Disability Index (ODI), and the Edmonton Frail Scale (EFS). Median survey responses were compared between patients with any cognitive impairment (MoCA score < 26) and no cognitive impairment (MoCA score ≥ 26) using the Mann-Whitney U-test. Associations between survey responses were tested using Spearman’s rank correlation analysis. Multivariate logistic regression analysis was performed to identify predictors of severe pain catastrophizing (PCS score ≥ 30). RESULTS A total of 210 patients (61.4% female, median age 66.5 years) were included in the study. Of these, 123 (58.6%) had normal cognition and 87 (41.4%) had mild or moderate cognitive impairment. Patients with cognitive impairment had greater median PCS scores compared with patients with normal cognition (total PCS score 25.0 vs 19.0, p = 0.01). Lower MoCA scores were significantly correlated with higher PCS (ρ = −0.23, p = 0.0007) and EFS (ρ = −0.21, p = 0.0074) scores, but not ODI and total SRS-22r scores. In the multivariate logistic regression analysis, lower MoCA and SRS-22r scores were associated with greater odds of having severe pain catastrophizing (MoCA: OR 0.82 [95% CI 0.68–0.98], p = 0.03; SRS-22r: OR 0.05 [95% CI 0.01–0.19], p < 0.0001), while ODI score, EFS score, age, and sex were not associated. CONCLUSIONS There was a high prevalence (41.4%) of cognitive impairment in patients with ASD. In both the correlation and multivariate logistic regression analyses, cognitive impairment was associated with pain catastrophizing and thus might contribute to pain perception and frailty in a way that is not consistently captured by traditional PROMs.

INTRODUCTION Seizures are made up of the coordinated activity of networks of neurons. It follows that control of neurons in the pathologic circuits of epilepsy could allow for control of the disease. In non-human disease models of epilepsy, optogenetics has been effective at stopping seizure-like activity by increasing inhibitory tone or decreasing excitation. However, this has not been shown in human brain tissue. Many of the genetic means for achieving channelrhodopsin expression in non-human models are not possible in humans, and vector-mediated methods are susceptible to species-specific tropism that may affect translational potential. There is currently no platform for testing the effects of these potentially disease-modifying tools on network activity in human brain tissue. METHODS Human hippocampus resected from patients with refractory epilepsy were collected, cut to 300um and plated at the air-fluid interface on cell-culture inserts. AAV transduction with channelrhodopsins driven by a glutamatergic promoter took place on the day of collection. Slices were plated on high-density micro-electrode arrays. Hyperactivity was promoted via bicuculline, low-magnesium media and kainic acid. Slices were illuminated by LED fiberoptics positioned over the slice using a custom recording chamber. RESULTS Neuronal transduction ranged from 12 – 54% (median 23%). Illumination of slices expressing the depolarizing channelrhodopsin HcKCR1 caused reduction in network firing rates in 8/8 slices expressing HcKCR1. Reductions in network firing rates were significant in all conditions, physiologic media, GABAaR blockade, low-magnesium media and low-magnesium media with kainic acid. CONCLUSIONS Here, we demonstrate AAV-mediated, optogenetic reductions in network firing rates of human hippocampal slices recorded on high-density microelectrode arrays under several hyperactivity provoking conditions. This platform can serve to bridge the gap between human and animal studies by exploring genetic interventions on network activity human brain tissue.

Seizures are made up of the coordinated activity of networks of neurons, suggesting that control of neurons in the pathologic circuits of epilepsy could allow for control of the disease. Optogenetics has been effective at stopping seizure-like activity in non-human disease models by increasing inhibitory tone or decreasing excitation, although this effect has not been shown in human brain tissue. Many of the genetic means for achieving channelrhodopsin expression in non-human models are not possible in humans, and vector-mediated methods are susceptible to species-specific tropism that may affect translational potential. Here we demonstrate adeno-associated virus–mediated, optogenetic reductions in network firing rates of human hippocampal slices recorded on high-density microelectrode arrays under several hyperactivity-provoking conditions. This platform can serve to bridge the gap between human and animal studies by exploring genetic interventions on network activity in human brain tissue.

Despite advancements in cancer immunotherapy, solid tumors remain formidable challenges. In glioma, profound inter- and intra-tumoral heterogeneity of antigen landscape hampers therapeutic development. Therefore, it is critical to consider alternative sources to expand the repertoire of targetable (neo-)antigens and improve therapeutic outcomes. Accumulating evidence suggests that tumor-specific alternative splicing (AS) could be an untapped reservoir of antigens. In this study, we investigated tumor-specific AS events in glioma, focusing on those predicted to generate major histocompatibility complex (MHC)-presentation-independent, cell-surface antigens that could be targeted by antibodies and chimeric antigen receptor-T cells. We systematically analyzed bulk RNA-sequencing datasets comparing 429 tumor samples (from The Cancer Genome Atlas) and 9166 normal tissue samples (from the Genotype-Tissue Expression project), and identified 13 AS events in 7 genes predicted to be expressed in more than 10% of the patients, including PTPRZ1 and BCAN, which were corroborated by an external RNA-sequencing dataset. Subsequently, we validated our predictions and elucidated the complexity of the isoforms using full-length transcript amplicon sequencing on patient-derived glioblastoma cells. However, analyses of the RNA-sequencing datasets of spatially mapped and longitudinally collected clinical tumor samples unveiled remarkable spatiotemporal heterogeneity of the candidate AS events. Furthermore, proteomics analysis did not reveal any peptide spectra matching the putative antigens. Our investigation illustrated the diverse characteristics of the tumor-specific AS events and the challenges of antigen exploration due to their notable spatiotemporal heterogeneity and elusive nature at the protein levels. Redirecting future efforts toward intracellular, MHC-presented antigens could offer a more viable avenue.

Background The TERT promoter mutation (TPM) is acquired in most IDH-wildtype glioblastomas (GBM) and IDH-mutant oligodendrogliomas (OD) enabling tumor cell immortality. Previous studies on TPM clonality show conflicting results. This study was performed to determine whether TPM is clonal on a tumor-wide scale. Methods We investigated TPM clonality in relation to presumed early events in 19 IDH-wildtype GBM and 10 IDH-mutant OD using three-dimensional comprehensive tumor sampling. We performed Sanger sequencing on 264 tumor samples and deep amplicon sequencing on 187 tumor samples. We obtained tumor purity and copy number estimates from whole exome sequencing. TERT expression was assessed by RNA-seq and RNAscope. Results We detected TPM in 100% of tumor samples with quantifiable tumor purity (219 samples). Variant allele frequencies (VAF) of TPM correlate positively with chromosome 10 loss in GBM (R = 0.85), IDH1 mutation in OD (R = 0.87), and with tumor purity (R = 0.91 for GBM; R = 0.90 for OD). In comparison, oncogene amplification was tumor-wide for MDM4- and most EGFR-amplified cases but heterogeneous for MYCN and PDGFRA, and strikingly high in low-purity samples. TPM VAF was moderately correlated with TERT expression (R = 0.52 for GBM; R = 0.65 for OD). TERT expression was detected in a subset of cells, solely in TPM-positive samples, including samples equivocal for tumor. Conclusion On a tumor-wide scale, TPM is among the earliest events in glioma evolution. Intercellular heterogeneity of TERT expression, however, suggests dynamic regulation during tumor growth. TERT expression may be a tumor cell-specific biomarker.

Background: Despite advancements in cancer immunotherapy, solid tumors remain formidable challenges. In glioma, profound inter- and intra-tumoral heterogeneity hamper therapeutic development. Therefore, it is critical to consider alternative sources to expand the repertoire of targetable (neo-)antigens and improve therapeutic outcomes. Accumulating evidence suggests that tumor-specific alternative splicing (AS) could be an untapped reservoir of neoantigens. Results: In this study, we investigated tumor-specific AS events in glioma, focusing on those predicted to generate major histocompatibility complex (MHC)-presentation-independent, cell-surface neoantigens that could be targets for antibodies and chimeric antigen receptor (CAR)-T cells. We systematically analyzed bulk RNA-sequencing datasets comparing 429 tumor samples (from The Cancer Genome Atlas [TCGA]) and 9,166 normal tissue samples (from the Genotype-Tissue Expression project [GTEx]), and identified 13 AS events in 7 genes predicted to be expressed in more than 10% of the patients, such as PTPRZ1 and BCAN. External RNA-sequencing dataset and full-length amplicon sequencing corroborated these findings. However, investigation of the RNA-sequencing datasets of spatially-mapped and longitudinally-collected clinical tumor samples revealed remarkable spatiotemporal heterogeneity of the candidate AS events. Moreover, proteomics analysis failed to detect peptide spectra matching the putative neoantigens. Conclusions: In conclusion, exploring tumor-specific AS-derived cell-surface neoantigens is challenging due to the low expression levels and spatiotemporal heterogeneity. Redirecting future efforts toward intracellular, MHC-presented antigens could offer a more viable avenue.

The thalamus plays a central coordinating role in the brain. Thalamic neurons are organized into spatially distinct nuclei, but the molecular architecture of thalamic development is poorly understood, especially in humans. To begin to delineate the molecular trajectories of cell fate specification and organization in the developing human thalamus, we used single-cell and multiplexed spatial transcriptomics. We show that molecularly defined thalamic neurons differentiate in the second trimester of human development and that these neurons organize into spatially and molecularly distinct nuclei. We identified major subtypes of glutamatergic neuron subtypes that are differentially enriched in anatomically distinct nuclei and six subtypes of γ-aminobutyric acid–mediated (GABAergic) neurons that are shared and distinct across thalamic nuclei.

Background Central nervous system (CNS) WHO grade 2 low-grade glioma (LGG) patients are at high risk for recurrence and with unfavorable long-term prognosis due to the treatment resistance and malignant transformation to high-grade glioma. Considering the relatively intact systemic immunity and slow-growing nature, immunotherapy may offer an effective treatment option for LGG patients. Methods We conducted a prospective, randomized pilot study to evaluate the safety and immunological response of the multipeptide IMA950 vaccine with agonistic anti-CD27 antibody, varlilumab, in CNS WHO grade 2 LGG patients. Patients were randomized to receive combination therapy with IMA950 + poly-ICLC and varlilumab (Arm 1) or IMA950 + poly-ICLC (Arm 2) before surgery, followed by adjuvant vaccines. Results A total of 14 eligible patients were enrolled in the study. Four patients received pre-surgery vaccines but were excluded from postsurgery vaccines due to the high-grade diagnosis of the resected tumor. No regimen-limiting toxicity was observed. All patients demonstrated a significant increase of anti-IMA950 CD8+ T-cell response postvaccine in the peripheral blood, but no IMA950-reactive CD8+ T cells were detected in the resected tumor. Mass cytometry analyses revealed that adding varlilumab promoted T helper type 1 effector memory CD4+ and effector memory CD8+ T-cell differentiation in the PBMC but not in the tumor microenvironment. Conclusion The combinational immunotherapy, including varlilumab, was well-tolerated and induced vaccine-reactive T-cell expansion in the peripheral blood but without a detectable response in the tumor. Further developments of strategies to overcome the blood-tumor barrier are warranted to improve the efficacy of immunotherapy for LGG patients.

The thalamus plays a central coordinating role in the brain. Thalamic neurons are organized into spatially-distinct nuclei, but the molecular architecture of thalamic development is poorly understood, especially in humans. To begin to delineate the molecular trajectories of cell fate specification and organization in the developing human thalamus, we used single cell and multiplexed spatial transcriptomics. Here we show that molecularly-defined thalamic neurons differentiate in the second trimester of human development, and that these neurons organize into spatially and molecularly distinct nuclei. We identify major subtypes of glutamatergic neuron subtypes that are differentially enriched in anatomically distinct nuclei. In addition, we identify six subtypes of GABAergic neurons that are shared and distinct across thalamic nuclei. One-Sentence Summary Single cell and spatial profiling of the developing thalamus in the first and second trimester yields molecular mechanisms of thalamic nuclei development.

Rubella virus is an important human pathogen that can cause neurological deficits in a developing fetus when contracted during pregnancy. Despite successful vaccination programs in the Americas and many developed countries, rubella remains endemic in many regions worldwide and outbreaks occur wherever population immunity is insufficient. Intense interest since rubella virus was first isolated in 1962 has advanced our understanding of clinical outcomes after infection disrupts key processes of fetal neurodevelopment. Yet it is still largely unknown which cell types in the developing brain are targeted. We show that in human brain slices, rubella virus predominantly infects microglia. This infection occurs in a heterogeneous population but not in a highly microglia-enriched monoculture in the absence of other cell types. By using an organoid-microglia model, we further demonstrate that rubella virus infection leads to a profound interferon response in non-microglial cells, including neurons and neural progenitor cells, and this response is attenuated by the presence of microglia.