Viviane Tabar

Objective: Aggressive pituitary neuroendocrine tumors (PitNETs) that progress despite surgery, radiotherapy (RT), and medical therapy are heterogenous; only a subset exhibits overtly malignant behavior. We examined genetic determinants of metastatic behavior and poor prognosis. Design: Retrospective genomic cohort study of 39 patients (62 tumor samples) with aggressive PitNETs. Methods: Next-generation sequencing was performed using the MSK-IMPACT platform on a sequencing protocol. Genetic alterations were correlated with metastatic status, and progression-free and overall survival (PFS and OS) from first progression following RT. Results: Cell-cycle alterations were found in 23.1% of patients and were more frequently identified in metastatic compared to non-metastatic tumors (42.1% vs 5.0%; OR=12.9; p=0.008). TP53, ATRX, DAXX, and/or SF3B1 alterations were associated with adverse clinical outcomes but did not differ by metastatic status (53% of metastatic vs 60% of non-metastatic tumors). Within the corticotroph lineage, gene-level alterations in TP53, ATRX, and/or DAXX captured corticotrophs with recurrent chromosomal loss-of-heterozygosity (rcLOH), and were associated with shorter PFS (10.1 vs 149.9 months; p=0.009) and OS (116.6 months vs not reached [NR]; p=0.022). A subset of corticotrophs lacking rcLOH harbored gain-of-function USP8 mutations and were associated with longer PFS (149.9 vs 11.5 months; p=0.022). In lactotroph PitNETs, TP53, ATRX, and/or SF3B1 mutations were associated with shorter PFS (4.6 vs 21.5 months; p=0.049). Conclusions: Cell-cycle alterations were enriched in metastatic PitNETs and may contribute to metastatic dissemination. Next-generation sequencing segregates aggressive PitNETs into prognostic subgroups.

Background: High-grade gliomas (HGGs) remain associated with high morbidity, mortality, and invariable recurrence after neurosurgical resection and chemoradiation. A major contributor to treatment failure is a population of stem-like glioma progenitor cells (GPCs) that depend on NOTCH-signaling for self-renewal and survival; clinical attempts to inhibit NOTCH with γ-secretase inhibitors failed due to systemic toxicity. Methods: Here, we investigate the anti-HGG therapeutic potential of a fully-human monoclonal antibody, 1H5, that selectively targets activated-ADAM10, an α-secretase required for NOTCH receptor processing, with high tumor specificity. We show that ADAM10 is markedly overexpressed and highly activated in primary and recurrent HGGs, using patient tissue, RNA-sequencing data, and early-passage HGG cell lines. Results: Across a panel of patient-derived HGG lines, sensitivity to 1H5 correlates with the cell-surface levels of activated-ADAM10, and shRNA-mediated ADAM10 knockdown recapitulates the growth-inhibitory effects of 1H5. ADAM10 inhibition suppresses NOTCH-pathway components and target genes, reduces expression of GPC markers, and impairs neurosphere formation while depleting Sox2- and Ki67-positive cells. We further find that 1H5 induces HGG cell death predominantly via necroptosis, characterized by survivin loss and increased phosphorylated MLKL and by transcriptional changes in necroptosis-related genes. In three-dimensional explants derived from primary HGG, 1H5 reduces growth and GPC marker expression while increasing TUNEL staining. Finally, convection-enhanced delivery of 1H5 in orthotopic xenograft models achieves high local intracranial concentrations, slows tumor growth, and prolongs survival, with regional pharmacodynamic effects consistent with NOTCH pathway suppression and necroptosis. Conclusions: These data support ADAM10-targeting 1H5 as a candidate for further preclinical development in HGG.

Parkinson’s disease (PD) is characterized by the progressive loss of midbrain dopaminergic (mDA) neurons ¹ . Stem cell–derived mDA neurons hold promise for disease modelling ^2,3 and are currently in clinical trials for cell replacement therapy ^4–6 . However, systematic benchmarking has been limited by the lack of a unified high-resolution reference and methods that quantify incomplete or mixed lineage specification in vitro ⁷ . We establish a single-cell and spatial atlas of the human developing diencephalon–midbrain–hindbrain axis resolving 93 cell subtypes, including 39 lacking prior single-cell characterization and 4 entirely novel populations. Using this atlas as a reference, we integrate 19 hPSC-derived mDA datasets, both published ^2,8–25 and unpublished, to build the Human Dopaminergic Neural Atlas (HDNA) spanning 2D, 3D, and graft models, including those used in clinical trials. To classify cells and quantify lineage fidelity, we develop CapybaraBrain, a marker-driven non-negative decomposition framework that assigns each cell continuous identity scores across all 93 developmental programs, enabling systematic discrimination of discrete, transitioning, and cross-lineage hybrid states ²⁶ . We uncover a pervasive landscape of off-target populations reflecting relaxed transcriptional boundaries in vitro, including a previously unrecognized TH–PITX2 midbrain neuronal population, and we validate atlas-predicted latent lineage plasticity through inducible genetic fate mapping in mouse models. We further define maturation-associated transcriptional programs by harmonizing adult mDA subtype atlases, revealing that dopaminergic identity and maturation are partially decoupled across protocols. Finally, projecting PD patient-derived tri-cultures onto the HDNA uncovers genotype- and cell-type-specific transcriptional dysregulation. Together, these integrated atlases and computational framework establish a unified standard for benchmarking differentiation fidelity, exposing off-target states, and guiding next-generation PD models and cell therapies.

Context: Patients with Cushing syndrome (CS) often experience impaired quality of life (QoL) and mood despite biochemical control. The relationship between late-night salivary cortisol (LNSC) and psychological outcomes remains poorly characterized. Objective: Assess QoL and mood outcomes in biochemically controlled CS patients based on circadian rhythm restoration. Design: Cross-sectional. Setting: Tertiary-care center. Participants: Ninety treated, biochemically controlled patients with CS (84 Cushing disease (CD); 6 adrenal CS), stratified into three groups: normal LNSC (Group A), abnormal LNSC (Group B), on long-term GC replacement (Group C). Main Outcome Measures: Hospital Anxiety and Depression Scale (HADS), CushingQoL questionnaire, Nottingham Health Profile (NHP). Results: Group A had lower HADS-Anxiety (4 vs 7, A vs B; p=0.006) and (HADS-Depression 2.5 vs 6 vs 9, A vs B; p=0.006, A vs C; p<0.001). QoL was better in group A vs C in psychosocial (67.5 vs 39.5; p<0.001) and physical (63.9 vs 44.3; p=0.005) domains. Group A had better Emotional Reaction (0 vs 24; p=0.002), A vs B; Energy Level (0 vs 63; p=0.001) and Sleep (13 vs 56; p<0.001), A vs C. In multivariable analyses excluding group C, LNSC normalization was consistently associated with better outcomes in all HADS domains, CushingQoL psychosocial, NHP Emotional Reaction, Social Isolation, Physical Abilities and Home Relationships. Group B had the highest diabetes rate. Among patients with surgically remitted CD, 18.6% had abnormal LNSC, characterizing a previously unrecognized clinical phenotype. Conclusions: In biochemically controlled CS, LNSC normalization correlates with QoL, mood, and metabolic outcomes, and could represent a therapeutic target.

While clinical trials of human pluripotent stem cell–derived midbrain dopamine (mDA) neuron precursor grafts for Parkinson’s disease (PD) are ongoing, current protocols remain suboptimal. In particular, the yield of TH+ mDA neurons after in vivo grafting and the expression of certain mDA neuron and subtype-specific markers require improvement. Single-cell transcriptomic analyses of grafts have revealed low proportions of mDA neurons and substantial off-target contamination. Here, we present an optimized mDA neuron differentiation strategy that builds on our clinical-grade (“Boost”) protocol by adding FGF18 and IWP2 treatment (“Boost+”) at the neurogenesis stage. Boost+ mDA neurons show higher expression of EN1, PITX3, and ALDH1A1. Improvements in mDA neuron yield and transcriptional similarity to primary mDA neurons are observed in vitro and following transplantation. Single-nucleus RNA sequencing demonstrates enrichment of A9 mDA neurons within Boost+ grafts. Functional studies in vitro demonstrate increased dopamine production and release and improved electrophysiological properties. In vivo analyses show higher percentages of TH+ mDA neurons, resulting in efficient rescue of amphetamine-induced rotation behavior in the 6-OHDA rat model and rescue of deficits in some nondrug-induced assays, including the ladder rung assay, which are not improved by Boost mDA neurons. The Boost+ conditions present an optimized differentiation protocol with advantages for disease modeling and mDA neuron grafting paradigms. This study introduces a refined method for producing dopamine neurons in vitro, providing a new protocol for applications in cell therapy for Parkinson's disease.

High-level copy-number (CN) amplification (HLAMP) is a major mechanism of oncogene activation in human cancer. Despite progress in therapeutically targeting HLAMPs, the processes underlying HLAMP evolution are incompletely understood, leaving critical knowledge gaps in their etiology and mechanisms of therapeutic response. To address this, we analyzed using novel computational approaches, the evolutionary trajectories of HLAMP in single-cell whole-genome sequencing of 86,239 cancer cells from 94 patients and 8 experimental systems. We found that two common etiologies of oncogene amplification - intrachromosomal amplification (ICamp) and extrachromosomal circular DNA (ecDNA) - lead to distinct CN distributions across cells in amplitude, variance and clone specificity. Notably, ICamp events exhibited widespread subclonal specificity, indicating their dynamic evolution through numeric or structural modulation associated with clonal expansions. In contrast, ecDNAs exhibited higher cell-to-cell CN variation linked to structural rearrangements, whereby complex ecDNA architectures were resolved at single-nucleotide resolution in individual cell genomes. Through joint analysis of ecDNA structure and their cellular abundance, we observed both divergent and convergent modes of ecDNA evolution shaped by DNA damage, selection, and chromosomal re-integration. Finally, modeling single-cell distributions of ecDNAs substantially improved bulk genome-graph based predictions, revealing that ecDNA prevalence is tissue-type specific, rather than broadly shared across cancer types, with implications for ecDNA-directed therapy.

Context: Cushing’s syndrome (CS) impairs quality of life (QoL) and mood. Prospective real-life data on posttreatment recovery and predictors of improvement are limited. Objectives: Evaluate changes in QoL, depression, and anxiety in patients with CS, before and after biochemical control, and identify predictors of clinically meaningful improvement. Design and Setting: Prospective observational study at a tertiary center. Patients: Sixty-seven patients with endogenous CS (60 pituitary, 7 adrenal) were assessed with active disease and again after achieving biochemical control through surgery and/or medication. Outcomes: Patient-reported outcomes included CushingQoL, Beck Depression Inventory-II (BDI-II), and State-Trait Anxiety Inventory (STAI). Results: Mean and longest follow-up was 2.3 and 11.5 years, respectively. Treatment led to improvements in mean scores across all domains (QoL: +18.2 ± 20.9, BDI-II: −6.8 ± 8.6, STAI-State: −9.6 ± 12.5, STAI-Trait: −8.6 ± 12.6; all P < .001). However, a minimal important difference was achieved in 64.6% for QoL, 67.9% for BDI-II, and 53.2% and 52.8% for STAI subscales. After multivariable analysis, QoL improvements were predicted by lower baseline body mass index, pretreatment symptoms 6 months, and normal follow-up late-night salivary cortisol (LNSC). Depression improvements were predicted by symptoms 6 months postoperative hydrocortisone. Depression improved more gradually than QoL and anxiety. Conclusion: Although effective treatment improves mood and QoL in CS, clinically meaningful recovery is variable and incomplete for some patients. Our findings highlight the need to limit diagnostic delay and provide comprehensive posttreatment care that includes normalization of cortisol circadian rhythm.

Background: Clinical outcomes in patients with glioblastoma and low Karnofsky Performance Scale (KPS) in the inpatient setting are rarely described. Limited survival data and high symptom burden complicate decision-making. This study describes patient-centered outcomes in functionally debilitated patients with newly diagnosed glioblastoma following cancer-directed treatment. Methods: We retrospectively reviewed charts of patients with newly diagnosed glioblastoma who received radiotherapy (RT) while hospitalized. Patient, tumor, and treatment characteristics were included in univariable Cox regression and logistic models to assess associations with overall survival (OS) and discharge home. Multivariable models included factors significant on univariable analysis. OS was estimated by the Kaplan-Meier method. Results: Of 52 patients (median age: 68 years, range: 21-85), median KPS was 50 (range: 30-60). All underwent RT and 50 (96%) received concurrent temozolomide; 12 (23%) received bevacizumab, and 46 (88%) completed planned RT. Median length of stay (LOS) was 29 days (range: 20-65). Only 40% of patients were able to participate in decision-making. Specialty palliative care consultation was provided to 16 (31%). Median OS was 5.82 months. In multivariable analysis, lower KPS remained the sole factor significantly associated with increased risk of death (P = .002) and decreased odd of discharge home (P = .02). Conclusion: Inpatient treatment for functionally disabled patients with glioblastoma was associated with prolonged LOS and poor outcomes. These findings highlight opportunities for early incorporation of specialty palliative care to multimodal cancer-directed therapies to optimize symptom management, elicit patient values and preferences, and facilitate goal-concordant decision-making. Prospective studies are necessary to identify predictors of functional recovery and treatment options.

Recurrent glioblastoma (rGBM) remains incurable. One barrier to the development of effective rGBM therapies is the difficulty in collecting posttreatment tumor tissue. Serial multiomic assays from longitudinal rGBM biopsies may uncover tumor responses to a treatment. Here, we obtained 97 serial rGBM biopsy cores over 4 months from the first two patients participating in a clinical trial of repeated intratumoral dosing of the immunotherapeutic agent CAN-3110. Multiomic analysis of the biopsy cores revealed therapeutic effects, including longitudinal and spatial reshaping of the rGBM’s microenvironment, expansion of new T cell tissue-resident effector memory clonotypes against CAN-3110 epitopes and other undetermined antigens, and expression of human leukocyte antigen (HLA)–presented immunopeptides, including cancer testis antigens. Moreover, serial integrated multimodal analyses provided evidence of therapeutic responses to CAN-3110 despite traditional magnetic resonance imaging indicating progression. Clinically, the two treated patients achieved a pathologic response or stable clinical disease, respectively. These results show the value of longitudinal tissue sampling to understand rGBM’s evolution during administration of an investigational therapy.