Citations

Abstract Introduction Sleep inertia (characterized by profound difficulty waking up with grogginess, disorientation, and cognitive impairment immediately after awakening) is a core idiopathic hypersomnia symptom and may also be experienced by individuals with narcolepsy. Sleep inertia in individuals with narcolepsy is not well studied. This post-hoc analysis of the DUET study (NCT05875974) evaluated sleep inertia in narcolepsy type 1 (NT1) and narcolepsy type 2 (NT2) subgroups of the DUET narcolepsy cohort. Methods DUET was a phase 4, prospective, open-label study with baseline (BL), titration/optimization, stable-dose, and end-of-treatment (EOT) periods. Exploratory outcomes included Psychomotor Vigilance Test (PVT; objective performance impairment) and Karolinska Sleepiness Scale (KSS; subjective sleepiness); both administered at BL and EOT, in the morning after overnight polysomnogram. The 10-minute PVT was completed at 10, 40, and 90 minutes after awakening. The KSS, a single-item, 9-point scale, was completed upon awakening and immediately before and after the PVT assessments. Separate nonlinear, mixed-effects regression models were applied to PVT lapses and KSS ratings to estimate objective and subjective sleep inertia magnitudes, respectively, for NT1 and NT2, along with sleep inertia dissipation time constants for the overall cohort. Treatment-emergent adverse events (TEAEs) were assessed. Results Of 55 enrolled participants with narcolepsy, 26 and 29 had NT1 and NT2, respectively. Most (NT1/NT2) were female (73.1%/72.4%) and White (73.1%/86.2%); 16/26 and 18/29 completed the study. Based on PVT lapses, objective sleep inertia ±SE (NT1/NT2) at BL was 33.1±6.2/14.9±3.2, and at EOT was 8.3±1.7/3.2±1.1 (BL-to-EOT changes [95% CI]: −24.8 [−35.5, −14.1] and −11.8 [−17.4, −6.1]). Based on KSS ratings, subjective sleep inertia ±SE (NT1/NT2) at BL was 1.1±0.6/1.1±0.5, and at EOT was 0.8±0.5/1.3±0.5 (BL-to-EOT changes [95% CI]: −0.3 [−1.9, 1.2] and 0.2 [−1.3, 1.6]). Sleep inertia dissipation time constants ±SE were 1.6±0.3h (PVT) and 0.4±0.1h (KSS) for the overall cohort. TEAEs were consistent with the known LXB safety profile. Conclusion Participants with narcolepsy, especially NT1, demonstrated objective sleep inertia at BL. Participants with NT1 or NT2 showed substantial improvements in objective sleep inertia with LXB treatment. Support (if any) Jazz Pharmaceuticals

Chimeric antigen receptor (CAR) T-cell therapy is an effective treatment for hematologic malignancies. However, it is limited by high costs, risk of severe toxicities such as cytokine release syndrome and neurotoxicity, and heterogeneous patient responses. The current therapy monitoring depends largely on subjective symptom assessment, routine laboratory tests, and basic vital signs, without real-time, quantitative evaluation of CAR T-cell expansion or activation in clinical practice. This lack of timely immune monitoring hampers individualized care and contributes to increased treatment costs. To address this need, we present a proof-of-concept, label-free rapid optical imaging (ROI) biosensor with automated machine learning analysis for direct quantification of CAR T-cells from whole blood. This microfluidic platform integrates red blood cell (RBC) removal, CAR T-cell capture, and imaging-based quantification on a single chip, eliminating the need for centrifugation, staining, and operator-dependent interpretation. For validation, 50 μL whole blood samples spiked with Jurkat cells expressing CD19 CARs underwent RBC depletion by agglutination and microfiltration. The remaining blood components were then incubated on a sensor chip functionalized with recombinant CD19 protein. Captured CAR T-cells were imaged by brightfield microscopy and automatically enumerated using a machine learning algorithm trained on fluorescence-validated cells. The CD-19 cells’ capture performance was validated by flow cytometry and fluorescence imaging. The trained machine learning model validated at 88% sensitivity and 96% specificity. Buffer and whole blood calibration curves were established across clinically relevant concentrations (1–1000 cells/µL) with triple replicates. The results showed high correlation (0.975 and 0.990 R2) between the spiked concentration and the detected CAR T-cells, with a 95% certainty limit of detection (LOD) and quantification (LOQ) of 0.6 and 1.1 cells/µL for spiked buffer, and 14 and 67 cells/µL for spiked whole-blood, respectively.

Abstract Intercellular communication between multiple myeloma (MM) cells and osteoclast precursor cells (pre-OCs) contributes extensively to the occurrence and development of myeloma-related bone destruction. However, key interacting modes and the exchanged substances involved in this communication remain unclear. In this study, we discover that tunneling nanotubes (TNTs) directly connect MM and pre-OCs. Using the stable isotope labeling with amino acids in cell culture assay and a positive-negative double selection strategy, we identify filamin A (FLNA) as a major protein transported from MM to pre-OCs. FLNA acts as a molecular clutch linking extracellular matrix–bound MAC1 to the cytoskeleton, activating Rho and MAPK signaling pathways and promoting F-actin polymerization, which subsequently enhances osteoclast differentiation by modulating cellular stiffness, traction force, and deformability. In addition, FLNA directly binds vinculin and promotes its recruitment to podosomes, thereby enhancing the functions of podosomes and the bone resorption capacity of osteoclasts. The conditional depletion of Flna in mice suppresses podosome activity and reduces stiffness, traction force, and deformability in pre-OCs, leading to significantly impaired osteoclast differentiation and increased bone mass. In the Vk∗MYC mouse model of myeloma, the administration of the TNT inhibitor latrunculin B disrupts FLNA transport to osteoclasts and alleviates osteolytic bone disease. These findings highlight the critical role of MM-transferred FLNA in osteoclastogenesis and suggest that targeting TNTs may represent a therapeutic strategy to limit pathological bone resorption associated with MM.

Introduction: Calibrated automated thrombogram assay parameters have been shown to assess hypercoagulation state and, therefore, may independently predict posttrauma, symptomatic venous thromboembolism (VTE). Yet, calibrated automated thrombogram lacks high throughput and standardization, limiting diagnostic potential. This study analyzed thrombin generation (TG) profiles in trauma patients using a high-throughput assay, processing up to 10 samples at once, providing batch testing capabilities. We hypothesized that trauma patients who developed VTE would show distinct TG profiles compared with those who did not. Methods: Trauma patients presenting to a Level I Trauma Center had samples collected within 12 hours postinjury prospectively. Follow-up was conducted to 90 days, time to symptomatic VTE diagnosis or death, confirmed via autopsy or imaging. TG profiles were measured from the ST Genesia assay. Data were presented as median (interquartile ranges) or n (%), with the Wilcoxon rank-sum or chi-squared test. Associations between TG parameters and clinical risk factors were assessed using Spearman correlation analysis or the Wilcoxon rank-sum test. Results: Two hundred and fifty-four trauma patients were analyzed [48.5 years (31.0, 62.0), 72.0% male]: 64 patients with VTE (25.2%) to 190 with non-VTE. VTE pts had a median time to VTE of 8 days, with 29 developing deep venous thrombosis, 24 with pulmonary embolism, and 11 with deep venous thrombosis/pulmonary embolism. No significant differences were found in age, sex, or body mass index; significantly more VTE patients underwent surgery and blood transfusions within 24 hours. TG profiles demonstrated accelerated TG in patients who developed VTE compared with non-VTE, with significant differences in peak height ( P = 0.001), time to peak ( P = 0.021), endogenous thrombin potential ( P < 0.001), and velocity index ( P = 0.001). Conclusion: TG profiles using ST Genesia can differentiate trauma patients at high risk of developing VTE. The platform suits clinical labs needing high throughput by providing batch testing capabilities. Study type: Prospective study with less than large effect and no negative criteria. Level of evidence: II.

Abstract Pancreatic ductal adenocarcinoma (PDAC) is an aggressive malignancy due in part to metabolic rewiring and a high rate of metastasis. Pancreatic tumors are particularly dependent upon glutamine (Gln) as a nutrient source for multiple metabolic pathways. To adapt to low levels of glutamine, PDAC cells upregulate the Rac1 dependent process of macropinocytosis, which internalizes extracellular macromolecules to scavenge them as a glutamine source. The proto oncogene Vav1 is an activator of the potent Rac/Cdc42 signaling cascades that regulate actin dynamics. While Vav1 is normally restricted to hematopoietic cells, Vav1 expression is significantly increased in PDAC, with high Vav1 correlating with worse overall survival. Here we uncover a novel Vav1 mediated regulation of glutamine metabolism in PDAC cells. While Vav1 is a potent activator of Rac1, surprisingly, Vav1 expression inhibited macropinocytosis in PDAC cells. Intriguingly, genomic and metabolomic analysis revealed that high Vav1 expression correlates with increased dependence on glutamine and glutamate (Glu) metabolism. Indeed, Vav1 dependent macropinocytosis was rescued by cell permeable glutamate, confirming disrupted Gln/Glu balance in Vav1 knockdown cells. RPPA screening indicated a Vav1 dependent effect on glutaminase 1 (GLS1, KGA isoform), which converts glutamine to glutamate. Mechanistically, we discovered that Vav1 modulates GLS1 localization to the mitochondria through regulation of GLS1 posttranslational modification. Disrupting GLS1 localization by deleting its mitochondrial targeting motif phenocopied loss of Vav1 in reducing the invasive potential of tumor cells. Investigation of Vav1 dependent metabolic consequences showed that knockdown of Vav1 alters 13C Gln derived metabolic flux into the TCA cycle, as well as a deficiency in the level of the antioxidant glutathione (GSH), which utilizes glutamate for its synthesis. Consequently, targeting Vav1 resulted in increased levels of cellular and mitochondrial reactive oxygen species (ROS) and increased sensitivity to lipid peroxidation. Notably, dual targeting of both glucose and glutamine pathways showed significant synergistic effects on cellular viability in Vav1 expressing cells. Here we established a new tumor promoting role for Vav1 via Gln/Glu regulation and uncover a novel subcellular regulation of mitochondrial glutamine metabolism. Our findings establish Vav1 as a critical regulator of metabolic rewiring in PDAC and orchestrates glutamine metabolism by controlling the KGA isoform of GLS1 mitochondrial localization. Importantly, Vav1 functions as a signaling adapter in glutamine metabolism, providing a potential therapeutic vulnerabilities for PDAC patients with high Vav1 expression. Citation Format: Mustafa Emre Gedik, Omar L. Gutierrez-Ruiz, Katherine M. Johnson, Ella Rose D. Chianis, Jing Chen, Alex Bittner, Adam Zahm, Ankit Chhoda, Taro Hitosugi, Mark A. McNiven, Gina Razidlo. Vav1 promotes pancreatic tumor cell invasion via GLS1-dependent glutamine metabolism [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2026; Part 1 (Regular Abstracts); 2026 Apr 17-22; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2026;86(7 Suppl):Abstract nr 7322.

Abstract Introduction/Objectives: The CARMA1-BCL10-MALT1 (CBM) signalosome plays crucial role in mediating antigen receptor-induced activation of NF-κB transcription factor and subsequent lymphocyte activation. The effector protein of the signalosome, MALT1, functions both as a scaffold to recruit downstream NF-κB signaling machinery, and as a protease to cleave and inactivate multiple substrates including negative regulators of NF-κB. Constitutive MALT1 activation, which can occur as a result of gain-of-function mutations of the B cell receptor, CARMA1, BCL10 or chromosomal translocation involving the MALT1 gene, underlies the pathogenesis of multiple lymphoid malignancies including activated B-cell type-diffuse large B-cell lymphoma (ABC-DLBCL) and MALT lymphoma. We previously identified G-protein-coupled receptor kinase 2 (GRK2) as a tumor suppressor in MALT1-dependent lymphomas which binds MALT1 and inhibits MALT1 scaffolding and proteolytic activities. Accordingly, GRK2 mRNA levels are markedly lower in a subset of DLBCL tumors compared to normal B cells and lower GRK2 level in ABC-DLBCL is associated with reduced patient survival. In current study, we sought to investigate the mechanism of how GRK2 levels are regulated in ABC-DLBCL. Methods/Results: Using DICER1 depleted cells, we showed that globally impaired miRNA processing leads to increased GRK2 protein levels. We then conducted bioinformatic analyses of patient tumor samples and performed screening via microRNA Data Integration Portal to identify microRNAs that target GRK2 in ABC-DLBCL. Further GRK2 3’UTR reporter assay confirmed direct targeting of GRK2 by candidate miRNAs. ABC-DLBCL cell lines, OCI-LY3 and TMD8, which demonstrate reduced GRK2 mRNA expression compared to primary B cells, show elevated expression of candidate GRK2-targeting microRNAs. Stable overexpression of miR-125a, 125b, or 148b in OCI-LY3 or TMD8 cells led to reduced GRK2 expression, enhanced MALT1 activity and increased cell proliferation. Conversely, in vitro inhibition of candidate miRNAs using anti-miRNA Locked Nucleic Acids (LNA) inhibitors led to enhanced GRK2 expression, reduced MALT1 activity and decreased cell proliferation. Importantly, treatment of mice with inhibitors of miR-125b and miR-148b abrogates the growth of ABC-DLBCL xenograft tumors in vivo. Conclusion: We have identified miRNAs that negatively regulate GRK2 expression in ABC-DLBCL. These GRK2-targeting miRNAs serve a pro-tumorigenic role by promoting MALT1 oncoprotein’s scaffold and protease activity. Furthermore, inhibitors of these miRNAs can enhance GRK2 expression and thereby suppress MALT1 activity and MALT1-dependent tumor cell proliferation, both in vitro and in vivo. These studies suggest that microRNA inhibitors which enhance GRK2 expression could represent a novel approach for restraining MALT1-dependent lymphomagenesis. Citation Format: Jing Cheng, Mei Smyers, Matt Trotta, Neil M. Carleton, Lisa M. Maurer, Uma R. Chandran, Min Xia, Ari M. Melnick, Peter C. Lucas, Linda M. McAllister-Lucas. MicroRNAs targeting tumor suppressor GRK2: Novel potential therapeutic strategies for restraining MALT1-dependent DLBCL [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2026; Part 1 (Regular Abstracts); 2026 Apr 17-22; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2026;86(7 Suppl):Abstract nr 2061.