Citations

Abstract Porous nanofibrous microspheres (PNMs) present a versatile and minimally invasive strategy for tissue regeneration, combining biomimetic morphology, tunable structure, and injectability. While self‐assembly and co‐axial electrospray are explored for PNM fabrication, these methods are limited in compositional versatility and production scalability. Here, a 3D‐printed microfluidic platform is presented that enables large‐scale fabrication of PNMs with precise control over size, pore architecture, and morphology. PNMs can be functionalized with bioactive molecules through UV crosslinking, enhancing their regenerative potential by promoting osteogenesis in human bone marrow stromal cells (hBMSCs), angiogenesis in human umbilical vein endothelial cells (HUVECs), and exerting anti‐inflammatory effects on macrophages. Subcutaneous implantation in rats demonstrates that PNMs support cell infiltration, minimize fibrosis, and facilitate tissue integration, achieving complete cell penetration and tissue incorporation within 14 days. These findings establish PNMs as versatile, scalable, and customizable platforms, ideal for applications as injectable drugs or cell carriers, as well as powders, offering promising solutions for wound healing and tissue regeneration.

PURPOSE The combination of the Bruton tyrosine kinase inhibitor ibrutinib with bendamustine-rituximab for first-line treatment of mantle cell lymphoma (MCL) prolonged progression-free survival (PFS), but without improvement in overall survival (OS), likely because of toxicity. Acalabrutinib was shown to be efficacious and less toxic than ibrutinib in a head-to-head trial in chronic lymphocytic leukemia and therefore might lead to better outcomes in MCL. METHODS Patients 65 years and older with previously untreated MCL received acalabrutinib (100 mg twice daily) or placebo (until disease progression or unacceptable toxicity), plus six cycles of bendamustine (90 mg/m 2 once daily; days 1 and 2) and rituximab (375 mg/m 2 as a single dose; day 1) followed by rituximab maintenance in responding patients for 2 years. Crossover to acalabrutinib at disease progression was permitted. The primary end point was PFS per the independent review committee; overall response rate and OS were secondary end points. RESULTS In total, 598 patients were randomly assigned, with 299 in each arm. At a median follow-up of 49.8 months using the reverse Kaplan-Meier method, the median PFS was 66.4 months in the acalabrutinib arm and 49.6 months in the placebo arm (hazard ratio [HR], 0.73 [95% CI, 0.57 to 0.94]; P = .0160). Benefit was seen across all subgroups, including those with high-risk features. Overall response/complete response rates were 91.0%/66.6% and 88.0%/53.5% in the acalabrutinib and placebo arms, respectively. OS was not significantly different (HR, 0.86 [95% CI, 0.65 to 1.13]; P = .27). Grade 3 or greater adverse events were reported in 88.9% and 88.2% in the acalabrutinib and placebo arms, respectively. CONCLUSION The combination of acalabrutinib with bendamustine-rituximab significantly improved PFS. Clinical benefit of acalabrutinib with bendamustine-rituximab was achieved with manageable toxicity.

Abstract Gliomas are highly malignant brain tumors with poor prognosis and short survival. Nicotinamide adenine dinucleotide (NAD+) plays a critical role in cancer progression, yet anti-cancer therapies targeting NAD+ synthesis have had limited clinical success due to an incomplete understanding of its mechanisms. Here, we demonstrate the essential role of the NAD+ synthase nicotinamide mononucleotide adenylyltransferase (NMNAT) in glioma progression and its novel contribution to nuclear atypia. Analysis of the patient glioma transcriptomics dataset (GAPIA) indicated a strong correlation between high NMNAT transcription levels and poorer disease-free survival in low grade glioma (LGG) and glioblastoma multiforme (GBM). Utilizing quantitative statistical analysis to assess the size and circularity of about 3 million cells in glioma tissue samples from 19 patients, we identified the significant association between NMNAT1 protein level and increased nuclear atypia in different types of gliomas. Mechanistic studies in human glioma cells and a Drosophila glial neoplasia model revealed that NMNAT enhances glioma growth by promoting NAD+-dependent post-translational modifications poly(ADP-ribosyl)ation (PARylation) and deacetylation of p53, thereby attenuating the pro-apoptotic activity of p53. Additionally, NMNAT1 disrupts nuclear lamina integrity by altering lamin A/C distribution, further contributing to nuclear atypia and glioma progression. These findings uncover a novel tumorigenic mechanism involving NMNAT's regulation of nuclear architecture and NAD+-mediated metabolic signaling pathways, highlighting its roles in glioma growth and nuclear atypia. This study underscores NMNAT as a potential therapeutic target in glioma treatment. Citation Format: Jiaqi Liu, Xianzun Tao, Yi Zhu, Xiang Gao, Tijana Canic, Kai Ruan, Zoraida Diaz-Perez, Sakir Humayun Gultekin, Priyamvada Rai, Qunfeng Dong, Grace Zhai. NMNAT-mediated NAD+ metabolism promotes glioma growth by modulating p53 post-translational modifications and nuclear atypia [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2025; Part 2 (Late-Breaking, Clinical Trial, and Invited Abstracts); 2025 Apr 25-30; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2025;85(8_Suppl_2):Abstract nr LB060.

Abstract Background: Leukocyte immunoglobulin-like receptor B2 (LILRB2) suppresses immune cell activation, particularly within myeloid cells in the tumor microenvironment (TME), while the PD-1/PD-L1 axis serves as a key immune checkpoint that inhibits T-cell function. SPX-303 is a novel bispecific antibody designed to simultaneously block LILRB2 and PD-L1, thereby revitalizing antitumor immune responses. Here, we present initial findings from the ongoing first-in-human Phase 1 trial of SPX-303 in patients with advanced solid tumors, including safety, pharmacokinetics (PK), and receptor occupancy (RO). Methods: This is an open-label, multicenter Phase 1 study for patients with advanced solid malignancies with 3+3 dose-escalation design, followed by an expansion cohort at the recommended Phase 2 dose (RP2D). SPX-303 is administered intravenously every three weeks and safety is assessed through CTCAE v5.0 criteria. Primary Objectives of this study are to determine the safety, tolerability, and RP2D of SPX-303. Secondary endpoints include antitumor activity as well as characterization of the PK and PD of SPX-303. This study is currently open and enrolling in escalation cohorts. Citation Format: Justin Moser, Hao Xie, Yujie Zhao, Yousef Zakharia, Dongdong Li, Brandon William, Dinh-Duc Nguyen, Jack Hovey, Tiffany Li, Robert Cai, Jichun Ma, Jingdong Qin, Martin Siekierzycki, Anthony Haight, Hua Jin, Weiwei Mao, Kevin Bin Li, Gui-Dong Zhu. A first-in-human phase 1 study on the safety, PK, receptor occupancy, and pharmacodynamic markers of SPX-303, a first-in-class LILRB2×PD-L1 bispecific antibody [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2025; Part 2 (Late-Breaking, Clinical Trial, and Invited Abstracts); 2025 Apr 25-30; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2025;85(8_Suppl_2):Abstract nr CT116.

Abstract The International League Against Epilepsy (ILAE) has updated the operational classification of epileptic seizures, building upon the framework established in 2017. This revision, informed by the implementation experience, involved a working group appointed by the ILAE Executive Committee. Comprising 37 members from all ILAE regions, the group utilized a modified Delphi process, requiring a consensus threshold of more than two thirds for any proposal. Following public comments, the Executive Committee appointed seven additional experts to the revision task force to address and incorporate the issues raised, as appropriate. The updated classification maintains four main seizure classes: Focal, Generalized, Unknown (whether focal or generalized), and Unclassified. Taxonomic rules distinguish classifiers, which are considered to reflect biological classes and directly impact clinical management, from descriptors, which indicate other important seizure characteristics. Focal seizures and those of unknown origin are further classified by the patient's state of consciousness (impaired or preserved) during the seizure, defined operationally through clinical assessment of awareness and responsiveness. If the state of consciousness is undetermined, the seizure is classified under the parent term, that is, the main seizure class (focal seizure or seizure of unknown origin). Generalized seizures are grouped into absence seizures, generalized tonic–clonic seizures, and other generalized seizures, now including recognition of negative myoclonus as a seizure type. Seizures are described in the basic version as with or without observable manifestations, whereas an expanded version utilizes the chronological sequence of seizure semiology. This updated classification comprises four main classes and 21 seizure types. Special emphasis was placed on ensuring translatability into languages beyond English. Its aim is to establish a common language for all health care professionals involved in epilepsy care, from resource‐limited areas to highly specialized centers, and to provide accessible terms for patients and caregivers.

Background: Cholangiocarcinoma is a molecularly heterogenous cancer arising from the biliary epithelium. Individual omic techniques have identified clinically relevant, targetable subtypes but most tumors have no targetable alterations based on exome or transcriptomic evaluation. Integrated multiomics approaches incorporating transcriptomic, proteomic, and phosphoproteomic characterization provide deeper understanding and identify non-mutated, activated pathways that can be targeted. To optimize the potential clinical utility of these approaches, minimal classifier features must be defined. Methods: Approximately 60,000 RNA, protein, and phosphoprotein features were extracted from 35 cholangiocarcinomas treated at the primary US study site. 206 patients treated and profiled at an institution in China were also included. Using the Multiomics Factor Analysis clustering method, patients were sorted into 3 distinct molecular profiles. Important molecular features were selected using competing feature selection (FS) methods: Boruta, RreliefF, variable selection using random forests (RF), recursive factor elimination (RFE), median decrease accuracy in RF, and Concrete Autoencoders. Machine learning classification prediction models including Extreme Gradient Boosting, gradient-boosted trees, multinomial logit, and support vector machine (SVM) were tested on each FS. Each combination of FS and predictive model was tested with 10 replicates of 10-fold cross-validation of a randomly-selected balanced test set and an oversampled, balanced training subset. Accuracy, F1 score, AUC, precision, and recall were assessed for each FS and predictive model combination. A custom function of physical assay cost and classification accuracy was optimized to find the best assay. KEGG pathway overrepresentation analysis was used to analyze feature subsets. Results: A consistent set of 50 important RNA and protein features was identified, including previously reported prognostic factors, like SpryD4 protein expression, as well as previously unreported pathway surrogates, such as Slc2A2 or Pipox protein. Pathway overrepresentation analysis found insulin signaling and resistance and amino acid biosynthesis/metabolism pathways were overrepresented in the discriminative feature set. The gradient-boosted trees algorithm combined with the RFE FS method had the best performance, with a multiclass macro average (±sd) cross-validation accuracy 89% (±6%), F1 score 89% (±8%), AUC 0.98 (±0.02), precision 90% (±10%), recall 89% (±6%). Conclusions: Feature selection methods and predictive machine learning models work synchronously to develop accurate molecular subtype assays based on just 50 protein and RNA levels. This strategy has the potential to make molecular subtyping for cholangiocarcinoma and other molecularly complex cancers fast and widely available. Citation Format: Ellen Larson, Erik Jessen, Dong-Gi Mun, Jennifer Tomlinson, Amro Abdelrahman, Danielle Carlson, Hojjat Salehinejad, Rory Smoot. Feature selection and machine learning strategies optimize an affordable molecular assay for cholangiocarcinoma subtype [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2025; Part 1 (Regular Abstracts); 2025 Apr 25-30; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2025;85(8_Suppl_1):Abstract nr 1107.

Immunosuppression within the tumor microenvironment (TME) poses a significant challenge to the efficacy of Chimeric Antigen Receptor (CAR) T-cell therapy in solid tumors, including glioblastoma (GBM), the most common and deadly primary brain tumor. Tumor and TME-residing immunosuppressive cells can hijack program death 1(PD1) pathway to evade antitumor T-cell immunity by upregulating its ligand, PD-L1. Thus, targeting PD-L1 emerges as a promising therapeutic strategy to simultaneously eliminate tumor cells and immunosuppressive components within the TME. We developed a novel PD-L1-targeting CAR T-cell therapy, MC9999, and investigated its therapeutic effects against GBM. The antigen-specific cytotoxicity of MC9999 CAR T-cells was firmly validated using a LN229 GBM cell line and its PD-L1-deficient variant, as well as patient-derived brain tumor-initiating cells (BTICs). Furthermore, in vivo therapeutic antitumor effects were confirmed in orthotopic GBM models through intracranial administration of MC9999 CAR T-cells. To pave the way to the translation of MC9999 CAR T-cell therapy to patient bedside, we established a unique experimental system using patient-specific CAR T-cells against their own tumor and immunosuppressive cells. High-quality, patient-specific MC9999 CAR T-cells were manufactured using T cells isolated from peripheral blood, while paired autologous BTICs were generated to serve as target cells. Autologous immunosuppressive cells include monocyte-derived M2 macrophages from patient’s peripheral blood and tumor-associated macrophages (TAMs) isolated from patient’s tumor. Data from multiple patients’ samples consistently demonstrated that these patient-specific MC9999 CAR T-cells elicited robust cytotoxicity against autologous GBM tumor and TME-residing immunosuppressive cells, supporting our hypothesis that targeting PD-L1 effectively address both tumor and its TME. Immunohistochemistry (IHC) staining of our GBM patients’ tissues showed that PD-L1 expression was restricted to tumor cells and macrophages /microglia within the tumor, with no expression detected in neurons, astrocytes, oligodendrocytes, or blood vessels. These findings suggest that MC9999 CAR T-cells can be safely delivered to the brain without compromising normal cell populations. Ongoing studies are underway to further validate the safety profile of MC9999 CAR T-cell therapy. Collectively, the data obtained from this preclinical study support the effectiveness, feasibility and safety of local administration of MC9999 CAR T-cells as a promising new approach for GBM treatment. These encouraging results have motivated us to prepare an Investigational New Drug application to initiate a phase I clinical trial. Citation Format: Yan Luo, Yaqing Qie, Maria J. Ulloa-Navas, Jesus E. Sanchez-Garavito, Vanessa K. Jones, Martha E. Gadd, Mieu M. Brooks, Tanya Hundal, Andrew D. Brim, Isas K. Vazquez-Rosario, Haidong Dong, Alfredo Quinones-Hinojosa, Hong Qin. Intracranial delivery of MC9999 CAR T-cell therapy: a novel strategy to simultaneously eliminate GBM tumors and immunosuppressive components in the tumor microenvironment [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2025; Part 1 (Regular Abstracts); 2025 Apr 25-30; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2025;85(8_Suppl_1):Abstract nr 4812.

Abstract Background: Cholangiocarcinoma (CCA) is a diverse type of cancer. For several years, platinum-based chemotherapy (Gemcitabine and Cisplatin) has been the standard treatment and serves as the foundation for current immunotherapy combinations. The typical response rates are generally between 20% and 30%. In this study, we present the results of preclinical efficacy trials conducted on CCA patient-derived xenograft (PDX) models, correlating them with the baseline multiomics profiling of these models to identify molecular characteristics linked to treatment response. Methods: Tumor samples from twenty-eight CCA PDX were used to isolate RNA and protein to be included in multiomics analysis (Phosphoproteomics, Proteomics, and Transcriptomics). A subset of PDX models were treated with various doses of Gemcitabine and Cisplatin. The Median Efficacy Index (MEI) was calculated as the ratio of the difference in median of delta in tumor volume at the last day of the study between the treatment and the control arms divided by the median of delta in tumor volume of the control at the last day of the treatment (positive= growth, negative=inhibition, and 0=no efficacy). A Hierarchical All-Against-All algorithm was utilized to compare multiomics features to the MEI and pathway analysis completed for identified molecular features. Pathway enrichment was calculated using the KEGG database, and random-forest variable selection chose the pathways most influential in predicting sensitivity or resistance. Results: After conducting preclinical efficacy trials, the 11 tested CCA PDX models were classified into sensitive, moderately sensitive, and resistant models based on the MEI on each treatment arm. The top up-regulated genes were identified for Gemcitabine response (SRPK2, GLK2, and RIOK2) and Cisplatin response (NTRK1, PRPF4B, and SRPK2). The top up-regulated genes related to Gemcitabine resistance were NTRK1, PIP5K1C, and CSNK2A2, while those related to Cisplatin resistance were SRPK1, PRPF4B, and CSNK2A1. Downregulation of the anti-folate resistance pathway is a strong predictor of gemcitabine sensitivity, and the lysine degradation pathway is a strong predictor of cisplatin sensitivity. Conclusion: Multiomics signatures of cholangiocarcinoma can forecast responses to standard chemotherapy in preclinical models. New markers of resistance to Gemcitabine and Cisplatin were discovered, linked to NTRK1, SRPK1, and SRPK2. These resistance profiles will be tested using suitable targeted therapies on the resistant CCA patient-derived xenograft (PDX) models. Citation Format: Amro M. Abdelrahman, Danielle M. Carlson, Ellen L. Larson, Erik Jessen, Dong-Gi Mun, Isaac Lynch, Alessandro Fogliati, Stella Konadu Adjei Antwi, Aushinie Abeynayake, Akhilesh Pandey, Mark J. Truty, Gregory J. Gores, Rory L. Smoot. Baseline multiomics and chemotherapy efficacy in preclinical trials using patient-derived xenograft models of cholangiocarcinoma can reveal distinct signatures of chemotherapy response [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2025; Part 1 (Regular Abstracts); 2025 Apr 25-30; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2025;85(8_Suppl_1):Abstract nr 6762.

Abstract Cholangiocarcinoma (CCA), an aggressive cancer of the bile ducts, is classified into intrahepatic (iCCA), perihilar (pCCA), and distal (dCCA) anatomic subtypes. Each subtype has a distinct epidemiology and pathogenesis. The tumor immune microenvironment (TIME) of each subtype is poorly understood. CCAs are poorly immunogenic tumors and systemic therapy has had subpar efficacy in CCA subtypes. Thus, we hypothesized that each anatomic subtype has a distinct TIME. To address this hypothesis, we completed single cell sequencing of tumor, liver, and bile duct from patients with either iCCA or pCCA/dCCA. 182,054 cells from paired liver (n=16), tumor (n=16) (9 iCCA, 7 pCCA/dCCA), and normal bile duct (n=9) samples were obtained from treatment-naïve patients undergoing resection from June 2020 to August 2023. The tissues were homogenized and flow sorted for immune (CD45+) populations immediately following resection. The cells then underwent single cell RNA sequencing (scRNA-seq) and cellular indexing of transcriptomes and epitopes by sequencing (CITE-seq) with antibodies for 157 proteins. Multiple immunocompetent, syngeneic/orthotopic murine models of iCCA with distinct genetic drivers were used to assess immunotherapeutics (PMID: 38458319). Immune cells of interest were isolated by flow sorting and functionally validated via suppression assays and multiparameter flow cytometry. Additionally, a novel pCCA murine model was established by injection of an Pik3caH1047R/p27Kip1-derived extrahepatic biliary cancer cell line into the porta/hilar tissue plane. Compared to non-involved liver, the bile duct immune microenvironment composition closely resembled that of the tumor. Multimodal analysis demonstrated both human and murine iCCA tumors have a significant upregulation of regulatory T cells (Treg) expressing T cell immunoreceptor with Ig and ITIM domain (TIGIT) compared to controls. Suppression assay with intratumoral TIGIT+ Treg from mouse compared with TIGIT- Treg indicate TIGIT+ Treg are more potent in inhibiting proliferation of CD8+ T cells. Moreover, TIGIT blockade using a monoclonal antibody resulted in a significant reduction in tumor burden in multiple iCCA murine models, including SB1, a model resistant to anti-PD-(L)1 monotherapy. Meanwhile, human pCCA tumors had a significant increase in IgA+ plasma cells compared to iCCA tumors. An innovative genetically and anatomically relevant pCCA murine model was developed to interrogate this unique plasma cell population. In summary, these data suggest that iCCA and pCCA have distinct immune microenvironments which likely impacts the differences in response to ICB between the subtypes observed in phase III clinical trials. Additionally, the uninvolved, tumor-adjacent bile duct immune microenvironment resembles that of the tumor, suggesting a pre-malignant niche that facilitates tumor development. Citation Format: Hannah E. Stumpf, Jingchun Yang, Haidong Dong, Rory L. Smoot, Sumera I. Ilyas. Multimodal single cell analysis of cholangiocarcinoma subtypes reveals distinct immune microenvironments [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2025; Part 1 (Regular Abstracts); 2025 Apr 25-30; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2025;85(8_Suppl_1):Abstract nr 6476.