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AI‐Optimized 3D‐Printed Dual‐Layer Implant: Resolving Chemo‐Immunotherapy Sequential Administration Dilemma for Colorectal Cancer Peritoneal Metastasis
ABSTRACT The synergistic integration of chemotherapy and immunotherapy represents the most promising strategy for enhancing therapeutic efficacy in cancer treatment. Chemotherapy initiates the therapeutic cascade by inducing immunogenic cell death (ICD), thereby releasing tumor antigens and prime immune sensitization. Subsequently, immunotherapy blocks immune evasion pathways, resulting in a coordinated relay‐like antitumor response. This temporally coordinated sequence maximizes synergistic therapeutic efficacy. However, current clinical practice cannot support the sequential and sustained administration of chemotherapy and immunotherapy. This study innovatively integrates artificial intelligence (AI) with 3D printing technology to develop a dual‐layer drug‐loaded implant (LEH@OG) to achieve precise spatiotemporally controlled sequential drug release. The AI model precisely predicted exposure time of the inner gel layer in advance by optimizing parameters such as outer shell thickness and concentration, thereby realizing an on‐demand sequential release process. This study demonstrates that combining AI with 3D printing enables precise sequential delivery of chemotherapy‐immunotherapy agents, providing a core solution for establishing personalized colorectal cancer peritoneal metastasis (CCPM) therapeutic platforms, while also offering a new paradigm for synergistic treatment of other solid tumors.
ViSENet: Joint Sequence and Structure Embeddings of Spike Proteins for Predicting Viral Dynamics 2310110
Abstract Introduction Predicting which viral strains will become predominant is challenging for vaccine design and therapeutic development. Viral evolution is driven by interactions between sequence variation, molecular structure, receptor binding, and immune escape, yet most computational methods consider these factors in isolation. A unified representation integrating them is needed to better anticipate viral evolutionary dynamics. We introduce ViSENet (Viral Sequence Evolution Network), a multimodal framework that jointly embeds viral spike protein sequence and structure in a chronologically organized latent space. Methods ViSENet integrates complementary neural architectures. Spike protein sequences are encoded using a transformer-based encoder that learns temporally organized embeddings. Structural information is captured using a geometric scattering encoder applied to AlphaFold-predicted structures, extracting multiscale features of key spike domains. Sequence and structure embeddings are fused into a shared latent space and trained with supervision from sequence reconstruction, emergence time, receptor binding affinity, and immune escape. Evolutionary dynamics are modeled using a neural ODE to enable continuous time forecasting. Results Applied to COVID-19 and influenza datasets, ViSENet learns latent representations that organize viral variants by temporal emergence and lineage, with related strains clustering by sequence similarity. The model accurately predicts binding affinity and outperforms unimodal baselines. Time split evaluations show that latent trajectories capture meaningful evolutionary trends, and the neural ODE enables projection of viral evolution several weeks into the future. Conclusion ViSENet provides a unified framework for modeling viral evolution by integrating sequence, structure, and functional properties within a temporally organized latent space, enabling interpretation of past trends and forecasting of emergent viral variants for anticipatory vaccine development. Funding Source n/a Topic Categories Computational and Systems Immunology (COMP)
Targeting mannose receptor (CD206)+, pro-fibrotic macrophages to halt lung fibrosis progression in chronic graft-versus-host disease 2251444
Abstract Introduction TGFβ−secreting, CD206+ profibrotic macrophages (macs) are key chronic graft-versus-host disease (cGVHD) drivers. Global depletion of macs could compromise protective immunity; therefore, we targeted CD206 + (mannose receptor) macs with mannosylated albumin nanoparticles (MANPs) containing TGFβ siRNA or non-targeting (NT) siRNA for control. A published bleomycin pulmonary fibrosis model showed disease prevention post TGFβ-MANPs internalization by CD206+ monocyte-derived alveolar macs. CD206+ TGFβ+ macs are also critical in sclerodermatous (Scl) GVHD (B10D2>Balb/c). We show that mice with established multiorgan cGVHD/bronchiolitis obliterans (BO) exhibit a progressive increase in lung tissue profibrotic macs (d49 vs. d28) highlighting their pathogenic role. Methods In our cGVHD/BO model, B10.BR recipients receive B6 bone marrow (BM) and a low dose of T cells (control = BM only). Co-culture of CD206 + (M2-like) and CD206neg (M1-like) macs confirmed that MANPs were preferentially internalized by CD206+ macs and effectively reduced TGFβ protein and mRNA with no effect on CD206neg macs. In vivo, MANPs are delivered intravenously 2x/week starting at d28 post transplantation. Results Compared to BM only controls, d49 cGVHD/BO mice had a significant increase (1.5-fold) in lung CD206+ macs frequency (p = 0.0010) and these cells had a 2.5-fold increase in TGFβ expression (p = 0.0070) by flow cytometry fluorescent intensity. TGFβ siRNA vs NT siRNA—loaded MANPs delivered i.v. to cGVHD/BO mice improved lung function to levels comparable to BM controls (n = 10/group): resistance (p < 0.0001), elastance (p < 0.05), compliance (p < 0.05). Early studies using MANP-based delivery of TGFβ siRNA in Scl GVHD improved skin scores (p < 0.0001) compared to NT siRNA delivery. Further studies with MANPs in Scl GVHD are ongoing. Conclusion These results identify CD206+ macs as a tractable therapeutic target with high selectivity and highlight MANP-based siRNA delivery as a promising strategy to limit cGVHD-driven pulmonary fibrosis. Funding Source NIH P01 HL158505 Topic Categories Transplantation Immunology (TRAN)
Harnessing the homeostatic proliferation capacity of memory CD8+ T cells to restore immune memory after radiation exposure 2253074
Abstract Introduction While lymphopenia elicits vigorous homeostatic proliferation and memory phenotype acquisition among naive CD8+ T cells, its impact on pre-existing memory CD8+ T (TMEM) cells remains unclear. We sought to define the T cell—intrinsic and —extrinsic factors governing the homeostatic proliferation of TMEM cells following radiation-induced lymphopenia and to assess whether adoptive transfer of cryopreserved TMEM cells could restore CD8+ T cell-mediated immune memory after acute radiation-induced lymphoablation. Methods Specific pathogen free mice were subject to 0 or 5Gy of whole-body irradiation (WBI) followed by co-adoptive transfer of equal numbers of allelically-distinct memory P14 cells of different age. The numbers, tissue distribution and phenotype of the donor cells were longitudinally analyzed. In parallel, LCMV-experienced P14 chimeric mice that had recently undergone WBI were given varying doses of cryopreserved Thy1-disparate memory P14 cells to compare the homeostatic proliferation and antigen-dependent functionality between WBI-surviving and post-WBI transferred P14 TMEM cell. Results Post-WBI transferred P14 TMEM cells numerically expanded in lymphopenic hosts and maintained stable numbers over time, with late P14 TMEM cells displaying greater proliferation capacity than early P14 TMEM cells. The extent of proliferation was directly proportional to the number of input cells and the duration of lymphopenia in the recipients. In addition to populating blood and secondary lymphoid organs, the progeny also resided in the brain tissue. Compared with WBI-surviving P14 TMEM cells, post-WBI transferred P14 TMEM cells exhibited enhanced homeostatic proliferation and secondary expansion upon antigen re-encounter while maintaining similar cytokine production capacity. Conclusion Transfer of small numbers of radiation-inexperienced TMEM cells into recently irradiated hosts partially restores both the number and functionality of the TMEM compartment otherwise depleted by radiation exposure. Funding Source n/a Topic Categories Lymphocyte Differentiation and Peripheral Maintenance (LYM)
Reels in Classroom Teaching: Enhancing Engagement and Learning 2256286
Abstract Introduction The integration of short-form videos, known as reels, into classroom teaching has transformed how educators deliver instructional content. Reels provide a dynamic and engaging way to present complex concepts, making them an effective tool for capturing students’ attention and encouraging active learning. Methods We at Chicago State University used appropriate surveillance methods, such as direct observation, digital monitoring, biometric sensors, and self-reporting tools. Ensured ethical practices by obtaining informed consent, protecting privacy, and maintaining transparency. Used tools such as LMS analytics, screen-monitoring software, and video analysis platforms to collect data. Analyzed the data using qualitative or quantitative methods. Results At Chicago State University, videos were used to implement a flipped classroom approach, where students engage with instructional content outside of class, freeing up class time for more interactive and discussion-based activities. Reels supplement traditional lectures by providing additional explanations, examples, and illustrations to support student understanding. The findings suggest that incorporating reels into classroom teaching offers several benefits, including increased student engagement, improved retention, and greater flexibility. Reels are created and shared to allow us to tailor content to meet the diverse needs of students. Additionally, reels promote active learning, spark discussions, and encourage critical thinking. Conclusion In conclusion, reels have the potential to transform classroom teaching by offering a lively and engaging way to deliver instructional content. Using reels enables educators to create a more interactive, student-centered learning environment, fostering more profound understanding, retention, and the application of complex ideas. As educators continue to explore innovative methods to engage students, reels are likely to become an increasingly important part of the future of classroom instruction. Funding Source n/a Topic Categories Immunology Education and Communication (EDU)
Gamma-Delta T cell—microglia crosstalk drives CD8+ T cell infiltration and promotes Alzheimer’s-like pathology in APP/PS1 mice 2257327
Abstract Introduction Neuroinflammation is a central feature of Alzheimer’s disease (AD), driving neuronal injury, synaptic dysfunction, and the accumulation of amyloid plaques and neurofibrillary tangles. In AD, microglia transition from a homeostatic (M0) to a neurodegenerative (MGnD) phenotype that promotes recruitment of peripheral immune cells to the brain. We identified γδ T cells as key modulators of this process. Methods We used flow cytometry, IF, and behavioral testing in APP/PS1Jucker and APP/PS1:γδ⁻/⁻ mice to assess immune infiltration, cytokine production, microglial phenotypes, and cognition; photoconversion and cell tracking in APP/PS1:Kaede mice to trace gut- and lymphoid-derived γδ T cells; and single-cell RNA sequencing of PBMCs and brain tissues from AD patients to profile γδ T cell transcriptional programs. Results In the APP/PS1Jucker model, γδ T cells infiltrated the brain by 3 months of age - preceding CD8+ T cell recruitment - and correlated with the emergence of an MGnD microglial signature. Infiltrating Vγ1 cells predominantly produced IFN-γ, while Vγ4 cells secreted IL-17. γδ T cell deficiency improved cognition, reduced amyloid deposition, shifted microglia toward a homeostatic phenotype, and diminished CD8+ T cell infiltration. Using photoconvertible APP/PS1:Kaede mice, we demonstrated that gut-derived Vγ4 γδ T cells preferentially migrate to the brain, supporting a microbiota-dependent gut—brain axis, whereas Vγ1 cells originate from lymphoid organs. Single-cell RNA sequencing of PBMCs and brains from AD patients revealed γδ T cell enrichment for IFN-γ and IL-17 transcriptional programs, mirroring mouse data. Conclusion Together, these findings uncover a pathogenic γδ T cell—microglia—CD8+ T cell axis linking gut immunity to neuroinflammation, identifying γδ T cells and their cytokines as promising therapeutic targets for AD. Funding Source Alzheimer’s Association Topic Categories Neuroimmunology (NEUR)
High-Throughput Immunomodulator Screening Reveals Subtype-Specific Responses in Low-Abundance Immune Cell Populations Using Scalable scRNA-seq 2260031
Abstract Introduction Understanding immune cell variability including rare populations is essential for dissecting inflammatory mechanisms and therapeutic response. However, functional screening of immunomodulators is often limited by precious sample availability and cell loss during processing. Methods We present a fixation-compatible combinatorial barcoding workflow optimized for low-input immune samples (< 20,000 PBMCs), enabling high-resolution profiling of rare subsets such as classical monocytes, regulatory T cells, and B-cell subtypes. This approach supports 96-condition parallel perturbation while preserving transcriptomic integrity and subtype recovery (>65% retention), even in fragile or low-abundance populations. Results In a screen of 88 immunomodulatory compounds in cultured PBMCs, including PDE4 inhibitors and glucocorticoids, we identified both drastic and subtle shifts in gene expression within classical monocytes, alongside pathway-specific perturbations in B and T cell compartments. Notably, compounds targeting the same pathway elicited concordant transcriptional programs, enabling comparative pharmacodynamic insights at single cell resolution. Conclusion This scalable low-input single-cell approach enables detailed investigation of immune cell behavior under perturbation, capturing both robust and subtle transcriptional responses across rare and abundant PBMC subsets. By accommodating limited sample availability, it supports exploration of how immunomodulators influence cell states, lineage stability, and inflammatory programs at high resolution. Such screens enable efficient discoveries of drug efficacy and detection of secondary effects, offering a powerful experimental framework for uncovering immune mechanisms, comparative pathway responses, and context-dependent variability within complex immune systems. Funding Source n/a Topic Categories Technological Innovations in Immunology (TECH)
B Cells Under Pressure: How Losing Desmosterol Fuels Atherosclerosis 2260473
Abstract Introduction Atherosclerosis is an inflammatory disease characterized by plaque buildup in vessels. Cholesterol homeostasis is crucial to prevent toxic cholesterol levels, but it is often dysregulated in hyperlipidemic states. Desmosterol, a cholesterol precursor, inhibits macrophage activation. While the impact of cholesterol metabolism on macrophage functions is well-studied, its role in regulating B cell functions remains unclear. Methods We generated atherosclerosis-prone Dhcr24fl/flCd19cre/+Ldlr-/- mice, in which B cells overexpress 24-dehydrocholesterol reductase (DHCR24), an enzyme that converts desmosterol into cholesterol. DHCR24 overexpression depleted desmosterol levels in B cells from Dhcr24fl/flCd19cre/+Ldlr-/- vs Cd19cre/+Ldlr-/- control mice. To test the impact of desmosterol depletion in atherogenesis, Dhcr24fl/flCd19cre/+Ldlr-/- and Cd19cre/+Ldlr-/- mice were fed a high-fat diet (HFD) for 12-16 weeks. Results Desmosterol-depleted B cells from HFD-fed Dhcr24fl/flCd19cre/+Ldlr-/- show increased mitochondrial superoxide production, elevated Ca2+ flux and pSYK in the response to anti-IgM-induced BCR activation vs B cells from Cd19cre/+Ldlr-/- mice. Desmosterol depletion in B cells increased lipid loading and shifted GC formation, which was accompanied by a decline in mitochondrial function of B cells from HFD-fed Dhcr24fl/flCd19cre/+Ldlr-/- mice. Importantly, Dhcr24fl/flCd19cre/+Ldlr-/- mice also increased atherogenesis vs Cd19cre/+Ldlr-/- mice. Conclusion Together, this suggests that desmosterol acts as a suppressive regulator of B cell activation, influencing their functions. This highlights a previously unrecognized role of desmosterol in modulating B cell responses in atherosclerosis. Funding Source n/a Topic Categories Lymphocyte Differentiation and Peripheral Maintenance (LYM)
WDFY1 is a key adaptor protein in Axl receptor tyrosine kinase signaling 2310082
Abstract Introduction The WD repeat and FYVE-domain containing protein 1 (WDFY1) encodes a phosphatidylinositol 3-phosphate (PI3P) binding protein localized to early endosomes. WDFY1 involves in endosomal trafficking, cell growth signaling, and autophagy and serves as a key adaptor protein in immune signaling. Axl receptor tyrosine kinase is critical in eliminating apoptotic debris and regulating cell proliferation/survival. We found a close association between Axl and WDFY1 expression. We wondered if WDFY1 serves as an adaptor protein for Axl signal transduction. Methods Mesangial cells were cultured and treated with siRNAs targeting WDFY1 and Axl expression. Cells were then stimulated with Axl agonists. Protein-protein interactions were investigated with co-immunoprecipitation and verified with immunofluorescence co-localization and proximity ligation assay (PLA). Axl signaling cascades were studied by Western blotting with anti-phospho antibodies. WDFY1 C347S point mutation were constructed and transfected into mesangial cells to study the importance of the WDFY1 FYVE domain in the process. Results We found that Axl activation leads to its internalization and merge with early endosomes, where PI3P is generated. WDFY1 binds to PI3P upon Axl activation and then recruits Akt to the site transducing downstream signal cascades. WDFY1 depletion and C347S mutation resulted in reduced Akt phosphorylation and disrupted PI3P-WDFY1 interaction. Conclusion Our data suggest a key role of WDFY1 in bridging the PI3P-Akt interaction, which is necessary for Axl signaling. Funding Source n/a Topic Categories Immune Response Regulation: Molecular Mechanisms (IRM)
Impaired IFNγ transcription and translation drive tumor-specific CD8 T cell dysfunction 2252067
Abstract Introduction Tumor-specific CD8 T cells (TST) are often dysfunctional and fail to produce IFNγ, a cytokine essential for tumor elimination. How transcriptional and translational mechanisms contribute to this defect remains unclear. Methods We analyzed Ifng transcription and translation in adoptively transferred antigen-specific CD8 T cells in acute infection and liver tumors. Using an Ifng fluorescent reporter together with intracellular cytokine staining, we measured Ifng mRNA and protein at the single-cell level by flow cytometry. Results Early after antigen encounter, both TST and Listeria-primed effector T cells (Teff) expressed Ifng mRNA. In contrast to Teff, TST did not produce IFNγ protein, suggesting an early translational block. At later time points, Teff rapidly and robustly upregulated Ifng mRNA and IFNγ protein upon antigen restimulation. However, with continued tumor exposure, TST failed to upregulate Ifng mRNA in response to antigen, demonstrating a TST-specific Ifng transcriptional block. Consequently, IFNγ production in TST is impaired at both the transcriptional and translational level. Although the Ifng locus underwent similar chromatin remodeling in TST and Teff upon activation, chromatin accessibility diverged over time, suggesting epigenetic mechanisms may engender the Ifng transcriptional defect. Conclusion In future studies, we will epigenetically target the Ifng locus to restore IFNγ mRNA expression and further study and target the Ifng translational block to improve TST IFNγ production. Understanding these regulatory mechanisms in TST will potentially lead to new strategies to strengthen antitumor T cell immunity. Funding Source n/a Topic Categories Tumor Immunology: Cellular Responses and Tumor Microevironment (TIME)
Intrinsically disordered sequence in the ZMIZ1 drives leukemic activity, regulates protein stability and biomolecular condensate formation 2253537
Abstract Introduction T-cell acute lymphoblastic leukemia (T-ALL) is an aggressive blood cancer in children and adults, marked by the uncontrolled growth of immature T cells infiltrating the bone marrow, with frequent peripheral blood, mediastinal, and central nervous system involvement. Our lab found that Zinc Finger MIZ-containing 1 (ZMIZ1), a transcription cofactor in the PIAS-like family, drives NOTCH1-mutated and early precursor T-ALL. ZMIZ1 binds various transcription factors, influencing development and disease. We identified somatic mutations in the alanine-rich domain (ARD) of ZMIZ1 in T-ALL patients and found these mutations are mutually exclusive with germline ARD mutations seen in neurodevelopmental disorder (NEDDFSA). The ARD’s function is unknown. Methods We aim to uncover ARD’s role in T-ALL by studying these mutations. We cloned WT and mutant ZMIZ1 into retroviruses and transduced them into ZMIZ1-deficient 8946 T-ALL cells. Results T-ALL mutants showed increased ZMIZ1 protein levels compared to WT, while a NEDDFSA mutant showed decreased levels, with mRNA unchanged, indicating effects on protein stability. Deleting ARD (ΔARD) raised ZMIZ1 abundance 2.5-fold, suggesting ARD acts as a degron; cycloheximide treatment supported higher stability in ΔARD cells. Preliminary data link ARD mutations to faster cell growth versus WT. To further study ARD, we made GFP-ARD chimeras with T-ALL and NEDDFSA mutations, finding T-ALL mutants had higher expression than NEDDFSA ones. Proteasome inhibition increased GFP-ARD levels, indicating ARD-induced degradation is proteasome-dependent. Our data suggest adjacent ARD mutations can either drive T-ALL or neurodevelopmental disorders by affecting ZMIZ1 stability. Conclusion Understanding ARD regulation of ZMIZ1 may open therapies for ZMIZ1-dependent cancers, as these mutations also appear in brain, breast, B-cell, and myeloid cancers. Funding Source Post-Doctoral Fellowship NIH NIGMS K12 GM 111725 Topic Categories Hematopoiesis and Immune System Development (HEM)
SARS-CoV-2-induced IL-6 paralyzes DC function through maturation impairment and immunosuppressive DC3 expansion 2256903
Abstract Introduction Severe COVID-19 is characterized by profound dendritic cell (DC) dysfunction, yet the molecular drivers remain poorly defined. Methods To evaluate DC heterogeneity during COVID-19, we longitudinally immunophenotyped circulating DC subsets in mild and severe cases. We then performed immune correlation analyses across 23 antigen-specific readouts, T cell responses, antibody titers, and inflammatory markers. High-parameter flow cytometry was used to identify drivers of SARS-CoV-2—associated DC dysfunction. Single-cell RNA sequencing (scRNA-seq) profiled peripheral blood DC subsets, and transcriptional analyses evaluated downstream mediators. Results We demonstrated that IL-6—enriched patient plasma and supernatant from Delta-infected lung epithelial cells reduce CD86 expression on cDC2s and expand the immunosuppressive CD163+ DC3 subset, a phenotype reversible with IL-6 receptor blockade. Single-cell RNA-sequencing revealed the expanded CD163+ DC3 population drives immunosuppression in severe COVID-19 patients. Mechanistically, we implicate the transcription factor C/EBPβ as a critical downstream mediator, based on its disease-associated expression and established role in myeloid cell programming. Conclusion Overall, this study unveils a novel immune-evasion pathway wherein SARS-CoV-2-induced IL-6 expands immunosuppressive DC3s via C/EBPβ, highlighting IL-6 and C/EBPβ as promising therapeutic targets for ameliorating DC dysfunction in severe COVID-19 and related hyperinflammatory disorders. Funding Source Hong Kong Collaborative Research Fund (CRF): C7156-20G Topic Categories Innate Immune Responses and Host Defense: Molecular Mechanisms (INM)
Elevated type I interferon levels during Influenza virus infection profoundly suppress pulmonary innate and adaptive immunity and exacerbate disease 2257518
Abstract Introduction Most mouse studies aimed at investigating the roles of specific cytokines in disease states use mice deficient for the cytokine or its receptor, or antibody treatment to block its signaling. While such analyses have found relatively minor impacts for type I interferon (IFN-I) signals during Influenza A (IAV) infection, elevated IFN-I expression has recently been correlated with worsened disease outcomes, though the underlying mechanisms are not clearly defined. Here, we investigated how increasing IFN-I availability during the first week of sublethal IAV infection impacts diverse aspects of the inflammatory landscape and disease severity. Methods Mice were systemically treated with exogenous IFN-I or not from days 2-6 after IAV infection followed by analysis at day 7, which marks the peak of the cellular immune response in the lungs. Results IFN-I treatment caused dramatic reductions of all innate and adaptive immune cell subsets assayed in the lungs, but not in the spleens, of both C57BL/6 and BALB/C mice. This impact was seen in unprimed mice responding to primary IAV infection, and in IAV-primed mice responding to heterosubtypic infection. Mechanistically, this broad suppressive impact requires expression of the IFN-I receptor on hematopoietic cells, but not the key downstream IFN-I signaling mediator, STAT1, or transcriptional regulators linked to antiviral Th1 immunity. Finally, we find that increasing IFN-I availability does not impact maximal weight loss or viral titers, but that it does promote altered expression of key inflammatory mediators, correlating with enhanced morbidity and mortality. Conclusion Our findings provide novel insights into how dysregulated IFN-I levels can impact IAV infection. More broadly, they reveal that testing how elevated cytokine levels affect outcomes can provide potentially valuable translational insights that could go unnoticed when relying on loss-of-function studies alone. Funding Source NIH Topic Categories Viral Immunology (VIR)
Short-term rapamycin treatment of an older human cohort alters immune and inflammatory markers but fails to re-set the epigenetic biological clock 2259884
Abstract Introduction Pharmacological inhibition of the mTOR pathway with rapamycin (RAPA) extends lifespan and improves aspects of healthspan in mice. Enhanced longevity was achieved even when treatment was initiated later in life, suggesting that RAPA could be used to slow progression of age-associated pathologies in older humans. Moreover, in some animal models, intermittent treatment with mTOR inhibitors elicited long-term beneficial outcomes while obviating adverse effects. Methods To determine whether short-term RAPA treatment was able to reverse age-associated deficits in humans, we undertook a placebo-controlled study of 1mg/d RAPA in older human subjects (PMID: 29408453). Using specimens available from this pilot trial, we have now characterized potential changes in immune, epigenetic, and microbiome parameters known to be negatively impacted by aging. Results RAPA effects on immune cells were observed, including increases in a unique CD11b+ myeloid subset which shares some features with MDSCs and in foxp3+ TREGS. An increase in suppressive cell subsets may be beneficial in preventing autoimmunity. Indeed, the 8-week RAPA treatment slightly decreased autoantibody levels in subjects with detectable titers. Treatment was also associated with concomitant decreases in sICAM-I and increases in sRAGE. Given the interplay between the microbiome, inflammation, and aging, it was of interest that the gut microbiota became more diverse within the first 6 weeks of RAPA treatment. On the other hand, short-term RAPA treatment did not alter the “epigenetic age” in PBMCs. Conclusion Although many RAPA effects could be elicited by short-term inhibition of mTOR, other parameters will likely require a longer treatment. To explore this possibility, we were recently funded for a PK/PD analysis of mTOR inhibition in older humans. The goal is to identify the optimal drug (RAPA vs. Everolimus), dose, and dosing frequency (daily vs. intermittent) that can be administered safely, tolerably, and long-term in humans (ages 65-90 yrs). Funding Source NIA [Pepper Center pilot grants; NIA/NIH R21; and NIA/NIH U01] Topic Categories Immune Mechanisms of Human Disease (HUM)
Unbiased Transcriptomic Clustering Reveals an Inflammatory PEC Niche Associated with Cytokine and RAAS Signaling in Glomerular Disease 2260566
Abstract Introduction Kidney diseases are a major cause of morbidity and mortality worldwide. Glomerular pathologies constitute a substantial subset and are typically diagnosed based on histopathological features from kidney biopsy. However, patients with the same pathological diagnosis often present with heterogeneous molecular signatures and clinical outcomes. Because the kidney is a compact organ composed of highly specialized cell types, bulk RNA sequencing has limited ability to resolve cell type—specific molecular alterations. Recent advances in single-nucleus RNA sequencing have enabled transcriptome-level analysis at cellular resolution, and this technology has already revealed novel molecular subtypes in several diseases. Methods In this study, we generated a single-nucleus glomerular disease atlas from treatment-naïve human kidney biopsies and applied a unified pseudobulk-based clustering strategy across major kidney cell types. Results Unbiased transcriptomic clustering identified a previously unrecognized sample cluster characterized by an enrichment of inflammatory parietal epithelial cells (PECs) expressing high levels of chemokines and CD44. These “activated PECs” were spatially localized to the glomerular tip region and displayed strong STAT1 activity, implicating convergent inflammatory signaling. In parallel, macula densa and juxtaglomerular cells within the same cluster exhibited RAAS pathway activation, suggesting a coordinated niche-level inflammatory-RAAS response. Conclusion This inflammatory PEC—centered niche defines a molecular program not aligned with conventional pathological categories, underscoring the limitation of histology-based classification. Our findings propose an inflammation—RAAS—PEC activation axis as a shared pathogenic mechanism and nominate STAT1 signaling as a potential therapeutic target in glomerular disease. Funding Source n/a Topic Categories Immune Mechanisms of Human Disease (HUM)
Reliable and Reusable All‐Solid‐State Contact‐Type Pre‐Lithiation Platform for High‐Performance All‐Solid‐State Batteries
ABSTRACT All‐solid‐state batteries represent a promising approach for achieving high energy density and enhanced safety by utilizing bipolar cell architectures and nonflammable solid electrolytes. However, lithium loss during initial cycling occurs primarily due to the limited electrochemical stability of solid electrolytes and the irreversibility of anode materials with high theoretical capacities, resulting in a lower achievable energy density than theoretically expected. Additionally, all‐solid‐state batteries are highly sensitive to interfacial phenomena, which makes addressing these challenges crucial for maximizing their electrochemical performance. In this study, we introduce a reliable and reusable contact‐type pre‐lithiation platform specifically designed for all‐solid‐state systems. This platform, consisting of solid electrolytes and electron‐conductive agents, exhibits balanced ionic and electronic transport characteristics, enabling uniform pre‐lithiation of all‐solid‐state electrodes through simple, reversible physical contact. Even under a low stack pressure of 8 MPa, pre‐lithiation can be effectively controlled by contact time and operating temperature, while the low stack pressure effectively suppresses the formation of highly resistive decomposition products at the interface. Consequently, the precisely pre‐lithiated anodes with optimized interfacial characteristics significantly enhance Coulombic efficiency during initial cycles and overall cycling performance, contributing to the realization of high‐performance all‐solid‐state batteries with genuinely high energy density.
Stk11 loss in lung adenocarcinoma increases tumor CXCL16 and is associated with an expansion of Areg+ CXCR6+ IL-17-producing γδ T cells 2310280
Abstract Introduction gd T cells have been shown to both promote tumor growth/metastasis and to efficiently kill tumor cells. These contrasting roles have been attributed to their different functional subsets, with gdT1 cells being associated with anti-tumor immunity, while gdT17 cells are associated with tumor growth and metastasis. However, the factors that regulate the activation and expansion of these functional subsets, as well as the mechanisms through which they regulate tumor growth are still unclear. Methods To investigate these questions, we developed a novel inducible model of Scgb1a1-linked Kras-driven lung adenocarcinoma with or without an Stk11 co-mutation and used multi-color flow cytometry to characterize the tumor-immune microenvironment and single-cell RNAseq to examine both the gd T cells and the tumor lung epithelium. Results Upon tumor induction KrasG12D/Stk11fl (KS) mice exhibited significantly higher tumor burden and faster disease progression than KrasG12D (K) mice. The more rapid disease progression in these mice was associated with a significant increase in Vg4 and Vg6 gdT17 cells, but not Vg1 gdT1 cells. scCITEseq revealed significant changes in the transcriptional diversity of the KS lung gdT17 cells that was associated with an expansion of an Areg+ Vg6 gdT17 subset. There was no significant difference, however, in gd T cell proliferation. Investigation of the Scgb1a1+ lung epithelium revealed that KS tumors upregulate the expression of numerous immune-relevant chemokines and cytokines, including Cxcl16. When cultured in vitro, tumor cells but not healthy epithelial cells from KS mouse lungs produced significant levels of CXCL16. Finally, we demonstrated that lung gdT17, but not gdT1 cells, express the CXCL16 receptor CXCR6. Conclusion Taken together, these data show that tumor-intrinsic Stk11 loss in lung adenocarcinoma is associated with an expansion of lung gdT17 cells and supports a model in which these pro-tumor cells are recruited to the KS mouse lung via a CXCL16-CXCR6 axis. Funding Source n/a Topic Categories Tumor Immunology: Cellular Responses and Tumor Microevironment (TIME)
A novel engineered cytokine immunotherapy reduces clinical features of disease in a mouse model of steroid-resistant asthma 2253062
Abstract Introduction Asthma is a disease of chronic airway inflammation associated with heterogenous immune phenotypes. While current immunotherapies have shown efficacy in treating allergic asthma, steroid resistant asthma critically lacks a druggable target, making it a major contributor of asthma morbidity and mortality. Here, we developed a cytokine immunotherapy to ameliorate clinical and pathological features of asthma in the steroid-resistant A/J mouse strain. Methods A/J mice subjected to house dust mite extract-induced allergy to induce asthma pathology. Mice were then subjected to methacholine induced airway hyperresponsiveness challenges to evaluate asthma physiology. The immunological makeup of the lungs and bronchoalveolar space were also evaluated with flow cytometry. Results Mice treated with the engineered cytokine exhibited reduced airway hyperresponsiveness to methacholine challenges, in contrast to those treated with dexamethasone. Notably, dexamethasone-treated mice presented with worse airway hyperresponsiveness than untreated controls. Histology demonstrated a reduction of airway inflammation and pathologic markers of disease whereas cytometry analysis revealed suppressed airway eosinophilia and neutrophilia. Cytokine immunotherapy reduced features associated with CD4+ T cell activation while increasing molecular markers of regulatory T cell activity. Conclusion Taken together, this cytokine immunotherapy has demonstrated promising results at ameliorating disease features in a pre-clinical model of steroid-resistant asthma which could lead towards a scalable and translatable treatment for patients with asthma. Funding Source NIH T32 (Medical Scientist Training Program), AHA Predoctoral Fellowship Topic Categories Immediate Hypersensitivity, Asthma, and Allergic Responses (HYP)
Meta-Data Informed Microphysiological System Replicates in vivo Macrophage Regulation of CD8+ T Cell Migration in Renal Cell Carcinoma 2254287
Abstract Introduction CD8+ T cells are abundant in clear cell renal cell carcinoma (ccRCC) yet are often retained at the tumor margin, a hallmark of immune exclusion that limits immunotherapy efficacy. Tumor-associated macrophages (TAMs) may regulate this trafficking, but the specific myeloid subsets affecting post-extravasation T-cell migration remain poorly defined. Standard in vitro assays rarely capture the full sequence from T cell transendothelial migration to directional movement toward tumor and macrophage-mediated rejection, and these dynamics are hard to monitor in vivo. Improved in vitro models that are physiologically relevant, are needed to dissect macrophage control of T-cell homing in ccRCC. Methods To build a patient metadata-informed ccRCC microphysiological system (MPS),we performed the first ccRCC single-cell meta-analysis (>1 million cells) and applied Monocle3 pseudotime to define myeloid states and trajectories, identifying a transitional macrophage population. Guided by these insights, we engineered a T cell migration MPS consisting of a perfusable HUVEC-lined lumen in 3D collagen, primary ccRCC spheroids, ex vivo generated transitional macrophages, and CD8+ T cells. The platform quantifies CD8+ T-cell transendothelial and interstitial migration. Results Our myeloid meta-analysis identified an IL10/IL1B-high transitional state between monocytes and pro-inflammatory TAMs, upstream of all other immunosuppressive TAMs by pseudotime. We generated IL10/IL1B-high—like TAMs through tumor co-culture. When incorporated into the MPS, these TAMs reduced CD8+ T-cell extravasation and curtailed interstitial migration, suggesting differentiation towards TREM2+ immunosuppressive TAMs. Conclusion We defined and recreated a novel IL10/IL1B-high transitional state between monocytes and TAMs. Integrating meta-analysis with ex vivo MPS, we enable trajectory reconstruction from tumor-recruited monocytes toward immunosuppressive TAMs for mechanistic study and therapeutic targets discovery in ccRCC. Funding Source Carbone Cancer Center Cancer Center Support Grant NIH P30CA014520 Topic Categories Tumor Immunology: Cellular Responses and Tumor Microevironment (TIME)
Characterizing Host-Viral Interaction Interfaces for Insights into Viral Transmission and Infections 2257274
Abstract Introduction Viruses hijack human cellular machinery through protein—protein interactions (PPIs). Since interaction interfaces play a critical role in modulating PPIs, studying human-viral interaction interfaces can provide a novel perspective on viral immunomodulatory mechanisms underlying disease pathogenesis. Methods We used AlphaFold-Multimer (AFM) to structurally resolve 11,667 experimentally validated human—viral PPIs spanning 33 viral families to create comprehensive 3D interactomes. Despite the absence of paired multiple-sequence alignments, AFM outperformed other methods in accurately identifying interaction interfaces. We then systematically analyzed these 3D interaction models to reveal signatures of selection, conservation, and molecular mimicry at human—viral interfaces. Results Using 1000 Genomes data, we found that variants at interfaces exhibited population-specific selection patterns, particularly for long-coevolving viruses such as HIV. Interestingly, when the evolutionary patterns of interface and non-interface residues were compared, interface residues at human PPIs were more conserved, as expected. However, this pattern was reversed for viral residues. Systematic analyses of human interface mimicry by viral proteins revealed mutation rate-dependent differences, with higher viral mutation rates being associated with greater mimicry. Similar mutation rate-dependent differences were seen in the interface sharing between viral proteins binding to a host protein. Moreover, we identified a novel example of herpes simplex virus protein UL37 mimicking the TRAF6-binding domain on MAVS, which potentially inhibits interferon signaling by disrupting MAVS—TRAF6 interactions. Conclusion Overall, our comprehensive 3D viral interactomes provide a rich resource for exploring viral adaptation, immune evasion, and pathogenesis mechanisms, empowering researchers to generate testable hypotheses and ultimately accelerate the discovery of novel therapeutic targets and intervention strategies. Funding Source n/a Topic Categories Computational and Systems Immunology (COMP)