In-Press Preview
Abstract
Tuberous sclerosis complex (TSC) is an inherited tumor predisposition syndrome caused by pathogenic variants in TSC1 or TSC2, yet no animal model faithfully recapitulates the cardiac rhabdomyomas (CR) that are among its earliest manifestations. We generated a heterozygous TSC2-mutant Yucatan minipig carrying the recurrent pathogenic TSC2 p.Arg505* variant and characterized its cardiac phenotype using histopathology, immunohistochemistry, and RNA sequencing. TSC2-mutant minipigs consistently developed multiple CR that closely resembled human lesions, including enlarged glycogen-rich spider cells and activation of mechanistic target of rapamycin complex 1 (mTORC1) signaling. Comparative transcriptomic analyses demonstrated enrichment of a conserved TSC-associated gene expression program and activation of microphthalmia/transcription factor E (MITF/TFE)-regulated lysosomal pathways in both human and minipig tumors. Although other manifestations of TSC were not observed during the first year of life, this large-animal model faithfully reproduced the principal histopathologic and molecular features of human CR. These findings have established the TSC2-mutant Yucatan minipig as what we believe to be the first large-animal model of TSC-associated CR and provide a physiologically relevant platform for investigating tumor pathogenesis, mechanisms of spontaneous regression, and therapeutic strategies targeting mTORC1- and MITF/TFE-dependent signaling.
Authors
Elias Bou Farhat, Heng Du, Chrystalle Katte Carreon, Jad Halawi, Sophie Kudler, Magdalena Losko, Zachary T. Herbert, Adrienne L. Watson, Daniel Carlson, Dennis A. Webster, Derek M. Korpela, David M. Meredith, Elizabeth P. Henske, David J. Kwiatkowski
Abstract
Osteoarthritis (OA) is a heterogeneous disease in which synovial inflammation drives structural progression and pain, yet the molecular diversity of synovitis remains poorly defined. Here, we performed a multimodal characterization of synovial tissues from 48 dogs with naturally occurring OA, integrating histopathology, immunohistochemistry, transcriptomics, MRI, and soluble mediator profiling. This analysis identified three discrete synovial pathotypes, pauci-immune, diffuse myeloid, and lympho-myeloid, defined by distinct immune cell composition and transcriptional programs reflecting biologically distinct endotypes. The lympho-myeloid pathotype, characterized by ectopic lymphoid structures and adaptive immune enrichment, was independently associated with greater structural joint damage on MRI despite comparable clinical scores, demonstrating that tissue stratification captures heterogeneity invisible to clinical assessment alone. Composite soluble mediator signatures correlated with disease severity across pathotypes. Cross-species analyses integrating a large human synovial cohort revealed conserved pathotype architecture and shared gene networks, including tyrosine kinase signaling and B cell receptor activation, validating the translational relevance of the canine model. Together, these findings provide a framework for patient stratification and therapeutic target discovery in OA, with broad implications for precision medicine within a One Medicine perspective.
Authors
Anaëlle Drieu-Vincent, Julien De Lima, Floriane M. Etienne, Romain Guiho, Noémie Podevin, Richard Danger, Florence Lezin, Nicolas Gaigeard, Sophie Allain, Jérôme Abadie, Thomas Rousseau, Pierre Maitre, Olivier Gauthier, Jérôme Guicheux, Marion S. Fusellier, Marie-Astrid Boutet
Abstract
Alveolar microcysts, a hallmark of Idiopathic Pulmonary Fibrosis (IPF), are basal cell (BC)-lined structures that develop near terminal airways and correlate with poor clinical outcomes. Using primary-cell organoids and spatial transcriptomics, we identified striking epithelial similarities between respiratory bronchioles (RBs) and IPF alveolar microcysts, both containing AT0 (ABCA3+/SCGB3A2+), SCGB3A2+, and basaloid/BC populations, although basaloid/BCs were markedly expanded in IPF microcysts. A CEACAM6+ subset of normal RB epithelial cells, termed respiratory airway secretory cells (RASCs), consisted predominantly of SCGB3A2+ cells with only rare BCs (1–5%) that by RNA trajectory analysis suggested BCs arise from SCGB3A2+ progenitors. Accordingly, in organoid cultures nearly all RASC-derived colonies converted to BCs within 9 days, resembling colonies generated by purified BCs. BCs derived from either RASCs or normal alveolar type II (AT2) cells strongly expressed the collagenase matrix metalloproteinase-1 (MMP1), recapitulating the phenotype of BCs lining IPF microcysts. MMP1high BCs degraded type I collagen ex vivo and generated microcyst-like structures in vivo following xenotransplantation, both abolished by MMP activity inhibition or specific MMP1 deletion by sh-RNAi. Finally, a Frizzled-5 WNT agonist antibody reversed the MMP1high phenotype, identifying AT2- and RB-derived basal cell transdifferentiation as a previously unrecognized driver of alveolar destruction and microcyst formation in IPF and a potential therapeutic target.
Authors
Marylene Toigo, Kiana Garakani, Alexis N. Brumwell, Ying Wei, Tsung Che Ho, Paul J. Wolters, Michael A. Matthay, Stefano A. Iantorno, Vincent C. Auyeung, Harold A. Chapman, Claude Jourdan Le Saux
Abstract
In alcohol use disorder (AUD), duodenal macrophage number are reduced, but the underlying mechanisms and consequences for gut permeability remain unknown. Actively drinking AUD patients undergoing standardized inpatient detoxification and healthy controls were enrolled. Fasting blood samples and duodenal biopsies were analyzed using proteomics, histology, multiplex immunohistofluorescence, flow cytometry, and quantitative polymerase chain reaction. Circulating monocytes were differentiated ex vivo into macrophages to assess migration, differentiation, phagocytosis, and cytokine release following stimulation with bacterial products. Gut permeability was evaluated using urinary 51Chromium- ethylenediaminetetraacetic acid excretion, and plasma surrogate markers of microbial translocation. AUD patients showed markedly reduced duodenal macrophages despite increased circulating monocytes and tissue monocyte-attracting chemokines. AUD monocytes exhibited reduced CCR2 expression and downregulation of tissue proteins involved in adhesion and trans-endothelial migration, thereby decreasing mucosal infiltration. Early macrophage differentiation associated was impaired, with reduced CSF1, CSF2, IL-34, and CSF1 receptor expression. Despite preserved phagocytic capacity, residual tissue macrophages displayed a mixed immune phenotype with reduced expression of scavenger receptors involved in microbial responses and cytokine hyporesponsiveness to lipopolysaccharide and peptidoglycan. Reduced macrophage numbers correlated with increased intestinal permeability and markers of microbial translocation. These results link disruption of the duodenal monocyte-macrophage axis to intestinal barrier failure in AUD.
Authors
Yifan Hu, Jerome Ambroise, Luca Maccioni, Laure-Alix Clerbaux, Caroline Bouzin, Bernd Schnabl, Peter Stärkel
Abstract
Children with cystic fibrosis (CwCF) have impaired linear growth that correlates with poor clinical outcomes. Growth-stunted CwCF also exhibit a marked fecal dysbiosis, and lower serum levels of the gut microbiome-regulated growth factor IGF-1, relative to those without stunting. We hypothesized that the CF gut microbiome negatively impacts IGF-1 and skeletal growth in CwCF. To test this hypothesis, we compared bone phenotypes, IGF-1 and linear growth trajectories of C57BL/6 Cftrtm1Unc mutant mice (CF) to their wild-type littermates (WT) either colonized with (specific pathogen free, SPF) or without (germ-free, GF) normal microbiota. At 4 weeks of age, CF mice had reduced femur length and diminished serum IGF-1 compared to WT mice under SPF, but not GF, conditions. In contrast, by 14 weeks femur lengths and IGF-1 were similar in CF mice raised under both SPF and GF conditions. Accordingly, we observed significantly slower growth in CF mice under SPF, but not GF, conditions only from 1 to 4 weeks. CF mice also exhibited more pronounced relative taxonomic and functional dysbiosis at 4 compared with 14 weeks. These results suggest that gut microbiota play a critical role in early postnatal CF skeletal growth impairment, while microbe-independent mechanisms likely predominate in later life.
Authors
Xiaomeng You, Jisun Paik, Stacey Meeker, Kevin S. Mears, Christopher E. Pope, Hillary S. Hayden, Kelsi Penewit, Adam Waalkes, Sruthi Sridevan, Stephen J. Salipante, Lucas R. Hoffman, Julia F. Charles
Abstract
Basal-like double-negative prostate cancer (DNPC) and triple-negative breast cancer (TNBC) are aggressive tumors with few targeted treatment options, raising the question of whether a shared, druggable cell-surface antigen can be leveraged for effective immunotherapy. Here, we integrated analyses of independent bulk and single-cell transcriptomic datasets with mechanistic and preclinical therapeutic studies to evaluate CDH3 (P-cadherin) as a candidate target in basal-like disease. CDH3 expression was consistently enriched in basal-like subtypes across prostate and breast cancer cohorts and was preferentially expressed in basal epithelial tumor compartments. Mechanistically, CDH3 expression was governed by a YAP1–WNT5A/ROR2 signaling axis, linking basal subtype gene expression to non-canonical Wnt signaling. To exploit this target, we evaluated both a CDH3-directed antibody-drug conjugate (ADC) and CDH3-specific chimeric antigen receptor (CAR)-T cells. The ADC exhibited antigen-dependent internalization and cytotoxicity in vitro and reduced metastatic tumor burden in vivo. CDH3 CAR-T cells showed antigen-specific activation and tumor cell killing in vitro and improved tumor control and survival in metastatic prostate and breast cancer xenograft models, with additional benefit when combined with PD-1 blockade (nivolumab). Together, these findings establish CDH3 as a clinically relevant and therapeutically actionable target and support CDH3-directed ADC and CAR-T strategies for aggressive, basal-like DNPC and TNBC.
Authors
Guoqiang Liu, Shiyu Wang, Ziyu Zeng, Loan Duong, Jing Yang, Nipuni Barupala, Yun Zhao, Sharif Rahmy, Shan Feng, Xiaotian Chi, Ariana Arce, Linchuan Jia, Jun Wan, Liang Cheng, Xuemin Lu, Xin Lu
Abstract
Authors
Alejandra M. Marinelarena, Thao D. Pham, Jeffrey H. Becker, Apurba Majumder, Hidayatullah G. Munshi
Natural killer cells from children with obesity exhibit a persistent ‘trained’ response to influenza
Abstract
Children and adults living with obesity have more severe influenza A virus (IAV) infection, but the underlying mechanisms are unclear. Natural killer (NK) cells play a key role in the innate immune responses to IAV and clearance of IAV-infected cells. ‘Trained immunity’ of NK cells (and other innate immune cells) refers to an initial stimulus (i.e. obesity) inducing metabolic and epigenetic changes that make these cells hyperinflammatory upon restimulation (i.e. with IAV). Here, we tested whether obesity in children induces trained immunity in NK cells. We have show that NK cells from children with obesity have a hyperinflammatory ‘trained’ phenotype, with increased expression of activation markers TLR2, CCR2, TLR4, HLA-DR and CD11b compared to children without obesity. Furthermore, we have demonstrate that NK cells from children with obesity have a heightened ex vivo IFN-γ response to stimulation with IAV (A/Auckland/4/2009, H1N1 and HKx31, H3N2) and epigenetic changes consistent trained immunity. A decrease in BMI z-score following weight-management was not associated with a reduction in the IFN-γ response, suggesting that weight loss is not sufficient to normalise NK cell function. Better understanding of obesity-related immune modulation is critical for development of new therapeutic strategies and improving outcomes in future IAV pandemics
Authors
Ellesandra C. Noye, Megan Airey, Danielle K. Longmore, Hui Xu, David Chisanga, Christoph Saner, Meagan Carney, Meg Kaegi, Hiu On Man, M. Zeeshan Chaudhry, Liam C. Kealy, Julian D. J. Sng, Isabelle Hocking, Melanie Wu, Georgina McCallum, Gabrielle T. Belz, Kim L. Good-Jacobson, David Burgner, Siroon Bekkering, Kirsty Short
Abstract
Targeted injury to the biliary epithelium and impaired bile flow are central mechanisms underlying childhood cholangiopathies. In biliary atresia (BA), immune-mediated cholangiocyte injury drives progressive bile duct obstruction, yet epithelial-intrinsic programs regulating injury susceptibility remain incompletely defined. Here, we modeled virus-epithelium interactions using rhesus rotavirus (RRV) infection of human liver-derived cholangiocyte organoids from normal donors (NCO) and infants with BA at diagnosis (BACO-Dx) or transplantation (BACO-Tx). Compared with NCO, BA organoids displayed baseline defects in tight junction architecture and barrier function, with prominent epithelial disruption and apoptosis following RRV exposure. Transcriptomic profiling revealed that while NCO and BACO-Dx mounted predominantly interferon-associated antiviral responses, BACO-Tx exhibited an expanded inflammatory program characterized by enrichment of NF-κB (RELA/NFKB1) regulatory network at baseline and amplified by infection. Inhibition of NF-κB signaling with the hydrogen sulfide donor GYY4137 restored organoid growth, improved tight junction organization, reduced permeability, and attenuated RRV-induced apoptosis. In the murine BA model, GYY4137 improved disease severity, preserved bile duct architecture, and protected epithelial junctional integrity while limiting cholangiocyte apoptosis. These findings identify an epithelial NF-κB-predominant program as a key determinant of intrinsic susceptibility to viral injury in BA and support targeting cholangiocyte NF-κB signaling to suppress disease progression.
Authors
Wenqi Li, Ashley Fonseca-Mejia, Reena Mourya, Pavithra Sudhakara, Pranavkumar Shivakumar, Jorge A. Bezerra
Abstract
The increase in antimicrobial resistant infections has fueled a resurgent interest in bacteriophage (phage) therapy. We conducted a literature review of personalized phage therapy cases in the peer-reviewed, English-language literature reported between 1/1/2005, to 8/31/2025 for which individual patient data were available, identifying and synthesizing data from a total of 323 patients described in 83 studies. Most patients (68%, 221/323) were treated with ≥ 2 phages (a phage cocktail). The median concentration of phages was 108 PFU/mL (IQR 107–109). Most patients (76%, 246/323) received antibiotics concurrent with phage therapy. The median duration of therapy was 10 days (IQR 5–21). The most common indications for phage therapy were skin/soft tissue, pulmonary, orthopedic, osteoarticular, and genitourinary infections. 144 patients received fixed (off-the-shelf) phage preparations whereas 179 received bespoke phage preparations (i.e., personalized therapy). For cases where outcomes data were available, 78% (253/323), experienced clinical improvement while the targeted pathogen was eradicated in 61% (177/290) of cases. Nine patients had treatment withdrawn due to tolerability concerns. These data indicate that there is substantial heterogeneity in current personalized phage therapy clinical practices but that this approach can be successful.
Authors
Patrick M. Blankenberg, Robert Targ, Grace D. Cullen, Gina A. Suh, Krystyna Dąbrowska, Aarav Singh, Tiffany Luong, Daisuke Furukawa, Paul L. Bollyky
No posts were found with this tag.
