Biology subjects

Danner, R.

Publications and source records attributed to Danner, R..

5 recordsLinked to original sources

bioRxiv2026

Folic acid polyglutamylation triggers MAP2K5 signaling to induce adipocyte lipid catabolism

Adipocyte signaling adaptively responds to macronutrients, but how essential micronutrients impact lipid homeostasis remains poorly understood. Here, we demonstrate that folate polyglutamylation, the sequential conjugation of glutamate residues to folate, serves as a dynamic biochemical process regulating metabolic signaling. Using our folate metabolomics platform, we show that polyglutamylated folic acid accumulates in healthy adipose tissue, but is depleted in obesity across mice and humans, independent of circulating folate levels. Genetic ablation of the polyglutamylation enzyme folylpolyglutamate synthase (Fpgs) in adipocytes suppresses lipid catabolism to induce cell-autonomous lipid accumulation, operating independently of canonical adipogenesis or downstream one-carbon flux. Target-engagement proteomics identifies monoglutamylated folic acid as a Map2k5-interacting molecule that inhibits Map2k5 activity, whereas Fpgs-mediated folic acid polyglutamylation acts as a chemical switch that disrupts this interaction to promote lipolysis. In vivo, whole-body inhibition of Map2k5 or adipose-targeted genetic depletion of Fpgs increases body fat mass in the absence of dietary obesogenic triggers. Furthermore, single-cell and single-nuclear transcriptomic analyses establish the Fpgs-Map2k5 pathway as a core transcriptional signature of mouse and human adipose tissues. These findings uncover a non-canonical signaling role for folate that regulates adipocyte lipid homeostasis.

biochemistry↗

bioRxiv2026

Gut microbiome derived folate metabolite suppresses colorectal cancer progression

The gut microbiota influences colorectal cancer (CRC) progression, primarily through the secretion of small molecule metabolites. While numerous microbial products are known to drive CRC, endogenous protective mechanisms remain largely uncharacterized. Utilizing a folate metabolomics platform, we demonstrate that the healthy gut microbiota produces folinic acid (FA), a known chemotherapeutic adjuvant also known as leucovorin. This microbially derived folinic acid is progressively depleted in mouse models of colitis-associated CRC and in human clinical metagenomic cohorts with advancing disease severity. Mechanistically, folinic acid acts as a signaling molecule that directly binds and inhibits the intracellular protease calpain-2. This interaction stabilizes epithelial E-cadherin protein expression and suppresses CRC epithelial-to-mesenchymal transition driving metastasis. Genetically manipulating gut microbial production of FA is sufficient to modulate CRC in vivo, even in the presence of chronic inflammation. This study reframes folinic acid from a chemotherapeutic enhancer to an endogenous microbial metabolite that actively suppresses CRC progression.

cancer biology↗

bioRxiv2026

Microbial metabolism of methotrexate produces a STAT3 signaling molecule that alleviates gut inflammation

Methotrexate (MTX) therapy in inflammatory bowel disease (IBD) is often limited by inter-individual variability in clinical response and adverse effects. Gut microbiota contribute to MTX therapeutic response and toxicity by metabolizing MTX and altering its bioavailability. However, how inflammation alters microbial MTX metabolism and how its metabolites influence the host remain poorly understood. Here, we identify Clostridium asparagiforme as a potent and efficient metabolizer of methotrexate, producing deoxyaminopteroic acid (DAMPA) in the distal gastrointestinal tract. We demonstrate that DAMPA preserves mitochondrial integrity by promoting mitophagy in intestinal epithelial cells through mitochondrial STAT3 signaling. DAMPA administration attenuates intestinal inflammation in vivo, and improves metabolic dysfunction associated with IBD. Together, these findings reveal an unappreciated role for a gut microbial MTX metabolite in mediating epithelial homeostasis during intestinal inflammation, thus reframing microbial MTX metabolism from passive drug detoxification to active regulation of host mitochondrial and inflammatory homeostasis.

microbiology↗

bioRxiv2023

HLA-DR-expressing fibroblast-like synoviocytes are inducible antigen presenting cells that present autoantigens in Lyme arthritis

BackgroundHLA-DR-expressing fibroblast-like synoviocytes (FLS) are a prominent cell type in synovial tissue in chronic inflammatory forms of arthritis. We recently showed that peptides from several extracellular matrix (ECM) proteins, including fibronectin-1 (FN1), contained immunogenic CD4+ T cell epitopes in patients with postinfectious Lyme arthritis (LA). However, the role of FLS in presentation of these T cell epitopes remains uncertain. MethodsPrimary LA FLS and primary murine FLS stimulated with interferon gamma (IFN{gamma}), Borrelia burgdorferi, and/or B. burgdorferi peptidoglycan (PG) were assessed for properties associated with antigen presentation. HLA-DR-presented peptides from stimulated LA FLS were identified by immunopeptidomics analysis. OT-II T cells were cocultured with stimulated murine FLS in the presence of cognate ovalbumin antigen to determine the potential of FLS to act as inducible antigen presenting cells (APC). ResultsFLS expressed HLA-DR molecules within inflamed synovial tissue and tendons from patients with post-infectious LA patients in situ. MHC class II and costimulatory molecules were expressed by FLS following in vitro stimulation with IFN{gamma} and B. burgdorferi and presented both foreign and self MHC-II peptides, including T cell epitopes derived from two Lyme autoantigens fibronectin-1 (FN1) and endothelial cell growth factor (ECGF). Stimulated murine FLS induced proliferation of naive OT-II CD4+ T cells, particularly when FLS were stimulated with both IFN{gamma} and PG. ConclusionsMHC-II+ FLS are inducible APCs that can induce CD4+ T cell activation and can present Lyme autoantigens derived from ECM proteins, thereby amplifying tissue-localized autoimmune CD4+ T cell responses in LA. AUTHORS SUMMARYThis study demonstrates that IFN{gamma}-activated MHC-II+ fibroblast-like synoviocytes (FLS) stimulated with Borrelia burgdorferi present foreign and self MHC-II antigens, including Lyme autoantigens. Furthermore, IFN{gamma}-activated MHC-II+ FLS stimulated with B. burgdorferi peptidoglycan can induce activation and proliferation of naive CD4+ T cells in an MHC-II antigen-dependent manner, demonstrating that activated MHC-II+ FLS are inducible antigen presenting cells.

immunology↗

bioRxiv2023

Epitope spreading of Lyme autoantigen apoB-100 and CD4+ T cell responses to Borrelia burgdorferi Mcp4 are regulated by IL-10 in murine Lyme disease

Borrelia burgdorferi, the causative agent of Lyme disease (LD), has evolved immune evasion mechanisms to establish a persistent infection in their vertebrate hosts, resulting in chronic inflammation and autoimmune T and B cell reactivity in many B. burgdorferi-infected individuals. In this study, we used an unbiased immunopeptidomics approach to identify foreign and self MHC class II peptides isolated from inguinal and popliteal lymph nodes from B. burgdorferi- infected C57BL/6 (B6) mice, which develop mild, self-limiting LD; and from infected B6 Il10-/- mice, which develop severe, persistent LD. Nearly 10,000 MHC-II peptides were identified by LC-tandem MS analysis which included many peptides derived from proteins abundant in arthritic joints that are associated with inflammation, tissue repair, and extracellular matrix remodeling. Notably, the number and variety of unique peptides derived from apolipoprotein B- 100 (apoB-100); a validated autoantigen in human Lyme arthritis (LA), atherosclerosis, and liver disease; was greatly expanded in lymph nodes of infected mice, particularly in Il10-/- mice at 4 weeks (6-fold increase) and 16 weeks (15-fold increase) post-infection, compared with uninfected mice, indicating epitope spreading. One of the apoB-100 peptides identified in infected, but not uninfected, B6 and Il10-/- mice was APOB3500-3515, an immunogenic cryptic epitope in murine autoimmune atherosclerosis. No apoB-100 peptides had sequence homology to any B. burgdorferi antigens. Surprisingly, only six peptides derived from B. burgdorferi proteins were validated in this study. One of these B. burgdorferi epitopes, derived from methyl- accepting chemotaxis protein Mcp4 (BB0680), was an immunogenic target of CD4+ T cell responses in B. burgdorferi-infected Il10-/- mice, but not in B6 mice. In conclusion, this study has shed light on the importance of IL-10 in suppressing epitope spreading and limiting B. burgdorferi-specific CD4+ T cell responses. Furthermore, this study supports epitope spreading and exposure of cryptic antigens as likely mechanisms of infection-induced apoB-100 autoimmunity in LD. AUTHOR SUMMARYLyme disease is caused by infection with the spirochetal pathogen Borrelia burgdorferi, and affects [~]500,000 individuals in the U.S. annually. T cell responses to both host and pathogen are dysregulated during infection, resulting in chronic infection and frequent development of autoimmunity. To assess the immune-relevant CD4+ T cell epitopes presented during development of Lyme disease, we used an unbiased, immunopeptidomics approach to characterized the MHC class II immunopeptidome in mice infected with Borrelia burgdorferi. We identified nearly 10,000 unique peptides. Peptides derived from apoB-100, a known human Lyme autoantigen, were highly enriched in infected mice, compared with uninfected controls, and showed evidence of epitope spreading. Furthermore, we identified several peptides derived from Borrelia burgdorferi, including one immunogenic peptide from a methyl-accepting chemotaxis protein, Mcp4. Interestingly, both apoB-100 epitope spreading and immune responses to Mcp4 were observed in mice lacking the anti-inflammatory cytokine IL-10, indicating an important role of IL-10 in suppressing T cell responses to Mcp4 and epitope spreading of Lyme autoantigen apoB-100.

immunology↗