Biology subjects

Roewe, J.

Publications and source records attributed to Roewe, J..

3 recordsLinked to original sources

bioRxiv2025

NGN2 Expression and Regional Patterning Allow Rapid Differentiation from hiPSCs to DRG-Like Neurons Responsive to Type 2 Cytokines

Itch or pruritus, is a sensation that elicits scratching behaviour and is a major symptom and cause of morbidity in skin diseases such as atopic dermatitis (AD), allergic contact dermatitis (ACD), prurigo nodularis (PN), and urticaria. Itch is often triggered by inflammatory stimuli in the skin including type 2 cytokines such as IL-4, IL-13, and/or IL-31. Several therapies targeting type 2 immune pathways have been developed to treat pruritus; however, itch improvement in many patients remains to be improved. Thus, additional approaches to modulate sensory neuron activity are needed. Ex vivo or even in vitro study of the molecular mechanisms underlying primary sensory neuron activation is challenging since harvesting neurons from dorsal root ganglia (DRG) in patients can only be done from cadavers. Herein, we describe rapid human sensory neurons generation (2 days to precursor cells) by in vitro differentiation of human induced pluripotent stem cells (hiPSC) from simultaneous application of patterning factors with NGN2 overexpression. We show that these hiPSC-derived sensory neurons possess key characteristics of primary sensory neurons. They express key neuronal markers, such as TRKA receptors, TRPV1 and TRPA1 channels, and functionally respond to the TRPV1 agonist capsaicin. In addition, they express key type 2 cytokine receptors such as interleukin (IL)-4R and IL31-R, known to promote itch in AD and PN. Moreover, these cells are functional as our sensory neurons respond to IL-4, IL-13 and IL-31 stimulation. Collectively, these data demonstrate that our protocol generates a phenotypic profile consistent with native somatosensory neurons that can facilitate development of novel approaches to model and treat pruritic disease.

neuroscience↗

bioRxiv2022

Bacterial Polyphosphates Induce CXCL4 and Synergize with Complement Anaphylatoxin C5a in Lung Injury

Polyphosphates are linear polymers of inorganic phosphates that exist in all living cells and serve pleiotropic functions. Bacteria produce long-chain polyphosphates, which can interfere with host defense to infection. In contrast, short-chain polyphosphates are released from platelet dense granules and bind to the chemokine, CXCL4. Here, we report that long-chain polyphosphates induced the release of CXCL4 from mouse bone marrow-derived macrophages and peritoneal macrophages in a dose-/time-dependent fashion resulting from an induction of CXCL4 mRNA. This polyphosphate effect was lost after pre-incubation with recombinant exopolyphosphatase (PPX) Fc fusion protein, demonstrating the potency of long chains over monophosphates and ambient cations. In detail, polyphosphate chains longer than 70 inorganic phosphate residues were typically required to mediate robust CXCL4 release. Polyphosphates acted independently of the purinergic P2Y1 receptor and the MyD88 and TRIF adaptors of Toll-like receptors. On the other hand, polyphosphates augmented LPS/MyD88-induced CXCL4 release, which was explained by intracellular signaling convergence on PI3K/Akt. Polyphosphates alone induced phosphorylation of Akt at threonine-308. Pharmacologic blockade of PI3K (wortmannin, LY294002) antagonized polyphosphate-induced CXCL4 release from macrophages. Intra-tracheal polyphosphate administration to C57BL/6J mice caused histologic signs of lung injury, disruption of the endothelial-epithelial barrier, influx of Ly6G+ polymorphonuclear neutrophils, depletion of CD11c+SiglecF+ alveolar macrophages, and release of CXCL4. Long-chain polyphosphates synergized with the complement anaphylatoxin, C5a, which was partly explained by upregulation of the receptor, C5aR1, on myeloid cells. C5aR1-/- mice were protected from polyphosphate-induced lung injury. In conclusion, we demonstrate that polyphosphates govern immunomodulation in macrophages and are capable of inducing lung injury.

immunology↗

bioRxiv2021

Proteome Microarray Screening Identifies Human Polyphosphate-Binding Proteins in the Phosphatidylinositol Signaling Pathway

Polyphosphates are linear chains of orthophosphate residues that are present in all living cells. Polyphosphates are released from platelet d-granules and are also produced in bacteria. Polyphosphates are procoagulant in mammalian species and in bacteria are required for energy and phosphate storage, stress resistance, chelation of metal ions and escaping host immunity. Despite these pleiotropic effects, sparse information is available on molecular binding partners of polyphosphates. Here, we used a slide-based human proteome microarray screen for the search of polyphosphate-binding proteins. This approach suggested several novel proteins with relation to the phosphatidylinositol signaling pathway. The highest signals were obtained for Disabled-1 (DAB1) and phosphatidylinositol-5-phosphate 4-kinase 2B (PIP4K2B). Isothermal titration calorimetry was used for confirmation of DAB1 interactions with long-chain polyphosphates. These results offer new rationale to further investigate the interference of polyphosphates with intracellular signaling pathways.

immunology↗