Biology subjects

Reed, A. K.

Publications and source records attributed to Reed, A. K..

4 recordsLinked to original sources

bioRxiv2026

Gradient Multinozzle 3D Printing

Direct ink writing is compatible with an expansive materials palette. While enabling diverse applications, this materials versatility brings significant bottlenecks in ink formulation, often requiring the mixing, printing, and testing of dozens to hundreds of ink compositions over the course of a project. To accelerate ink-space exploration, we introduce gradient embedded multinozzle (GEM) printheads that combine the high-throughput parallelized printing of multinozzles with combinatorial ink mixing. These printheads allow simultaneous mixing of two-, three-, and four-input inks which are distributed to printer nozzles to create complex 3D structures with graded compositions of inks. Using a two-way GEM printhead, we vali-date cell compatibility by printing scaffolds containing various concentrations of fibroblasts and observing non-linear compaction behaviours. We next test a three-way GEM multinozzle to print ten compositions of di- and multi-functionalized poly(ethylene-glycol) diacrylate hydrogel tri-leaflet valves, optimizing for stiffness, swelling ratio, and toughness. Our GEM multinozzles are compatible with open-source printers and either pressure- or volume-driven extrusion systems and promise to accelerate iterative ink design and testing.

bioengineering↗

bioRxiv2026

Targeting AKAP13 RhoGEF activity ameliorates pro-fibrotic phenotypes driven by the IPF associated AKAP13 risk variant

RationaleIdiopathic pulmonary fibrosis (IPF) is a progressive, incurable scarring disease of the lung. A common genetic variant near AKAP13, a multifunctional scaffold protein that integrates intracellular signalling through its interactions with RhoA and protein kinase A (PKA), has been associated with IPF susceptibility and elevated AKAP13 mRNA expression in lung tissue from patients. However, its contribution to the pathogenesis of IPF remains unclear. ObjectiveThis study investigates how an AKAP13 variant alters epithelial signalling and evaluates the therapeutic potential of targeting AKAP13. Findingsrs62025270-bearing iHBECs exhibited selective upregulation of AKAP13 isoforms, accompanied by increased cell adhesion and reduced proliferation. Transcriptomic profiling revealed upregulated fibrosis-related genes in rs62025270-bearing iHBECs, including SAA1, FGF2, MMP1, CTSB, COL4A1, and CDKN1A. rs62025270-bearing iHBECs also displayed increased RhoA activation and SMAD2 phosphorylation following LPA stimulation. Furthermore, cells harbouring the AKAP13 variant showed reduced intracellular cAMP levels. Pharmacological inhibition of AKAP13 with A13 reversed the pro-adhesive phenotype and reduced RhoA activation in iHBECs. Moreover, in IPF-derived PCLS, A13 suppressed SERPINE1, CCN2, and MMP7 expression, reduced SMAD2 nuclear translocation, and decreased hydroxyproline levels. ConclusionsPresence of an AKAP13 variant disrupts epithelial homeostasis and promotes pro-fibrotic signalling. Inhibition of AKAP13s RhoGEF domain with A13 restores epithelial function and attenuates fibrotic activation, supporting AKAP13 as a therapeutic target in IPF.

genetics↗

bioRxiv2025

Selective targeting of IL-1RAP-dependent eosinophilic inflammation in allergic fungal airway disease

It is estimated that in excess of 10 million people around the globe are affected by severe asthma and fungal sensitisation (SAFS) or allergic bronchopulmonary aspergillosis (ABPA), severe asthma endotypes driven by hypersensitivity to environmentally ubiquitous fungal pathogens, primarily Aspergillus fumigatus. Here, we sought to define the immunological pathways underlying these allergic fungal airway diseases. To do so, we exploit the chronic exposure repeat challenge model using live A. fumigatus conidia to systematically define the key immunological pathways driving airway inflammation in allergic fungal airway disease. In response to daily intranasal challenge, we observed increased absolute numbers of neutrophils and eosinophils in bronchoalveolar lavage fluid (BALF), characteristic of human allergic fungal airway disease, with significant depletion of the alveolar macrophage population. Transcriptomic analysis of BALF cells identified increased expression of IL-1 family cytokines and receptors including IL-1{beta}, IL-1RL1, IL-1R2 and the IL-18 binding protein. Complementary proteomic analysis of BALF revealed increased levels of cell death related proteins calprotectin and IL-1 Receptor Accessory Protein (IL-1RAP). Targeting IL-1RAP, using knockout mice, led to selective reduction in eosinophilia, IL-5 and IL-13 in the airways without impairment of fungal killing. This study identifies a role for IL-1RAP in the generation of eosinophilia independent of neutrophil influx and highlights its potential as a novel immunotherapeutic target for the treatment of allergic fungal airway disease. Author SummaryAllergic bronchopulmonary aspergillosis (ABPA) is a form of lung disease primarily affecting those with asthma or cystic fibrosis induced by the ubiquitous pathogen Aspergillus fumigatus. Understanding the immunological mechanisms contributing to this airway disease is important for the development of novel treatments to improve quality of life and preserve lung function. In this study, we show that an Aspergillus fumigatus repeat challenge model phenocopies immunological features of allergic fungal airway disease and is hallmarked by increased IL-1 family signalling. We identify the IL-1RAP as a contributor to eosinophilia and the release of Type 2 cytokines during allergic fungal airway disease. This work represents major step in describing the molecular mechanisms of airway eosinophilia in response to fungal exposure and lays the groundwork for further dissection of inflammatory pathways to target with immunotherapeutic approaches.

immunology↗

bioRxiv2024

Multiscale single-cell assessment of the fibrotic niche in idiopathic pulmonary fibrosis

BackgroundIdiopathic pulmonary fibrosis (IPF) is a progressive, fatal disease characterised by excessive extracellular matrix deposition within the lung. Recent advances in single-cell RNA sequencing have identified distinct fibrotic populations, yet their origins and spatial relationships remain incompletely understood. MethodsUsing spatial transcriptomics and Hyperion imaging mass cytometry we compared the cellular composition in formalin fixed paraffin embedded fibrotic lesions (n=9 patients) with control lung (n=9), and cellular interactions were inferred using CellChat V2 ligand-receptor analysis. Monolayers of airway epithelial cells were used to identify changes in keratin (KRT) expression following cell detachment and cyclical mechanical stretch. ResultsSpatial multiomics profiling of human lung cells confirmed the in situ localisation of previously described IPF-enriched populations, and identified a previously unrecognized KRT5low/KRT17 epithelial population derived from airway basal cells that progressively acquiring molecular features of aberrant basaloid cells, forming a unique fibrotic niche enriched with the Secreted Phosphoprotein 1 (SPP1) positive macrophages. Functional studies demonstrated that epithelial detachment and cyclical mechanical stretch drive KRT5 reduction, providing a mechanism for the emergence of this transitional state. In addition, we also identified distinct immune-stromal niches enriched in lymphocytes and alveolar fibroblasts. ConclusionThese findings delineate distinct fibrotic epithelial niches in IPF and support a model in which epithelial loss induces aberrant basaloid differentiation and fibroblast activation, with subsequent airway traction and epithelial detachment generating a secondary niche enriched in basal-derived KRT5low/KRT17 cells and SPP1 macrophages.

molecular biology↗