Biology subjects

Pellicoro, A.

Publications and source records attributed to Pellicoro, A..

5 recordsLinked to original sources

bioRxiv2026

Beyond Navigation - Tissue-contacting Fluorescent Lifetime Imaging reveals a pathology-linked lung cancer phenotype at the point of biopsy

Background: Accurate sampling of suspected peripheral lung cancers depends on access to the lesion and confirmation that the biopsy tool is in contact with target tissue. Current bronchoscopic navigation and imaging techniques can guide instruments to a target but do not provide real-time biological confirmation at the point of sampling. Fluorescence lifetime imaging microscopy (FLIM) provides molecular contrast by measuring fluorescence decay - how long photons continue to be emitted from fluorescent molecules. In the Precision Lung clinical study (ISRCTN15093468), the Prothea Imaging System (Generation 1) identified a candidate tumour-associated phenotype of spatially overlapped low fluorescence lifetime and low intensity (LLLI) from in-vivo imaging. We used this observation as the basis for a reverse-translational study to determine whether the LLLI phenotype is linked to cancer pathology; reproducible with the Imaging System (Generation 2); and distinguishable from normal lung tissue. Methods: Previously reported Precision Lung findings were used as the clinical starting observation and were not re-analysed. Validation was then performed using: (i) pathology linked benchtop FLIM of early-stage non-small-cell lung tissue microarrays encompassing malignant cell clusters of approximately 300 um2, matched to the EoT imaging scale; (ii) five sequential fresh lung-cancer resections imaged at tumour and comparator regions, including visibly blood-rich contact sites, using the (Generation 2) Imaging System; and (iii) systematic mapping of two ventilated non-cancer donor lungs, one from a smoker and one from a non-smoker, across all available lobes. The LLLI phenotype was defined as spatial co-localisation of low intensity and short lifetime. Results: Using a real time fibre based FLIM system, capable of deployment through a working channel of a bronchoscope, the LLLI tumour phenotype was optically identified in freshly resected tumour tissue. The same phenotype was identified in fixed tissue samples with known pathology, and with images taken in the Precision Lung clinical study. Whole human lung controls did not show evidence of the tumour phenotype. Conclusions: This evidence forms a reverse-translational chain that supports the concept of the Prothea Imaging System - as a platform that confirms that the tool is in contact with a region of cancer in the lesion, while preserving continuous access for biopsy or intervention.

cancer biology↗

bioRxiv2026

Therapeutic signature mapping of paired direct and indirect LPS injury in an ex vivo human lung perfusion platform reveals injury-specific druggable programs

Acute Respiratory Distress Syndrome (ARDS) remains highly morbid and lacks approved disease-modifying pharmacotherapies. Direct (pulmonary) and indirect (extrapulmonary) insults may initiate biologically distinct early injury programs, but human tissue-level evidence from the first hours is scarce. Here we establish a paired, acellular ex vivo lung perfusion (EVLP) platform using human donor lungs unsuitable for transplantation to model direct (endobronchial) and indirect (perfusate) lipopolysaccharide (LPS) injury within the same donor. We profiled lung tissue proteomes at 4 h post-insult and performed therapeutic nomination by querying proteomics-derived injury signatures against the CLUE L1000 perturbational compendium with independent cross-platform validation. Both models developed histological injury and robust cytokine release. Direct injury preferentially enriched neutrophil degranulation, extracellular matrix remodelling and metabolic reprogramming modules, whereas indirect injury showed prominent complement/coagulation perturbation with greater endothelial activation markers in perfusate. Cross-platform prioritisation converged on tractable signalling and epigenetic axes, including JAK/STAT, PI3K/AKT/mTOR, SYK, CDK and HDAC inhibitor classes - yielding a tiered shortlist for EVLP intervention testing. This intact human lung perturbation platform enables injury-stratified mechanistic inference and therapeutic prioritisation in early lung injury relevant to ARDS.

immunology↗

bioRxiv2026

Endomicroscopic fluorescence lifetime imaging enables molecular detection and targeted sampling in the distal human lung

PurposeAccurate molecular characterisation of infection and inflammation within the distal human lung remains challenging, particularly in critically ill patients, due to limited access to the alveolar space and delayed diagnostic workflows. Molecular imaging approaches capable of real-time detection and targeted sampling could substantially improve diagnostic precision and the future translational development of molecular imaging probes and therapeutics. MethodsIn a preclinical setting, we evaluated a clinic-ready endomicroscopic fluorescence lifetime imaging microscopy (eFLIM) platform combined with molecularly targeted SmartProbes for in situ detection of bacteria and activated neutrophils in the distal human lung. A multifunctional 1.9-mm diameter imaging and sampling catheter (Eyes on Target; EoT) enabled real-time fluorescence intensity and lifetime imaging alongside directed alveolar microlavage via a 1.2-mm working channel. Fluorescence intensity and lifetime signatures of Gram-negative bacteria, Gram-positive bacteria, and activated neutrophils were characterised using three wash-free SmartProbes: NBD-PMX, Merocy-Van, and a neutrophil activation probe (NAP). Imaging and sampling performance were assessed in ventilated ex vivo human lungs. ResultsEoT reliably navigated to alveolar regions across all lung lobes in both phantom and ventilated human lung models. eFLIM distinguished alveolar microanatomy and enabled probe-specific molecular detection within the distal lung. Increased NBD-PMX signal was detected in Escherichia coli-instilled lobes, while Merocy-Van lifetime signatures selectively identified Staphylococcus aureus-instilled regions. Activated neutrophils were detected throughout lung tissue following NAP administration. Directed alveolar microlavage enabled recovery of cellular material and bacterial DNA from imaged regions for downstream analysis. ConclusioneFLIM using EoT combined with molecular SmartProbes enables real-time molecular imaging and targeted sampling within the distal human lung. This platform provides a translatable approach for evaluating infection and inflammation at the alveolar level and supports the clinical development of molecular imaging probes for pulmonary disease.

microbiology↗

bioRxiv2026

Persistent CCL3 expression sustains neutrophil recruitment and contributes to delayed wound healing in advanced age

Delayed wound healing in advanced age is associated with prolonged inflammation, but the mechanisms that impair inflammatory resolution in this context remain poorly defined. Neutrophils are essential for early repair, but their persistence is predicted to disrupt resolution and delay healing. Here we show that aged mice exhibit impaired wound closure accompanied by sustained neutrophil infiltration from days 7-14 post-injury, long after neutrophils have resolved in young wounds. In aged skin, macrophage abundance and neutrophil-macrophage interactions were reduced, consistent with reduced macrophage-mediated clearance. Instead, neutrophils preferentially localised to perivascular regions, consistent with ongoing recruitment. Transcriptomic profiling demonstrated that young wounds transitioned from inflammatory to reparative programs, whereas aged wounds retained inflammatory signatures, including enrichment for neutrophil chemotaxis and LPS-response pathways. Notably, Ccl3, Cxcl2 and Cxcl3 remained elevated in aged wounds. Analysis of human chronic ulcers identified macrophages as a major source of these transcripts, and aged mouse wounds possessed more CCL3+ macrophages than in young. Targeted blockade of CCL3 beginning at day 4 post-injury reduced neutrophil accumulation, accelerated wound closure, and rescued key features of age-impaired healing, when administered after the initial inflammatory phase. Together, these findings identify persistent CCL3 expression as a contributor to sustained neutrophil recruitment and impaired repair in advanced age. Therapeutically targeting this pathway offers a strategy to restore timely resolution and improve healing outcomes.

immunology↗

bioRxiv2022

MC1R drives cutaneous repair by promoting angiogenesis and lymphangiogenesis

Cutaneous healing results in scarring with significant functional and psychological sequelae, while chronic non-healing wounds represent repair failure often with devastating consequences, including amputation and death. Due to a lack of effective therapies, novel interventions addressing scarring and chronic wounds are urgently needed. Here, we demonstrate that harnessing melanocortin 1 receptor with a selective agonist (MC1R-Ag) confers multifaceted benefits to wound repair. MC1R-Ag accelerates wound closure and re-epithelialization while improving wound bed perfusion and lymphatic drainage by promoting angiogenesis and lymphangiogenesis. Concomitant reductions in oxidative stress, inflammation and scarring were also observed. To evaluate the therapeutic potential of targeting MC1R in pathological healing, we established a novel murine model that recapitulates the hallmarks of human non-healing wounds. This model combines advanced age and locally elevated oxidative stress. Remarkably, topical application of MC1R-Ag restored repair, whereas disrupting MC1R signalling exacerbated the chronic wound phenotype. Our study highlights MC1R agonism as a promising therapeutic approach for scarring and non-healing wound pathologies, and our chronic wound model as a valuable tool for elucidating ulcer development mechanisms.

pathology↗