Biology subjects

Lombardi, R.

Publications and source records attributed to Lombardi, R..

4 recordsLinked to original sources

bioRxiv2026

Inhibition of the Adenosine pathway activates the immune response against Mesothelioma.

BackgroundMesothelioma (Me) is an aggressive cancer with limited response to conventional therapies. The tumors harsh microenvironment contributes to immune escape and therapy resistance and the effects of ICIs on Me are still unclear. Adenosine, an immunosuppressive molecule produced from AMP by the enzyme CD73, accumulates in hypoxic tumor areas. Elevated CD73 and adenosine receptor A2B (A2Br) levels on Me cells are linked to worse patient outcomes, indicating their important role in disease progression and potential as targets for treatment. AimThis study characterizes the Me-ME (micro environment) and evaluates the efficacy of TT-4 (A2B inibitor) and AB680 (CD73 inibitor), alone or with aPD-1, using 3D models in vitro and in vivo. MethodsCD73 and A2B receptor levels were quantified in tumor and normal samples using qRT-PCR and IHC. Cells lines were treated with CoCl2 to mimic hypoxia, then CD73, A2Br and related markers were analyzed. MSTO-211H and REN cells were silenced for CD73, grown as spheroids and adenosine release was measured. Co-culture spheroids of MSTO-211H and Jurkat cells were treated with AMP and CD73 inhibitor, then analyzed for viability and immune markers. An orthotopic Me model was established by injecting AB1-B/c-LUC cells and monitored by in vivo imaging. Proteomic analysis of spheroids was conducted to identify proteins and pathways involved. ResultsHypoxia boosts CD73 and A2Br expression in Me cells, leading to adenosine production via CD73. In 3D co-cultures, AB680 lowered Me cell viability and enhanced activation of Jurkat T cells. In mice, combining aPD-1 therapy with A2Br or CD73 inhibitors strongly reduced tumor growth. Proteomics identified 93 proteins influenced by adenosine signaling through A2B. ConclusionTargeting the adenosine pathway alongside PD-1 blockade offers a promising new immunotherapy strategy for Me.

cancer biology↗

bioRxiv2025

Epidermal Langerhans Cells Drive Painful Diabetic Neuropathy in a Sex-dependent Manner

The interaction between non-neuronal cells and nerve endings in the epidermis significantly influences the development of various diseases, including Painful Diabetic Neuropathy (PDN). PDN is a common and challenging complication of diabetes, characterized by changes in skin innervation accompanied by neuropathic pain. While there is growing evidence that epidermal non-neuronal cells, such as resident immune cells, play a crucial role in the progression of PDN, the underlying mechanisms of this neuropathy remain poorly understood. In our studies, we utilized transgenic methods, pain behavioral assessments, and single-cell RNA sequencing (scRNAseq) in a clinically relevant high-fat diet (HFD) mouse model of PDN and skin biopsy samples from PDN patients, to investigate the role of epidermal Langerhans cells (LCs) in this condition. We observed an increased density of LCs in the skin of PDN male mice coinciding with the onset of mechanical allodynia. Furthermore, we found that LCs density correlated with small fiber degeneration in HFD mice and skin biopsies taken from well-characterized PDN patients compared to healthy controls. Importantly, the selective ablation of LCs using a diphtheria-toxin strategy was found to prevent nociceptive behavior in male mice. This indicates that LCs are both necessary and sufficient for the development of mechanical allodynia and spontaneous pain in HFD male mice. Interestingly, when LCs were ablated in HFD female mice, it did not prevent but rather promoted pain behavior, suggesting the existence of sex-specific mechanisms mediated by LCs. scRNAseq transcriptomic analysis of the paw epidermis from HFD mice, which included both males and females, revealed significant sex-mediated differences in the expression of specific target genes within this PDN model. In male mice, scRNAseq identified differentially expressed genes associated with axonal guidance and immune responses in LCs. Additionally, by integrating single-cell RNA data from the epidermis and dorsal root ganglia (DRG) of male mice, we uncovered altered communication between LCs and cutaneous afferents through Semaphorin-Plexin signaling pathways in PDN. These findings highlight LCs as key contributors to the development of PDN and suggest their potential as therapeutic targets for innovative treatments, particularly topical therapies aimed at modulating immune cell activity and neuroimmune communication in the skin. Our investigations of male and female mice indicate that LCs may play different roles in mechanical sensation under both normal and pathological conditions, such as PDN. This underscores the importance of considering sex differences when developing more effective treatments. BRIEF SUMMARYThe interaction between non-neuronal cells and nerve endings in the epidermis plays a significant role in the development of Painful Diabetic Neuropathy (PDN), a challenging complication of diabetes. Using transgenic methods, pain behavioral assessments, and single-cell transcriptomics in a clinically relevant high-fat diet (HFD) mouse model of PDN, and in skin tissues from PDN patients, we identified epidermal Langerhans cells (LCs) as key contributors to the development of PDN. In these studies, we revealed, for the first time, a dimorphic role for LCs in the development of mechanical allodynia and spontaneous pain in a HFD model of PDN. Single-cell RNA sequencing (scRNAseq) of the paw epidermis from both male and female HFD mice identified sex-mediated differences in the expression of specific target genes within this PDN model. In male mice, scRNAseq of LCs revealed differentially expressed genes related to immune responses and axonal guidance, including Semaphorin-Plexin signaling pathways.

neuroscience↗

bioRxiv2021

Ferries and environmental DNA: underway sampling from commercial vessels provides new opportunities for systematic genetic surveys of marine biodiversity

Marine environmental DNA (eDNA) is an important tool for biodiversity research and monitoring but challenges remain in scaling surveys over large spatial areas, and increasing the frequency of sampling in remote locations at reasonable cost. Here we demonstrate the feasibility of sampling from commercial vessels (Mediterranean ferries) while underway, as a strategy to facilitate replicable, systematic marine eDNA surveys in locations that would normally be challenging and expensive for researchers to access. Sixteen eDNA samples were collected from 4 fixed sampling stations, and in response to 4 cetacean sightings, across three cruises undertaken along the 300 km ferry route between Livorno (Tuscany) and Golfo Aranci (Sardinia) in the Ligurian/Tyrrhenian Seas, June-July 2018. Using 12SrDNA and 16SrDNA metabarcoding markers, we recovered diverse marine vertebrate Molecular Operational Taxonomic Units (MOTUs) from teleost fish, elasmobranchs, and cetaceans. We detected sample heterogeneity consistent with previously known variation in species occurrences, including putative species spawning peaks associated with specific sea surface temperature ranges, and increased night time abundance of bathypelagic species known to undertake diel migrations through the water column. We suggest commercial vessel based marine eDNA sampling using the global shipping network has potential to facilitate broad-scale biodiversity monitoring in the worlds oceans.

molecular biology↗

bioRxiv2019

Novel Universal Primers For Metabarcoding eDNA Surveys Of Marine Mammals And Other Marine Vertebrates

Metabarcoding studies using environmental DNA (eDNA) and high throughput sequencing (HTS) are rapidly becoming an important tool for assessing and monitoring marine biodiversity, detecting invasive species, and supporting basic ecological research. Several barcode loci targeting teleost fish and elasmobranchs have previously been developed, but to date primer sets focusing on other marine megafauna, such as marine mammals have received less attention. Similarly, there have been few attempts to identify potentially universal barcode loci which may be informative across multiple marine vertebrate Orders. Here we describe the design and validation of four new sets of primers targeting hypervariable regions of the vertebrate mitochondrial 12S and 16S rRNA genes, which have conserved priming sites across virtually all cetaceans, pinnipeds, elasmobranchs, boney fish, sea turtles and birds, and amplify fragments with consistently high levels of taxonomically diagnostic sequence variation. In silico validation using the OBITOOLS software showed our new barcode loci outperformed most existing vertebrate barcode loci for taxon detection and resolution. We also evaluated sequence diversity and taxonomic resolution of the new barcode loci in 680 complete marine mammal mitochondrial genomes demonstrating that they are effective at resolving amplicons for most taxa to the species level. Finally, we evaluated the performance of the primer sets with eDNA samples from aquarium communities with known species composition. These new primers will potentially allow surveys of complete marine vertebrate communities in single HTS metabarcoding assessments, simplifying workflows, reducing costs, and increasing accessibility to a wider range of investigators.

ecology↗