Ubisense

<bold>BACKGROUND:</bold> Chronic Obstructive Pulmonary Disease (COPD) is a global health concern, and accurate diagnosis is crucial for early intervention and improving patient quality of life, while also reducing the risk of exacerbations and healthcare costs. <bold>OBJECTIVE:</bold> To quantify diagnostic performance of N-Tidal Diagnose (NTD) machine learning models in a cohort of known COPD patients and healthy controls. <bold>METHODS:</bold> Capnography data was collected using 75 seconds of tidal breathing through TidalSense’s N-Tidal capnometer from a cross-sectional observational data set, which included 51 COPD subjects and 51 healthy controls. The performance of NTD 1 v1.0 software’s machine learning models, which perform diagnostic classification using 25 geometric capnography features, was measured. <bold>RESULTS:</bold> <fig><object-id>erj;66/suppl_69/PA1460/F1</object-id><object-id>F1</object-id><object-id>F1</object-id><caption> <italic>Figure 1.</italic> Confusion matrix evaluating the diagnostic performance of the NTD models. </caption><graphic></graphic></fig> The models were unable to diagnose eight patients who fell within the ‘indeterminate’ category with prediction probability close to 0.5. For all other participants, the NTD software yielded the following performance metrics: sensitivity 0.960, specificity 0.932, positive predictive value 0.953, and negative predictive value 0.941. <bold>CONCLUSIONS:</bold> The N-Tidal Diagnose ML models were able to accurately distinguish COPD subjects from healthy controls. This suggests that NTD could be used as a rapid and accessible alternative to spirometry for rapid point-of-care testing of COPD.

BackgroundEffective cancer immunotherapy is driven by robust T cell responses against neoantigens. A sufficient number of neoantigens is crucial to stimulate at least 2–3 cancer-specific T cell responses that can home into the tumor microenvironment and exert therapeutic activity. Current clinical trials support these principles, showing promising results with combinations of approved immunotherapies, including anti-PD1 antibodies. However, most existing delivery systems rely on single-platform approaches, such as long peptides, plasmid DNA or mRNA-LNPs, which often result in limited and short-lived immune responses.Materials and MethodsTo address the limitations of conventional delivery platforms, we investigated a synthetic approach combining the Hermes™ lipopolyplex nanoparticle platform encapsulating mRNA in combination with delivery of synthetic linear hairpin (hp)DNA™. Synthesis of hpDNA™ is fully enzymatic, facilitating fast production of DNA vaccine devoid of bacterial backbone with improved quality over traditional plasmid fermentation processes. Meanwhile, Hermes™ nanoparticles exploit a cationic ligand, combined with traditional lipids, to drive payload condensation and extend shelf life at refrigerator temperature.ResultsTo explore the efficacy of these technologies, vaccines targeting 20 neoantigens of the MC38 tumor model were generated and combined in an innovative heterologous prime-boost protocol. This strategy partners the potent but transient nature of mRNA with synthetic DNA to enhance immunogenicity and durability of the anti-tumor response. While hpDNA encoding a large number of neoantigens is effective in inducing significant immune responses and antitumor activity, its combination with the new mRNA-Hermes™ formulation provide a faster T cell induction and improved tumor responses. This new therapeutic protocol represents the first example of a completely synthetic heterologous prime-boost vaccination protocol for cancer therapy. The results from our study suggest that the heterologous prime-boost regimen induces a polyfunctional and polyspecific T cell response against neoantigens, offering several advantages in the clinical setting. Responses in the MC38 tumor model are characterized by long-lasting activity and significant anti-tumor effects. The heterologous synthetic platform demonstrates a potent ability to elicit sustained immune responses, surpassing the performance of individual delivery systems including plasmid DNA or mRNA-LNP-based systems.ConclusionsIn conclusion, our findings provide compelling evidence that a synthetic biology-based heterologous prime-boost strategy is feasible and may offer an alternative valid immunotherapy platform. This approach holds significant potential for developing more effective and durable cancer immunotherapies, offering new hope for patients with challenging malignancies.I. Esposito: None. C. Tonini: None. A. Bianchi: None. E. Young: None. A. Dhir: None. M. Esposito: None. L. Aurisicchio: None. A. Walker: None. F. Palombo: None.

3 D human epidermal equivalents (HEEs) are a state-of-the-art organotypic culture model in pre-clinical investigative dermatology and regulatory toxicology. Here, we investigated the utility of electrical impedance spectroscopy (EIS) for non-invasive measurement of HEE epidermal barrier function. Our setup comprised a custom-made lid fit with 12 electrode pairs aligned on the standard 24-transwell cell culture system. Serial EIS measurements for seven consecutive days did not impact epidermal morphology and readouts showed comparable trends to HEEs measured only once. We determined two frequency ranges in the resulting impedance spectra: a lower frequency range termed EISdiff correlated with keratinocyte terminal differentiation independent of epidermal thickness and a higher frequency range termed EISSC correlated with stratum corneum thickness. HEEs generated from CRISPR/Cas9 engineered keratinocytes that lack key differentiation genes FLG, TFAP2A, AHR or CLDN1 confirmed that keratinocyte terminal differentiation is the major parameter defining EISdiff. Exposure to pro-inflammatory psoriasis- or atopic dermatitis-associated cytokine cocktails lowered the expression of keratinocyte differentiation markers and reduced EISdiff. This cytokine-associated decrease in EISdiff was normalized after stimulation with therapeutic molecules. In conclusion, EIS provides a non-invasive system to consecutively and quantitatively assess HEE barrier function and to sensitively and objectively measure barrier development, defects and repair.