Work package 11
Local and systemic intervention studies to stimulate lung repair and prevent disease
In our search for novel, personalized drug targets, based on the integrated outcomes from the in vitro studies (WP10) and the multiomics studies in the PRIL cohort (WP8), we will modulate the identified therapeutic targets.
To this end we will use (repurposed) pharmacological inhibitors as well as CRISPR/(d)Cas techniques, depending on the origin of the identified target. The top treatable targets/genes will be assessed in the optimized 3D lung models (WP3) for different exposures to study effects on damage and repair responses, using cell lines, primary cells and/or lung tissue from animal models and human origin. In addition, biomarkers from extra-pulmonary tissues identified in WP8 & 10 will be evaluated as candidates for therapeutic intervention.
The first aim is to integrate the outcome of the RNAseq data from WP10 and data from WP2,3,4-8 and interfere with the identified genes/pathways involved in abnormal damage and repair responses using (repurposed) pharmacological inhibitors and/or the gene editing system CRISPR/(d)Cas9 in human lung cell lines (airway or alveolar, dependent on function/expression of the gene) as mentioned above. We will study consequences for damage (cellular stress responses, cell death by immunodetection/flow cytometry), repair read-outs, repair responses using scratch wounding and wounding by electroporation using real-time electric cell-substrate impedance sensing (ECIS) and organoid cultures to study regenerative responses. Pharmacological inhibitors will subsequently be tested ex vivo in the precision-cut lung slices.
The second aim is to evaluate soluble mediators (metabolites, growth factors, cytokines) derived from extra-pulmonary tissue sources identified in WP6/7/10 as potential determinants of lung damage and repair responses as candidates for therapeutic intervention. Using the in vitro models outlined in WP10, and relevant in vivo models (WP11, project 1), the aberrant secretome of skeletal muscle associated with an unhealthy lifestyle, will be modulated with targeted genetic (causality) and dietary/activity (translational) interventions, followed by evaluation of the protective or stimulatory effects on lung damage or repair.
The deliverable of this work package is novel treatment strategies to reduce tissue damage and to stimulate repair in chronic lung disease.
Figure 10 Set up of work package 11.