Leader: Luca Francioso (CNR-IMM); Other collaborator(s): Sanofi (UNIMIB), Corsonello, Antonio Cherubini (INRCA), Annamaria De Luca, Paola Pontrelli (UNIBA), (Pietro Siciliano, CNR, da spoke 9)
Polypharmacy, that usually describes the accumulation of 5, and often more medications for co-morbidities treatment frequently results in therapy problems related to drug toxicity and interactions, delirium, and clinical nonadherence. These effects are associated with resulting increased hospitalizations and higher costs of care for individuals and health care systems. The proposed MicroPhysiological Platforms (MPS or Organ-On-Chip) are miniaturized, live tissues-based devices with microfluidics and sensing capabilities that allows the development and study of preclinical models for drugs interactions predictions and optimization of drug candidates. Combined toxicity and efficacy testing can be performed with an higher experiment throughput; this approach unravels new biological pathways and targets and their clinical relevance at various stages of disease progression.
Brief description of the activities and of the intermediate results
The research activities within Task 5,1 are focused on two main research lines related to the application of Organ-On-Chip platforms for the polypharmacy management. UNIBA/CNR-IMM are working to an approvedy clinical study named DECORE 'Characterisation of the COgnitive DEclinic in chronic renal disease: study of specific functional and cognitive alterations' reserved for patients with chronic renal disease. This study has already been approved by the Ethics Committee and this study is in continuity with InTEMPO protocol because in the main protocol, patients with chronic disease are excluded but the evaluation of albuminuria is included, which will make it possible to identify a subcategory of patients who may progress to chronic renal disease, known to represent a condition of accelerated ageing. In order to therefore assess the predictive value of InTEMPO biomarkers also in the setting of the nephropathic patient, we will evaluate in the DECORE protocol both INTEMPO biomarkers and our other biomarkers, including alpha-klotho, for which the role in ageing processes is known. We will also evaluate sarcopenia markers in collaboration with Prof. De Luca group. The section of Nephrology at DiMePRe-J is actively collaborating with the National Research Council - Institute for Microelectronics and Microsystems (CNR-IMM) in Lecce on the development of a custom antibody-based sensor for assessing the levels of alpha-Klotho, a senescence marker, in the serum of patients enrolled in the DECORE clinical trial. Experimental conditions were established and the feasibility of initiating a study on Caco2/HUVEC co-culture, first in transwell and then in a custom microfluidic chip was assessed. The Caco2/HT-29/HUVEC co-cultures will first be incubated with culture media containing drugs of different dosages, such as glyphozines, branched-chain amino acids and taurine. A parallel activity is ongoing in cooperation with UNIBA partner active in 3D muscle chip, where miniaturizad pillars to allow direct transfer electric stimulation and/or to trigger muscle contraction and being able to evaluate precisely force production and maximal contraction ability of muscle bundle. A proprietary method from CNR-IMM will be also tested for in-situ detection of pillars deflection with an optical microprobe. The above proposed technology will support in particular the WP1 study IN-Tempo, correlating the results with standard biomarkers collected from population like BDNF and irisin and WP2 study “OPTIMAge-IT” in relation to sarcopenia prevention. Intermediate results are:
The research activity related to the definition of the experimental protocol for the study of the Diabetic Kidney Disease inter-organ signaling and Sodium/GLucose coTransporter 2 (SGLT2) modulation effects has been completed and consolidated with selected stimuli definition (high glucose, lipopolysaccharides, PCS). The experimental campaign will evaluate the effect of SGLT2 inhibitors on the gut microbiota and the endothelium together with the correlation with sarcopenia markers. The proposed Organ-On-Chip (OOC) platforms are miniaturized, live tissues-based devices with embedded microfluidics, mechanical stimulation and sensing capabilities, that allow the development and study of preclinical models for drugs interactions and dose optimization.
The implementation of the OOC platform has been consolidated with demonstrated multiple chamber perfusion. Mechanical simulation has been completely automatized allowing contemporary experimental tests on 8 independent chambers. The chosen SGLT2 inhibitors candidate drugs will be Empaglifozin, Canaglifozin and Dapaglifozin. To therefore assess the predictive value also in the setting of the nephropathic patient, we will dose the new α-klotho marker with a custom fabricated biosensor, under development.
During the reporting period, a co-culture model consisting of epithelial cells (Caco-2), mucus-producing goblet-like cells (HT29-MTX), and endothelial cells (HUVEC) was developed in conventional transwell plates as a physiologically relevant gastrointestinal model. The cell culture systems were characterized via transepithelial electrical resistance (TEER) measurements to monitor both the growth and permeability of the co-culture model. Selected stimuli, such as high glucose and lipopolysaccharide treatments, were used to simulate a Diabetic Kidney Disease pathological condition. Intestinal permeability alterations were evaluated using TEER measurements and apparent permeability values for fluorescein isothiocyanate-dextran (FITC-dextran) with different molecular weights as water-soluble marker compounds. Inflammatory cytokine responses were also evaluated. Following this, the selected SGLT2 inhibitors (Empagliflozin, Canagliflozin, and Dapagliflozin) were tested on the affected co-culture model to assess their effect on the altered gastrointestinal tract.
Prototypes
Publications