Further information at: IMI 10th anniversary scientific symposium
|HUMAN iPSC-DERIVED CELLS: A POWERFUL TOOL FOR NEUROPATHIC PAIN DISEASE MODELLING
Presenting author: Lucia Rutigliano1
Co-authors: uhn C1, Ricci F1, Nitzsche A2, Haupt S2, Faedo A1, Tarroni P1, Montag K1, Cainarca S1, Scarabottolo L1.
1Axxam SpA. Via A. Meucci 3 – 20091 Bresso. Milano, Italy; 2LIFE& BRAIN GmbH. Sigmund-Freud-Strasse 25 – 53127 Bonn. Germany
Chronic neuropathic pain (NP) is a condition with highly unmet medical need and social economic burden. This condition is triggered by inflammation and noxious stimuli, which induce lesions of the somatosensory system and increase neuronal activity in specific nociceptive pathways. Neuropathic pain strongly impairs quality of life. Current therapies generally suppress neuronal activity reducing the symptoms of the pain, but they are not curative and have limited efficacy with extensive adverse effects.
Understanding the exact pathophysiology of NP is fundamental to develop new therapies. Recent evidences in animal models report the activation of glial cells and neuro–glial interactions as key mechanisms underlying chronic pain. In fact, nerve injury provokes a peripheral immune response which leads to the release of inflammatory chemokines and neurotrophic factors, the recruitment and activation of microglia and astrocytes, and the proliferation of Schwann cells.
The majority of these mechanisms have been identified in animal models of NP. However, so far research based on preclinical animal models has failed to deliver truly novel treatment options, due to their limited translatability. Additionally, screening to isolate potential new drugs or to identify or validate molecular mechanisms would require standardized and scalable human systems. Recently, human induced pluripotent stem cell (hiPSC) technology is emerging as a potent tool to in vitro recapitulate the disease mechanisms and the response to therapeutic agents in pain research.
The aim of our IMI granted project, in partnership with academia (KCL, NMI), pharmaceutical industries (Esteve, Grünenthal) and biotech companies (Axxam, LIFE&BRAIN) is to in vitro recapitulate the in vivo mechanisms of chronic NP focusing on the development of hiPSC-derived neuron-glia co-culture system. Taking advantage of the CRISPR/Cas9 technology, we integrated neuronal and glial lineage-specific fluorescence reporters in hiPSCs, to monitor and optimize the differentiation process and set up of defined neuron-glia co-cultures. Then, to monitor functional perturbations of hiPSC-derived nociceptors in mono- and co-culture systems, we integrated, in the genomic “safe harbor” region, a Ca2+ biosensor, for measuring Ca2+-mediated signaling events. We expect this genome editing approach to deliver superior iPSC models for fundamental pain research and drug development. This will facilitate the establishment of a humanized in vitro high content/high throughput screening assay platform for the identification of novel therapeutic molecules based on NP-relevant neuron-glia interactions and pathways.