(1406-C) Scalable and Efficient Generation of iPSC-derived Human Sensory Neurons for CIPN and Pain Research
Tuesday, February 6, 2024
12:00 PM – 1:00 PM EST
Location: Exhibit Halls AB
Abstract: Chemotherapy-induced peripheral neuropathy (CIPN) is a common side effect of cancer treatment that results in damage to the peripheral nerves. This can significantly impact a patient's quality of life and can even lead to dose reductions or treatment discontinuation. In order to develop treatments and/or preventative measures for CIPN, there is a need for high-throughput, reproducible peripheral nerve disease models that give insight into the interplay between chemotherapeutics, axonal morphology, function, and potential neuroprotective compounds. In a phenotypic study, we treated human induced pluripotent stem cell-derived sensory neurons (hiPSC-SN) that are functionally and molecularly similar to primary DRG with four chemotherapy drugs (bortezomib, oxaliplatin, paclitaxel, and vincristine) in dose response with and without pre-treatment of the SARM1 inhibitor DSRM-3716. After 48 hours of treatment, axonal and mitochondria health was analyzed via beta-III tubulin (TUJ1) and tetramethylrhodamine, methyl ester (TMRM) staining. Paclitaxel altered soma morphology and axonal branching, while oxaliplatin showed minimal morphology changes. Both showed minimal mitochondrial membrane dysfunction. Bortezomib and Vincristine greatly degraded axons while also reducing mitochondrial activity. Vincristine-treated mitochondria exhibited a “fragmented” morphology. Using an AI-based high content analysis software, AutoHCS™, images from the study were automatically detected and scored to identify dose-dependent phenotypic responses to the drugs and the neuroprotective effects of DSRM-3716. Here, it is demonstrated that DSRM-3716 could have a protective effect on sensory neurons treated with paclitaxel and oxaliplatin. To study how sensory neuron function could be affected by chemotherapeutics, sensory neurons were cultured on microelectrode arrays for 21 days. Vincristine induced hyperactivation after one hour of application. When co-treated with gabapentin after 48 hours, there was a rescue effect where spike train parameters returned to control levels. Together, these findings demonstrate the ability to phenotypically and functionally screen CIPN-related and potential neuroprotective compounds in human nociceptors in high throughput systems.