Chemotherapy Induced Peripheral Neuropathy (CIPN) is a debilitating neurodegenerative disorder affecting more than half of patients treated with chemotherapeutics. Dying back of peripheral sensory nerve axons produces symptoms of allodynia, paresthesia, temperature sensitivity, and numbness in a stocking and glove distribution, which may lead to early termination of therapy in severe cases. To better understand underlying molecular mechanisms of CIPN and identify potential therapeutic targets, human induced sensory neurons (hiSNs) have been used as a model of chemotherapy related neurodegeneration. While these models have successfully demonstrated axonopathy with treatment of various chemotherapeutics, the cell lines utilized were commercially available or generated from subjects in good health. Given that several genomic studies have recently demonstrated distinct genetic mutations linked to the onset of CIPN, we have established new hiSN models using mononuclear blood cells (PBMCs) collected from 20 patients with ovarian cancer, treated with paclitaxel and carboplatin therapy, who either developed severe CIPN (Case, n=10) or did not (Controls, n=10). Case and Control subjects were matched based on BMI, age, sex, and ethnicity; subjects impacted by diabetic neuropathy were excluded. PBMCs were used to generate induced pluripotent stem cells (iPSCs) which were then differentiated into mature hiSNs using a novel co-culture system of matched Case and Control lines, eliminating environmental variability during terminal differentiation. iSN identity was confirmed using immunohistochemistry to visualize expression of neuronal maturation markers (TUJ1, Peripherin, BRN3a, TRPV1, TRKA/B/C), and iSN function was established using calcium imaging after application of capsaicin (TRPV1 agonist) and ATP (P2RX3 agonist). iPSCs were also differentiated into induced motor neurons (iMNs) to determine specificity of CIPN effects on neuronal subtype. This iPSC library and iSN models will serve as a tool to determine whether genomic, proteomic, and/or metabolomic changes occur in patients that develop severe CIPN versus those that do not. 

 

Funded by NIH NCI 2R01CA205255-06A1 and DFCI Men's Collaborative Supporting Women’s Cancers