源自人類誘導性多潛能幹細胞的感覺神經元之 神經毒性分析

Abstract

誘導性多潛能幹細胞（iPSC）是多年來具有潛力，且備受期待的技術，可以誘導為多種細胞類型，以克服體內實驗對某些疾病的局限性。在我們的研究中，我們將人類誘導性多潛能幹細胞分化為感覺神經元，通過辣椒素和長春新鹼模擬神經退化的模型，並進一步評估了神經毒性和神經退化的潛在機制。為了確定治療後細胞體和神經突的形態，使用peripherin（Peri），neurofilament heavy chain（NF）和βIII-tubulin（TUB）作為標記物以檢查神經元數量和神經突長度。結果指出，根據LC50，辣椒素對細胞體造成了嚴重損害；相反的，長春新鹼抑制神經突生長。此外，我們也利用LC50來確定辣椒素和長春新鹼作用於不同的感覺神經元表型：Peri（+）/ NF（-）和Peri（+）/ NF（+）神經元。我們進一步研究了由辣椒素和長春新鹼引起的神經退化機制，其中包括檢測神經退化的信號級聯反應之藥物實驗。總而言之，這項研究證明了源自於iPSC的感覺神經元進行神經毒性測試的可行性，有助於了解iPSC作為模型的特性以探索機制，並進一步地進行臨床治療和藥物篩選。

Induced pluripotent stem cells (iPSCs) have the potential and are highly anticipated technology in years, and it can be induced into various cell types to overcome the limitations of in vivo experiment for certain disorders. In our research, we differentiated human iPSCs into sensory neurons, mimicked neurodegenerative model by capsaicin and vincristine, and further assessed the neurotoxicity and underlying mechanisms of neurodegeneration. To determine the morphology of soma and neurite after treatment, peripherin (Peri), neurofilament heavy chain (NF) and βIII-tubulin (TUB) were used as markers to examine neuron number and neurite length. The result indicated that capsaicin caused severe damage on the soma. On the contrary, vincristine inhibited neurite outgrowth according on LC50. Moreover, we also calculated LC50 to determine that capsaicin and vincristine mainly acted on different sensory neuron phenotypes, Peri(+)/NF(-) and Peri(+)/NF(+) neurons. We further investigated the mechanism underlying neurodegeneration caused by capsaicin and vincristine. There included pharmaceutical experiment examine signaling cascades leading as neurodegeneration. In summary, this study demonstrated the feasibility of neurotoxicity testing of iPSCs-derived sensory neuron contribute to understand property of iPSC serving as a model to explore mechanism of degeneration for clinical treatment and drug screening.