探討 AXL 抑制劑 Dubermatinib 抑制神經母細胞瘤癌化進程及分子機轉

神經母細胞瘤是兒童常見的顱外實體瘤，來自於交感神經系統，主要好發於腹腔之神經節及腎上腺。現今研究採用最先進的治療方法，高危族群的神經母細胞瘤患者的生存率仍是低於 50%。表示現今臨床藥物仍不足有效治療神經母細胞瘤患者，因此，開發治療神經母細胞瘤的新藥是迫切需要的。之前文獻證實 AXL 抑制劑 Dubermatinib(TP0903)是一種新型多激?抑制劑且具有抗癌的活性。但是目前對於 TP0903 抑制神經母細胞瘤的分子機轉仍是未知。本實驗證實 TP-0903 顯著抑制神經母細胞瘤細胞(SH-SY5Y 和 Neuro-2a)的生長速率，但是對於正常細胞毒性不會產生毒性。我們也證實 TP-0903 誘導細胞凋亡、粒線體失衡、粒線體自噬、和 ROS 產生，伴隨著增加 c-PARP、ccaspase-9、BNIP3、PINK1、Parkin、BNIP3、NIX 及 LC3B，和抑制Bcl-2、Mcl-1 和 p62 表現。利用 RNA seq 方式證實 TP-0903 抑制 DKK1的蛋白和 mRNA 表現，同時也證實 DKK1 高度表達在神經母細胞瘤組織。此外我們也證實 TP-0903 誘導 miRNA-335 表現增加，過度表達 miRNA-335 抑制 DKK1 表現，同時促進 Neuro-2a 細胞走向凋亡路徑。動物實驗也證實 TP-0903 抑制神經母細胞瘤腫瘤生長能力且不會造成器官毒性。綜合以上說明 TP-0903 具備抑制神經母細胞瘤細胞的增生是透過細胞凋亡及 DKK1 表現，未來仍需要更加釐清 TP-0903抑制神經母細胞瘤是如何透過調控 DKK1 的分子機轉及角色。

Neuroblastoma is a common extracranial solid tumor in childrenoriginating from the sympathetic nervous system, predominantly affectingneural ganglia and adrenal glands in the abdomen. The survival rate forhigh-risk neuroblastoma patients remains below 50%. This indicates thatcurrent clinical drugs are still insufficient for effectively treatingneuroblastoma patients, indicating the urgent need for the development ofnew therapeutics for neuroblastoma. Previous reports has confirmed thatthe AXL inhibitor Dubermatinib (TP-0903) is a novel multiple tyrosinekinase inhibitor with anticancer activity in various tumor cells. However,the molecular mechanisms underlying TP-0903 inhibition ofneuroblastoma is still unknown. This study demonstrates that TP-0903significantly inhibits the growth rate of neuroblastoma cells (SH-SY5Y andNeuro-2a) without exhibiting toxicity to normal cells. We also suggest thatTP-0903 induces cell apoptosis, mitochondrial dysfunction, mitophagy,and ROS production, accompanied by increased levels of c-PARP, ccaspase-9, BNIP3, PINK1, Parkin, NIX, and LC3B, and decreased level ofBcl-2, Mcl-1, and p62 expression. Using RNA sequencing, we found thatTP-0903 inhibits the expression of DKK1 protein and mRNA, and we alsovalidate the high expression of DKK1 in neuroblastoma tissues inTNMplot database. Additionally, we also confirmed that TP-0903 inducesmiRNA-335 expression. Overexpression of miRNA-335 inhibits DKK1expression while signnificantly promoting the apoptotic pathway in Neuro2a cells. Furthermore, in vivo animal experiments demonstrate that TP0903 inhibits neuroblastoma tumor growth without causing organ toxicity. In summary, TP-0903 inhibits the proliferation of neuroblastoma cellsthrough induction of apoptosis and modulation of DKK1 expression.Further investigation is needed to clarify the molecular mechanisms androles of TP-0903 in inhibiting neuroblastoma tissue through the regulationof DKK1.