玻尿酸修饰的低毒性纳米药物对肝癌干细胞RNA甲基化的调控效果和机制研究

Hepatocellular carcinoma stem cells (HCSC) are the origin of malignant phenotypes such as proliferation, metastasis and recurrence of hepatocellular carcinoma. The increased expression of RNA-m6A Eraser (RME) in HCSC may be responsible for the increased stemness of HCSC. Previously, the applicant conducted a large number of studies using targeted nanotechnology (ACS Nano. IF = 13) to inhibit the activity of hepatocellular carcinoma stem cells in vivo (Nat Commun. IF = 12; Adv Sci. IF = 12). .In this project, we intend to further improve the low toxicity nanocarriers developed earlier (Adv Mat. IF = 21) . Targeted modification of nanocarriers with "hyaluronic acid" was carried out. The nanocarrier can recognize the HCSC surface marker CD44. Nano-drug delivery agent of RME inhibitors specifically to HCSC can increase the level of m6A and achieve therapeutic effect. .Pre-experiments have confirmed that the nanomedicine has low toxicity, targeting delivery ability of HCSC. And has RNA-m6A level elevation and stemness inhibition effect of hepatocellular carcinoma. This study aims to use this targeted nanotechnology to regulate the m6A level of HCSC and inhibit the proliferation and metastasis of hepatocellular carcinoma. We attempt to develop a new low toxicity targeted nanomedicine for hepatocellular carcinoma, and promote its clinical transformation and industrialization.

肝癌干细胞（HCSC）是肝癌增殖转移复发等恶性表型的根源。HCSC中RNA-m6A去甲基化酶（RME）表达升高，引起的RNA-m6A水平下降可能是其干性增强的原因。申请人前期进行了大量利用靶向化纳米技术（ACS Nano.IF=13），抑制体内肝癌干细胞活性的研究（Nat Commun.IF=12；Adv Sci.IF=12）。本课题拟进一步改进前期开发的低毒性药物载体（Adv Mat.IF=21），利用HCSC表面标志物CD44的配体“玻尿酸”进行靶向修饰。使其实现对HCSC特异性输送RME多靶点抑制剂，进而升高m6A水平，实现治疗效果。预实验已初步证实该药物在低毒性的前提下，具有预期的HCSC靶向递送、RNA-m6A水平升高、肝癌干性抑制效果。本研究拟利用该靶向化纳米技术调控HCSC的m6A水平，抑制肝癌的增殖、转移等恶性表型，开发新型低毒性肝癌靶向纳米药物，并促进其临床转化和产业化。

本项目在研究中获得的现有证据及前期研究结果，均证明了肝癌干细胞（HCSC）是肝癌增殖转移复发等恶性表型的根源。我们的研究结果显示，HCSC中RNA-m6A去甲基化酶（RME）表达升高，引起RNA-m6A水平下降，进而使得肝癌细胞干性增强。根据申请人前期掌握的靶向化纳米载体合成技术，以及体内肝癌干细胞活性抑制技术，我们在本项目执行期内，合成了针对HCSC的纳米药物。我们在研究中，利用HCSC表面标志物CD44的配体“玻尿酸”，成功地实现低毒性的纳米药物载体的靶向修饰。使纳米载体实现了对HCSC特异性输送RME多靶点抑制剂的功能。在应用该药物后，实现了升高HCSC的m6A水平，并实现了治疗效果。我们证实，该药物在低毒性的前提下，具有HCSC靶向递送药物、升高RNA-m6A水平、抑制肝癌干性的效果。我们验证，开发的靶向化纳米技术可以高效调控HCSC的m6A水平，进而抑制肝癌的增殖、转移等恶性表型。我们的研究结果已经发表IF＞10的SCI论文1篇，并获得发明专利授权1项，申请发明专利4项。这些成果可以促进新型低毒性肝癌靶向纳米药物的临床转化和产业化。