伊曲康唑对婴幼儿血管瘤增殖和血管生成的作用及其分子机制研究

Hemangioma, the most common tumor for infants, can present as a therapeutic challenge to the clinicians. Itraconazole is effective and safe in the treatment and prevention of fungal disease in infants. We occasionally found that itraconazole have a therapeutic effect for infantile hemangiomas accompanied with Candia albicans infection in the clinical, and later successfully repeated full therapeutic course in another 5 cases of infantile hemangiomas. We also found that itraconazole inhibits vascular endothelial cell proliferation and angiogenesis of infantile hemangiomas in vitro. The RNA was extracted to do microarray after itraconazole treated hemangioma endothelial cells, by which we selected a group of candidate proteins, DDIT4L, ADM2 and HDAC9, closely associated with vascular proliferation and tumor angiogenesis. Taking DDIT4L as a key regulatory protein for example, this study intends to use cell culture and animal models to address the following scientific queries: 1) Further clarifying the effects of itraconazole on cell proliferation and angiogenesis in infantile hemangioma; 2) Screening and verification of a key regulatory protein DDIT4L related with hemangioma proliferation and angiogenesis; 3) Regulatory role of protein DDIT4L in itraconazole inhibiting vascular proliferation and tumor angiogenesis of hemangioma; 4) Mechanism and key effector molecules of protein DDIT4L in itraconazole inhibiting vascular proliferation and tumor angiogenesis of hemangioma； 5） Investigating correlations between protein DDIT4L and tumor vascular proliferation and angiogenesis of hemangioma from the perspective of histopathology.

血管瘤是婴幼儿最常见肿瘤，治疗具挑战性。伊曲康唑在治疗和预防婴幼儿真菌病有效且安全。研究表明伊曲康唑能抑制血管生成和肿瘤生长。我们在临床上发现伊曲康唑对婴幼儿血管瘤有效；体外实验证实对血管瘤内皮细胞的增殖和血管生成也有抑制作用；将伊曲康唑与血管瘤内皮细胞作用后提取RNA做基因芯片分析筛选出能抑制血管瘤增殖和血管生成相关的候选蛋白DDIT4L、ADM2、HDAC9。以DDIT4L为例，本课题拟体内外实验解决以下科学问题：1）进一步明确伊曲康唑对婴幼儿血管瘤内皮细胞增殖和血管生成的作用；2）筛选并验证与血管瘤增殖和血管生成相关的关键调控蛋白DDIT4L；3）明确DDIT4L在伊曲康唑抑制血管瘤增殖和血管生成过程中的调控作用；4）研究DDIT4L在伊曲康唑抑制血管瘤增殖和血管生成过程中的作用机制及关键效应分子；5）从组织病理学角度探讨DDIT4L表达水平与血管瘤增殖和血管生成相关性。

血管瘤是婴幼儿最常见的良性肿瘤，常用的治疗药物包括糖皮质激素和普萘洛尔，但仍有很多血管瘤患儿的临床疗效不令人满意。本课题围绕伊曲康唑对婴幼儿血管瘤治疗有效这一临床发现，从前瞻性临床观察和体外细胞两个层面阐述了伊曲康唑治疗婴幼儿血管瘤的机制。前瞻性临床观察，进一步确认了伊曲康唑治疗婴幼儿血管瘤的临床有效性，没有发现严重副作用，30%可逆性腹泻。采用口服液剂型使临床用药更加方便和更准确易操作，应用皮肤镜观察治疗过程中皮损的变化使疗效评价更客观更精准。通过对伊曲康唑作用后差异表达基因的筛选验证，发现伊曲康唑可调控与血管生成密切相关的PDGF-D和DDIT4L基因表达。利用蛋白免疫印迹法证实伊曲康唑可能通过抑制PI3K/Akt/mTOR/p70S6K信号通路活性，抑制血管瘤内皮细胞增殖并诱导其凋亡。本课题为阐明伊曲康唑治疗婴幼儿血管瘤的具体分子机制提供了新的证据。