角化棘皮瘤进展为鳞状细胞癌的致癌关键突变及分子机制研究

The main cause of the patients' deaths is the progression events of the tumor cells, i.e. proliferation, invasion, metastasis and drug resistance. The acquisition of the key mutations in tumor cells is the driving force to promote the tumor progression. Therefore, these mutations are found to be of great significance for understanding the mechanism of tumor progression. However, currently, the samples of cancer studies are mostly sporadic, lacking the tumor samples that reflect the key progression events of the transforming from benign to malignant in the same individual, and there is no effective tool in the current studies to reveal the mechanism of the action of the mutations. These have hampered the discovery of the key mutations. Our previous work analyzed the key events in tumor cell drug resistance and studied the mechanism of the action of the important candidate genes in key tumor events. Our pilot work collected the samples of the benign skin tumors - keratoacanthoma and the malignant skin tumors - squamous cell carcinoma which are progressed from the keratoacanthoma, at consecutive time points and have completed the whole genome sequencing of these samples. In this project, we plan to use bioinformatic methods to identify key candidate mutations from benign to malignant tumor progression, and then use gene editing techniques to introduce the mutations to be examined to the human skin organ chips, to reveal the mechanism and the process that causes tumorigenesis and tumor progression, ultimately to provide new clues for understanding the malignant progression of cancer.

肿瘤导致患者死亡的主要原因在于肿瘤细胞的增殖、浸润、转移和耐药性的产生等进展事件。肿瘤细胞获得关键突变是推动肿瘤进展的驱动力。因此，发现这些突变对于理解肿瘤进展机制具有十分重要的意义。然而目前肿瘤研究的样本多为散发的，缺乏在同一个体上，从良性转化为恶性的能够体现关键进展事件的肿瘤样本，且当前研究中缺乏有效的工具揭示突变的作用机制。这些都妨碍了关键突变的发现。我们之前的工作分析了肿瘤细胞耐药性的关键事件，并研究了重要候选基因在肿瘤关键事件中的作用机制。我们的前期工作收集了从良性皮肤肿瘤-角化棘皮瘤进展为恶性皮肤肿瘤-鳞状细胞癌的连续时间点的样本，并完成了全基因组测序工作。在本项目中，我们拟采用生物信息学方法，确定从良性向恶性肿瘤进展过程中的关键候选突变，再使用基因编辑技术在人体皮肤器官芯片中引入待检验的突变，揭示其导致肿瘤发生和进展的机制和过程，最终为理解肿瘤恶性进展事件提供新的线索。

皮肤鳞状细胞癌是皮肤癌中第二大常见恶性肿瘤，如不及时治疗，会有极大机会发生转移导致病人死亡，与其病理表现非常相似的角化棘皮瘤是一种良性肿瘤，不需治疗可自愈，但部分角化棘皮瘤可进展为鳞状细胞癌，因此我们对49名患者的肿瘤样本和部分外周血样本进行了全基因组测序和生物信息学分析，获得了一批肿瘤细胞进展事件相关的关键基因，并采用基因敲除和细胞三维培养的方法发现了皮肤癌进展的关键基因（TP53、CKDN2A、CDH10、DYNC1H1、DPYSL2等）及关键突变位点，以期为角化棘皮瘤的鉴别诊断、皮肤肿瘤进展预防及治疗提供理论依据。