CD47+视网膜母细胞瘤干细胞自分泌SPINK1在自我更新中的作用和机制研究

Retinoblastoma (RB), a malignancy that arises in the retina, is always happened in young children. RB stem cells with unlimited proliferation and differentiation potential are responsible for the recurrence or metastasis. Our recent studies have found that CD47+ RB cells can exhibit strong cancer stem cell properties in vitro and in vivo. And SPINK1 autocrine by CD47+ RB cells plays the key role in maintaining the features of CSC by regulating WNT pathway. This project aims to confirm the CD47 as the marker of RB stem cells, and find out the influence and mechanism of SPINK1 in CD47+ RB stem cells at the levels of molecular, cell and animal models using by lentivirus system and clinical samples. We will evaluate the possibilities of using CD47 and SPINK1 as factors revealing RB diagnosis and prognosis by IHC. Test the SPINK1 in plasma by ELISA and make the model as the diagnostic and prognosis factor. This project aims to clarify the new understanding and concept of RB stem cells, and will help provide a new strategy for RB therapies.

视网膜母细胞瘤是婴幼儿最常见的眼内原发恶性肿瘤，危及患儿视力甚至生命。当前的治疗措施不能有效清除具有无限增殖和多向分化潜能的肿瘤干细胞是其复发转移的主要原因。前期通过基因芯片，体内外验证研究发现CD47+视网膜母细胞瘤细胞具有肿瘤干细胞特性。进一步，证实自分泌的SPINK1通过调控Wnt通路在维持其恶性行为中起关键作用。本项目拟通过原代培养，流式细胞仪等全面验明CD47作为视网膜母细胞瘤干细胞标记物的特异性；通过表达谱芯片，慢病毒干预、单克隆抗体等探讨SPINK1维持CD47+视网膜母细胞瘤恶性特征的分子机制，并在临床标本中逐级验证。免疫组化分析评估CD47及SPINK1与患者预后的关系；ELISA检测血清标本中SPINK1，建立诊断及预后评估模型。旨在阐明SPINK1调控视网膜母细胞瘤干细胞的机制，为治疗视网膜母细胞瘤提供新思路，最终为改善视网膜母细胞瘤的整体预后带来新的临床策略。

视网膜母细胞瘤是危害极大的恶性肿瘤，当前的治疗措施不能有效清除肿瘤干细胞是其复发转移的主要原因。通过基因芯片，我们于体内外验证研究发现CD47+视网膜母细胞瘤细胞具有肿瘤干细胞特性。通过过表达或抑制SPINK1，我们明确SPINK1在维持视网膜母细胞瘤干细胞特性起关键作用。进一步研究明确SPINK1可通过调控WNT/β-Catenin信号通路改变视网膜母细胞瘤干细胞特性。临床组织标本中验证发现CD47及SPINK1可作为视网膜母细胞瘤靶点及预后指标，视网膜母细胞瘤患者血清标本中检测肿瘤分泌SPINK1的浓度，同样发现SPINK1可以作为恶性肿瘤的临床诊断指标，但并不具有明显的视网膜母细胞瘤特异性。本项目明确SPINK1通过WNT/β-Catenin信号通路调控视网膜母细胞瘤干细胞的机制，为视网膜母细胞瘤的治疗、诊断及预后观察提供新思路。