靶向FGFR1新型复方DC疫苗抗肿瘤血管生成研究

Cell penetrating peptides 44（CPP44 ）is a kind of short peptide with the super membranepenetrability targeting only tumor cells and can carry large molecules into cells without any toxic side effect on normal cells. MIP3α, the most important specific chemokine, can effectively improve the antigens presenting ability of DC. We have previously proved that immunotherapy targeting FGFR1 has obvious anti tumor effect, and found that the oncolytic virus recombinant herpes simplex virus type 1 (HSV1)-MIP3α not only retains the oncolytic properties of HSV1, but also carries the information of MIP3α. On the basis of previous work, We propose here to use the virus recombinant transfected with MIP3α/CPP44, along with FGFR1 aptamer (aFR1) encapsulated by nano liposome and produce a final complex product: "aFR1/HSV1-MIP3α-CPP44" bioreactor. This bioreactor will generate high concentrated MIP3α combined with CPP44 at the targeted tumor site, and the aFR1 can also inhibit angiogenesis in tumor.This bioreacctor will greatly enhance specifically targeted killing effect on tumor cells and/or tumor angiogenesis. Accordingly, such "aFR1/HSV1-MIP3α-CPP44 bioreacctor" is very likely a promising in vivo DC vaccine. We will investigate the mechanism by in vivo and in vitro.

细胞穿膜肽CPP44是一类只针对肿瘤细胞具有超强细胞膜穿透能力的短肽，可携带大分子物质进入细胞且对正常细胞无毒副作用。MIP3α是DC特异性趋化因子，能有效提高DC递呈抗原能力。我们前期已证实靶向FGFR1免疫治疗具有显著抗肿瘤效果，且发现重组单纯疱疹病毒1型(HSV1)-MIP3α溶瘤病毒既保留了HSV1溶瘤特性，还携带有MIP3α信息。本项目拟在前期基础上将转染高效表达MIP3α/CPP44基因的病毒重组子与FGFR1适配体(aFR1)用纳米脂质体包裹偶联成"aFR1/HSV1-MIP3α-CPP44"生物反应器，该生物反应器能让肿瘤部位靶向富集高浓度MIP3α，加上HSV1溶瘤和aFR1靶向抗肿瘤血管生成作用，将可能极大增强它们对肿瘤细胞和/或肿瘤新生血管特异性靶向杀伤作用，并研究该多功效生物反应器是否是一种简单而有效的体内DC疫苗模式。我们将通过体内外实验研究其抗肿瘤作用和机理。

本研究利用基因工程技术成功制备了一种复方生物反应器aFR1/HSV1-MIP3α-CPP44（简称aFHMC）并在体内外进行抗肿瘤作用观察。研究发现，aFHMC复方生物反应器具有肿瘤组织特异性和血管靶向性，能够靶向抑制肿瘤血管生成作用；还发现该生物反应器具有较好体内DC趋化募集和一定程度的溶瘤作用。此外，增加了FGFR1/MIP3α-Fc融合蛋白制备及其用途研究，发现其不仅具有良好肿瘤组织特异性和血管靶向性，而且可以抑制肿瘤血管生成和趋化募集DC到达肿瘤部位进而发挥抗肿瘤功效，阶段性成果将为靶向FGFR1肿瘤生物治疗提供新的策略。