Development of human serine protease-based therapeutics targeting Fn14 and identification of Fn14 as a new target overexpressed in TNBC.
Development of human serine protease-based therapeutics targeting Fn14 and identification of Fn14 as a new target overexpressed in TNBC.
复制标题
DOI：
10.1158/1535-7163.mct-14-0346
复制
发表时间：
2014-11
影响因子：
5.7
通讯作者：
Rosenblum MG
中科院分区：
文献类型：
--
作者：
Zhou H;Mohamedali KA;Gonzalez-Angulo AM;Cao Y;Migliorini M;Cheung LH;LoBello J;Lei X;Qi Y;Hittelman WN;Winkles JA;Tran NL;Rosenblum MG
细胞因子TWEAK及其受体Fn14已成为癌症治疗的潜在有价值的靶点。产生含有粒酶B（GrB）的Fn14靶向构建体，其含有Fn14配体TWEAK（GrB-TWEAK）或抗Fn14人源化单链抗体（GrB-Fc-IT 4）作为靶向部分。两种构建体均显示出对一组Fn14表达的人肿瘤细胞（包括三阴性乳腺癌（TNBC）系）的高亲和力和选择性细胞毒性。GrB抑制剂PI-9在靶细胞中的细胞表达对任一构建体的细胞毒性作用没有影响。MDR1的细胞表达显示对融合构建体没有交叉抗性。GrB-TWEAK和GrB-Fc-IT4激活细胞内半胱天冬酶级联和细胞色素C相关的促凋亡途径，这与GrB在靶细胞中的已知细胞内功能一致。与单独的媒介物相比，用GrB-TWEAK处理携带已建立的HT-29异种移植物的小鼠显示出显著的肿瘤生长抑制（P <0.05）。GrB-TWEAK和GrB-Fc-IT 4在给予携带原位MDA-MB-231（TNBC）肿瘤异种移植物的小鼠时均显示出显著的肿瘤生长抑制作用。TCGA分析显示，与激素受体阳性乳腺癌相比，TNBC和HER2阳性疾病中的Fn14 mRNA表达显著更高（P <0.0001），与其他TNBC分子亚型相比，基底样2肿瘤中的Fn14 mRNA表达显著更高（P = 0.01）。对101名患者TNBC肿瘤微阵列的免疫组织化学分析显示，55/101（54%）的肿瘤对Fn14染色呈阳性，表明这可能是精确治疗方法的极好的潜在靶标。使用全人的、含GrB的融合构建体靶向Fn14可以形成用于靶向治疗应用的新型、有效和高效构建体的基础。
The cytokine TWEAK and its receptor, Fn14, have emerged as potentially valuable targets for cancer therapy. Granzyme B (GrB)-containing Fn14-targeted constructs were generated containing either the Fn14 ligand TWEAK (GrB-TWEAK) or an anti-Fn14 humanized single-chain antibody (GrB-Fc-IT4) as the targeting moieties. Both constructs showed high affinity and selective cytotoxicity against a panel of Fn14-expressing human tumor cells including triple-negative breast cancer (TNBC) lines. Cellular expression of the GrB inhibitor PI-9 in target cells had no impact on the cytotoxic effect of either construct. Cellular expression of MDR1 showed no cross-resistance to the fusion constructs. GrB-TWEAK and GrB-Fc-IT4 activated intracellular caspase cascades and cytochrome C-related pro-apoptotic pathways consistent with the known intracellular functions of GrB in target cells. Treatment of mice bearing established HT-29 xenografts with GrB-TWEAK showed significant tumor growth inhibition compared to vehicle alone (P < 0.05). Both GrB-TWEAK and GrB-Fc-IT4 displayed significant tumor growth inhibition when administered to mice bearing orthotopic MDA-MB-231 (TNBC) tumor xenografts. TCGA analysis revealed that Fn14 mRNA expression was significantly higher in TNBC and in HER2-positive disease (P<0.0001) compared to hormone receptor-positive breast cancer, and in basal-like 2 tumors (P=0.01) compared to other TNBC molecular subtypes. Immunohistochemistry analysis of a 101 patient TNBC tumor microarray showed that 55/101 (54%) of tumors stained positive for Fn14 suggesting that this may be an excellent potential target for precision therapeutic approaches. Targeting Fn14 using fully-human, GrB-containing fusion constructs may form the basis for a new class of novel, potent and highly effective constructs for targeted therapeutic applications.