Humanizing CXCL13 and CXCR5 in mice

摘要/摘要 血管免疫母细胞T细胞淋巴瘤(AITL)在两年内的存活率为47%，尽管 积极的细胞毒治疗，20多年来存活率没有显著增加。CXCR5是一种 趋化因子G蛋白偶联受体(GPCR)，定位于AITL细胞膜。我们有 发现了一种拮抗CXCR5的小分子化合物，并在AITL-PDX中显示了有利的作用 学习。我们还利用噬菌体展示技术和生物信息学从一种 约168,000个CXCL13变异体的文库。与耶鲁大学的合作者(塞缪尔·卡茨，医学博士)合作，我们还 以CXCR5为目标研制CAR-T。本应用程序旨在制作人性化的CXCL13和CXCR5 最终目标是测试小分子拮抗剂和以CXCL13为基础的 在肿瘤微环境的背景下进行生物治疗，这在PDX小鼠身上是不可能的。这个 人源化CXCL13和CXCR5的表达将被定量，并与正常动物体内水平进行比较 人源化小鼠和正常C57BL/6小鼠的细胞在功能上进行了比较 18F-拮抗剂的药效学将通过体内PET研究来确定，以获得更多 准确描述老鼠模型的特征。人源化的CXCL13和CXCR5小鼠不仅可以 对于未来的AITL研究以及皮肤T细胞淋巴瘤的小鼠模型，各种CXCR5- 表现为B细胞淋巴瘤、前列腺癌、结肠癌、肺癌和乳腺癌以及自身免疫性疾病。

Summary/Abstract Angioimmunoblastic T-cell lymphoma (AITL) has a 47% survival rate over a two-year period despite aggressive cytotoxic therapy, with no significant increase in survival for over two decades. CXCR5 is a chemokine G protein-coupled receptor (GPCR) that is localized on the membrane of AITL cells. We have identified a small molecule compound that antagonizes CXCR5 and shown beneficial effects in AITL-PDX study. We also used phage display technologies and bioinformatics to identify 102 potential antagonists from a library of ~168,000 CXCL13 variants. With a collaborator at Yale (Samuel Katz, MD/PhD), we are also developing a CAR-T by targeting CXCR5. This application intends to make a humanized CXCL13 and CXCR5 strain of C57Bl/6. The ultimate goal is to test the small molecule antagonist and the CXCL13-based biotherapeutic in the context of a tumor microenvironment, which is not possible with PDX mice. The expression of humanized CXCL13 and CXCR5 will be quantitated and compared to vivo levels of normal expression of C57Bl/6. Cells from humanized mice and normal C57Bl/6 mice will be functionally compared in vitro, and the pharmacodynamics of the 18F-antagonist will be determined by in vivo PET studies to more accurately characterize the mouse model. The humanized CXCL13 and CXCR5 mice will not only be available for future AITL studies but also for mouse models of cutaneous T-cell lymphoma, a variety of CXCR5- expressing B-cell lymphomas, cancers of the prostate, colon, lung, and breast, and autoimmune diseases.