Humanizing CXCL13 and CXCR5 in mice

摘要/摘要 血管免疫母细胞性T细胞淋巴瘤（AITL）在2年内的生存率为47%， 积极的细胞毒性治疗，在超过二十年的生存率没有显着增加。CXCR 5是一种 趋化因子G蛋白偶联受体（GPCR），其位于AITL细胞的膜上。我们有 鉴定了一种拮抗CXCR 5的小分子化合物，并在AITL-PDX中显示出有益效果 study.我们还使用噬菌体展示技术和生物信息学从一个细胞中鉴定了102种潜在的拮抗剂。 约168，000个CXCL 13变体的文库。与耶鲁大学的合作者（Samuel Katz，MD/PhD），我们还 通过靶向CXCR 5开发CAR-T。本申请旨在制备人源化CXCL 13和CXCR 5， 菌株C57 B1/6。最终目标是测试小分子拮抗剂和基于CXCL 13的药物组合物。 在肿瘤微环境的背景下，这是不可能的PDX小鼠。的 将定量人源化CXCL 13和CXCR 5的表达，并与正常人源化CXCL 13和CXCR 5的体内水平比较。 C57 B1/6的表达。来自人源化小鼠和正常C57 B1/6小鼠的细胞将在功能上进行比较。 18F-拮抗剂的药效学将通过体内PET研究确定， 准确描述小鼠模型。人源化CXCL 13和CXCR 5小鼠不仅可以用于 对于未来的AITL研究以及皮肤T细胞淋巴瘤的小鼠模型， 表达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.