Targeting of Doppel-axis to Control Lung Tumor Angiogenesis and Immunity

项目摘要 肿瘤，就像正常组织一样，需要血管来接收营养和氧气，并消除废物， 二氧化碳.为了确保这种血液供应，肿瘤从已建立的血管创建自己的血管床 一个叫做血管生成的过程。这一过程在肿瘤生长、存活和侵袭性中起着重要作用。 目前，各种单克隆抗体（mAb）和小分子量药物被用于抑制 血管生成并使肿瘤缺乏营养。例如，贝伐珠单抗和雷莫芦单抗是两种mAb， 血管生成-贝伐珠单抗通过与血管内皮生长因子（VEGF）结合，雷莫芦单抗通过 阻断VEGF受体（VEGFR）。当单独使用或与化疗联合使用时，抗血管生成药物 减缓转移，阻止疾病恶化，延长癌症患者的总体生存时间。然而，在这方面， 血管生成不仅驱动肿瘤中血管的生长，还执行许多重要的生理功能。 在身体中发挥作用。“好的”（生理性）血管生成调节血栓形成过程，维持血管 语气，并加快伤口愈合。不幸的是，目前的抗血管生成药物不加选择地抑制了这两个良好的 和“坏的”（肿瘤导向的）血管生成，从而导致血液凝固障碍，高血压， 甲状腺功能减退蛋白尿和肠穿孔原则上，应该可以减少或消除许多 目前的抗血管生成药物通过阻断仅在肿瘤中表达的信号分子的副作用 内皮细胞（TEC），但不是在正常内皮细胞（NEC）。最近，我们发现朊病毒样蛋白 一种叫做doppel的基因只在TEC中表达，而在NEC中不表达。在初步的研究中，我们表明（i）多普勒是 在人类和动物癌症中表达;（ii）TEC（Dplhi-TEC）中增加的多普勒表达与 肿瘤中血管密度增加;（iii）TEC和小鼠的多普勒消除（即，doppel knockout） 减少肿瘤中血管的数量;（iv）抗多普勒mAb减缓小鼠中的肿瘤生长;和（v）多普勒敲除 小鼠显示肿瘤内CD 8与调节性T细胞的比率更高。因此，我们认为Dplhi-TEC促进了 新血管生成并产生免疫抑制性TME，以及使用新的单克隆抗体靶向Dplhi-TEC 抗体增加了化学疗法和免疫疗法的功效。我们将在三个主要的假设下测试这个假设。 多组实验：（i）阐明Doppel激活新血管生成和肿瘤的分子机制 在非小细胞肺癌（NSCLC）自发模型中的进展;（ii）评估药代动力学， 抗Doppel mAb在KrasLSL-G12 D; p53 Frt/FRT小鼠中的药效学和抗肿瘤功效 （iii）评估Dplhi-TEC如何控制肺肿瘤免疫微环境。这个项目 因为我们将了解Dplhi-TEC在肿瘤中的生物学特性， 微环境以及使用构建新的单克隆抗体来阻断这种新的分子靶点，其在微环境中没有已知的作用。 生理性血管生成

Project summary Tumors, just like normal tissues, require blood vessels to receive nutrients and oxygen and to eliminate wastes and carbon dioxide. To ensure this blood supply, tumors create their own vascular beds from established blood vessels by a process called angiogenesis. This process plays a major role in tumor growth, survival, and invasiveness. Currently, various monoclonal antibodies (mAb) and small-molecular-weight drugs are used to restrain angiogenesis and starve tumors of nutrients. Bevacizumab and ramucirumab, for example, are two mAbs that inhibit angiogenesis-bevacizumab by binding with vascular endothelial growth factors (VEGF) and ramucirumab by blocking VEGF receptors (VEGFR). When used alone or in combination with chemotherapy, anti-angiogenic drugs slow down metastasis, stop disease deterioration, and extend the overall survival time of cancer patients. However, angiogenesis not only drives the growth of blood vessels in tumors, it also performs many important physiological functions in the body. The ‘good’ (physiological) angiogenesis regulates the thrombotic process, maintains vascular tone, and speeds up wound healing. Unfortunately, current anti-angiogenic drugs indiscriminately inhibit both good and ‘bad’ (tumor-directed) angiogenesis, thus resulting in blood-coagulation disorders, hypertension, hypothyroidism, proteinuria, and bowel perforation. In principle, it should be possible to reduce or eliminate many of the side effects of current anti-angiogenic drugs by blocking signaling molecules that are expressed only in tumor endothelial cells (TECs) but not in normal endothelial cells (NECs). Recently, we discovered that a prion-like protein called doppel is expressed only in TECs but not in NECs. In preliminary study, we showed that (i) doppel is expressed in both human and animal cancers; (ii) increased doppel expression in TECs (Dplhi-TECs) is associated with increased blood vessel density in tumors; (iii) doppel erasure from TECs and from mice (i.e., doppel knockout) reduces the number of vessels in tumors; (iv) anti-doppel mAb slows tumor growth in mice; and (v) doppel-knockout mice shows higher ratio of intratumoral CD8 versus regulatory T cells. Thus, we posit that Dplhi-TECs promote neoangiogenesis and create an immunosuppressive TME, and targeting of Dplhi-TECs using novel monoclonal antibodies increase the efficacy of chemo- and immune-therapies. We will test this assumption under three major sets of experiments: (i) elucidate the molecular mechanisms by which doppel activates neoangiogenesis and tumor progression in a spontaneous model of non-small cell lung carcinoma (NSCLC); (ii) assess the pharmacokinetics, pharmacodynamic and antitumor efficacy of anti-Doppel mAbs in KrasLSL-G12D;p53Frt/FRT mice in combination with chemotherapeutics; and (iii) assess how Dplhi-TECs control lung tumor immune microenvironment. This project has both basic and translational applicability, because we will understand the biology of Dplhi-TECs in tumor microenvironment as well use construct new mAbs to block this novel molecular target that has no known roles in physiological angiogenesis.