Oligomannose antigens as conserved immunological targets of Non-Hodgkin lymphomas

项目摘要 非霍奇金淋巴瘤（NHL）主要是B细胞起源的，但实质上是异质性的，这使得 开发肿瘤特异性免疫疗法的难度很大。免疫检查点抑制剂的研究进展 定向和泛B标记靶向免疫疗法是绕过肿瘤问题的替代策略， 异源性，并取得了良好的治疗效果。然而，靶向肿瘤细胞和正常 细胞仍然是一个固有的缺点，因为这种治疗可能会导致不希望的，甚至致命的副作用， 方面的影响.因此，对于新的靶向策略以进一步推进NHL的免疫疗法存在未满足的需求。 我们在SRI International的肿瘤糖组学团队研究免疫学上重要的聚糖标志物， 潜在地可用于诊断和/或治疗应用。最近，我们发现高甘露糖自身抗原 在弥漫性大B细胞淋巴瘤（DLBCL）（侵袭性NHL的主要类别）的组织中过表达。在 此外，我们发现几种新建立的Man 9特异性单克隆抗体识别细胞表面标记物， 由鼠B-淋巴瘤A20和Raji和拉莫斯人B-淋巴瘤细胞系表达（初步 结果）。因此，BMan 9自身抗原可能在进化上是保守的，并且可以作为BMan 9自身抗原的“泛”标记物。 B淋巴瘤靶向;因此，抗BMan 9 mAb是用于开发靶向的 免疫疗法来治疗主要NHL肿瘤亚型。 我们在加州大学洛杉矶分校的抗体工程团队已经开发了多个平台 重组抗体以改善治疗功效。值得注意的是，这些包括人源化聚合IgG和人源化聚合IgG。 免疫细胞因子聚合物IgG平台可用于捕获和根除循环肿瘤细胞， 癌症干细胞（CTC/CSC），免疫细胞因子被设计为指导有效的免疫- 调节因子，如干扰素-α（IFN α），对肿瘤微环境，以增强抗肿瘤 免疫应答和肿瘤杀伤活性。因此，我们计划探索这种技术组合， 使用我们新的BMan 9靶向mAb组开发NHL特异性免疫疗法。具体来说，我们将 确定人源化多聚IgGB-Man 9抗体是否鉴定组织中NHL的主要亚型，并检测 NHL患者血液样品中的CTC（目的1）。我们还将评估抗BMan 9 IgG抗体和 它们的IFN γ融合蛋白提供了对BMan 9免疫类型的NHL细胞的高效杀伤[体外和体内]。 A20小鼠模型]（Aim 2）。[此外，我们将努力探索患者的BMan 9免疫型， 用于评估针对人源性肿瘤的Man 9靶向策略的治疗潜力的衍生的异种移植物（PDX）模型 人NHL。]我们的团队已经确定了靶向单克隆抗体，并建立了生产 重组蛋白，我们希望在研究过程中开发出几种有前途的新型NHL靶向药物。 R21期间。我们的工作也可能揭示探索其他相对保守的NHL的可行性 亚型特异性聚糖标记物，可以推进下一代精确的NHL免疫治疗。

PROJECT SUMMARY Non-Hodgkin lymphoma (NHL) is primarily of B-cell origin but is substantially heterogeneous, which makes developing a tumor-specific immunotherapy for it difficult. Recent advances in immune check-point inhibitor- directed and pan-B marker-targeted immunotherapies are alternative strategies to bypass the problem of tumor heterogenicity and have achieved promising therapeutic effects. However, targeting both tumor cells and normal cells remains an intrinsic disadvantage because such treatments may cause undesired and even lethal side effects. Thus, there is an unmet need for new targeting strategies to further advance immunotherapy for NHL. Our tumor glycomics team at SRI International studies immunologically important glycan markers that are potentially useful in diagnostic and/or therapeutic applications. Recently, we found high-mannose autoantigens were overexpressed in tissues of diffuse large B-cell lymphomas (DLBCLs), a major class of aggressive NHL. In addition, we found that several newly established Man9-specific mAbs recognize the cell-surface markers co- expressed by a murine B-lymphoma A20 and the Raji and Ramos human B-lymphoma cell lines (Preliminary Results). Thus, the BMan9 autoantigens are likely evolutionarily conserved and may serve as “pan” markers for B-lymphoma targeting; anti-BMan9 mAbs are, therefore, promising candidates for use in developing targeted immunotherapies to treat major NHL tumor subtypes. Our antibody engineering team at University of California, Los Angeles has developed multiple platforms of recombinant antibodies to improve therapeutic efficacy. Notably, these include humanized polymeric IgGs and immunocytokines. The polymeric IgG platform can be used to trap and eradicate circulating tumor cells and cancer stem cells (CTCs/CSCs) in vivo, and the immunocytokines are designed to direct potent immune- modulating factors, such as interferon-alpha (IFN), to the tumor microenvironment to enhance anti-tumor immune responses and tumor-killing activities. We plan, therefore, to explore this combination of technologies to develop NHL-specific immunotherapies using our new panel of BMan9 -targeting mAbs. Specifically, we will determine if humanized polymeric IgGB-Man9 antibodies identify major subtypes of NHLs in tissues and detect CTCs in the blood samples of NHL patients (Aim 1). We will also assess whether anti-BMan9 IgG antibodies and their IFN fusion proteins offer highly efficient killing of the BMan9 - immune types of NHL cells [in vitro and in the A20 mouse models] (Aim 2). [Additionally, we will extend our effort to explore the BMan9-immunotypes of Patient- Derived xenograft (PDX) models for evaluating the therapeutic potential of the Man9-targeting strategy against human NHLs.] Our team has already identified targeting mAbs and established technologies for producing recombinant proteins, and we expect to develop several promising novel NHL-targeting agents during the proposed R21 period. Our work may also reveal the feasibility of exploring other relatively conserved NHL subtype-specific glycan markers that could advance next-generation precision NHL immunotherapy.