Antitumor potential of AvFc lectibody in non-small cell lung cancer

项目摘要/摘要 本项目的目标是开发一种针对非小细胞肺的新型肿瘤糖标记物靶向药物。 癌症(NSCLC)。非小细胞肺癌占所有肺部肿瘤的大多数，经常被诊断为晚期 分期，预后差。目前的治疗方法，包括新型靶向药物和免疫疗法，已经导致 改善了结果，但对晚期疾病的疗效较差，其中几种药物导致 抵抗。鉴于可能需要多种治疗策略的组合才能产生最佳的 治疗结果，针对非小细胞肺癌独特生物标志物的新型治疗药物的开发 这是正当的。特别是，可以增强抗肿瘤免疫反应的肿瘤靶向免疫疗法可能 增强免疫检查点抑制剂的疗效。有越来越多的证据表明， 癌细胞，包括非小细胞肺癌细胞的糖类中的高甘露糖聚糖。为了瞄准这个糖生物标记物， 申请人的实验室已经研制出高效的重组抗体样分子--Avaren-Fc(AVFC) 识别恶性细胞表面的高甘露糖聚糖。AVFC的抗肿瘤作用及安全性研究 已在人类A549和H460 NSCLC异种移植攻击模型中证明使用免疫缺陷 在小鼠和同基因B16F10黑色素瘤挑战模型中使用免疫活性C57BL/6小鼠。在……里面 在后一种模型中，AVFC治疗增加了非经典单核细胞和其他髓系细胞的浸润。 CD_4/CD_8 T淋巴细胞。此外，免疫组织化学分析显示，AVFC可以选择性地 在邻近非肿瘤肺组织上识别原发人类非小细胞肺癌肿瘤。基于这些发现，我们 AVFC可作为一种新型的肿瘤相关高甘露糖免疫治疗剂的假设 非小细胞肺癌中的葡聚糖。为了在更具临床相关性的非小细胞肺癌模型中证实这种可能性，这项研究的目标是 R21项目是为了揭示AVFC的免疫治疗效果和肿瘤靶向性 KRAS/P53突变驱动的非小细胞肺癌小鼠模型。在具体目标1中，我们将确定和描述 基于总生存期、肺肿瘤免疫组织化学和免疫组织化学的AVFC疗效 肺浸润性免疫细胞的免疫表型。此外，我们还将评估AVFC在 与免疫检查点阻滞剂抗PD-1抗体相结合。在具体目标2中，我们将分析 应用放射性标记的AVFC衍生物在NSCLC小鼠体内的生物分布和肿瘤检测 一种microPET/CT成像分析。在我们初步数据的基础上，我们将设计和优化无线电标记 AVFC衍生物具有最佳的药代动力学曲线和肿瘤检测灵敏度。特别是，我们将 评估放射性标记AVFC检测肿瘤进展和转移的能力。总体而言， 拟议的研究将提供关键信息，解决AVFC是否可行的问题 针对非小细胞肺癌的免疫治疗和/或诊断试剂。如果答案是肯定的，结果将是 极大地促进了AVFC向I期临床试验的进一步临床前开发。

PROJECT SUMMARY / ABSTRACT The goal of this project is to develop a novel tumor glycobiomarker-targeting agent against non-small cell lung cancer (NSCLC). NSCLC accounts for the majority of all lung tumors and is frequently diagnosed at an advanced stage with poor prognosis. Current therapies, including novel targeted drugs and immunotherapies, have led to improved outcomes but have had less efficacy in advanced disease, and several of these agents result in resistance. Given that a combination of multiple therapeutic strategies may be necessary to produce an optimal therapeutic outcome, development of novel therapeutic agents targeting a unique biomarker of NSCLC is warranted. Particularly, tumor-targeted immunotherapy that can boost an antitumor immune response might enhance the efficacy of immune checkpoint inhibitors. There is growing evidence for an aberrant increase of high-mannose glycans in the glycome of cancer cells, including those of NSCLC. To target this glycobiomarker, the applicant’s lab has developed Avaren-Fc (AvFc), a recombinant antibody-like molecule “lectibody” efficiently recognizing high-mannose glycans on the surface of malignant cells. The antitumor potential and safety of AvFc has been demonstrated in human A549 and H460 NSCLC xenograft challenge models using immunodeficient mice as well as in syngeneic B16F10 melanoma challenge models using immunocompetent C57bl/6 mice. In the latter model, AvFc treatment increased the infiltration of non-classical monocytes and other myeloid cells as well as CD4/CD8 T lymphocytes. Furthermore, immunohistochemical analysis revealed that AvFc can selectively recognize primary human NSCLC tumors over adjacent non-tumor lung tissues. Based on these findings, we hypothesize that AvFc may serve as a novel immunotherapeutic agent targeting tumor-associated high-mannose glycans in NSCLC. To substantiate the possibility in a more clinically relevant NSCLC model, the goal of this R21 project is to reveal the immunotherapeutic efficacy and tumor-targeting profile of AvFc in a conditional Kras/p53 mutation-driven NSCLC mouse model. In Specific Aim 1, we will determine and characterize the therapeutic effects of AvFc based on overall survival, immunohistochemistry of lung tumors, and immunophenotyping of lung-infiltrated immune cells. Additionally, we will assess the efficacy of AvFc in combination with the immune checkpoint blocker anti-PD-1 antibody. In Specific Aim 2, we will analyze biodistribution and tumor detection profiles of AvFc in the NSCLC mice, using a radiolabeled AvFc derivative in a microPET/CT imaging analysis. Building on our preliminary data, we will design and optimize radiolabeled AvFc derivatives to have an optimal pharmacokinetic profile and tumor-detection sensitivity. In particular, we will assess the capability of radiolabeled AvFc to detect tumor progression and metastasis. Collectively, the proposed research will provide critical information addressing the question of whether AvFc could be a viable immunotherapeutic and/or diagnostic agent against NSCLC. Should the answer be positive, the results will significantly facilitate further preclinical development of AvFc towards a Phase I clinical trial.