Preclinical Evaluation of a Novel ADAM10 Modulator to Treat ColorectalCancer

摘要 结直肠癌（CRC）是美国第三大诊断癌症， 全球每年的死亡人数，主要是由于高度侵袭性、耐药性疾病的复发 其特征在于具有干细胞样性质的低分化癌细胞。一个共同的签名， 化疗耐药肿瘤是Notch受体信号转导失调，以及其 金属蛋白酶激活剂，ADAM 10。尽管Notch或ADAM 10的小分子抑制已经被证实是有效的。 这些治疗策略显示出有效的抗肿瘤作用，但在临床试验中失败，主要是由于 全身毒性，特别是对胃肠道和肌肉骨骼系统的细胞毒性作用， 强调需要制定更有针对性的办法。我们之前已经证明， ADAM 10主要以抑制状态存在于正常组织中，但在肿瘤细胞中通过激活ADAM 10来激活ADAM 10。 细胞外结构域的构象变化，从而为肿瘤特异性调节提供了潜在的靶点 ADAM 10活性在这里，我们希望表征一种新的人单克隆抗体试剂（1H 5）， 选择性靶向肿瘤细胞上活化的ADAM 10的暴露的细胞外区域。我们之前 证明了用鼠型抗体（8 C7）处理，其特异于两种抗体的活化形式， 小鼠和人的ADAM 10，在人CRC细胞系中赋予肿瘤负荷的显著降低， 在异种移植模型中进行细胞培养和移植，并且当8 C7与 化疗本提案的目标是进行深入的临床前“进行/不进行”实验， 提供了一个明确的评估ADAM 10单克隆抗体作为一种治疗方法。在这里，我们期待 在分子、细胞和生理水平上研究我们的鼠和人mAb试剂， 生物化学，通过转移性CRC病变的体内评估表征其作用机制 在同系免疫活性小鼠模型（8 C7）和通过使用人肿瘤的异种移植模型（1H 5）中。 我们假设，通过使用遗传工具结合体外和体内模型，我们可以获得 重要的信息，显着降低风险的临床开发的1H 5为基础的治疗剂，因为我们 推进临床试验，以治疗广泛的CRC患者。我们的研究不仅能帮助我们 1H 5朝着成功的临床试验，它也将开创一个新的标准，全面研究新的 本提案的总体目标是充分表征 一种独特的治疗剂，在CRC模型中选择性地损害Notch途径活化， 但毒性较小的疗法，只通过临床试验推进更有可能成功的候选药物。

Abstract Colorectal cancer (CRC) is the third most diagnosed cancer in the USA and accounts for more than 600,000 deaths annually worldwide, primarily due to relapse with highly aggressive, chemo-resistant disease characterized by poorly differentiated cancer cells with stem cell-like properties. A common signature of these chemo-resistant tumors is dysregulation of Notch receptor signaling, as well as upregulation of its metalloprotease activator, ADAM10. Although small molecule inhibition of either Notch or ADAM10 has been shown to produce potent anti-tumor effects, these therapeutic strategies have failed in clinical trials primarily due to systemic toxicities, especially cytotoxic effects on the gastrointestinal tract and musculoskeletal system, highlighting the need for development of more targeted approaches. We have previously demonstrated that ADAM10 predominantly exists in an inhibited state in normal tissues but is activated in tumor cells through a conformational change in the extracellular domain, thus providing a potential target for tumor-specific modulation of ADAM10 activity. Here, we look to characterize a novel human monoclonal antibody agent (1H5) that selectively targets an exposed extracellular region of activated ADAM10 on tumor cells. We previously demonstrated that treatment with a murine version of the antibody (8C7), specific for the activated form of both mouse and human ADAM10, conferred a significant reduction in tumor burden against human CRC cell lines in cell culture and transplants in xenograft models, and relapse was prevented when 8C7 was combined with chemotherapy. The goal of this proposal will be perform in-depth preclinical “go/no-go” experimentation that will provide a definitive evaluation of ADAM10 monoclonal antibodies as a therapeutic approach. Here, we look to investigate our murine and human mAb agents at the molecular, cellular, and physiological levels by biochemically, characterizing their mechanism of action through in vivo assessment of metastatic CRC lesions in syngeneic immune-competent mouse models (8C7) and through xenograft models using human tumors (1H5). We hypothesize that through the use of genetic tools combined with in vitro and in vivo models, we can gain important information that significantly de-risks the clinical development of 1H5-based therapeutic agents as we move forward clinical trials to treat a broad population of CRC patients. Our research will not only help us move 1H5 toward successful clinical trials, it will also pioneer a new standard to comprehensively study novel therapeutic agents and identify potential failures early on. The overall goal of this proposal is to fully characterize a unique therapeutic agent to selectively impair Notch pathway activation in models of CRC to produce potent but less toxic therapies, advancing only candidates with a greater chance of success through clinical trials.