Characterizing ALCAM as an oncoprotein and immunotherapeutic target in neuroblastoma

项目摘要 神经母细胞瘤是一种儿童颅外实体恶性肿瘤，起源于发展中的交感神经系统， 系统高危神经母细胞瘤患者面临的五年生存率只有50%，尽管密集的 化疗、手术、放疗和免疫疗法的干预。新疗法的开发 由于缺乏靶向致癌突变，神经母细胞瘤一直具有挑战性。但最近的 靶向GD 2的单克隆抗体疗法的批准使免疫疗法成为治疗糖尿病的新途径。 神经母细胞瘤的治疗进展相反，抗GD 2治疗导致显著的靶向、脱瘤 由于GD 2在伤害感受神经元上的存在而引起的毒性以及由于GD 2表达丧失而引起的复发仍然存在 共同为了颠覆作为耐药机制的抗原下调，我们的实验室专注于开发 免疫治疗的目标是差异表达和肿瘤生存所必需的。我们最近 确定了细胞粘附分子ALCAM作为免疫疗法开发的潜在靶点。ALCAM是一个 细胞粘附分子参与视网膜神经节的发育，ALCAM高表达是阴性的。 是多种癌症的预后标志物。我们的RNA测序数据显示，ALCAM在大多数人中高度表达。 神经母细胞瘤患者。此外，靶向ALCAM的抗体-药物缀合物CX-2009目前正在开发中。 晚期转移性或局部不可切除的成人癌症的临床试验。该项目旨在发现 ALCAM在神经母细胞瘤中过度表达的机制，以及确定ALCAM的贡献 过度表达对肿瘤生长和转移的影响。初步数据表明shRNA介导的ALCAM 敲低在体外诱导神经突形成，这是与分化相关的表型。此外，ChIP- 来自两个神经母细胞瘤细胞系的测序数据显示，ALCAM的表达可能受到一个网络的调控， 转录因子称为核心调控回路（CRC）。CRC是一种前馈转录 每个转录因子调节自身和其他转录因子表达的调节回路 定义了以蛋白质如酪氨酸羟化酶的表达为特征的去甲肾上腺素能表型， 多巴胺β羟化酶通过整合生物信息学和分子生物学方法，我们将揭示 ALCAM在神经母细胞瘤中过表达的机制。更广泛地说，该项目将认证ALCAM 作为免疫治疗的可行靶点，以及肿瘤存活所必需的癌蛋白。

Project Summary Neuroblastoma is a pediatric extracranial solid malignancy that arises from the developing sympathetic nervous system. Patients with high risk neuroblastoma face five-year survival rates of only 50%, despite intensive intervention with chemotherapy, surgery, radiation, and immunotherapy. Development of new therapies for neuroblastoma has been challenging due to the paucity of targetable oncogenic mutations. However, recent approval of monoclonal antibody therapy targeting GD2 has credentialed immunotherapy as a new avenue of therapeutic development in neuroblastoma. Conversely anti-GD2 therapy causes significant on-target, off-tumor toxicity due to the presence of GD2 on nociceptive neurons, and relapse due to loss of GD2 expression remains common. To subvert antigen downregulation as a mechanism of resistance, our lab focuses on development of immunotherapy targets that are both differentially expressed and necessary for tumor survival. We recently identified the cell adhesion molecule ALCAM as a potential target for immunotherapy development. ALCAM is a cellular adhesion molecule involved in retinal ganglion development, and high ALCAM expression is a negative prognostic marker in a variety of cancers. Our RNA-sequencing data shows ALCAM is highly expressed in most patient neuroblastomas. Furthermore, an ALCAM-targeted antibody-drug conjugate, CX-2009, is currently in clinical trials for advanced metastatic or locally unresectable adult cancers. This project aims to discover the mechanism of ALCAM overexpression in neuroblastoma, as well as define the contribution of ALCAM overexpression to tumor growth and metastasis. Preliminary data suggests shRNA-mediated ALCAM knockdown induces neurite formation in vitro, a phenotype associated with differentiation. Additionally, ChIP- sequencing data from two neuroblastoma cell lines shows expression ALCAM may be regulated by a network of transcription factors termed the core regulatory circuit (CRC). The CRC is a feed-forward transcriptional regulatory circuit in which each transcription factor regulates the expression of itself and the others, together defining a noradrenergic phenotype characterized by expression of proteins such as tyrosine hydroxylase and dopamine beta hydroxylase. By integrating bioinformatics and molecular biology approaches, we will uncover the mechanism of ALCAM overexpression in neuroblastoma. More broadly, this project will credential ALCAM as a viable target of immunotherapy, as well as an oncoprotein necessary for tumor survival.