Elucidating the therapeutic tractability and functional role of SHP2 in ALK-driven high-risk neuroblastoma

项目摘要：神经母细胞瘤(NB)是一种主要起源于神经脊细胞的儿科癌症，是一种 由于基因组和肿瘤内的异质性，临床上具有挑战性的疾病。有三个风险 疾病特征的分层--低风险、中风险和高风险--基于几个 突变负担和确诊时的年龄等因素。高危NB患者预后较差，5- 尽管接受了剂量密集的放化疗治疗，一年的存活率约为50%。为数不多的共同点 高危NB的基因组异常包括40%的病例MYCN癌基因扩增，异常 在受体酪氨酸激酶(RTK)、间变性淋巴瘤激酶(ALK)等病例中有14%为ALK基因 F1174、F1245和R1275残基的扩增和三个热点突变- 独立的激酶活性和编码SHP2蛋白的PTPN11基因突变，在3.4%的 案子。高危NB治疗的唯一靶向治疗是ALK抑制，尽管事实是第三种 新一代ALK抑制剂劳拉替尼比前一代ALK抑制剂更有效和更优越，复发 在服用劳拉替尼的患者中发生。因此，新的靶向治疗和给药计划的发现需要克服 诊所所经历的挑战是最重要的。 一种新的治疗药物靶点，含Src同源2的酪氨酸磷酸酶(SHP2)是一种非受体 蛋白酪氨酸磷酸酶与努南综合征、豹子等多种疾病状态有关 综合症，以及各种癌症。Shp2在RAS/MAPK信号的完全激活中起关键作用 级联，以及从几个RTK到下游通路的信号转导。研究的目的是 对SHP2的作用的了解主要集中在信号的催化功能上 转导。我假设SHP2在ALK驱动的高危NB中是一个易处理的治疗漏洞。 最近，针对SHP2的变构小分子抑制剂SHP099被发现。而我们的 初步数据显示，通过SHP099抑制SHP2的催化活性不足以阻止NB肿瘤细胞 可行性，最近的研究表明，SHP2抑制与其他靶向基因结合显示出翻译前景 碱性磷酸酶抑制剂等治疗方法。确定SHP2突变体的功能效应势在必行 Nb是由于PTPN11基因突变所致。因此，我将表征PTPN11的突变，并研究 SHP2在高危NB中的磷酸酶非依赖性作用。这些研究将揭示这一被忽视的潜力。 SHP2的作用，并允许更合理的针对SHP2的药物开发，以产生临床效果 治疗选项。除了定义SHP2的功能作用外，我还将研究劳拉替尼和 SHP099以确定ALK驱动的高危NB在联合治疗中的抗肿瘤活性和存活率。 这些研究将为未来哪些患者群体将从这种组合中受益提供证据 临床试验设计并转化为临床。

PROJECT SUMMARY: Neuroblastoma (NB), a predominantly pediatric cancer of neural crest cell origin, is a clinically challenging disease due to the genomic and intratumoral heterogeneity. There are three risk stratifications to characterize the disease - low-risk, intermediate-risk, and high-risk - which are based on several factors such as mutation burden and age at diagnosis. High-risk NB patients have a poor prognosis, with a 5- year survival rate of roughly 50% despite dose-intensive chemoradioimmunotherapy treatment. The few common genomic aberrations in high-risk NB include amplification of the MYCN oncogene in 40% of cases, aberrations in the receptor tyrosine kinase (RTK) Anaplastic Lymphoma Kinase (ALK) in 14% of cases such as ALK gene amplification and three hot spot mutations at the F1174, F1245, and R1275 residues conferring ligand- independent kinase activity, and mutations in the PTPN11 gene, which encodes the SHP2 protein, in 3.4% of cases. The only targeted therapy for high-risk NB treatment is ALK inhibition, and despite the fact that the third generation ALK inhibitor lorlatinib is more potent and superior than previous generation ALK inhibitors, relapse in patients on lorlatinib occurs. Thus, the discovery of new targeted therapies and dosing schedules to overcome the challenges endured in the clinic is paramount. A novel therapeutic drug target, the Src homology 2-containing tyrosine phosphatase (SHP2), is a non-receptor protein tyrosine phosphatase implicated in several disease states including Noonan Syndrome, LEOPARD Syndrome, and a variety of cancers. SHP2 plays a critical role in the full activation of the RAS/MAPK signaling cascade, as well as signal transduction from several RTKs to downstream pathways. Research aimed at understanding the role of SHP2 has largely been focused on the catalytic function with respect to signal transduction. I hypothesize that SHP2 is a tractable therapeutic vulnerability in ALK-driven high-risk NB. Recently, an allosteric small molecule inhibitor which targets SHP2, SHP099, has been discovered. While our preliminary data show that inhibiting SHP2 catalytic activity via SHP099 is not sufficient to prevent NB tumor cell viability, recent studies reveal that SHP2 inhibition shows translational promise in combination with other targeted therapies such as ALK inhibitors. It is imperative to define the functional effect of the SHP2 mutants observed in NB due to the mutations in PTPN11. Therefore, I will characterize the mutations in PTPN11 and investigate the phosphatase-independent role of SHP2 in high-risk NB. These studies will shed light on this overlooked potential role of SHP2 and allow for more rational drug development targeting SHP2 to result in a clinically impactful treatment option. In addition to defining the functional role of SHP2, I will study the combination of lorlatinib and SHP099 to determine anti-tumor activity and survival of ALK-driven high-risk NB upon combination treatment. These studies will provide evidence for which patient populations would benefit from this combination for future clinical trial design and translation to the clinic.