Advancing RAS and RASopathy Therapies

在2019财年，CCR和DCEG的一组调查人员成功地为Rasopathies的新NCI计划竞争资金。有了这笔资金，我们在2019年2月的一个项目启动会上召集了一个由国际RA专家组成的多学科小组。作为这次会议的结果，我们为非NF1风湿病患者进行了一项纵向队列研究，这项研究将通过DCEG进行。该议定书最近得到了CIRB的批准，目前正在登记。到目前为止，已有60多名参与者参加了现场队列，6名患者访问了临床中心。这项试验的目标是前瞻性地研究这些患者的癌症发生情况，并确定可用于监测介入试验成功或失败的特定可测量终点。这包括使用专业人士，由POB的行为健康核心带头。已经确定了可能的第一个介入试验，并进行了适当的行业接触，包括与库拉肿瘤科就替普法尼在Costello综合征中的使用进行了CRADA。除了临床方案，我们还启动了几项临床前研究，以评估RAS靶向药物在Rasopathies中的疗效。我们计划在Costello综合征的HRASG12S模型中研究替法尼布，并在CFCMEKY130C模型中研究米达米尼。这些研究将与CAPR(Zoe Weaver-Ohler)和Lino Tessarollo博士合作进行。HRASG12S模型项目将在RASopathiesNET专题讨论会上以抽象形式介绍。我们已经开始与Raj Chiari和Roackie AwDisi合作创建新的Noonan综合征小鼠模型。已经鉴定出转基因不能进入生殖系的创始小鼠，现在将用b-肌动蛋白Cre小鼠培育这些小鼠来表达转基因。最后，在道格拉斯·斯图尔特博士领导的人口基因组学工作中，我们的目标是识别新的Rasathy变种和基因。该项目已经开始，新确定的RAS变种将在Yohe实验室进行功能验证。这种验证包括使用纯化蛋白的生化评估(与罗斯曼实验室合作)，细胞中RAS-GTP及PERK和PARK水平的评估(Yohe实验室)，细胞定位的评估(Turbyville实验室)，与下游效应物相互作用的评估(Turbyville实验室)，对成肌分化的影响(Yohe实验室)，对细胞增殖的影响(Yohe实验室)，以及对斑马鱼胚胎形态的影响(LCDS水产核心Christine Kettenhofen)。HRAS变种的鉴定结果将在RASopathiesNet研讨会上公布，新的RAF变种将在莫里森实验室进行功能鉴定。新的LZTR1变种也将在Yohe实验室进行评估，这些变种是通过Rasathy人群基因组学工作以及通过与Michael Sargen博士(黑色素瘤易发家族)的合作而确定的。

In FY 2019, a group of investigators from CCR and DCEG successfully competed for funding for a new NCI initiative for the RASopathies. With this funding, we assembled a multi-disciplinary group of international RASopathy experts at a project kick-off meeting in February 2019. As a result of this meeting, we have produced a longitudinal cohort study for patients with non-NF1 RASopathies that will be conducted through DCEG. This protocol was recently approved by the CIRB and is currently enrolling. Over 60 participants have enrolled in the field cohort and 6 patients have visited the clinical center to date. The goals of this trial are to prospectively study the incidence of cancer development in these patients but also to identify specific measurable endpoints that can be used for monitoring the success or failure of interventional trials. This includes the use of PROs, being spearheaded by the behavioral health core in the POB. Potential first interventional trials were identified, and appropriate industry contacts have been made, including a CRADA with Kura Oncology for the use of tipifarnib in Costello syndrome. In addition to the clinical protocol, we also have initiated several preclinical studies to evaluate the efficacy of RAS-targeting agents in RASopathies. We plan a study of tipifarnib in an HRASG12S model of Costello syndrome and a study of mirdametinib in a MEKY130C model of CFC. These studies will be conducted in collaboration with CAPR (Zoe Weaver-Ohler)and Dr. Lino Tessarollo. The HRASG12S model project will be presented in abstract form at the RASopathiesNET symposium. We have begun the process of creating new mouse models of Noonan syndrome in collaboration with Raj Chiari and Roackie Awasthi. Founder mice with incorportation of the transgene into the germline have been identified, and these mice will now be bred with b-actin Cre mice to express the transgene. Finally, in a population genomics effort headed by Dr. Douglas Stewart, we aim to identify new RASopathy variants and genes. This project has begun, and newly identified RAS variants will be functionally validated in the Yohe lab. This validation includes biochemical assessment with purified protein (collaboration with the Rossman lab), assessment of RAS-GTP and pERK and pAKT levels in cells (Yohe lab), assessment of cellular localization (Turbyville lab), assessment of interactions with downstream effectors (Turbyville lab), impact on myogenic differentiation (Yohe lab), impact on cell proliferation (Yohe lab), and impact on zebrafish embryo morphology (Christine Kettenhofen in the LCDS aquatics core). Results from the characterization of HRAS variants will be presented at the RASopathiesNet symposium, Novel RAF variants will be functionally characterized in the Morrison lab. Novel LZTR1 variants identified through the RASopathy population genomics effort and through collaborations with Dr. Michael Sargen (melanoma prone families) will also be assessed in the Yohe lab.