Development of novel protein-based therapeutics for lung cancer

肺癌是全球癌症死亡的主要原因。最常见的肺癌类型是非 小细胞肺癌(NSCLC)。这个应用程序的目标是实现一个协作工作，包括 临床前模型、生物工程和功能基因组学进一步表征和验证一种新的 肺癌治疗的“类”治疗策略。我们之前发表的工作和广泛的 初步数据表明，阻断CLCF1-CNTFR信号代表着一种独特的和以前的 未探索的肺癌治疗方法。斯威特-科德罗和科克伦实验室 在过去的几年里，为了验证这个信号轴，我们进行了广泛的合作，首先是利用shRNA基因 方法，现在通过开发一种经过改造的CNTFR受体诱饵(ECNTFR)。在目标1中， 我们将通过测量eCNTFR的热稳定性和蛋白分解稳定性来进一步开发eCNTFR作为治疗候选药物。 潜在的免疫原性和毒性、可制造性和药代动力学，并将优化这些 根据需要设置属性。我们还将与CLCF1进行eCNTFR复合体的结构分析，以定义 高亲和力结合特性。最后，我们将测试eCNTFR在广泛的 非小细胞肺癌的临床前模型，包括人类细胞系和患者来源的异种移植。在目标2中，我们将 进行体外生化分析以充分确定eCNTFR的细胞自主作用机制 封锁。为了进一步了解eCNTFR在体内的作用机制，我们将对异种移植进行补充 具有良好特征的肺癌基因工程小鼠模型。重要的是，这种模式 将使我们能够研究eCNTFR不仅对肿瘤细胞而且对微环境的影响，并 评估eCNTFR是否具有免疫调节作用。在目标3中，我们将确定最有效的 加强eCNTFR治疗的联合方法。这些努力将在理性和 不偏不倚地利用CRISPR/Cas9，使用专门针对FDA目标的库 批准的药物。候选的联合疗法随后将在动物模型中进行测试。我们的研究将 阐明这一配体/受体信号轴的关键生物学基础，并将提供临床前 验证了针对肺癌的创新战略，以保证其进一步的临床开发。

Lung cancer is the leading cause of cancer deaths worldwide. The most prevalent type of lung cancer is Non- Small Cell Lung Cancer (NSCLC). The goal of this application is to implement a collaborative effort involving preclinical models, bioengineering and functional genomics to further characterize and validate a novel “first in class” therapeutic strategy for the treatment of lung cancer. Our prior published work and extensive preliminary data indicates that blockade of CLCF1-CNTFR signaling represents a unique and previously unexplored approach for lung cancer therapy. The Sweet-Cordero and Cochran laboratories have collaborated extensively over the past several years to validate this signaling axis, first with shRNA genetic approaches, and now through the development of an engineered CNTFR receptor decoy (eCNTFR). In Aim 1, we will further develop eCNTFR as a therapeutic candidate by measuring its thermal and proteolytic stability, potential for immunogenicity and toxicity, manufacturability, and pharmacokinetics, and will optimize these properties as needed. We will also perform structural analysis of eCNTFR in complex with CLCF1 to define high affinity binding characteristics. Lastly, we will test eCNTFR for therapeutic efficacy across a wide array of preclinical models of NSCLC including human cell lines and patient-derived xenografts. In Aim 2, we will perform in vitro biochemical assays to fully define the cell-autonomous mechanism of action of eCNTFR blockade. To further understand the mechanism of action of eCNTFR in vivo, we will complement xenograft models with a well-characterized genetically engineered mouse model of lung cancer. Importantly, this model will allow us to study the effects of eCNTFR not only on tumor cells but also on the microenvironment, and to assess whether eCNTFR has immunomodulatory effects. In Aim 3, we will identify the most effective combination approaches to enhance eCNTFR therapy. These efforts will be carried out in a rational and unbiased manner by leveraging CRISPR/CAS9 using a library directed specifically at the targets of FDA approved drugs. Candidate combination therapies will then be tested in animal models. Our studies will elucidate critical biological underpinnings of this ligand/receptor signaling axis, and will provide preclinical validation of an innovative strategy for targeting lung cancer to warrant its further clinical development.