A Predictive In Vitro Model for Screening Personalized Responses to CFTR-directed Therapeutics

摘要 本研究的目的是开发一种新的、可预测的体外模型，用于个性化对CFTR的反应。 定向治疗学。该建议响应了RFA-HL-15-027(用于预测的人体细胞模型 囊性纤维化跨膜电导调节剂指导的治疗的个体反应)。 囊性纤维化是由囊性细胞功能丧失突变引起的一种缩短生命的遗传性疾病。 纤维化跨膜电导调节基因编码对离子至关重要的阴离子通道 和液体运输。对某些个体(例如，G551D杂合子)的良好临床反应 使用一种新的cftr导向的调节剂药物iVacaftor，但对大多数患者来说，好处是 更不用说实质性的了。为了改善所有CF患者的生活，至关重要的是体内条件，包括 多种药物治疗的特定突变的多样性和复杂性(以及药代动力学相互作用) 在体外环境中准确重现，可用于快速准确地预测药物疗效。 我们在微流体平台上提出了一种非常新颖的体外个性化预测工具，利用患者的 自己的细胞，针对个人复杂的遗传背景制定治疗策略，并全面评估 对CFTR型药物的生理反应。这种型号将在我们的商业产品的基础上开发。 SynVivo®系列基于细胞的分析，将模拟CF肺的复杂呼吸道结构，包括 血液、上皮细胞和内皮细胞之间的规模、形态和细胞相互作用。我们会 将其与CFTR功能和呼吸道生理的新颖、综合评估相结合，包括 在体外环境中通过显微光学相干断层扫描清除粘液的各个方面 生物现实主义研究。 第一阶段将以清楚演示用于生理反应的微流控平台而告终 观察具有G551D门控突变的CF患者。在第二阶段，我们将通过 多药物治疗的CFTR靶向治疗的评估和详细的临床验证。一个多- 在所有领域拥有专业知识的学科、行业和学术合作伙伴关系对成功 项目目标的完成已经完成，包括研究微流控电池的熟练调查人员- 基础分析、肺生理学、药物发现和开发、治疗筛选和临床 学习。最终产品将商业化给制药公司、药物研究实验室和 大学/非营利性中心从事精确治疗、药物发现和药物输送。这个 主要目的是开发一种用作临床工具的分析方法，以先验地确定对 为CF患者提供个性化依据。

Abstract The objective of this study is to develop a novel, predictive in vitro model for personalized responses to CFTR- directed therapeutics. This proposal responds to RFA-HL-15-027 (Human Cellular Models for Predicting Individual Responses to Cystic Fibrosis Transmembrane Conductance Regulator- Directed Therapeutics). Cystic fibrosis (CF) is a life-shortening genetic disease caused by loss-of-function mutations of the Cystic Fibrosis Transmembrane conductance Regulator (CFTR) gene that encodes an anion channel critical for ion and fluid transport. Excellent clinical responses for some individuals (e.g., G551D heterozygotes) have been seen with ivacaftor, a new CFTR-directed modulator drug, but for the majority of patients, benefit has been much less substantial. To improve the lives of all CF patients, it is crucial that in vivo conditions, including the variety of specific mutations and complexity of multi-drug therapy (as well as pharmacokinetic interactions) are faithfully reproduced in an in vitro environment that can be used to rapidly and accurately predict drug efficacy. We propose a highly novel in vitro personalized predictive tool on a microfluidics platform, utilizing a patient’s own cells, to target the therapeutic strategy to an individual’s complex genetic background and assess full physiological responses to CFTR-directed drugs. This model will be developed on our commercially available SynVivo® family of cell based assays and will mimic the complex airway structure of the CF lung, including scale, morphology, and cellular interactions between the blood, the epithelium and the endothelium. We will couple this with a novel, integrative assessment of CFTR function and airway physiology including multiple aspects of mucus clearance via micro-optical coherence tomography in an in vitro environment enabling biologically realistic studies. Phase I will culminate with a clear demonstration of the microfluidic platform for physiological responses observed in CF patients with the G551D gating mutation. During Phase II, we will expand the platform by the evaluation of CFTR-targeted therapeutics with multi-agent therapy and detailed clinical validation. A multi- disciplinary, industry-academic partnership with expertise in all areas essential to the successful accomplishment of project goals has been assembled including skilled investigators studying microfluidics cell- based assays, CF lung physiology, drug discovery and development, therapeutic screening and clinical studies. The end-product will be commercialized to pharmaceutical firms, drug research labs and universities/non-profit centers engaged in precision therapeutics, drug discovery, and drug delivery. The primary endpoint is to develop an assay for use as a clinical tool to a priori determine efficacy on a personalized basis for CF patients.