3D High Throughput Model to Predict Drug Efficacy in Fibrosis Progression vs Reversal

3D 高通量模型预测纤维化进展与逆转的药物疗效

基本信息

批准号：
9975675
负责人：
LOUISE HECKER
金额：
$ 23.5万
依托单位：
GEORGIA INSTITUTE OF TECHNOLOGY
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-07-15 至 2022-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9975675
关键词：
3-Dimensional Accounting Affect Age Aging Apoptosis Biological Assay Biomedical Engineering Blood Vessels Cells Cellular Assay Cessation of life Cicatrix Collagen Contracts Development Disease Disease Progression Drug Combinations Drug usage Elderly Epidemic Exposure to FDA approved Fibroblasts Fibrosis Gel Heart Human Hydrogen Peroxide Individual Injury Kidney Liver Lung Lung diseases Modeling Monitor Myofibroblast Organ Oxidation-Reduction Pathologic Pathway interactions Patients Pharmaceutical Preparations Pharmacotherapy Phase Phenotype Pirfenidone Population Process Production Psychological reinforcement Quality of life Resistance Risk System Technology Testing Therapeutic age related aged aqueous base bioprinting body system clinically relevant cytokine drug candidate drug efficacy high throughput screening idiopathic pulmonary fibrosis improved inhibitor/antagonist novel pre-clinical preclinical development response screening senescence success therapeutic evaluation therapeutic target

项目摘要

ABSTRACT: 3D High Throughput Model to Predict Drug Efficacy in Fibrosis Progression vs Reversal Idiopathic pulmonary fibrosis (IPF) is the most relentlessly progressive and fatal fibrotic lung disorder, which disproportionately affects the elderly. Although two drugs have recently gained FDA-approval for IPF, these drugs only moderately slow the progression of lung decline and do not improve quality of life for patients. There are no available therapies that can `reverse' fibrosis. Despite efforts by numerous groups to develop IPF treatments, progress has been aggravatingly slow. This proposal focuses on two possible reasons for these difficulties: (1) Current pre-clinical screening models fail to reliably predict the success of drug candidates in humans, and (2) Although IPF is widely regarded as an age-related disease, drug treatments have not targeted age-associated pathologic mechanisms. The existing paradigm, that pathologic fibrosis is a “fibro-proliferative” process, has not led to effective IPF treatments. This proposal integrates expertise in fibroblast aging and novel IPF therapeutics in development (Hecker lab) with cutting edge technologies for microscale bioprinting and 3D cell assays (Takayama lab) to develop a high throughput phenotypic cellular screening assay to determine efficacy for fibrosis reversal. The proposed studies will utilize normal “control”, aged “senescent”, and IPF human lung fibroblasts in small numbers to bioengineer a high-throughput phenotypic assay that will evaluate fibrosis over a 21 day period. An aqueous two phase system (ATPS) bioprinting of these cells will be used to create microscale contraction assays that are several order of magnitude smaller in volume compared to conventional assays. Importantly, the project will repeatedly micro-print fresh collagen around already contracted cell-laden gels to enable repeated contractions over 21 days. The proposed model will enable the first high-throughput phenotypic screening assay with the capability to determine a drug candidate's efficacy for fibrosis progression and reversal. The new cellular assay will be validated for its ability to identify fibrosis reversal drugs using “Noxindoline” a highly selective Nox4 inhibitor that is currently in preclinical development by the Hecker lab. Noxindoline was identified by the Hecker lab through studies of age-dependent alterations in Nox4 that results in a sustained redox imbalance, and promotes senescence and apoptosis-resistance of myofibroblasts. The proposal hypothesizes that current therapies (Nintedanib and Pirfenidone) will inhibit the progression of pro-fibrotic phenotypes (but not reversal), whereas treatment with Noxindoline will promote the reversal of established pro-fibrotic phenotypes. The aims are: Aim1: Develop high throughput bioprinted cellular assay for fibrosis progression using non-senescent cells Aim2: Monitor fibrosis progression and reversal of senescent cells and IPF patient cells

摘要：3D 高通量模型预测纤维化进展与逆转的药物疗效特发性肺纤维化 (IPF) 是最严重的进行性和致命性纤维化肺部疾病，尽管最近有两种药物获得 FDA 批准用于治疗 IPF，但这些药物对老年人的影响尤为严重。药物只能适度减缓肺功能衰退的进展，并不能改善患者的生活质量。尽管许多团体都在努力开发能够“逆转”纤维化的疗法。 IPF 治疗进展极其缓慢，该提案主要集中在两个可能的原因上。这些困难：（1）当前的临床前筛选模型无法可靠地预测药物的成功 (2) 尽管 IPF 被广泛认为是一种与年龄相关的疾病，但药物治疗尚未针对与年龄相关的病理机制。病理性纤维化是一种“纤维增殖”过程的现有范式并未导致有效的 IPF 该提案整合了成纤维细胞衰老方面的专业知识和正在开发的新型 IPF 疗法。（Hecker 实验室）拥有用于微型生物打印和 3D 细胞测定的尖端技术（Takayama 实验室）开发高通量表型细胞筛选测定法以确定纤维化逆转的功效。拟议的研究将利用正常“对照”、老年“衰老”和 IPF 人肺成纤维细胞数字生物工程高通量表型测定将评估 21 天期间的纤维化。这些细胞的水性两相系统（ATPS）生物打印将用于产生微尺度收缩与传统检测相比，检测体积小几个数量级。该项目将在已经收缩的充满细胞的凝胶周围重复微打印新鲜胶原蛋白，以实现所提出的模型将实现第一个高通量表型。筛选试验能够确定候选药物对纤维化进展的功效，以及新的细胞测定法将验证其识别纤维化逆转药物的能力。 “Noxindoline”是一种高度选择性的 Nox4 抑制剂，目前由 Hecker 实验室进行临床前开发。 Hecker 实验室通过对 Nox4 中年龄依赖性变化的研究发现了 Noxindoline，导致持续的氧化还原失衡，并促进衰老和细胞凋亡抵抗该提案主张当前的疗法（尼达尼布和吡非尼酮）将抑制肌成纤维细胞。促纤维化表型的进展（但不是逆转），而 Noxindoline 治疗会促进逆转已确定的促纤维化表型。目的是：目标 1：使用非衰老细胞开发用于纤维化进展的高通量生物打印细胞测定法目标 2：监测衰老细胞和 IPF 患者细胞的纤维化进展和逆转