Engineering Dynamic 3D-Bioprinted Models of Pulmonary Vascular Disease

肺血管疾病的工程动态 3D 生物打印模型

• 批准号: 10514521
• 负责人: Duncan J Davis-Hall
• 金额: $ 3.83万
• 依托单位: UNIVERSITY OF COLORADO DENVER
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-02-01 至 2024-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10514521
• 关键词: 3-Dimensional; 3D Print; Bathing; Biocompatible Materials; Biological Assay; Biological Models; Biomanufacturing; Biomedical Engineering; Blood Pressure; Blood Vessels; Cardiovascular Diseases; Cardiovascular system; Cell Culture Techniques; Cell Survival; Cells; Chronic; Clinical; Collagen; Colorado; Complex; Crosslinker; Cues; Deposition; Development; Dimensions; Disease; Disease Progression; Elasticity; Encapsulated; Engineering; Environment; Exhibits; Extracellular Matrix; Extracellular Matrix Proteins; Fellowship; Fibroblasts; Fibrosis; Foundations; Gelatin; Gene Expression; Genes; Geometry; Goals; Human; Hydrogels; Immunohistochemistry; In Vitro; Laboratories; Learning; Light; Lung; Lung diseases; Matrix Metalloproteinases; Measures; Medical; Mentors; Methacrylates; Methods; Modeling; Modulus; Myofibroblast; Pathogenesis; Pathologic; Pathology; Patients; Peptides; Phenotype; Physicians; Physiological; Physiology; Population; Production; Proteins; Pulmonary Hypertension; Pulmonary arterial remodeling; Quantitative Reverse Transcriptase PCR; Reaction; Research; Research Personnel; Research Training; Right Ventricular Dysfunction; Scientist; Smooth Muscle Actin Staining Method; Solid; Structure; Symptoms; Syringes; System; Techniques; Technology; Testing; Therapeutic; Time; Tissue-Specific Gene Expression; Tissues; Training; Translational Research; Ultraviolet Rays; Universities; Vascular Diseases; Vascular remodeling; base; bioink; bioprinting; cell type; ethylene glycol; experience; experimental study; human disease; human model; human study; immunocytochemistry; insight; mechanical properties; mortality; new therapeutic target; novel; polymerization; pre-doctoral; pulmonary arterial hypertension; pulmonary vascular disorder; response; right ventricular failure; success; symposium; three dimensional cell culture; three dimensional structure; three-dimensional modeling; time use; tool

PROJECT SUMMARY/ABSTRACT Pulmonary arterial hypertension (PAH) is a progressive and incurable disease, characterized by elevated pulmonary blood pressure, remodeling of the pulmonary arteries, and ultimately the development of right ventricular failure. Unfortunately, the only clinically available therapeutic treatments mitigate symptoms but do not cure the disease. Existing cell culture techniques represent a significant barrier to discovering new therapeutic targets for PAH because these systems do not adequately reproduce key aspects of human physiology, specifically, the complex 3D structure of the pulmonary vasculature and the time-dependent changes in extracellular matrix (ECM) mechanical properties that occur during disease progression. Therefore, an urgent need remains to develop new tools and technologies that enable us to study the pathogenesis of PAH over time. We propose to develop a new class of phototunable poly(ethylene glycol) (PEG)-based hydrogel biomaterials and biomanufacturing techniques that allow investigators to control the mechanical properties of the local microenvironment (i.e., stiffen) on-demand around patient-derived fibroblasts encapsulated within 3D-printed vascular models using focused light, with the goal of emulating PAH pathogenesis in vitro. The advanced biomaterial platform implemented here will provide the foundation for biological models of increasing complexity comprising multiple cell types that are cultured under flow under development in the sponsor's laboratory that reveal novel mechanistic insights into reduction of human disease. This project will bring together the applicant and a diverse mentoring team made up of bioengineers and clinician-scientists specializing in cardiovascular and pulmonary diseases to further develop the pulmonary workforce. Fellowship training will include clinical experiences through the Pulmonary Hypertension Breakthrough Initiative, presentations at research conferences and grand rounds to aid professional development, and hands-on training in Dr. Kurt Stenmark's Cardiovascular Pulmonary Research Lab to learn experimental techniques essential to understanding PAH. The completed model will provide a platform for testing and validating therapies for pulmonary hypertension, advancing translational research in this field. We propose two specific aims to demonstrate the feasibility of this approach. AIM I: Engineer a dynamic 3D- bioprinted cell culture platform with controllable modulus of elasticity. AIM II: Investigate the influence of dimensionality and material modulus on fibroblast activation using patient-derived cells. The success of Aim I will be measured through rheological characterization and cell viability assays. Aim II will measure fibroblast activation through immunohistochemistry and a concise qRT-PCR array to compare phenotypic changes among healthy patient-derived cells grown in 3D-bioprinted blood vessel mimics that emulate pathological ECM, healthy cells grown on 2D hydrogel substrates, and freshly isolated PAH patient cells.

项目摘要/摘要 肺动脉高压（PAH）是一种进行性疾病，其特征是 肺血压，肺动脉的重塑以及最终的右发育 心室衰竭。不幸的是，唯一可以缓解症状的临床治疗治疗方法 无法治愈疾病。现有的细胞培养技术代表了发现新的障碍 PAH的治疗靶标，因为这些系统不能充分再现人类的关键方面 生理学，特别是肺脉管系统的复合体3D结构和时间依赖性 疾病进展过程中发生的细胞外基质（ECM）机械性能的变化。所以， 迫切需要开发新工具和技术，使我们能够研究 随着时间的流逝。我们建议开发一类新的光电聚（乙二醇）（PEG） 水凝胶生物材料和生物制造技术允许研究人员控制机械 局部微环境的特性（即，僵硬）在患者衍生的成纤维细胞周围点播 使用聚焦光封装在3D打印的血管模型中，以模拟PAH 体外发病机理。这里实施的高级生物材料平台将为 增加复杂性的生物学模型，包括多种细胞类型，这些细胞类型在流动下培养 赞助商的实验室的发展，揭示了对人类减少的新机械见解 疾病。该项目将汇集由生物工程组成的申请人和多样化的指导团队 和临床医生科学家专门研究心血管和肺部疾病，以进一步发展肺部 劳动力。奖学金培训将包括通过肺动脉高压 突破性倡议，在研究会议上的演讲和盛大的回合以帮助专业 Kurt Stenmark博士的心血管肺部研究实验室的开发和动手培训 实验技术对于理解PAH至关重要。完整的模型将为 测试和验证肺动脉高压的疗法，在该领域进行转化研究。我们 提出了两个具体的目的，以证明这种方法的可行性。 AIM I：设计动态的3D- 具有可控弹性模量的生物打印细胞培养平台。 AIM II：调查 使用患者衍生的细胞激活成纤维细胞激活的尺寸和材料模量。目标的成功 将通过流变特征和细胞活力测定法进行测量。 AIM II将测量成纤维细胞 通过免疫组织化学和简洁的QRT-PCR阵列激活以比较表型变化 在3D-生物包裹的血管中生长的健康患者衍生细胞中，模仿病理学 ECM，在2D水凝胶底物上生长的健康细胞和新鲜分离的PAH患者细胞。