Embedded Printing of Human Respiratory Model with Air-Liquid Interface for COVID-19 Research

用于 COVID-19 研究的具有气液界面的人体呼吸模型的嵌入式打印

• 批准号: 10353655
• 负责人: Yong Huang
• 金额: $ 17.66万
• 依托单位: UNIVERSITY OF FLORIDA
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-08-01 至 2024-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10353655
• 关键词: 2019-nCoV; 3-Dimensional; ACE2; Adipose tissue; Air; Alveolar; Alveolus; Animal Model; Antiviral Agents; Bathing; Biology; Blood Vessels; Blood capillaries; Bronchial Tree; COVID-19; COVID-19 outbreak; COVID-19 pandemic; Cells; Cellular Stress; Cellular biology; Cessation of life; Characteristics; Clinical; Cytoskeleton; Detection; Development; Dextrans; Diffuse; Diffusion; Disease; Drug Modelings; Endothelial Cells; Endothelium; Enzymes; Epithelial Cells; Exposure to; Fibroblasts; Fluorescein; Gelatin; Goals; Human; Immune response; Immunofluorescence Immunologic; In Vitro; Infection; Ink; Investigation; Isothiocyanates; Liquid substance; Lower respiratory tract structure; Lung; Mediating; Microcirculation; Modeling; Patients; Peptide Hydrolases; Perfusion; Permeability; Pharmaceutical Preparations; Physiological; Polymerase Chain Reaction; Printing; Process; Proteins; Research; Research Personnel; Respiratory System; Reverse Transcription; SARS-CoV-2 infection; Serine Protease; Solid; Solid Neoplasm; Stains; Stress; Stromal Cells; Study models; Supporting Cell; Surface; System; TMPRSS2 gene; Therapeutic; Thick; Time; Tissue constructs; Tissues; Transcript; Vaccines; Viral; Virus; airway epithelium; alveolar epithelium; base; bioprinting; crosslink; design; innovation; lung injury; membrane model; model development; novel; receptor; respiratory; screening; self assembly; stem; success; systems research; vaccine evaluation

Project Summary The overarching goal of this research is to fabricate an in vitro three-dimensional (3D) human respiratory model with an air-liquid interface (ALI), which can serve as a platform for COVID-19 related biomedical investigations. A recent COVID-19 outbreak, which is due to the infection of a severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), results in more than 100 million confirmed cases and about 2.3 million deaths with numbers increasing everyday globally. For COVID-19 related research, there remains an urgent need for a 3D fully heterogeneous, cellular respiratory model, which can enhance our understanding of how SARS-CoV-2 induces lung injury and facilitate the development of new treatments as complementary to animal models and clinical patients. We therefore propose to develop such a human respiratory model, which 1) expresses angiotensin-converting enzyme 2 (ACE2) protein and transmembrane protease serine type 2 (TMPRSS2) that allow viral entry to study the SARS-CoV-2 infection process, and 2) has the diffusional permeability between the airway and primary fluid channel to enable the mass transport across ALI, permitting the screening of clinically proven drugs for COVID-19 use. We hypothesize that an in vitro human respiratory model can be realized by embedded printing and further developing a perfusable construct in a yield-stress matrix bath containing primary human stromal, endothelial, and fibroblast cells. Accordingly, two specific aims are proposed as follows: Aim 1: Embedded printing and perfusion of a 3D in vitro human respiratory model in a stromal cell-based cross-linkable yield-stress matrix bath. Aim 2: Characterization of the human respiratory model via detection of ACE2 receptor and TMPRSS2 protease in the airway epithelium and determination of the diffusional permeability of ALI. For the first time, a gelatin microgels and gelatin solution-based cross-linkable yield-stress cellular composite matrix bath will be innovated for embedded printing of perfusable tissue constructs in it, enabling human respiratory model fabrication. This model development study will result in an in vitro 3D human respiratory model with ALI and cellular stroma by utilizing the novel cross-linkable yield-stress cellular matrix bath for embedded printing, and no other models with this physiological complexity exist. Such a human respiratory model will provide a versatile in vitro platform for studying the COVID-19 pandemic-related infection process and screening related drugs.

项目摘要 这项研究的总体目标是制造体外三维（3D）人类呼吸 具有空气界面（ALI）的模型，该模型可以用作COVID-19相关生物医学的平台 调查。最近的Covid-19爆发是由于严重急性呼吸道的感染 综合征冠状病毒2（SARS-COV-2），导致超过1亿例确认病例，约230万例 每天的死亡人数在全球范围内增加。对于COVID-19相关研究，仍然是紧急的 需要3D完全异质的细胞呼吸模型，这可以增强我们对如何的理解 SARS-COV-2诱导肺损伤，并促进新疗法的发展作为互补的动物 模型和临床患者。因此，我们建议开发这样的人类呼吸模型，1） 表达血管紧张素转换酶2（ACE2）蛋白和跨膜蛋白酶2型 （TMPRSS2）允许病毒进入研究SARS-COV-2感染过程，2）具有扩散 气道和初级流体通道之间的渗透性使质量运输能够穿越ALI，从而允许 筛查临床证明的药物以供1900次使用。我们假设一种体外人类呼吸道 模型可以通过嵌入式印刷和进一步开发屈服于压力的灌注构建体来实现 含有原发性人基质，内皮和成纤维细胞的基质浴。 因此，提出了两个具体目标，如下所示： AIM 1：在基于基于基于细胞的基于基质细胞中的3D体外人类呼吸模型的嵌入式印刷和灌注 可以交叉连接的产量压力矩阵浴。 AIM 2：通过检测ACE2受体和TMPRSS2来表征人呼吸模型 气道上皮中的蛋白酶和ALI扩散渗透性的测定。 首次，明胶微凝胶和明胶溶液的交叉连接屈服压力细胞 复合基质浴将进行创新，以嵌入其中的灌注式组织构建体的嵌入 人类呼吸模型制造。这项模型开发研究将导致体外3D人类 通过使用新型的交叉连接屈服压细胞基质，具有ALI和细胞基质的呼吸模型 用于嵌入式印刷的浴室，没有其他具有这种生理复杂性的模型。这样的人 呼吸模型将提供一个多功能的体外平台，用于研究COVID-19与大流行有关的感染 过程和筛查相关的药物。