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.

项目概要 这项研究的总体目标是制造体外三维 (3D) 人体呼吸系统 具有气液界面（ALI）的模型，可作为 COVID-19 相关生物医学的平台 调查。最近爆​​发的 COVID-19 疫情是由严重急性呼吸道感染引起的 冠状病毒 2 综合征 (SARS-CoV-2)，导致确诊病例超过 1 亿例，确诊病例约 230 万例 全球死亡人数每天都在增加。对于 COVID-19 相关研究，仍然有一个紧迫的任务 需要一个 3D 完全异构的细胞呼吸模型，这可以增强我们对如何进行呼吸的理解 SARS-CoV-2 会引起肺损伤，并促进新疗法的开发，作为对动物的补充 模型和临床患者。因此，我们建议开发这样一种人体呼吸模型，其中 1) 表达血管紧张素转换酶 2 (ACE2) 蛋白和 2 型跨膜蛋白酶丝氨酸 (TMPRSS2) 允许病毒进入以研究 SARS-CoV-2 感染过程，2) 具有扩散性 气道和主要流体通道之间的渗透性，以实现穿过 ALI 的质量运输，从而允许 筛选经过临床证明可用于 COVID-19 的药物。我们假设体外人类呼吸道 模型可以通过嵌入式打印并进一步开发屈服应力下的可灌注结构来实现 基质浴含有原代人基质细胞、内皮细胞和成纤维细胞。 据此，提出两个具体目标如下： 目标 1：基于基质细胞的 3D 体外人体呼吸模型的嵌入式打印和灌注 可交联屈服应力基质浴。 目标 2：通过检测 ACE2 受体和 TMPRSS2 表征人类呼吸模型 气道上皮中的蛋白酶和 ALI 扩散渗透性的测定。 首次开发出基于明胶微凝胶和明胶溶液的可交联屈服应力细胞 复合基质浴将进行创新，用于在其中嵌入打印可灌注组织结构，从而使 人体呼吸模型制作。该模型开发研究将产生体外 3D 人体 利用新型可交联屈服应力细胞基质建立 ALI 和细胞基质呼吸模型 用于嵌入式打印的浴，并且不存在具有这种生理复杂性的其他模型。这样的人 呼吸模型将为研究 COVID-19 大流行相关感染提供多功能体外平台 工艺和筛选相关药物。