Development of a Three-Dimensional Bioengineering Technology to Generate Lung Tissue for Personalized Disease Modeling

Abstract

Stem cell technologies, especially patient‐specific, induced stem cell pluripotency and directed differentiation, hold great promise for changing the landscape of medical therapies. Proper exploitation of these methods may lead to personalized organ transplants, but to regenerate organs, it is necessary to develop methods for assembling differentiated cells into functional, organ‐level tissues. The generation of three‐dimensional human tissue models also holds potential for medical advances in disease modeling, as full organ functionality may not be necessary to recapitulate disease pathophysiology. This is specifically true of lung diseases where animal models often do not recapitulate human disease. Here, we present a method for the generation of self‐assembled human lung tissue and its potential for disease modeling and drug discovery for lung diseases characterized by progressive and irreversible scarring such as idiopathic pulmonary fibrosis (IPF). Tissue formation occurs because of the overlapping processes of cellular adhesion to multiple alveolar sac templates, bioreactor rotation, and cellular contraction. Addition of transforming growth factor‐β1 to single cell‐type mesenchymal organoids resulted in morphologic scarring typical of that seen in IPF but not in two‐dimensional IPF fibroblast cultures. Furthermore, this lung organoid may be modified to contain multiple lung cell types assembled into the correct anatomical location, thereby allowing cell‐cell contact and recapitulating the lung microenvironment. Our bottom‐up approach for synthesizing patient‐specific lung tissue in a scalable system allows for the development of relevant human lung disease models with the potential for high throughput drug screening to identify targeted therapies. Stem Cells Translational Medicine 2017;6:622–633

3D pulmospheres serve as a personalized and predictive multicellular model for assessment of antifibrotic drugs

“…Growing cells through several passages alters cellular phenotype. Others have developed lung 3D culture systems from IPF lung biopsies (22,23). However, because their goals in examining 3D cultures were different, they used other techniques to develop them.…”

“…The aim of the study was to regenerate stem cells for the treatment of IPF, not to provide a tool for personalized precision therapy for unique individual patients, which is the goal of the present study (22). Another method, that of Wilkinson et al used multiple alginate beads as a scaffold to grow multicellular structures on the surface of the beads using STEMCCA Cre-Excisable Constitutive Polycistronic Lentivirus-transduced cells to reprogram them as pluripotent cells (23). This method of preparation does not recapitulate the physical or the cell-autonomous characteristic of an individual patient lung.…”

Proceedings: Regenerative Medicine for Lung Diseases: A CIRM Workshop Report

“…Dr. Brigitte Gomperts (UCLA), reviewed the diverse lung tissues that have been differentiated from cell lines, including tracheospheres, bronchiolar organoids, bronchioalveolar spheres, alveolospheres, proximal and distal airway, and distal airway scaffolds onto which many cell types can be patterned. To date, organoid‐based disease models have been developed for CF, lung fibrosis , and BPD . Targeted correction of the CFTR gene in organoids derived from CF patients has also been reported .…”

“…Combining organoids with microfluidics devices has created the “lung on a chip,” consisting of epithelial and endothelial chambers in which breathing motions simulate cyclic stretch . Multiple pathologic states have been modeled this way, including asthma, COPD, airway response to injury, sepsis, and pulmonary edema .…”

Rising influence of synthetic biology in regenerative medicine

“…This structure was obtained within three weeks as opposed to the typical three months, thereby allowing the development of clinically functioning, fully in vivo tissue engineered airway replacements that can be obtained within a clinically useful time [23,46]. In saying this, research is still underway to develop improved human tissue structures through the use of novel state-of-the art technologies and methods, for instance three-dimensional (3D) printing, 3D-biofabrication approaches (a combination of 3D-bioplotting and thermally-assisted forming) [47,48], and the generation of orgainoids by bioreactor-assisted self-assembly methods [49].…”