Applications and Approaches for Three-Dimensional Precision-Cut Lung Slices. Disease Modeling and Drug Discovery

Alsafadi, Hani N.; Uhl, Franziska E.; Pineda, Ricardo; Bailey, Kolene E.; Rojas, Mauricio; Wagner, Darcy E.; Königshoff, Mélanie

doi:10.1165/rcmb.2019-0276tr

Cited by 91 publications

(88 citation statements)

References 84 publications

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“…The use of PCLS has recently gained attention as a promising model for studying lung physiology and pathology and as a possible means to screen for novel therapeutics (1). However, the use of PCLS in medical research has been limited by difficulties in obtaining RNA of sufficient quality for non-biased and high through-put analysis methods such as RNA-seq.…”

Section: Discussionmentioning

confidence: 99%

“…The previous study, utilizing rat tissue, (25), used a low agarose concentration (0.75 %) for filling the rat lungs, which most likely facilitated the isolation of high quality RNA. However, the usage of an equivalent percentage of agarose is usually not a possibility for generating PCLS from larger species, such as pig or human (especially from diseased lung explants), where an agarose concentration of 3 % is most commonly used (1). Additionally, a recent study by Niehof et al reported improved RNA isolation from human PCLS containing 1.5 % agarose for RNA microarray analysis, using a magnetic bead-based isolation approach (8).…”

Section: Discussionmentioning

confidence: 99%

“…Precision-cut lung slices (PCLS) have received increased attention as a relevant model for studying and modeling lung diseases (1). Most lung diseases involve several cell types and derangements in their surrounding environment, such as the extracellular matrix.…”

Section: Introductionmentioning

confidence: 99%

“…They thus require complex disease models to explore disease pathogenesis and novel therapeutics. PCLS excel at retaining the native architecture of the lung and keeping the spatial composition of cells and their surrounding environment intact, making it possible to study cell-cell and cell-matrix interactions in health or disease (1).…”

Section: Introductionmentioning

confidence: 99%

“…The complexity of PCLS and their ability to recapitulate the native environment of the lung makes it well-suited for advanced drug screening (5). Additionally, PCLS can provide readouts from multiple levels, such as gene/protein expression and imaging, which allows for both phenotypic and genotypic profiling after exposure to a specific drug, including small molecules and biologics (1).…”

Section: Introductionmentioning

confidence: 99%

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Isolation of high yield and quality RNA from human precision-cut lung slices for RNA-sequencing and computational integration with larger patient cohorts

Stegmayr

Alsafadi

Langwiński

et al. 2020

Preprint

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Precision-cut lung slices (PCLS) have gained increasing interest as a model to study lung biology and disease, as well as for screening novel therapeutics. In particular, PCLS derived from human tissue can better recapitulate some aspects of lung biology and disease as compared to PCLS derived from animals (e.g. clinical heterogeneity), but access to human tissue is limited. A number of different experimental readouts have been established for use with PCLS, but obtaining high yield and quality RNA for downstream gene expression analysis has remained challenging. This is particularly problematic for utilizing the power of next-generation sequencing techniques, such as RNA-sequencing (RNA-seq), for non-biased and high through-put analysis of PCLS human cohorts. In the current study, we present a novel approach for isolating high quality RNA from a small amount of tissue, including diseased human tissue, such as idiopathic pulmonary fibrosis (IPF). We show that the RNA isolated using this method is of sufficient quality for both RT-qPCR and RNA-seq analysis. Furthermore, the RNA-seq data from human PCLS was comparable to data generated from native tissue and could be used in several established computational pipelines, including deconvolution of bulk RNA-seq data using publicly available single-cell RNA-seq data sets. Deconvolution using Bisque revealed a diversity of cell populations in human PCLS derived from distal lung tissue, including several immune cell populations, which correlated with cell populations known to be present and aberrant in human disease, such as IPF.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Isolation of high yield and quality RNA from human precision-cut lung slices for RNA-sequencing and computational integration with larger patient cohorts

Stegmayr

Alsafadi

Langwiński

et al. 2020

Preprint

Self Cite

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Microphysiological Models of Lung Epithelium‐Alveolar Macrophage Co‐Cultures to Study Chronic Lung Disease

Lagowala,

Wally,

Wilmsen

et al. 2023

Advanced Biology

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The interactions between immune cells and epithelial cells influence the progression of many respiratory diseases, such as chronic obstructive pulmonary disease (COPD). In vitro models allow for the examination of cells in controlled environments. However, these models lack the complex 3D architecture and vast multicellular interactions between the lung resident cells and infiltrating immune cells that can mediate cellular response to insults. In this study, three complementary microphysiological systems are presented to delineate the effects of cigarette smoke and respiratory disease on the lung epithelium. First, the Transwell system allows the co‐culture of pulmonary immune and epithelial cells to evaluate cellular and monolayer phenotypic changes in response to cigarette smoke exposure. Next, the human and mouse precision‐cut lung slices system provides a physiologically relevant model to study the effects of chronic insults like cigarette smoke with the dissection of specific interaction of immune cell subtypes within the structurally complex tissue environment. Finally, the lung‐on‐a‐chip model provides an adaptable system for live imaging of polarized epithelial tissues that mimic the in vivo environment of the airways. Using a combination of these models, a complementary approach is provided to better address the intricate mechanisms of lung disease.

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Hydrogel‐Embedded Precision‐Cut Lung Slices Model Lung Cancer Premalignancy Ex Vivo

Blomberg,

Sompel,

Hauer

et al. 2023

Adv Healthcare Materials

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Lung cancer is the leading global cause of cancer‐related deaths. Although smoking cessation is the best prevention, 50% of lung cancer diagnoses occur in people who have quit smoking. Research into treatment options for high‐risk patients has been constrained to rodent models, which are time‐consuming, expensive, and require large cohorts. Embedding precision‐cut lung slices (PCLS) within an engineered hydrogel and exposing this tissue to vinyl carbamate, a carcinogen from cigarette smoke, creates an in vitro model of lung cancer premalignancy. Hydrogel formulations are selected to promote early lung cancer cellular phenotypes and extend PCLS viability to six weeks. Hydrogel‐embedded PCLS are exposed to vinyl carbamate, which induces adenocarcinoma in mice. Analysis of proliferation, gene expression, histology, tissue stiffness, and cellular content after six weeks reveal that vinyl carbamate induces premalignant lesions with a mixed adenoma/squamous phenotype. Putative chemoprevention agents diffuse through the hydrogel and induce tissue‐level changes. The design parameters selected using murine tissue are validated with hydrogel‐embedded human PCLS and results show increased proliferation and premalignant lesion gene expression patterns. This tissue‐engineered model of human lung cancer premalignancy is the foundation for more sophisticated ex vivo models that enable study of carcinogenesis and chemoprevention strategies.This article is protected by copyright. All rights reserved

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Applications and Approaches for Three-Dimensional Precision-Cut Lung Slices. Disease Modeling and Drug Discovery

Cited by 91 publications

References 84 publications

Isolation of high yield and quality RNA from human precision-cut lung slices for RNA-sequencing and computational integration with larger patient cohorts

Isolation of high yield and quality RNA from human precision-cut lung slices for RNA-sequencing and computational integration with larger patient cohorts

Microphysiological Models of Lung Epithelium‐Alveolar Macrophage Co‐Cultures to Study Chronic Lung Disease

Hydrogel‐Embedded Precision‐Cut Lung Slices Model Lung Cancer Premalignancy Ex Vivo

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