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A high-throughput 3D cantilever array to model airway smooth muscle hypercontractility in asthma

Asthma is often characterized by tissue-level mechanical phenotypes that include remodeling of the airway and an increase in airway tightening, driven by the underlying smooth muscle. Existing therapies only provide symptom relief and do not improve the baseline narrowing of the airway or halt progr...

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Autores principales: Beri, Pranjali, Plunkett, Christopher, Barbara, Joshua, Shih, Chien-Cheng, Barnes, S. Whitney, Ross, Olivia, Choconta, Paula, Trinh, Ton, Gomez, Datzael, Litvin, Bella, Walker, John, Qiu, Minhua, Hammack, Scott, Toyama, Erin Quan
Formato: Online Artículo Texto
Lenguaje:English
Publicado: AIP Publishing LLC 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10191677/
https://www.ncbi.nlm.nih.gov/pubmed/37206658
http://dx.doi.org/10.1063/5.0132516
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author Beri, Pranjali
Plunkett, Christopher
Barbara, Joshua
Shih, Chien-Cheng
Barnes, S. Whitney
Ross, Olivia
Choconta, Paula
Trinh, Ton
Gomez, Datzael
Litvin, Bella
Walker, John
Qiu, Minhua
Hammack, Scott
Toyama, Erin Quan
author_facet Beri, Pranjali
Plunkett, Christopher
Barbara, Joshua
Shih, Chien-Cheng
Barnes, S. Whitney
Ross, Olivia
Choconta, Paula
Trinh, Ton
Gomez, Datzael
Litvin, Bella
Walker, John
Qiu, Minhua
Hammack, Scott
Toyama, Erin Quan
author_sort Beri, Pranjali
collection PubMed
description Asthma is often characterized by tissue-level mechanical phenotypes that include remodeling of the airway and an increase in airway tightening, driven by the underlying smooth muscle. Existing therapies only provide symptom relief and do not improve the baseline narrowing of the airway or halt progression of the disease. To investigate such targeted therapeutics, there is a need for models that can recapitulate the 3D environment present in this tissue, provide phenotypic readouts of contractility, and be easily integrated into existing assay plate designs and laboratory automation used in drug discovery campaigns. To address this, we have developed DEFLCT, a high-throughput plate insert that can be paired with standard labware to easily generate high quantities of microscale tissues in vitro for screening applications. Using this platform, we exposed primary human airway smooth muscle cell-derived microtissues to a panel of six inflammatory cytokines present in the asthmatic niche, identifying TGF-β1 and IL-13 as inducers of a hypercontractile phenotype. RNAseq analysis further demonstrated enrichment of contractile and remodeling-relevant pathways in TGF-β1 and IL-13 treated tissues as well as pathways generally associated with asthma. Screening of 78 kinase inhibitors on TGF-β1 treated tissues suggests that inhibition of protein kinase C and mTOR/Akt signaling can prevent this hypercontractile phenotype from emerging, while direct inhibition of myosin light chain kinase does not. Taken together, these data establish a disease-relevant 3D tissue model for the asthmatic airway, which combines niche specific inflammatory cues and complex mechanical readouts that can be utilized in drug discovery efforts.
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spelling pubmed-101916772023-05-18 A high-throughput 3D cantilever array to model airway smooth muscle hypercontractility in asthma Beri, Pranjali Plunkett, Christopher Barbara, Joshua Shih, Chien-Cheng Barnes, S. Whitney Ross, Olivia Choconta, Paula Trinh, Ton Gomez, Datzael Litvin, Bella Walker, John Qiu, Minhua Hammack, Scott Toyama, Erin Quan APL Bioeng Articles Asthma is often characterized by tissue-level mechanical phenotypes that include remodeling of the airway and an increase in airway tightening, driven by the underlying smooth muscle. Existing therapies only provide symptom relief and do not improve the baseline narrowing of the airway or halt progression of the disease. To investigate such targeted therapeutics, there is a need for models that can recapitulate the 3D environment present in this tissue, provide phenotypic readouts of contractility, and be easily integrated into existing assay plate designs and laboratory automation used in drug discovery campaigns. To address this, we have developed DEFLCT, a high-throughput plate insert that can be paired with standard labware to easily generate high quantities of microscale tissues in vitro for screening applications. Using this platform, we exposed primary human airway smooth muscle cell-derived microtissues to a panel of six inflammatory cytokines present in the asthmatic niche, identifying TGF-β1 and IL-13 as inducers of a hypercontractile phenotype. RNAseq analysis further demonstrated enrichment of contractile and remodeling-relevant pathways in TGF-β1 and IL-13 treated tissues as well as pathways generally associated with asthma. Screening of 78 kinase inhibitors on TGF-β1 treated tissues suggests that inhibition of protein kinase C and mTOR/Akt signaling can prevent this hypercontractile phenotype from emerging, while direct inhibition of myosin light chain kinase does not. Taken together, these data establish a disease-relevant 3D tissue model for the asthmatic airway, which combines niche specific inflammatory cues and complex mechanical readouts that can be utilized in drug discovery efforts. AIP Publishing LLC 2023-05-16 /pmc/articles/PMC10191677/ /pubmed/37206658 http://dx.doi.org/10.1063/5.0132516 Text en © 2023 Author(s). https://creativecommons.org/licenses/by/4.0/All article content, except where otherwise noted, is licensed under a Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) ).
spellingShingle Articles
Beri, Pranjali
Plunkett, Christopher
Barbara, Joshua
Shih, Chien-Cheng
Barnes, S. Whitney
Ross, Olivia
Choconta, Paula
Trinh, Ton
Gomez, Datzael
Litvin, Bella
Walker, John
Qiu, Minhua
Hammack, Scott
Toyama, Erin Quan
A high-throughput 3D cantilever array to model airway smooth muscle hypercontractility in asthma
title A high-throughput 3D cantilever array to model airway smooth muscle hypercontractility in asthma
title_full A high-throughput 3D cantilever array to model airway smooth muscle hypercontractility in asthma
title_fullStr A high-throughput 3D cantilever array to model airway smooth muscle hypercontractility in asthma
title_full_unstemmed A high-throughput 3D cantilever array to model airway smooth muscle hypercontractility in asthma
title_short A high-throughput 3D cantilever array to model airway smooth muscle hypercontractility in asthma
title_sort high-throughput 3d cantilever array to model airway smooth muscle hypercontractility in asthma
topic Articles
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10191677/
https://www.ncbi.nlm.nih.gov/pubmed/37206658
http://dx.doi.org/10.1063/5.0132516
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