Differential gene expression induced by different TLR agonists in A549 lung epithelial cells is modulated by CRISPR activation of TLR10

Knez, Špela (author), Narat, Mojca (author), Ogorevc, Jernej (author)

Toll-like receptor 10 (TLR10) is the only member of the TLR family whose function and ligand have not been clearly described. Literature reports on its function are contradictory and suggest a possible immunomodulatory role that depends on the cell type, the pathogen, and the level of TLR10 expression. To investigate the regulatory role of TLR10 in A549 lung epithelial cells, we overexpressed TLR10 using CRISPRa technology and examined the differential expression of various genes involved in TLR signaling activated by different TLR ligands, namely dsRNA, LPS, and Pam3Cys. The expression of proinflammatory cytokines, such as IL1β, IFNβ, TNFα, IL8, CXCL10, and CCL20, decreased in the challenged cells overexpressing TLR10, whereas the expression of the anti-inflammatory cytokine IL10 and the antimicrobial peptide hβD-2 increased. For several of the regulated inflammatory markers, we were able to show the change in gene expression was translated to the protein level. It appears that TLR10 can function as an anti-inflammatory in A549 cells, depending on its expression level and that the mode of action may be virulence factor-specific. The potential suppression of inflammation by regulating expression of TLR10 in lung epithelial cells may allow the development of new approaches to balance an inflammatory response and prevent extensive tissue damage in respiratory diseases.

• Language: English
• Keywords: CRISPR/dCas9, cytokines, inflammation, TLR10, lung epithelium, innate immunity
• Work type: Article
• Typology: 1.01 - Original Scientific Article
• Organization: BF - Biotechnical Faculty
• Publication status: Published
• Publication version: Version of Record
• Year: 2023
• Number of pages: 15 str.
• Numbering: Vol. 13, iss. 1, art. 19
• PID: 20.500.12556/RUL-143572
• UDC: 577.27
• ISSN on article: 2218-273X
• DOI: 10.3390/biom13010019
• COBISS.SI-ID: 135026947
• Publication date in RUL: 28.12.2022

Record is a part of a journal

• Title: Biomolecules
• Shortened title: Biomolecules
• Publisher: MDPI
• ISSN: 2218-273X
• COBISS.SI-ID: 519952921

Licences

• License: CC BY 4.0, Creative Commons Attribution 4.0 International
• Link: http://creativecommons.org/licenses/by/4.0/
• Description: This is the standard Creative Commons license that gives others maximum freedom to do what they want with the work as long as they credit the author.

Secondary language

• Language: Slovenian
• Keywords: imunologija, pljuča, epitelne celice, vnetje, TLR10, genetika, izražanje genov

Projects

• Funder: ARRS - Slovenian Research Agency
• Project number: P4-0220
• Name: Primerjalna genomika in genomska biodiverziteta

• Funder: ARRS - Slovenian Research Agency
• Funding programme: Young researchers