Effects of anti-TSLP treatment on the bronchial epithelial viral immune transcriptome and airway epithelial–smooth muscle cell crosstalk
(2024) MOBM02 20241Degree Projects in Molecular Biology
- Abstract
- Background: Asthma, a prevalent chronic lung disease, affecting millions worldwide, is characterised by airway inflammation and increased smooth muscle mass and constriction, leading to the narrowing of the airway walls. Recent biologics have emerged as promising treatments for asthma. This study focuses on Tezepelumab, a monoclonal antibody targeting TSLP, which has been shown to improve airway hyperresponsiveness, as well as reduce asthma exacerbations in patients with asthma. We aimed to investigate Tezepelumab's immunomodulatory effects on unstimulated and virally stimulated HBECs in asthma patients, exploring epithelial-smooth muscle cell cross-talk differences between treatments.
Methods: We performed a gene expression analysis,... (More) - Background: Asthma, a prevalent chronic lung disease, affecting millions worldwide, is characterised by airway inflammation and increased smooth muscle mass and constriction, leading to the narrowing of the airway walls. Recent biologics have emerged as promising treatments for asthma. This study focuses on Tezepelumab, a monoclonal antibody targeting TSLP, which has been shown to improve airway hyperresponsiveness, as well as reduce asthma exacerbations in patients with asthma. We aimed to investigate Tezepelumab's immunomodulatory effects on unstimulated and virally stimulated HBECs in asthma patients, exploring epithelial-smooth muscle cell cross-talk differences between treatments.
Methods: We performed a gene expression analysis, using nanoString (multiplex) and RT-qPCR, in human bronchial epithelial cells (HBECs), unstimulated or poly (I:C) stimulated, obtained from patients with asthma, following treatment with Tezepelumab or placebo. Furthermore, we investigated the effects of epithelial-derived conditioned media (CM) and viral stimuli on the expression of IL-33, TSLP, IFN-β and TGF-β, in bronchial smooth muscle cells (BSMCs). BSMCs were cultured and stimulated with various epithelial-derived CM, and poly (I:C) alone and in combination. Gene expressions were measured using RT-qPCR and protein release was measured by ELISA.
Results: Gene expression analysis in HBECs initially revealed downregulation of S100A8 and S100A9 through nanoString technology, which was subsequently confirmed by qPCR, indicating a consistent reduction. Additionally, further analysis indicated that Tezepelumab treatment also downregulated MUC5AC gene expression. Moreover, exposure to epithelial-derived CM elicited upregulation of IL-33, IFN-β, and TGF-β3 gene expression alongside increased IL-8 protein release in BSMCs. Notably, Tezepelumab treatment did not significantly change the expression of these cytokines.
Conclusion: In conclusion, this study suggests that Tezepelumab downregulates inflammatory mediators S100A8, S100A9, and MUC5AC, reducing airway inflammation and mucus production. It highlights the role of epithelial cells and their mediators in asthma, impacting airway smooth muscle cells, and underscores Tezepelumab's therapeutic potential in asthma management. (Less) - Popular Abstract (Swedish)
- Hur påverkar anti-TSLP cellkommunikation och inflammation i lungan?
Astma är en kronisk andningssjukdom som påverkar miljontals människor runt om i världen. Sjukdomen kännetecknas av luftvägsinflammation, överkänslighet och obstruktion, vilket leder till symtom som väsande andning, andnöd och hosta. Ny forskning visar att läkemedlet Tezepelumab, ett anti-TSLP läkemedel som blockerar TSLP, kan minska symptomen och förbättra patienters livskvalite.
Projektet gick ut på att utforska Tezepelumabs immunomodulatoriska effekt på bronkiala epitelceller, som både blivit ostimulerade och stimulerade med en virusliknande substans. Tezepelumab blockerar TSLP, ett viktigt inflammatoriskt protein, från att interagera med sin receptor (Figur 1),... (More) - Hur påverkar anti-TSLP cellkommunikation och inflammation i lungan?
Astma är en kronisk andningssjukdom som påverkar miljontals människor runt om i världen. Sjukdomen kännetecknas av luftvägsinflammation, överkänslighet och obstruktion, vilket leder till symtom som väsande andning, andnöd och hosta. Ny forskning visar att läkemedlet Tezepelumab, ett anti-TSLP läkemedel som blockerar TSLP, kan minska symptomen och förbättra patienters livskvalite.
Projektet gick ut på att utforska Tezepelumabs immunomodulatoriska effekt på bronkiala epitelceller, som både blivit ostimulerade och stimulerade med en virusliknande substans. Tezepelumab blockerar TSLP, ett viktigt inflammatoriskt protein, från att interagera med sin receptor (Figur 1), och därmed minskar inflammation. Utöver det utforskade projektet hur Tezepelumab påverkar samspelet mellan bronkiala epitelceller och bronkiala glatta muskelceller.
För att undersöka hur Tezepelumab påverkar bronkiala epitelceller från astmapatienter, odlades dessa celler före och efter behandling med läkemedlet eller placebo. Sedan utsattes cellerna för en virusliknande substans. Vi analyserade därefter genuttrycket i dessa celler för att se förändringar. Likaså odlades bronkiala glatta muskelceller och utsattes för supernatanten från de epitelceller som behandlas med Tezepelumab, före och efter behandling. Genom att undersöka genutryck och proteinmängder av olika signalsubstanser, försökte vi förstå hur Tezepelumab påverkar kommunikationen mellan cellerna i luftvägarna hos personer med astma.
Resultat
Efter behandling med Tezepelumab visade våra resultat en trend mot nedreglering av tre gener i bronkiala epitelceller, involverade i immunförsvaret i asthma. Dessa gener är involverade i inflammation och slemproduktion. Deras nedreglering överensstämmer med Tezepelumabs kända fördelar för att minska astmasymtom och exacerbationer. Även om dessa trender inte var konsekvent statistiskt signifikanta, belyser de behovet av ytterligare forskning för att förstå de exakta mekanismerna och den terapeutiska effekten av Tezepelumab vid astma.
Vår studie fann att bronkiala glatta muskelceller svarar på virusliknande substanser genom att uppreglera viktiga inflammatoriska gener, vilket indikerar en aktiv roll i astma-relaterad inflammation. Utöver det visade våra resultat på ett betydande samspel mellan bronkiala glatta muskelceller och bronkiala epitelceller. Våra resultat visar trender mot modulering av inflammatoriska vägar efter behandling med Tezepelumab, men dessa effekter var inte signifikanta. Vår studie understyrker vikten av samspel mellan bronkiala epitelceller och bronkiala glatta muskel celler i asthma och visar behovet av ytterligare forskning för att förstå detta samspelet samt den potentiella terapeutiska effekten Tezepelumab har på detta samspel.
Masterexamensprojekt i Molekylärbiologi 30 hp 2024
Biologiska institutionen, Lunds universitet
Handledare: Lena Uller, Sangeetha Ramu & Mandy Menzel
Biomedicinskt Centrum, D12, Respiratorisk Immunofarmakologi, Lunds universitet (Less)
Please use this url to cite or link to this publication:
http://lup.lub.lu.se/student-papers/record/9176955
- author
- Glans Lindörn, Philippa
- supervisor
-
- Lena Uller LU
- Sangeetha Ramu LU
- Mandy Menzel LU
- organization
- course
- MOBM02 20241
- year
- 2024
- type
- H2 - Master's Degree (Two Years)
- subject
- language
- English
- id
- 9176955
- date added to LUP
- 2024-10-23 11:06:59
- date last changed
- 2024-10-23 11:06:59
@misc{9176955,
abstract = {{Background: Asthma, a prevalent chronic lung disease, affecting millions worldwide, is characterised by airway inflammation and increased smooth muscle mass and constriction, leading to the narrowing of the airway walls. Recent biologics have emerged as promising treatments for asthma. This study focuses on Tezepelumab, a monoclonal antibody targeting TSLP, which has been shown to improve airway hyperresponsiveness, as well as reduce asthma exacerbations in patients with asthma. We aimed to investigate Tezepelumab's immunomodulatory effects on unstimulated and virally stimulated HBECs in asthma patients, exploring epithelial-smooth muscle cell cross-talk differences between treatments.
Methods: We performed a gene expression analysis, using nanoString (multiplex) and RT-qPCR, in human bronchial epithelial cells (HBECs), unstimulated or poly (I:C) stimulated, obtained from patients with asthma, following treatment with Tezepelumab or placebo. Furthermore, we investigated the effects of epithelial-derived conditioned media (CM) and viral stimuli on the expression of IL-33, TSLP, IFN-β and TGF-β, in bronchial smooth muscle cells (BSMCs). BSMCs were cultured and stimulated with various epithelial-derived CM, and poly (I:C) alone and in combination. Gene expressions were measured using RT-qPCR and protein release was measured by ELISA.
Results: Gene expression analysis in HBECs initially revealed downregulation of S100A8 and S100A9 through nanoString technology, which was subsequently confirmed by qPCR, indicating a consistent reduction. Additionally, further analysis indicated that Tezepelumab treatment also downregulated MUC5AC gene expression. Moreover, exposure to epithelial-derived CM elicited upregulation of IL-33, IFN-β, and TGF-β3 gene expression alongside increased IL-8 protein release in BSMCs. Notably, Tezepelumab treatment did not significantly change the expression of these cytokines.
Conclusion: In conclusion, this study suggests that Tezepelumab downregulates inflammatory mediators S100A8, S100A9, and MUC5AC, reducing airway inflammation and mucus production. It highlights the role of epithelial cells and their mediators in asthma, impacting airway smooth muscle cells, and underscores Tezepelumab's therapeutic potential in asthma management.}},
author = {{Glans Lindörn, Philippa}},
language = {{eng}},
note = {{Student Paper}},
title = {{Effects of anti-TSLP treatment on the bronchial epithelial viral immune transcriptome and airway epithelial–smooth muscle cell crosstalk}},
year = {{2024}},
}