Biology subjects

Woldhuis, R. R.

Publications and source records attributed to Woldhuis, R. R..

4 recordsLinked to original sources

bioRxiv2023

The lung extracellular matrix protein landscape in severe early-onset and moderate chronic obstructive pulmonary disease.

Extracellular matrix (ECM) remodeling has been implicated in the irreversible obstruction of airways and destruction of alveolar tissue in chronic obstructive pulmonary disease (COPD). Studies investigating differences in the lung ECM in COPD have mainly focused on some collagens and elastin, leaving an array of ECM components unexplored. We investigated the differences in the ECM landscape comparing severe-early onset (SEO-) COPD and moderate COPD to control lung tissue for collagen type I chain 1 (COL1A1), COL6A1, COL6A2, COL14A1, fibulin 2 and 5 (FBLN2, FBLN5), latent transforming growth factor-beta binding protein 4 (LTBP4), lumican (LUM), versican (VCAN), decorin (DCN), and elastin (ELN) using image analysis and statistical modelling. Percentage area and/or mean intensity of expression of LUM in the parenchyma, and COL1A1, FBLN2, LTBP4, DCN, and VCAN in the airway walls, was proportionally lower in COPD compared to controls. Lowered levels of most ECM proteins were associated with decreasing FEV1 measurements, indicating a relationship with disease severity. Furthermore, we identified six unique ECM signatures where LUM and COL6A1 in parenchyma and COL1A1, FBLN5, DCN, and VCAN in airway walls appear essential in reflecting the presence and severity of COPD. These signatures emphasize the need to examine groups of proteins to represent an overall difference in the ECM landscape in COPD, that are more likely to be related to functional effects, than individual proteins. Our study revealed differences in the lung ECM landscape between control and COPD and between SEO and moderate COPD signifying distinct pathological processes in the different subgroups. NEW & NOTEWORTHYOur study identified COPD-associated differences in the lung ECM composition. We highlight the compartmental differences in the ECM landscape in different subtypes of COPD. The most prominent differences were observed for severe-early onset COPD. Moreover, we identified unique ECM signatures that describe airway walls and parenchyma providing insight into the intertwined nature and complexity of ECM changes in COPD that together drive ECM remodeling and may contribute to disease pathogenesis.

molecular biology↗

bioRxiv2023

Strong paracrine effects of SASP from senescence-induced severe early-onset COPD-derived fibroblasts

BackgroundLung fibroblasts from Severe Early-Onset (SEO-)COPD patients exhibit increased cellular senescence with higher levels of senescence-associated secretory phenotype (SASP) protein secretion. Yet, the impact of senescent fibroblasts, and their SASP, on surrounding fibroblasts in SEO-COPD lungs remains unclear. AimTo identify the effect of the SASP secreted by senescent SEO-COPD-derived fibroblasts on surrounding lung fibroblasts. MethodsCellular senescence was induced in lung fibroblasts derived from seven SEO-COPD patients (age[≤]53 years, FEV1<40% predicted), and conditioned medium (CM) containing the SASP, was collected (senescent CM). CM from untreated fibroblasts was used as control. Fibroblasts were stimulated with senescent and control CM, and with tissue plasminogen activator (t-PA), a previously identified COPD-associated SASP protein. Effects on paracrine senescence, inflammation, and extracellular matrix (ECM) regulation were assessed. ResultsStimulation with senescent CM increased the percentage of Senescence-associated beta-galactosidase positive fibroblasts and decreased p16, p21 & LMNB1 expression (p<0.05). T-PA did not affect these markers. Senescent CM increased IL-8 gene expression, increased IL-6 secretion, and strongly increased IL-8 secretion compared to control CM. T-PA slightly decreased IL-6 and IL-8 secretion. Additionally, senescent CM and t-PA stimulation both reduced decorin secretion. Senescent CM reduced FN1 gene expression, while DCN and MMP2 expression remained unaffected. T-PA did not affect ECM gene expression. ConclusionThe SASP from senescence-induced SEO-COPD-derived fibroblasts has a strong paracrine effect on untreated fibroblasts, suggesting that senescent lung fibroblasts contribute to chronic inflammation and ECM dysregulation. These findings imply involvement of senescent fibroblasts in abnormal lung ageing and possibly disease pathology in COPD.

cell biology↗

bioRxiv2023

E-cigarette vapour induces cellular senescence in primary lung fibroblasts and may contribute to lung pathology

Cellular senescence has been recognized to play a role in COPD pathophysiology. Non-aerosolized E-liquid treatment of a lung fibroblast foetal cell line resulted in cellular senescence induction, but this has not been assessed using E-vapour exposure and using primary lung fibroblasts. Therefore, we investigated whether E-vapour exposure induces cellular senescence in primary human lung fibroblasts and whether this affects their tissue repair function. Primary human lung fibroblasts were stimulated with E-cigarette vapour extracts, and cigarette smoke extract (CSE) and Paraquat (PQ) as positive controls. IL-8 secretion was measured to confirm a stimulatory response. Multiple senescence markers (p16, p21, SA-{beta}-gal, and proliferation) were assessed and the tissue repair function was assessed using a scratch assay. Finally, we tried to validate our findings in an E-cigarette-exposed mouse model. Upon stimulation with CSE, PQ, and E-vapour extracts, cellular senescence was induced, which seemed dose-dependent and nicotine-independent. E-vapour-induced senescence resulted in an impaired tissue repair function. No significant difference was observed in the E-cigarette-exposed mouse model. In this study, we identify E-cigarette vapours potential to induce cellular senescence in primary human lung fibroblasts and that this affects their tissue repair function, which further adds to the identified risks of E-cigarette use.

cell biology↗

bioRxiv2023

Cellular Senescence Affects ECM Regulation in COPD Lung Tissue

IntroductionHigher levels of cellular senescence have been demonstrated in COPD patients, including severe early-onset (SEO)-COPD. Recently, we demonstrated that senescence induces extracellular matrix (ECM) dysregulation in lung fibroblasts. However, this in vitro observation has not been demonstrated in vivo yet. Therefore, we investigated whether cellular senescence can contribute to COPD-associated ECM changes in parenchymal lung tissue. MethodsTranscriptomic and proteomic analyses were performed on parenchymal lung tissue from 60 COPD patients (including 18 SEO-COPD patients) and 32 controls. Differential expression of ECM-related genes and proteins was compared between (SEO-)COPD and controls, followed by correlations with six senescence markers and four senescence signature scores. Significant ECM-senescence correlations were verified using histology and primary lung fibroblasts. ResultsWe identified 12 COPD- and 57 SEO-COPD-associated ECM genes and 4 COPD- and 9 SEO-COPD-associated ECM proteins of which the majority, 45 genes and 5 proteins, correlated with senescence. The correlations for COL6A1, COL6A2 and FBLN5 were confirmed in situ and correlations for 21 ECM genes were confirmed in primary lung fibroblasts at baseline. Four genes were successfully functionally validated in our senescence-induced lung fibroblast model, including increased protein levels of ADAMST1 and a non-functional FBLN5 protein. ConclusionsWe confirm a strong link between (SEO-)COPD-associated ECM changes and senescence in vivo in peripheral lung tissue from COPD patients. The strongest and most consistent senescence-associated ECM components include proteases, elastogenesis genes, and collagens 6. These results indicate a contributing role for senescence in disturbed ECM and elastic fiber organization, and protease-antiprotease imbalance in COPD.

pathology↗