# ELN regulates cellular senescence: Emerging hypothesis for a non-canonical role

Joanna Czarnecka-Herok, Kexin Zhu, Emmanuelle Leblanc, Kristell Lazou, Carine Nizard, Anne-Laure Bulteau, Nadine Martin, David Bernard

► To cite this version:

Joanna Czarnecka-Herok, Kexin Zhu, Emmanuelle Leblanc, Kristell Lazou, Carine Nizard, et al. ELN regulates cellular senescence: Emerging hypothesis for a non-canonical role. Mechanisms of Ageing and Development, 2025, 225, pp.112055. 10.1016/j.mad.2025.112055. hal-05064921

HAL Id: hal-05064921

https://hal.science/hal-05064921v1

Submitted on 17 Nov 2025

HAL is a multi-disciplinary open access
archive for the deposit and dissemination of sci-
entific research documents, whether they are pub-
lished or not. The documents may come from
teaching and research institutions in France or
abroad, or from public or private research centers.

L'archive ouverte pluridisciplinaire HAL, est
destinée au dépôt et à la diffusion de documents
scientifiques de niveau recherche, publiés ou non,
émanant des établissements d'enseignement et de
recherche français ou étrangers, des laboratoires
publics ou privés.

<figure><img src="image_1.png" /></figure>

Distributed under a Creative Commons Attribution 4.0 International License
<figure><img src="image_2.png" /></figure>

<figure><img src="image_3.png" /></figure>

# ELN regulates cellular senescence: Emerging hypothesis for a non-canonical role

Joanna Czarnecka-Herok<sup>a,b</sup>, Kexin Zhu<sup>a,b</sup>, Emmanuelle Leblanc<sup>c</sup>, Kristell Lazou<sup>c</sup>, Carine Nizard<sup>c</sup>, Anne-Laure Bulteau<sup>c,*,1</sup>, Nadine Martin<sup>a,b,**,1</sup>, David Bernard<sup>a,b,**,1</sup>

## ARTICLE INFO

**Keywords:**

Cellular senescence

ELN

HMOX1

ROS

Mitochondria

Aging

## ABSTRACT

Elastic fibers are well-known extracellular matrix components that are essential for elastic properties and thus function of many tissues. Tropoelastin is encoded by the *ELN* gene which is the main component of the elastic fibers. Elastic fibers decrease with aging and this decrease is proposed to contribute to this process. Senescent cells, cells that stop to proliferate and that instruct their microenvironment, accumulate with aging and promote it. Until recently, whether *ELN* expression and function is linked to cellular senescence was unknown. Here we will comment and extend recent results supporting a function of the *ELN* gene in protecting cells from cellular senescence. We will also discuss hypotheses on mechanisms by which *ELN* could regulate cellular senescence, and especially a hypothesis that involves a non-canonical function of *ELN* regulating the mitochondrial respiratory chain activity, thereby controlling oxidative stress and cellular senescence. These findings provide critical insights into the molecular and cellular processes potentially underlying the phenotypes driven by *ELN* deletion in the context of aging.

## 1. Introduction

Tropoelastin, also named elastin, is an essential protein as its depletion induces embryonic lethality (Li et al., 1998). It is encoded by the *ELN* gene as a secreted monomer. In the extracellular matrix, it undergoes complex and organized post-translation modifications, mainly crosslinking, to form mature elastic fibers. Elastic fibers, together with some other types of fibers, are required for elasticity allowing tissues to stretch and recoil, maintaining tissue structural integrity and functionality under mechanical stress, including for lung during breathing, skin during distortion or vessels for blood circulation (Mecham, 2018; Schmelzer and Duca, 2022). To date, research on *ELN* has been mainly focused on elastic fibers. Whether *ELN* also exerts non-canonical elastic fiber-independent functions, which could contribute to *ELN* role in physiological and pathological conditions, is barely known.

Elastic fibers are mainly produced during development and at young

age, and then *ELN* expression decreases and largely ceases in the adulthood. With aging, or during exposures accelerating aging, elastic fibers are damaged without possibility of proper repair. This deterioration is thought to contribute to physiological tissue and organism aging as well as to pathological aging, altering parameters such as breath capacity during chronic obstructive pulmonary disease or blood circulation in some cardiovascular diseases (Schmelzer and Duca, 2022; Heinz, 2021). In addition to exerting mechanical effects on tissues, the degradation of elastic fibers during the aging process or the development of age-related diseases also results in the production of elastin-derived peptides (EDP), also known as elastokines. These peptides have the potential to exert biological activity and have been linked to a range of detrimental effects, including those observed in pathologies associated with senescent cells (Antonicelli et al., 2007; Gayral et al., 2014; Houghton et al., 2006; Maclay et al., 2012; Houssaini et al., 2018; Beaulieu et al., 2021).

Mainly induced by a broad range of stresses (e.g. telomere

<footnote><sup>a</sup> Centre de Recherche en Cancérologie de Lyon, Inserm U1052, CNRS UMR 5286, Centre Léon Bérard, Université de Lyon, Lyon, France</footnote>

<footnote><sup>b</sup> Equipe Labellisée la Ligue Contre le Cancer, Lyon, France</footnote>

<footnote><sup>c</sup> LVMH Recherche, 185, avenue de Verdun, Saint Jean de Braye, 45804, France</footnote>

<footnote>* Corresponding author.</footnote>

<footnote>** Corresponding authors at: Centre de Recherche en Cancérologie de Lyon, Inserm U1052, CNRS UMR 5286, Centre Léon Bérard, Université de Lyon, Lyon, France.</footnote>

<footnote>E-mail addresses: abulteau@research.lvmh-pc.com (A.-L. Bulteau), nadine.martin@lyon.unicancer.fr (N. Martin), david.bernard@lyon.unicancer.fr (D. Bernard).</footnote>

<footnote n="1">¹ contributed equally</footnote>
dysfunction, DNA damage, reactive oxygen species (ROS), replicative stress), cellular senescence is a cellular phenotype characterized by a stable proliferation arrest and a senescence-associated secretory phenotype (SASP). Cellular senescence is one of the cellular mechanisms contributing to aging and is functionally linked, in a bidirectional way, to other mechanisms contributing to aging such as chronic inflammation, deregulated nutrient-sensing, mitochondrial dysfunction and epigenetic alterations (López-Otín et al., 2013, 2023). Aging or exposure to toxics such as cigarette smoke or UV induce accumulation of senescent cells. These senescent cells contribute to many age-related diseases and overall aging as their depletion ameliorates many diseases and improves healthy and overall aging (Zhang et al., 2023; Di Micco et al., 2021). Mechanistically, many types of triggers induce senescence and they often share some downstream mechanisms such as induction of DNA damage and change in mitochondrial activity and function (Di Micco et al., 2021). During cellular senescence, increased ROS, especially produced by the mitochondria, have been consistently reported, which could among others lead to DNA damage, another largely common feature of senescent cells (Correia-Melo and Passos, 2015).

The links of *ELN* or cellular senescence with the processes of aging and age-related diseases have prompted the suggestion that the two may be functionally connected. A key role of *ELN* in the regulation of cellular senescence has recently been demonstrated (Czarnecka-Herok et al., 2024; Lee et al., 2024; Huang et al., 2023). Interestingly, this new function has been found to probably be independent of elastic fibers (Czarnecka-Herok et al., 2024). Still many questions, especially on the mechanistic side, remain unanswered. Here we will review these recent findings, expand them and propose new hypothesis on the molecular and cellular mechanisms underlying this non-canonical role of *ELN*.

## 2. ELN protects from cellular senescence

### 2.1. ELN loss induces premature senescence

As reported in our article by Czarnecka-Herok and collaborators (Czarnecka-Herok et al., 2024), a reduction in *ELN* mRNA levels can be observed in several datasets comparing senescent cells to non-senescent cells, including in human fibroblasts during replicative senescence and upon oncogenic stress, in mouse embryonic fibroblasts (MEFs) upon oncogenic activation or in astrocytes after an oxidative stress. This downregulation of *ELN* might play a role in those models of senescence as *ELN* knockdown in normal human embryonic lung fibroblasts, MRC5, and *ELN* knockout in MEFs induce premature senescence (Czarnecka-Herok et al., 2024). This effect is not specific to embryo-derived cells as similar observations can be made in cells at later stage. Indeed, we observed that *ELN* mRNA levels were decreased in late passage human skin-derived fibroblasts (Fig. 1A) and in this cell type as in aforementioned ones, *ELN* knockdown (Fig. 1B) provoked the induction of all the hallmarks of premature senescence examined: up-regulation of the cyclin-dependent kinase inhibitor p21 (Fig. 1C) and of the SASP factor GDF15 (Fig. 1D) as well as increase in senescence-associated-β-galactosidase (SA-β-Gal) activity and decrease in cell number (Fig. 1E). *ELN* knockdown was also suggested to trigger premature senescence in human retinal pigment epithelium cells (ARPE-19), evidenced by increased SA-β-Gal activity (Huang et al., 2023). Together these data support that *ELN* expression protects from cellular senescence in human and mouse cells, irrespective of whether they were derived from embryos or later in life.

### 2.2. Extracellular tropoelastin protein delays mesenchymal stem cell senescence

To assess potential functional links between *ELN* and cellular senescence, Lee and collaborators (Lee et al., 2024) took a different approach than the one in the Czarnecka-Herok et al. study. Indeed, they used recombinant tropoelastin proteins, either soluble or fixed to the

substrate, to investigate its impact on mesenchymal stem cell (MSC) senescence. They first observed a decrease in tropoelastin expression in late passage MSC when compared to early passage ones, in line with our results in skin fibroblasts (Fig. 1A) and several other senescence transcriptomic datasets (Czarnecka-Herok et al., 2024). Treating late passage MSC with recombinant tropoelastin proteins decreased features of senescence. Indeed tropoelastin-treated MSC had an extended replicative potential together with diminished SA-β-Gal activity, p21 or p16 expression as well as SASP factors expression (Lee et al., 2024).

Altogether, the recent findings described in this section support a role for *ELN* in protecting cells from cellular senescence in different cell types, with an involvement of p21 in this process in all these contexts.

## 3. Potential mechanisms of action of ELN to protect from cellular senescence

### 3.1. Potential mechanisms involving extracellular tropoelastin

Lee and collaborators added recombinant tropoelastin and observed decreased cellular senescence in MSC (Lee et al., 2024), but no detailed mechanism of action was proposed to explain this observation. For instance, the biophysical properties of this recombinant protein are not reported and whether it can enter the cells or be maturated by cross-linking to form pseudo-elastic fibers and eventually EDP when degraded is unknown. In addition, this multistep maturation process is quite long and difficult to be performed in classical 2D models (Halm et al., 2016), particularly in the time frame of the experiments (Lee et al., 2024).

In another study, this recombinant tropoelastin has been proposed to stimulate MSC attachment and proliferation via αV integrins (Yeo and Weiss, 2019), nevertheless the direct mechanisms behind these observations are unknown (Yeo and Weiss, 2019) as well as whether this effect is linked to the decreased senescence recently reported. Integrins have been reported to display pro- or anti-senescence activity pending on the studies (Tragoonlugkana et al., 2024; Rapisarda et al., 2017). Even if at this stage we cannot rule out an effect of pseudo-elastic fibers, it is more likely that the recombinant tropoelastin protein acts independently of its function on elastic fibers and of its elastokine sub-products. Of note, there is no report so far documenting the impact of ectopic expression of *ELN* on cellular senescence, which would be an interesting approach to improve our understanding of *ELN* protein localization and action in this context.

### 3.2. Potential non-canonical mechanisms

In our study by Czarnecka-Herok and coauthors, we did not observe pseudo-elastic fibers in MRC5 fibroblasts, although the cells enter into premature senescence following the knockdown of *ELN*. Our findings support the hypothesis that *ELN* loss-induced senescence is independent of tropoelastin role in the fibers or its degradation and the production of elastokines (Czarnecka-Herok et al., 2024). Quite unexpectedly, *ELN* downregulation by siRNA resulted in some molecular changes as early as 24 hours after siRNA transfection as evidenced by transcriptome analyses. This finding lends further support to the hypothesis that *ELN* plays a non-canonical function in cellular senescence.

Strikingly, expression of the heme oxygenase 1 (HMOX1) is quickly upregulated after *ELN* knockdown in MRC5 human lung fibroblasts (Czarnecka-Herok et al., 2024). This induction seems to be a general effect of *ELN* loss as it was also observed in human dermal fibroblasts (Fig. 2). The HMOX1 protein cleaves heme, generating biliverdin, carbon monoxide and Fe²⁺ and this enzyme can display complex effects including both anti- and pro-oxidant activities (Porto et al., 2007; Baranano et al., 2002; Galaris et al., 2019) and pro- and anti-senescent effects (Bansal et al., 2014; Görg et al., 2019; Shan et al., 2019) depending on the studies. Upregulation of HMOX1 during *ELN* downregulation-induced senescence participates in the induction of cellular senescence, as demonstrated by the fact that inhibiting HMOX1
A

ELN

A

B

ELN

C

CDKN1A

D

GDF15

E

#113

#718

<figure><img src="image_4.png" /></figure>

<figure><img src="image_5.png" /></figure>

<figure><img src="image_6.png" /></figure>

<figure><img src="image_7.png" /></figure>

<figure><img src="image_8.png" /></figure>

<figure><img src="image_9.png" /></figure>

<figure><img src="image_10.png" /></figure>

<figure><img src="image_11.png" /></figure>

**Fig. 1.** ELN downregulation triggers senescence in human dermal fibroblasts. A. ELN mRNA level assessed by RT-qPCR in five different strains of normal human dermal fibroblasts (HDFs) at early (<9) or later (P15) passage. Mean +/- SEM for 5 independent HDF strains. Un-paired Student t-test. B-D. mRNA level of ELN (B), CDKN1A, encoding p21 (C) and GDF15 (D) assessed by RT-qPCR 4 days after transfection of HDFs with siCTRL or siELN. Mean +/- SEM of 3 independent experiments for 2 independent HDF strains. Un-paired Welch's t-test. E. Representative micrographs (left) and quantification (right) of SA-β-galactosidase (SA-β-gal) positive cells and total cell number 4 days after transfection of HDFs with siCTRL or siELN. Mean +/- SEM of 3 independent experiments for 2 independent HDF strains (#113 and #718). Un-paired Welch's t-test. Skin samples were collected from anonymous adult patients who have undergone plastic surgeries performed by independent surgeons. Surgical residues were harvested according to French regulations (agreement DC-2023-5824) and after obtaining written informed consent from each donor. Skin cell culture fibroblasts (HDFs) were established individually from abdominal skin biopsies obtained from healthy Caucasian female. They were cultured in DMEM containing 1 g/L glucose (Fisher Scientific), with 10% fetal bovine serum and 1% penicillin-streptomycin antibiotics under standard conditions (37°C, 5% CO₂). Cells were transfected as described in (Czarnecka-Herok et al., 2024) using a final siRNA concentration of 45 nM. RNA extraction, reverse transcription, real-time quantitative PCR and senescence-associated-β-galactosidase assay were performed as documented in (Czarnecka-Herok et al., 2024). Graphs were created with GraphPad Prism 9 and statistical analysis was also performed with this software. Parametric tests were two-tailed, unpaired: Student's t-test (for equal variance) or Welch's t-test (for non-equal variance). P-values are indicated in the graphs.
HMOX1

<figure><img src="image_12.png" /><figcaption>**Fig. 2.** ELN downregulation induces HMOX1 expression. HMOX1 mRNA level assessed by RT-qPCR 4 days after transfection of HDFs with siCTRL or siELN. Mean +/- SEM of 3 independent experiments for 2 independent HDF strains. Un-paired Welch's t-test. Cells were transfected as described in (Czarnecka-Herok et al., 2024) using a final siRNA concentration of 45 nM. RNA extraction, reverse transcription and real-time quantitative PCR were performed as documented in (Czarnecka-Herok et al., 2024). Graph was created with GraphPad Prism 9 and statistical analysis was also performed with this software. P-value is indicated in the graph.</figcaption></figure>

using siRNA or chelating its downstream product, iron, largely rescued the senescent phenotype. The HMOX1-dependent increase in intracellular iron concentration observed upon ELN downregulation was shown to contribute to senescence-associated gene expression reprogramming through the iron-dependent histone demethylase plant homeodomain finger protein 8 (PHF8) (Czarnecka-Herok et al., 2024). Indeed, PHF8 binds to and induces the expression of the cell cycle inhibitor p21-encoding gene CDKN1A and binds as well to several other genes which critically promote senescence implementation and are upregulated upon ELN loss. However, it is very likely that increased iron levels in this context also have other functional impacts triggering senescence, yet to be determined.

4. ELN: a novel regulator of mitochondrial activity?

4.1. ELN loss triggers mitochondrial ROS production and may alter electron transport chain (ETC) activity

Transcriptome and Gene Set Enrichment Analysis (GSEA) of fibroblasts upon ELN knockdown revealed a significant enrichment of a gene set associated with response to oxidative stress. An increase in mitochondrial ROS was indeed detected early after ELN downregulation (Czarnecka-Herok et al., 2024), supporting that ELN loss impacts mitochondria. Antioxidant prevented both HMOX1 upregulation and senescence induction in this context. Moreover, p53 and NRF2, two transcription factors known to be activated by ROS, were found to play a key role in the induction of HMOX1 expression and senescence upon ELN loss (Czarnecka-Herok et al., 2024).

As change in ETC activity is an important mechanism leading to mitochondrial ROS production (Zhao et al., 2019; Nolfi-Donegan et al., 2020), we measured whether ETC activity change upon ELN loss. We conducted comparative analyses of mitochondrial bioenergetic profiles using Seahorse technology to measure respiration (oxygen consumption rate (OCR)). By incorporating various mitochondrial inhibitors (Oligomycin as ATP synthase inhibitor, FCCP to disrupt the proton gradient and uncouple oxygen consumption from ATP production, and Rotenone/ Antimycin A to inhibit complex I and III) we assessed different bioenergetic parameters: basal OCR, maximal OCR and ATP-linked OCR. When compared to control cells, cells with downregulated ELN had significantly lower basal OCR (Fig. 3A), maximal OCR (Fig. 3B) as well as ATP-linked OCR (Fig. 3C). Together these results support that ELN loss leads to mitochondrial uncoupling and dysfunction and it is well known that perturbing ETC activity can lead to premature senescence (Stöckl et al., 2006, 2007; Miwa et al., 2014). Mitochondrial consequences of ELN loss will need to be further characterized as impaired ETC activity may not only trigger mitochondrial ROS production but also other alterations such as an impairment of TCA cycle activity (Fig. 4).

4.2. Directions for further investigating the link between ELN and mitochondria

Given that the impact of ELN loss on ROS production and HMOX1

<figure><img src="image_13.png" /><figcaption>**Fig. 3.** ELN downregulation triggers mitochondrial OCR alterations. A-C. Basal OCR (A), maximal OCR (B) and ATP-linked OCR (C) data obtained by Seahorse analysis 2 days after transfection of normal human dermal fibroblasts (HDFs) with siCTRL or siELN. Mean +/- SEM of 10 technical replicates for 2 independent HDF strains. Un-paired Student t-test. OCR was measured on a Seahorse (Agilent Technologies) under basal conditions and after the sequential addition of 1.5 µM oligomycin, 3 µM carbonyl cyanide p-trifluoromethoxyphenylhydrazone (FCCP) and 1.5 µM antimycin A + 1.5 µM rotenone (Agilent Technologies). Data were normalized to cell number and calculations were performed as follows: Basal OCR= rate measurement before oligomycin injection - minimum rate measurement after Rotenone/Antimycin A injection; Maximal OCR= maximum rate measurement after FCCP injection - minimum rate measurement after Rotenone/Antimycin A injection; ATP-linked OCR= basal OCR - rate measurement after oligomycin injection. Graphs were created with GraphPad Prism 9 and statistical analysis was also performed with this software. P-values are indicated in the graphs.</figcaption></figure>
<figure><img src="image_14.png" /><figcaption>**Fig. 4.** Scheme depicting *ELN* impact on mitochondrial function and cellular senescence. *ELN* downregulation triggers a decrease in mitochondrial respiration and an increase in the production of mitochondrial ROS, which activate *HMOX1* expression through NRF2 and p53. *HMOX1* induction causes iron accumulation, which leads to a reprogramming in gene expression involving the iron-dependent histone demethylase PHF8. Further investigation is needed to understand the mechanisms through which *ELN* loss impacts mitochondria. The scheme was created using BioRender.</figcaption></figure>

upregulation is detectable early after siRNA transfection, it seems plausible to suggest that the link between *ELN* and mitochondria is quite direct. Most of the articles on *ELN* focus on its extracellular form, the elastic fibers and eventually its bioactive subproducts, the elastokines. However, whether *ELN* can be localized and have a role inside the cells or even inside the mitochondria is still unknown. Indeed, we have not identified studies suggesting this localization in the mitochondria at this stage. Nevertheless, analysis of prediction of *ELN* localization did not rule out a mitochondrial localization (using Genecards and WoLF PSORT II). In both cases, the extracellular localization was identified as the primary prediction, followed by a potential localization in the mitochondria.

In order to determine whether *ELN* localizes in the mitochondria and, if so, to identify its role and mechanism of action within this organelle, it will be necessary to develop new specific tools. Isolating mitochondria and examining the presence of *ELN* in this fraction would be a first approach. Should *ELN* be identified in this compartment, a subsequent step would be to investigate its role in the mitochondria, which could start by defining its local interactome. Even if this hypothesis may initially appear improbable, a growing list of proteins, which were not initially observed in the mitochondria, have been now shown to localize in this organelle (p53 and other transcription factors being well-known examples) (Psarra and Sekeris, 2008). Of course, impact of *ELN* on ETC activity without being localized in the mitochondria is also a valuable
hypothesis but still it has to be a quite direct effect given the reported
kinetic of effect of ELN loss.

5. Conclusion and perspectives

Together, the data of several recently published studies (Czarnecka-Herok et al., 2024; Lee et al., 2024; Huang et al., 2023) support a novel function of ELN in regulating cellular senescence, although the mechanisms involved might be different depending on the context, as explained above. All the studies concluded that ELN protects from cellular senescence, nevertheless the use of different tools and cells make more advanced direct comparison of the results hazardous. The reported findings point out an elastic fiber-independent role of ELN in the context of cellular senescence. Here we further propose that loss of ELN results in alterations of the mitochondrial ETC activity, which is a strong candidate to mediate ROS production and senescence induction. Still, we can speculate that elastic fibers, by impacting elasticity and mechano-transduction, and/or when degraded by releasing elastokines and by inducing specific signaling, could also regulate cellular senescence through other mechanisms than those proposed by Czarnecka-Herok and coll, Lee and coll (Czarnecka-Herok et al., 2024; Lee et al., 2024) and this article.

Importantly, the discussed findings suggest that cellular senescence and elastic fiber-independent functions of ELN could potentially play a role in the phenotypes driven by ELN deletion. This could be the case for instance in Williams-Beuren syndrome (WBS), a rare developmental disease caused by a hemizygous deletion of 26-28 genes including ELN on chromosome 7. WBS patients display an array of symptoms comprising intellectual disability and cardiovascular disease as well as, in the adulthood, diabetes, premature vascular and skin aging and osteoporosis (Pober, 2010). These signs of mild accelerated aging and the fact that MEFs harboring a heterozygous deletion of the WBS critical region display a premature cell proliferation arrest (Segura-Puimedon et al., 2014) suggest an involvement of premature senescence in this syndrome. ELN downregulation is considered as responsible for the cardiovascular disease and soft skin observed in WBS (Pober, 2010) due to its impact on elastic fibers and tissue elasticity. However, the findings discussed in the present article suggest that ELN loss could also drive or contribute to some of the disorders observed in WBS by acting independently of elastic fibers (potentially impacting mitochondria) and triggering cellular senescence. More globally, we can speculate that during aging, decreased expression of ELN could perturb mitochondrial respiration and activity, leading to ROS production and cellular senescence, further participating to other aging hallmarks and aging as a whole (Miwa et al., 2014) (Fig. 4).

In conclusion, the discovery of an unexpected role of ELN in the regulation of cellular senescence provides new insights to understand the physio-pathological effects of ELN modulation. Further work will allow to test hypotheses on the underlying mechanisms of action and to reveal how loss of ELN remodels mitochondrial activity, which would have strong implications especially, but likely not only, in the context of cellular senescence.

Author contribution

JCH, KZ, KL and EL performed the experiments and results were analyzed by all the co-authors. CN, ALB, NM and DB designed the study. ALB, NM and DB supervised the work and wrote the manuscript with inputs from all the co-authors.

CRediT authorship contribution statement

Lazou Kristell: Writing – review & editing, Resources, Methodology.
Leblanc Emmanuel: Writing – review & editing, Resources, Meth-
odology. Bulteau Anne-Laure: Writing – review & editing, Writing –
original draft, Supervision, Conceptualization. Nizard Carine: Writing –

review & editing, Resources, Methodology. Martin Nadine: Writing – review & editing, Writing – original draft, Supervision, Funding acquisition, Conceptualization. Czarnecka-Herok Joanna: Writing – review & editing, Visualization, Resources, Methodology. Bernard David: Writing – review & editing, Writing – original draft, Supervision, Funding acquisition, Conceptualization. Zhu Kexin: Writing – review & editing, Resources, Methodology.

Declaration of Competing Interest

This study was partially supported by LVMH Recherche. ALB, CN, KL and EL are employees of LVMH Recherche. No other competing interest to declare.

Acknowledgements

We thank Brigitte Manship for critical reading of the manuscript. This study was supported by research grants from “Autour des Williams” association to NM and INCA, IReSP (IRESP-19-TABAC-V3-01), Fonda- tion ARC and LVMH to DB. We acknowledge the contribution of the imaging facilities of the CRCL and of LVMH Recherche and the Seahorse facility of SFR Biosciences, Laurence Canaple, Lyon (UAR 3444 ANIRA ImmOs).

Data availability

Data will be made available on request.

References

Antonicelli, F., Bellon, G., Debelle, L., Hornebeck, W., 2007. Elastin-elastases and
inflamm-aging. Curr. Top. Dev. Biol. 79, 99-155. https://doi.org/10.1016/S0070-
2153(06)79005-6

Bansal, S., Biswas, G., Avadhani, N.G., 2014. Mitochondria-targeted heme oxygenase-1
induces oxidative stress and mitochondrial dysfunction in macrophages, kidney
fibroblasts and in chronic alcohol hepatotoxicity. Redox Biol. 2, 273-283. https://
doi.org/10.1016/j.redox.2013.07.004

Baranano, D.E., Rao, M., Ferris, C.D., Snyder, S.H., 2002. Biliverdin reductase: a major
physiologic cytoprotectant. Proc. Natl. Acad. Sci. USA 99, 16093-16098. https://
doi.org/10.1073/pnas.252626999

Beaulieu, D., Attwe, A., Breau, M., Lipskaia, L., Marcos, E., Born, E., Huang, J., Abid, S., Derumeaux, G., Houssaini, A., Maitre, B., Lefevre, M., Vienney, N., Bertolino, P., Jaber, S., Noureddine, H., Goehrig, D., Vindrieux, D., Bernard, D., Adnot, S., 2021. Phospholipase A2 receptor 1 promotes lung cell senescence and emphysema in obstructive lung disease. Eur. Respir. J. 58, 2000752. https://doi.org/10.1183/
13993003.00752-2020

Correia-Melo, C., Passos, J.F., 2015. Mitochondria: are they causal players in cellular senescence? Biochim Biophys. Acta 1847, 1373-1379. https://doi.org/10.1016/j.bbio.2015.05.017

Czarnecka-Herok, J., Zhu, K., Flaman, J.-M., Goehrig, D., Vernier, M., Makulyte, G.,
Lamboux, A., Dragic, H., Rhinn, M., Médard, J.-J., Faury, G., Bertolino, P., Balter, V.,
Debret, R., Adnot, S., Martin, N., Bernard, D., 2024. A non-canonical role of ELN
protects from cellular senescence by limiting iron-dependent regulation of gene
expression. Redox Biol. 73, 103204. https://doi.org/10.1016/j.redox.2024.103204

Di Micco, R., Krizhanovsky, V., Baker, D., d'Adda di Fagagna, F., 2021. Cellular senescence in ageing: from mechanisms to therapeutic opportunities. Nat. Rev. Mol. Cell Biol. 22, 75-95. https://doi.org/10.1038/s41580-020-00314-w

Galaris, D., Barbouti, A., Pantopoulos, K., 2019. Iron homeostasis and oxidative stress: an intimate relationship. Biochim Biophys. Acta Mol. Cell Res. 1866, 118535. https://doi.org/10.1016/j.jbamcr.2019.118535

Gayral, S., Garnotel, R., Castaing-Berthou, A., Blaise, S., Fougerat, A., Berge, E.,
Montheil, A., Malet, N., Wymann, M.P., Maurice, P., Debelle, L., Martiny, L.,
Martinez, L.O., Pshezhetsky, A.V., Duca, L., Laffargue, M., 2014. Elastin-derived
peptides potentiate atherosclerosis through the immune Neu1-PI3Kγ pathway.
Cardiovasc Res. 102, 118-127. https://doi.org/10.1093/cvr/cvt336

Görg, B., Karababa, A., Schütz, E., Paluschinski, M., Schrimpf, A., Shafigullina, A., Castoldi, M., Bidmon, H.J., Häussinger, D., 2019. O-GlcNAcylation-dependent upregulation of HO1 triggers ammonia-induced oxidative stress and senescence in hepatic encephalopathy. J. Hepatol. 71, 930-941. https://doi.org/10.1016/j.jhep.2019.06.020

Halm, M., Schenke-Layland, K., Jaspers, S., Wenck, H., Fischer, F., 2016. Visualizing tropoelastin in a long-term human elastic fibre cell culture model. Sci. Rep. 6, 20378. https://doi.org/10.1038/srep20378

Heinz, A., 2021. Elastic fibers during aging and disease. Ageing Res. Rev. 66, 101255.
https://doi.org/10.1016/j.arr.2021.101255

Houghton, A.M., Quintero, P.A., Perkins, D.L., Kobayashi, D.K., Kelley, D.G.,
Marconcini, L.A., Mecham, R.P., Senior, R.M., Shapiro, S.D., 2006. Elastin fragments
drive disease progression in a murine model of emphysema. J. Clin. Investig. 116, 753-759. https://doi.org/10.1172/JCI25617.

Houssaini, A., Breau, M., Kebe, K., Abid, S., Marcos, E., Lipskaia, L., Rideau, D., Parpaleix, A., Huang, J., Amsellem, V., Vienney, N., Validire, P., Maitre, B., Attwe, A., Lukas, C., Vindrieux, D., Boczkowski, J., Derumeaux, G., Pende, M., Bernard, D., Meiners, S., Adnot, S., 2018. mTOR pathway activation drives lung cell senescence and emphysema, 93203 JCI Insight 3, e93203. https://doi.org/10.1172/jci.insight.93203.

Huang, L., Ye, L., Li, R., Zhang, S., Qu, C., Li, S., Li, J., Yang, M., Wu, B., Chen, R., Huang, G., Gong, B., Li, Z., Yang, H., Yu, M., Shi, Y., Wang, C., Chen, W., Yang, Z., 2023. Dynamic human retinal pigment epithelium (RPE) and choroid architecture based on single-cell transcriptomic landscape analysis. Genes Dis. 10, 2540-2556. https://doi.org/10.1016/j.gendis.2022.11.007.

Lee, S.S., Al Halawani, A., Teo, J.D., Weiss, A.S., Yeo, G.C., 2024. The matrix protein tropoelastin prolongs mesenchymal stromal cell vitality and delays senescence during replicative aging. Adv. Sci. (Weinh. ) 11, e2402168. https://doi.org/10.1002/advs.202402168.

Li, D.Y., Brooke, B., Davis, E.C., Mecham, R.P., Sorensen, L.K., Boak, B.B., Eichwald, E., Keating, M.T., 1998. Elastin is an essential determinant of arterial morphogenesis. Nature 393, 276-280. https://doi.org/10.1038/30522.

López-Otín, C., Blasco, M.A., Partridge, L., Serrano, M., Kroemer, G., 2013. The hallmarks of aging. Cell 153, 1194-1217. https://doi.org/10.1016/j.cell.2013.05.039.

López-Otín, C., Blasco, M.A., Partridge, L., Serrano, M., Kroemer, G., 2023. Hallmarks of aging: an expanding universe. Cell 186, 243-278. https://doi.org/10.1016/j.cell.2022.11.001.

Maclay, J.D., McAllister, D.A., Rabinovich, R., Haq, I., Maxwell, S., Hartland, S., Connell, M., Murchison, J.T., van Beek, E.J.R., Gray, R.D., Mills, N.L., Macnee, W., 2012. Systemic elastin degradation in chronic obstructive pulmonary disease. Thorax 67, 606-612. https://doi.org/10.1136/thoraxjnl-2011-200949.

Mecham, R.P., 2018. Elastin in lung development and disease pathogenesis. Matrix Biol. 73, 6-20. https://doi.org/10.1016/j.matbio.2018.01.005.

Miwa, S., Jow, H., Baty, K., Johnson, A., Czapiewski, R., Saretzki, G., Treumann, A., von Zglinicki, T., 2014. Low abundance of the matrix arm of complex I in mitochondria predicts longevity in mice. Nat. Commun. 5, 3837. https://doi.org/10.1038/ncomms4837.

Nolfi-Donegan, D., Braganza, A., Shiva, S., 2020. Mitochondrial electron transport chain: oxidative phosphorylation, oxidant production, and methods of measurement. Redox Biol. 37, 101674. https://doi.org/10.1016/j.redox.2020.101674.

Pober, B.R., 2010. Williams-Beuren syndrome. N. Engl. J. Med. 362, 239-252. https://doi.org/10.1056/NEJMra0903074.

Porto, B.N., Alves, L.S., Fernández, P.L., Dutra, T.P., Figueiredo, R.T., Graça-Souza, A.V., Bozza, M.T., 2007. Heme induces neutrophil migration and reactive oxygen species

generation through signaling pathways characteristic of chemotactic receptors. J. Biol. Chem. 282, 24430-24436. https://doi.org/10.1074/jbc.M703570200.

Psarra, A.-M.G., Sekeris, C.E., 2008. Nuclear receptors and other nuclear transcription factors in mitochondria: regulatory molecules in a new environment. Biochim Biophys. Acta 1783, 1-11. https://doi.org/10.1016/j.bbamcr.2007.10.021.

Rapisarda, V., Borghesan, M., Miguela, V., Encheva, V., Snijders, A.P., Lujambio, A., O'Loghlen, A., 2017. Integrin Beta 3 regulates cellular senescence by activating the TGF-β pathway. Cell Rep. 18, 2480-2493. https://doi.org/10.1016/j.celrep.2017.02.012.

Schmelzer, C.E.H., Duca, L., 2022. Elastic fibers: formation, function, and fate during aging and disease. FEBS J. 289, 3704-3730. https://doi.org/10.1111/febs.15899.

Segura-Puimedon, M., Sahún, I., Velot, E., Dubus, P., Borralleras, C., Rodrigues, A.J., Valero, M.C., Valverde, O., Sousa, N., Herault, Y., Dierssen, M., Pérez-Jurado, L.A., Campuzano, V., 2014. Heterozygous deletion of the Williams-Beuren syndrome critical interval in mice recapitulates most features of the human disorder. Hum. Mol. Genet 23, 6481-6494. https://doi.org/10.1093/hmg/du368.

Shan, H., Li, T., Zhang, L., Yang, R., Li, Y., Zhang, M., Dong, Y., Zhou, Y., Xu, C., Yang, B., Liang, H., Gao, X., Shan, H., 2019. Heme oxygenase-1 prevents heart against myocardial infarction by attenuating ischemic injury-induced cardiomyocytes senescence. EBioMedicine 39, 59-68. https://doi.org/10.1016/j.ebiom.2018.11.056.

Stöckl, P., Hütter, E., Zwerschke, W., Jansen-Dürr, P., 2006. Sustained inhibition of oxidative phosphorylation impairs cell proliferation and induces premature senescence in human fibroblasts. Exp. Gerontol. 41, 674-682. https://doi.org/10.1016/j.exger.2006.04.009.

Stöckl, P., Zankl, C., Hütter, E., Unterluggauer, H., Laun, P., Heeren, G., Bogengruber, E., Herndler-Brandstetter, D., Breitenbach, M., Jansen-Dürr, P., 2007. Partial uncoupling of oxidative phosphorylation induces premature senescence in human fibroblasts and yeast mother cells. Free Radic. Biol. Med. 43, 947-958. https://doi.org/10.1016/j.freeradbiomed.2007.06.005.

Tragoonlugkana, P., Pruksapong, C., Ontong, P., Kamprom, W., Supokawej, A., 2024. Fibronectin and vitronectin alleviate adipose-derived stem cells senescence during long-term culture through the AKT/MDM2/P53 pathway. Sci. Rep. 14, 14242. https://doi.org/10.1038/s41598-024-65339-z.

Yeo, G.C., Weiss, A.S., 2019. Soluble matrix protein is a potent modulator of mesenchymal stem cell performance. Proc. Natl. Acad. Sci. USA 116, 2042-2051. https://doi.org/10.1073/pnas.1812951116.

Zhang, L., Pitcher, L.E., Prahalad, V., Niedernhofer, L.J., Robbins, P.D., 2023. Targeting cellular senescence with senotherapeutics: senolytics and senomorphics. FEBS J. 290, 1362-1383. https://doi.org/10.1111/febs.16350.

Zhao, R.-Z., Jiang, S., Zhang, L., Yu, Z.-B., 2019. Mitochondrial electron transport chain, ROS generation and uncoupling (Review). Int J. Mol. Med. 44, 3-15. https://doi.org/10.3892/ijmm.2019.4188.