Identification of a Repair-Supportive Mesenchymal Cell Population during Airway Epithelial Regeneration

Moiseenko, Alena; Vazquez‐Armendariz, Ana Ivonne; Kheirollahi, Vahid; Chu, Xuran; Tata, Aleksandra; Rivetti, Stefano; Günther, Stefan; Lebrigand, Kévin; Herold, Susanne; Braun, Thomas; Mari, Bernard; Langhe, Stijn De; Kwapiszewska, Grażyna; Günther, Andreas; Chen, Chengshui; Seeger, Werner; Tata, Purushothama Rao; Zhang, Jin‐San; Bellusci, Savério; Agha, Elie El

doi:10.1016/j.celrep.2020.108549

Cited by 33 publications

(44 citation statements)

References 36 publications

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“…Our knowledge about the sources of FGF10 in this context has been evolving. FGF10 was first described to be expressed by airway smooth muscle cells (ASMCs) ( Volckaert et al, 2011 ), whereas more recent work identified a peribronchiolar mesenchymal population capable of producing FGF10 during the repair process, which is not derived from the ASMCs ( Moiseenko et al, 2020 ).…”

Section: Discussionmentioning

confidence: 99%

“…In the context of the repair process, Fgf10 deletion in peribronchial mesenchymal cells leads to impaired repair following injury to the bronchial epithelium using naphthalene ( Volckaert et al, 2011 ; Moiseenko et al, 2020 ). On the other hand, overexpression of Fgf10 reduces the severity of lung fibrosis in bleomycin-induced mice ( Gupte et al, 2009 ).…”

Section: Introductionmentioning

confidence: 99%

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Validation of a Novel Fgf10Cre–ERT2 Knock-in Mouse Line Targeting FGF10Pos Cells Postnatally

Chu

Taghizadeh

Vazquez‐Armendariz

et al. 2021

Front. Cell Dev. Biol.

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Fgf10 is a key gene during development, homeostasis and repair after injury. We previously reported a knock-in Fgf10Cre–ERT2 line (with the Cre-ERT2 cassette inserted in frame with the start codon of exon 1), called thereafter Fgf10Ki–v1, to target FGF10Pos cells. While this line allowed fairly efficient and specific labeling of FGF10Pos cells during the embryonic stage, it failed to target these cells after birth, particularly in the postnatal lung, which has been the focus of our research. We report here the generation and validation of a new knock-in Fgf10Cre–ERT2 line (called thereafter Fgf10Ki–v2) with the insertion of the expression cassette in frame with the stop codon of exon 3. Fgf10Ki−v2/+ heterozygous mice exhibited comparable Fgf10 expression levels to wild type animals. However, a mismatch between Fgf10 and Cre expression levels was observed in Fgf10Ki–v2/+ lungs. In addition, lung and limb agenesis were observed in homozygous embryos suggesting a loss of Fgf10 functional allele in Fgf10Ki–v2 mice. Bioinformatic analysis shows that the 3′UTR, where the Cre-ERT2 cassette is inserted, contains numerous putative transcription factor binding sites. By crossing this line with tdTomato reporter line, we demonstrated that tdTomato expression faithfully recapitulated Fgf10 expression during development. Importantly, Fgf10Ki–v2 mouse is capable of significantly targeting FGF10Pos cells in the adult lung. Therefore, despite the aforementioned limitations, this new Fgf10Ki–v2 line opens the way for future mechanistic experiments involving the postnatal lung.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Validation of a Novel Fgf10Cre–ERT2 Knock-in Mouse Line Targeting FGF10Pos Cells Postnatally

Chu

Taghizadeh

Vazquez‐Armendariz

et al. 2021

Front. Cell Dev. Biol.

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“…Fgf10 inactivation leads to lung agenesis [19] while Fgf7 null mice are viable and display no obvious lung phenotype [20]. Changes in endogenous Fgf10 expression has been correlated with disease progression and/or repair after lung injury both in mice and humans [21][22][23]. Evidence for such a role for Fgf7 in the embryonic or adult lung in human or mice are still lacking in spite of the fact that Fgf7 was discovered 7 years before Fgf10 [24,25].…”

Section: Discussion Rmc-sca1 Pos Fgf10 Pos Are Different From the Rmc-sca1 Pos Axin2 Posmentioning

confidence: 99%

Characterization in mice of the stromal niche maintaining AT2 stem cell self-renewal in homeostasis and disease

Taghizadeh

Heiner

Wilhelm

et al. 2021

Preprint

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Resident mesenchymal cells (rMCs defined as Cd31NegCd45NegEpcamNeg) control the self-renewal and differentiation of alveolar epithelial type 2 (AT2) stem cells in vitro. The identity of these rMCs is still elusive. Among them, Axin2Pos mesenchymal alveolar niche cells (MANCs), which are expressing Fgf7, have been previously described. We propose that an additional population of rMCs, expressing Fgf10 (called rMC-Sca1PosFgf10Pos) are equally important to maintain AT2 stem cell self-renewal.The alveolosphere model, based on the AT2-rMC co-culture in growth factor reduced Matrigel, was used to test the efficiency of different rMC subpopulations isolated by FACS from adult murine lung to sustain the self-renewal and differentiation of AT2 stem cells.We demonstrate that rMC-Sca1PosFgf10Pos cells are efficient to promote the self-renewal and differentiation of AT2 stem cells. Co-staining of adult lung for Fgf10 mRNA and Sftpc protein respectively, indicate that 28% of Fgf10Pos cells are located close to AT2 cells. Co-ISH for Fgf7 and Fgf10 indicate that these two populations do not significantly overlap. Gene arrays comparing rMC-Sca1PosAxin2Pos and rMC-Sca1PosFgf10Pos support that these two cell subsets express differential markers. In addition, rMC function is decreased in diabetic and obese ob/ob mutant compared to WT mice with a much stronger loss of function in males compared to females.In conclusion, rMC-Sca1PosFgf10Pos cells play important role in supporting AT2 stem cells self-renewal and differentiation. This result sheds a new light on the subpopulations of rMCs contributing to the AT2 stem cell niche in homeostasis and in the context of COVID-19 pathogenesis.Key messageWhat is already known about the subject?Resident mesenchymal cells (rMCs defined as Cd31NegCd45NegEpcamNeg) control the self-renewal and differentiation of alveolar epithelial type 2 (AT2) stem cells in vitro. The identity of these rMCs is still elusive. Among them, Axin2Pos mesenchymal alveolar niche cells (MANCs), which are expressing Fgf7, have been previously described.What does this study add?Our study shows that an additional population of rMCs, expressing Fgf10 (called rMC-Sca1PosFgf10Pos) is equally important to maintain AT2 stem cell self-renewal. rMC-Sca1PosFgf10Pos are LipidToxHigh and are located close to AT2s. In addition, rMC-Sca1PosFgf10Pos cells support AT2 stem cell self-renewal and differentiation thereby identifying these cells as bone fide functional lipofibroblasts (LIFs). We have previously reported that LIF can transdifferentiate into activated MYF in the context of bleomycin-induced fibrosis in mice [1] and that activated MYF isolated from the lungs of end stage idiopathic fibrosis human patients can respond to Metformin to undergo transdifferentiation back to the LIF phenotype [2]. We also show that the function of rMCs-Sca1Pos is negatively impacted by gender and obesity, which represent two major aggravating factors for COVID-19 pathogenesis, leading to either death or major complications after infection recovery such as lung fibrosis.How might this impact on clinical practice and future development?By establishing that rMC-Sca1PosFgf10Pos are different from the MANCs, our study opens the way for a new key mesenchymal cell population that should be targeted to either prevent or reverse fibrosis. In addition, as this population maintains the AT2 stem cells self-renewal and differentiation, such targeting will also allow to progressively recover the loss in respiratory function.

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“…Here, we provide a detailed protocol for culturing airway organoids, or bronchiolospheres, which provide an assessment of the ability of mesenchymal cells to support club-cell growth. For complete details on the use and execution of this protocol, please refer to Moiseenko et al. (2020) .…”

mentioning

confidence: 99%

“…For complete details on the use and execution of this protocol, please refer to Moiseenko et al. (2020) .…”

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confidence: 99%

Protocol for the generation of murine bronchiolospheres

et al. 2021

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Summary Organoid models have been shown to be valuable tools for studying epithelial-mesenchymal crosstalk during biological and pathological settings. Our data identified ACTA2+ PDGFRα+ repair-supportive mesenchymal cells as an important component of the conducting airway niche. Here, we provide a detailed protocol for culturing airway organoids, or bronchiolospheres, which provide an assessment of the ability of mesenchymal cells to support club-cell growth. For complete details on the use and execution of this protocol, please refer to Moiseenko et al. (2020) .

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Identification of a Repair-Supportive Mesenchymal Cell Population during Airway Epithelial Regeneration

Cited by 33 publications

References 36 publications

Validation of a Novel Fgf10Cre–ERT2 Knock-in Mouse Line Targeting FGF10Pos Cells Postnatally

Validation of a Novel Fgf10Cre–ERT2 Knock-in Mouse Line Targeting FGF10Pos Cells Postnatally

Characterization in mice of the stromal niche maintaining AT2 stem cell self-renewal in homeostasis and disease

Protocol for the generation of murine bronchiolospheres

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