Human embryonic stem-cell derivatives for full reconstruction of the pluristratified epidermis: a preclinical study

Guénou, Hind; Nissan, Xavier; Larcher, Fernando; Feteira, Jessica; Saidani, Manoubia; Río, Marcela Del; Barrault, C.; Bernard, François‐Xavier; Peschanski, Marc; Baldeschi, Christine; Waksman, Gilles

doi:10.1016/s0140-6736(09)61496-3

Cited by 229 publications

(221 citation statements)

References 31 publications

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“…BMP-4-based protocols have been established to obtain keratinocytes from pluripotent cells (12,14,25) and very recently, Itoh et al used BMP-4 combined with retinoic acid for a limited time to generate keratinocytes from iPSC from dystrophic epidermolysis bullosa patients reaching 30-40% of K14 + cells that could be enriched only after passaging or cell sorting (26). Despite possible enrichment and satisfactory 3D potential (14,26), improved protocols for highly homogenous production of keratinocytes from pluripotent cells are still needed to consider cell therapy. We observed that BMP-4-induced epidermal commitment of iPSC was significantly low compared with what was reported with hESC (14), apparently because of limited spontaneous neuroectoderm commitment that led to the generation of BMP-4-induced trophoectoderm.…”

Section: δNp63mentioning

confidence: 99%

“…To define whether EECiPSC lines could mimic these defects, we optimized protocols to differentiate human iPSC into epidermal cells. Epidermal commitment of embryonic stem cells can be induced by BMP-4, which inhibits neural differentiation and promotes epidermal fate of neuroectodermal cells (12)(13)(14). Prolonged treatment with BMP-4 and ascorbic acid combined with keratinocyte growth conditions have been shown to promote efficiently hESC differentiation into mature keratinocytes (14).…”

mentioning

confidence: 99%

“…Epidermal commitment of embryonic stem cells can be induced by BMP-4, which inhibits neural differentiation and promotes epidermal fate of neuroectodermal cells (12)(13)(14). Prolonged treatment with BMP-4 and ascorbic acid combined with keratinocyte growth conditions have been shown to promote efficiently hESC differentiation into mature keratinocytes (14). However, applying this protocol to iPSC lines appeared inefficient because most of the cells differentiate in a heterogenous manner, with few ectodermal-like cells and with large cystic-like structures (Fig.…”

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confidence: 99%

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Impaired epithelial differentiation of induced pluripotent stem cells from ectodermal dysplasia-related patients is rescued by the small compound APR-246/PRIMA-1 ^MET

Shalom-Feuerstein

Serror

Aberdam

et al. 2013

Proc. Natl. Acad. Sci. U.S.A.

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Ectodermal dysplasia is a group of congenital syndromes affecting a variety of ectodermal derivatives. Among them, ectrodactyly, ectodermal dysplasia, and cleft lip/palate (EEC) syndrome is caused by single point mutations in the p63 gene, which controls epidermal development and homeostasis. Phenotypic defects of the EEC syndrome include skin defects and limbal stem-cell deficiency. In this study, we designed a unique cellular model that recapitulated major embryonic defects related to EEC. Fibroblasts from healthy donors and EEC patients carrying two different point mutations in the DNA binding domain of p63 were reprogrammed into induced pluripotent stem cell (iPSC) lines. EEC-iPSC from both patients showed early ectodermal commitment into K18 + cells but failed to further differentiate into K14 + cells (epidermis/limbus) or K3/K12 + cells (corneal epithelium). APR-246 (PRIMA-1 MET ), a small compound that restores functionality of mutant p53 in human tumor cells, could revert corneal epithelial lineage commitment and reinstate a normal p63-related signaling pathway. This study illustrates the relevance of iPSC for p63 related disorders and paves the way for future therapy of EEC.

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Section: δNp63mentioning

confidence: 99%

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confidence: 99%

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confidence: 99%

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Impaired epithelial differentiation of induced pluripotent stem cells from ectodermal dysplasia-related patients is rescued by the small compound APR-246/PRIMA-1 ^MET

Shalom-Feuerstein

Serror

Aberdam

et al. 2013

Proc. Natl. Acad. Sci. U.S.A.

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“…RHE was prepared as previously described [61]. Suspensions of cultured primary human keratinocytes (see above) were further cultured on polycarbonate culture inserts (Millipore) in Epilife medium (Thermo Fisher Scientific) supplemented with 1.5 mM calcium chloride and 50 µg/mL ascorbic acid, and then transferred to the air–liquid interface for 10 days.…”

Section: Methodsmentioning

confidence: 99%

Pseudomonas aeruginosa flagellum is critical for invasion, cutaneous persistence and induction of inflammatory response of skin epidermis

Garcia

Morello

Garnier³

et al. 2018

Virulence

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Pseudomonas aeruginosa, an opportunistic pathogen involved in skin and lung diseases, possesses numerous virulence factors, including type 2 and 3 secretion systems (T2SS and T3SS) and its flagellum, whose functions remain poorly known during cutaneous infection. Using isogenic mutants deleted from genes encoding each or all of these three virulence factors, we investigated their role in induction of inflammatory response and in tissue invasiveness in human primary keratinocytes and reconstructed epidermis. Our results showed that flagellum, but not T2SS and T3SS, is involved in induction of a large panel of cytokine, chemokine, and antimicrobial peptide (AMP) mRNA in the infected keratinocytes. Chemokine secretion and AMP tissular production were also dependent on the presence of the bacterial flagellum. This pro-inflammatory effect was significantly reduced in keratinocytes infected in presence of anti-toll-like receptor 5 (TLR5) neutralizing antibody. Bacterial invasion of human epidermis and persistence in a mouse model of sub-cutaneous infection were dependent on the P. aeruginosa flagellum. We demonstrated that flagellum constitutes the main virulence factor of P. aeruginosa involved not only in early induction of the epidermis inflammatory response but also in bacterial invasion and cutaneous persistence. P. aeruginosa is mainly sensed by TLR5 during the early innate immune response of human primary keratinocytes.

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“…28 Skin fibroblasts from two EEC patients carrying R304W and R204W mutations in TP63 and one healthy control were reprogrammed into iPSC by lentivirus-mediated over-expression of Oct3/4, Sox2, Klf4 and c-myc. Protocol for epidermal differentiation was adapted from Aberdam et al 38 and Guenou et al 39 (Petit et al, manuscript in preparation).…”

Section: Methodsmentioning

confidence: 99%

Diverse p63 and p73 isoforms regulate Δ133p53 expression through modulation of the internal TP53 promoter activity

et al. 2011

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In response to stress, p53 binds and transactivates the internal TP53 promoter, thus regulating the expression of its own isoform, D133p53a. Here, we report that, in addition to p53, at least four p63/p73 isoforms regulate D133p53 expression at transcriptional level: p63b, DNp63a, DNp63b and DNp73c. This regulation occurs through direct DNA-binding to the internal TP53 promoter as demonstrated by chromatin immunoprecipitation and the use of DNA-binding mutant p63. The promoter regions involved in the p63/p73-mediated transactivation were identified using deleted, mutant and polymorphic luciferase reporter constructs. In addition, we observed that transient expression of p53 family members modulates endogenous D133p53a expression at both mRNA and protein levels. We also report concomitant variation of p63 and D133p53 expression during keratinocyte differentiation of HaCat cells and induced pluripotent stem cells derived from mutated p63 ectodermal dysplasia patients. Finally, proliferation assays indicated that D133p53a isoform regulates the anti-proliferative activities of p63b, DNp63a, DNp63b and DNp73c. Overall, this study shows a strong interplay between p53, p63 and p73 isoforms to orchestrate cell fate outcome. The TP53 family, composed of TP53, TP63 and TP73 genes, presents a strong homology of structures and expression patterns. 1,2 The three genes encode several protein isoforms carrying distinct N-termini (TA or D forms) and C-termini (a, b, g, y), because of the use of alternative promoters, splicing sites and translational initiation sites. 3,4 The full-length proteins also share a similar protein structure with a N-terminal transactivation domain, a central DNA-binding domain (DBD) and a C-terminal oligomerisation domain. 4,5 A strong interplay has been described between p53 family members. They all bind specifically to DNA response elements (p53RE), modulate gene expression and thus cellfate outcome. 6 Moreover, the p53-mediated apoptosis is severely impaired in the absence of p63 and p73 in response to DNA damage. 7 The interplay between p53 family members is not limited to transcriptional modulation of common target genes; they also regulate each other's expression and activity. 6 p53, p73 and DNp73 transactivate the promoter of DNp73 isoform, which in turn, inhibits the p53-and p73-mediated transcriptional activity and apoptosis through direct competition for DNAbinding. [8][9][10][11][12] Similarly, p53, DNp63 and p73g modulate the activity of the internal TP63 promoter regulating DNp63 expression, which inhibits p53, p63 and p73 transcriptional activities. 2,6,[13][14][15] Recently, it has been reported that p53 regulates the transcription of D133p53 isoform, which lacks the entire transactivation domain and part of the DBD. 16,17 p53 directly binds to p53REs located within the internal TP53 promoter leading to an increase of D133p53 mRNAs. Those transcripts generate two different N-terminal p53 isoforms, D133p53 and D160p53, through the use of two alternative translational initiation sites (co...

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Human embryonic stem-cell derivatives for full reconstruction of the pluristratified epidermis: a preclinical study

Cited by 229 publications

References 31 publications

Impaired epithelial differentiation of induced pluripotent stem cells from ectodermal dysplasia-related patients is rescued by the small compound APR-246/PRIMA-1 ^MET

Impaired epithelial differentiation of induced pluripotent stem cells from ectodermal dysplasia-related patients is rescued by the small compound APR-246/PRIMA-1 ^MET

Pseudomonas aeruginosa flagellum is critical for invasion, cutaneous persistence and induction of inflammatory response of skin epidermis

Diverse p63 and p73 isoforms regulate Δ133p53 expression through modulation of the internal TP53 promoter activity

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Human embryonic stem-cell derivatives for full reconstruction of the pluristratified epidermis: a preclinical study

Cited by 229 publications

References 31 publications

Impaired epithelial differentiation of induced pluripotent stem cells from ectodermal dysplasia-related patients is rescued by the small compound APR-246/PRIMA-1 MET

Impaired epithelial differentiation of induced pluripotent stem cells from ectodermal dysplasia-related patients is rescued by the small compound APR-246/PRIMA-1 MET

Pseudomonas aeruginosa flagellum is critical for invasion, cutaneous persistence and induction of inflammatory response of skin epidermis

Diverse p63 and p73 isoforms regulate Δ133p53 expression through modulation of the internal TP53 promoter activity

Contact Info

Product

Resources

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Impaired epithelial differentiation of induced pluripotent stem cells from ectodermal dysplasia-related patients is rescued by the small compound APR-246/PRIMA-1 ^MET

Impaired epithelial differentiation of induced pluripotent stem cells from ectodermal dysplasia-related patients is rescued by the small compound APR-246/PRIMA-1 ^MET