Lgr6-expressing functional nail stem-like cells differentiated from human-induced pluripotent stem cells

PLoS ONE, Journal Year: 2024, Volume and Issue: 19(5), P. e0303260 - e0303260

Published: May 14, 2024

The nail matrix containing stem cell populations produces nails and may contribute to fingertip regeneration. Nails are important tissues that maintain the functions of hand foot for handling objects locomotion. Tumor chemotherapy impairs growth and, in many cases, loses them, although not permanently. In this report, we have achieved successful differentiation (NS)-like cells from human-induced pluripotent (iPSCs) via digit organoids by stepwise stimulation, tracing molecular processes involved limb development. Comprehensive mRNA sequencing analysis revealed organoid global gene expression profile fits human finger NS-like expressed Lgr6 protein produced type-I keratin, KRT17, type-II KRT81, which abundant nails. Furthermore, succeeded producing functional Lgr6-reporter iPSCs. reporter iPSC-derived Lgr6-positive also KRT17 KRT81 proteins percutaneously transplanted region. To best our knowledge, is first report Our method construct enable discovery drugs repair possibly fingertip-regenerative therapy.

Language: Английский

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Sall4 regulates downstream patterning genes during limb regeneration

Developmental Biology, Journal Year: 2024, Volume and Issue: 515, P. 151 - 159

Published: July 26, 2024

Many salamanders can completely regenerate a fully functional limb. Limb regeneration is carefully coordinated process involving several defined stages. One key event during the patterning of blastema to inform cells what they must differentiate into. Although it known that many genes involved in initial development limb are re-used regeneration, exact molecular circuitry this not understood. Several large-scale transcriptional profiling studies axolotl have identified transcription factors up-regulated after amputation. Sall4 factor has been play essential roles maintaining an undifferentiated state and also plays unique role development. Inactivation bud results defects anterior-posterior found be both Xenopus salamanders, but date function untested. We confirmed using qRT-PCR present skin within blastema. Using CRISPR technology we microinjected gRNAs specific for complexed with cas9 protein into specifically knockout only. This resulted defects, including missing digits, fusion digit elements, radius ulna. suggests may similar regulating specification anterior-proximal skeletal elements.

Language: Английский

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Unveiling axolotl transcriptome for tissue regeneration with high-resolution annotation via long-read sequencing

Computational and Structural Biotechnology Journal, Journal Year: 2024, Volume and Issue: 23, P. 3186 - 3198

Published: Aug. 23, 2024

Highlights•Long-read sequencing was used to overcome annotation limitations, improving coverage and accuracy of axolotl genome.•The study identified 4997 novel transcribed regions 583 sequences that encoded functional proteins.•Potential proteins were discovered with key regenerative functions.•Alternative splicing variants detected across critical regeneration pathways.AbstractAxolotls are known for their remarkable ability. Exploring transcriptome provides insight into mechanisms. However, the current is limited, leaving role unannotated transcripts in unknown. To discourse this challenge, we exploited long-read technology, which enables direct observation full-length RNA transcripts, greatly enhancing annotation. By utilizing method, 222 gene loci 4775 quantified using short-read data. Through inclusive analysis, homologs, potential proteins, noncoding RNAs, alternative events pathways. In particular, high protein-coding implicated cell cycle regulation musculoskeletal development, identified. Interestingly, splice also diverse pathways regeneration. This specifies these potentially play vital roles underpinning robust capacities axolotls. Single-cell transcriptomic analysis further revealed isoforms predominantly exist limb chondrocytes mature tissue populations. Overall, findings significantly advanced consideration provided a new perspective understanding mechanisms abilities axolotls.Graphical abstract

Language: Английский

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Cellular senescence promotes progenitor cell expansion during axolotl limb regeneration

bioRxiv (Cold Spring Harbor Laboratory), Journal Year: 2022, Volume and Issue: unknown

Published: Sept. 2, 2022

SUMMARY Axolotl limb regeneration is accompanied by the transient induction of cellular senescence within blastema, structure which nucleates regeneration. The precise role this blastemal senescent cell (bSC) population, however, remains unknown. Here, through a combination gain- and loss-of-function assays, we elucidate functions molecular features in vivo . We demonstrate that plays positive during axolotl regeneration, creating pro-proliferative niche supports progenitor expansion blastema outgrowth. Senescent cells impact on their microenvironment via Wnt pathway modulation. Further, uncover link between signalling induction, propose bSC-derived signals facilitate proliferation neighbouring part preventing into senescence. This work defines roles complex structures.

Language: Английский

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Material properties in regenerating axolotl limbs using inverse finite element analysis

Journal of the mechanical behavior of biomedical materials/Journal of mechanical behavior of biomedical materials, Journal Year: 2023, Volume and Issue: 150, P. 106341 - 106341

Published: Dec. 24, 2023

Language: Английский

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Gene expression analysis of the Xenopus laevis early limb bud proximodistal axis

Developmental Dynamics, Journal Year: 2022, Volume and Issue: 251(11), P. 1880 - 1896

Published: July 9, 2022

Abstract Background Limb buds develop as bilateral outgrowths of the lateral plate mesoderm and are patterned along three axes. Current models proximal to distal patterning early amniote limb suggest that two signals, a organizing signal from apical epithelial ridge (AER, Fgfs) an opposing (retinoic acid [RA]) act on pattern this axis. Results Transcriptional analysis stage 51 Xenopus laevis hindlimb sectioned proximal‐distal axis showed region is distinct rest limb. Expression capn8.3 , novel calpain, was located in cells immediately flanking AER. The Wnt antagonist Dkk1 AER‐specific limbs. Two transcription factors, sall1 zic5 were expressed mesenchyme. Zic5 has no described association with development. We also describe expression genes, gata5 tnn not previously associated Differentially genes Fgf, Wnt, RA signaling well differential cell adhesion proliferation. Conclusions identify new candidate for proximodistal patterning. Our RA‐regulated supports role transient gradients bud proximal‐to‐distal anamniote model organism.

Language: Английский

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Conserved Genes in Highly Regenerative Metazoans Are Associated with Planarian Regeneration

Genome Biology and Evolution, Journal Year: 2024, Volume and Issue: 16(5)

Published: April 23, 2024

Abstract Metazoan species depict a wide spectrum of regeneration ability which calls into question the evolutionary origins underlying processes. Since with high are widely distributed throughout metazoans, there is possibility that metazoan ancestor had an common molecular mechanism. Early metazoans like sponges possess regenerative ability, but, due to large differences they have Cnidaria and Bilateria regarding symmetry neuronal systems, it can be inferred this different. We hypothesized last possessed remarkable was lost during evolution. separated three classes possessing whole-body regenerating, low ability. Using multiway BLAST gene phylogeny approach, we identified genes conserved in regenerating labeled them Bilaterian genes. Through transcription factor analysis, were associated overabundance homeodomain regulatory elements. RNA interference resulted loss phenotype for HRJDa, HRJDb, DUF21, DISP3, ARMR observed DUF21 knockdown highly lethal early stages indicating potential role wound response. Also, showed after second amputation. The results strongly correlate their respective RNA-seq profiles. propose play major across Bilateria.

Language: Английский

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The Impact of Senescent Cells on Limb Regeneration

Cellular Reprogramming, Journal Year: 2024, Volume and Issue: 26(3), P. 91 - 92

Published: May 8, 2024

Cellular senescence is a state in which cells enter cell cycle arrest. However, senescent have the ability to secrete signaling molecules such as chemokines, cytokines, and growth factors. This secretory activity an important feature of cells, since secreted factors impact surrounding cellular microenvironment. Indeed, their secretome play crucial role during limb development. whether process regeneration also relies on remains unclear. Creation novel targeted depletion strategy that can eliminate regenerating has now demonstrated for regeneration. linked cell-derived Wnt signaling. These findings reveal previously unknown through

Language: Английский

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Cartilage mechanobiology during limb growth

Published: Jan. 1, 2024

The mechanobiology of cartilage during limb growth represents a complex interplay between mechanical forces and biological processes. However, the fundamental processes that involve formation from mesenchymal stem cells are not fully understood. Further, cellular level response in its native environment under physiological load is characterized. This thesis aims to bridge critical gaps our understanding development regeneration by investigating nuanced interactions mechanics within cartilaginous tissues. Identifying these research gaps, this encompasses three specific aims. Aim 1 focuses on characterizing viscoelastic material properties growing limbs, employing axolotls as an animal model. Our results reveal significant increases both instantaneous equilibrium shear moduli regeneration, coupled with notable changes short- long-term stress relaxation times. glycosaminoglycan content also development. 2 explores calcium signaling in-situ chondrocytes physiologically relevant cyclic loads dynamic hydrostatic pressure. findings underscore strain rate-dependent increase percentage responsive compressive loads, non-distinct time characteristics across loading conditions. Conversely, low magnitude pressure showed no impact chondrocytes. 3 investigates expression mechanosensitive ion channels (TRPV4, PIEZO1, PIEZO2) axolotl regeneration. study unveils presence TRPV4 PIEZO2 blastemal early late heightened condensing mesenchyme These taken together shed light intricate responses. implications abnormal mechanobiological profound, contributing developmental disorders musculoskeletal diseases. Understanding mechanisms conditions opens avenues for therapeutic strategies aimed at promoting proper mitigating skeletal abnormalities. Future will focus elucidating functional roles further expanding interconnections biology.--Author's abstract

Language: Английский

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Putative epithelial–mesenchymal transitions during salamander limb regeneration: Current perspectives and future investigations

Annals of the New York Academy of Sciences, Journal Year: 2024, Volume and Issue: unknown

Published: Sept. 13, 2024

Abstract Previous studies have implicated epithelial–mesenchymal transition (EMT) in salamander limb regeneration. In this review, we describe putative roles for EMT during each stage of regeneration axolotls and other salamanders. We hypothesize that EMT‐like gene expression programs may regulate three main cellular processes regeneration: (1) keratinocyte migration wound closure; (2) transient invasion the stump by epithelial cells undergoing EMT; (3) use non‐epithelial blastemal progenitor to escape confines their niches. Finally, propose nontraditional warrant further investigation, including alternative regulators, stem cell activation, fibrosis induced aberrant EMT.

Language: Английский

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