Опубликована: Янв. 1, 2024
Язык: Английский
Trends in Molecular Medicine, Год журнала: 2024, Номер 30(10), С. 950 - 964
Опубликована: Май 14, 2024
Duchenne muscular dystrophy (DMD) is caused by mutations in the X-linked DMD gene, resulting absence of dystrophin, progressive muscle degeneration, and heart failure. Genetically tailored pig models resembling human recapitulate biochemical, clinical, pathological hallmarks with an accelerated disease progression compared to patients. pigs have been used evaluate therapeutic concepts such as gene editing reframe a disrupted reading frame or delivery artificial chromosome vectors carrying complete gene. Moreover, instrumental validating new diagnostic modalities multispectral optoacoustic tomography (MSOT) for non-invasive monitoring progression. may thus help bridge gap between proof-of-concept studies cellular rodent clinical
Язык: Английский
Процитировано
4
bioRxiv (Cold Spring Harbor Laboratory), Год журнала: 2024, Номер unknown
Опубликована: Июнь 8, 2024
Abstract Human embryonic development is generally slower compared with mouse, and one of the model systems for such inter-species differences in developmental tempo segmentation clock. The oscillation period human clock, as measured induced presomitic mesoderm (iPSM) cells, two times longer than that mouse. While core clock gene HES7 known to show protein degradation iPSM mouse iPSM, it remains unclear whether concept species-specific stability generalizable other genes. Here we systematically rates approximately 5000 genes between by using dynamic SILAC-based proteomics, demonstrating a pervasive trend cells. difference was observed not only proteasome-mediated but also lysosome-mediated degradation. We further investigated effect metabolism on profile. Treating cells glycolysis inhibitor extended period, aligning profile more closely though less These results highlight universality suggest modulators.
Язык: Английский
Процитировано
4
Nature Communications, Год журнала: 2025, Номер 16(1)
Опубликована: Янв. 20, 2025
Abstract Numerous cellular and molecular processes during embryonic development prompt the fundamental question of how their tempos are coordinated whether a common global modulator exists. While segmentation clock tempo scales with kinetics gene expression degradation core Hes7 across mammals, coordination these remains unclear. This study examines metabolic activities serve as for clock, finding them to be selective instead. Several inhibitions extend period but affect key differently: glycolysis inhibition slows protein production delay without altering intron delay, while electron transport chain extends influencing other processes. Combinations distinct exhibit synergistic effects. We propose that scaled species may result from combined modulators shaped by evolutionary constraints, rather than single modulator.
Язык: Английский
Процитировано
0
Nature Reviews Genetics, Год журнала: 2025, Номер unknown
Опубликована: Март 4, 2025
Язык: Английский
Процитировано
0
Stem Cell Reports, Год журнала: 2025, Номер unknown, С. 102425 - 102425
Опубликована: Март 1, 2025
Outer retinal degenerative diseases (RDDs) and injuries leading to photoreceptor (PR) loss are prevailing causes of blindness worldwide. While significant progress has been made in the manufacture human pluripotent stem cell (hPSC)-derived PRs, robust production (PSC)-PRs from swine, a popular preclinical large animal model, would provide an avenue collect conspecific functional safety data complement xenograft studies. Toward this goal, we describe highly efficient generation PR-dominant porcine induced PSC (piPSC)-derived organoids (ROs) using modifications our established hPSC-RO differentiation protocol. Porcine iPSC-ROs were characterized immunocytochemistry (ICC) single-cell RNA sequencing (scRNA-seq), which revealed presence maturation major neural retina types, including PRs ganglion cells, possess molecular signatures akin those found hPSC-ROs. In late piPSC-ROs, organized outer neuroepithelium was observed with rods cones possessing segments axon terminals expressing pre-synaptic markers adjacent dendritic bipolar cells. The existence piPSC lines protocols that support reproducible, scalable female male ROs will facilitate transplant studies models injury RDDs unconfounded by immunological evolutionary incompatibilities inherent xenografts.
Язык: Английский
Процитировано
0
Current Opinion in Genetics & Development, Год журнала: 2024, Номер 84, С. 102149 - 102149
Опубликована: Янв. 9, 2024
The rate of development is highly variable across animal species. However, the mechanisms regulating developmental tempo have remained elusive due to difficulties in performing direct interspecies comparisons. Here, we discuss how pluripotent stem cell-based models can be used investigate cell- and tissue-autonomous temporal processes. These systems enable quantitative comparisons different species under similar experimental conditions. Moreover, constantly growing cell zoo collection allows extension studies a great number unconventional We argue that constitutes powerful platform perform comparative tempo, as well study other forms biological time control such species-specific lifespan, heart rate, circadian clocks.
Язык: Английский
Процитировано
3
Frontiers in Cell and Developmental Biology, Год журнала: 2024, Номер 11
Опубликована: Янв. 29, 2024
Somitogenesis is a hallmark feature of all vertebrates and some invertebrate species that involves the periodic formation block-like structures called somites. Somites are transient embryonic segments eventually establish entire vertebral column. A highly conserved molecular oscillator segmentation clock underlies this event pace regulates somite formation. Although signaling pathways govern in most vertebrates, mechanisms underlying species-specific divergence various characteristics remain elusive. For example, classical model such as zebrafish, chick, mouse embryos tick with periodicity ∼30, ∼90, ∼120 min respectively. This enables them to form number vertebrae during their overall timespan somitogenesis. Here, we perform systematic review features keen focus on embryos. We using three different perspectives: Notch-responsive genes, ligand-receptor dynamics, synchronization between neighboring oscillators. further reports use non-classical organisms
Язык: Английский
Процитировано
2
Current Protocols, Год журнала: 2024, Номер 4(5)
Опубликована: Май 1, 2024
Abstract Although protocols to generate authentic transgene‐free mouse and human induced pluripotent stem cells (iPSCs) are now well established, standard methods for reprogramming porcine somatic still suffer from low efficiency transgene retention. The Basic Protocol describes procedures establish iPSCs (PiPSCs) fibroblasts. This method uses episomal plasmids encoding POU5F1 , SOX2 NANOG KLF4 SV40LT c‐MYC LIN28A microRNA‐302/367 combined with an optimized medium, PiPSC lines. Support describe the establishment characterization of clonal lines, as preparation feeder EBNA1 mRNA. optimized, step‐by‐step approach tailored this species enables efficient derivation PiPSCs in ∼4 weeks. provides a new valuable model studies larger mammalian species’ development, disease, regenerative biology. © 2024 Authors. Current Protocols published by Wiley Periodicals LLC. : Reprogramming fibroblasts 1 Preparation embryonic layer 2 vitro –transcribed mRNA 3 Establishment cell (PiPSC) lines 4 characterization: Genomic DNA PCR RT‐PCR 5 Immunostaining
Язык: Английский
Процитировано
2
bioRxiv (Cold Spring Harbor Laboratory), Год журнала: 2024, Номер unknown
Опубликована: Июнь 6, 2024
Abstract A sequence of cellular and molecular processes unfolding during embryonic development prompts fundamental questions how the tempo multiple is coordinated whether a common global modulator exists. The oscillation segmentation clock well-studied model developmental tempo. While period known to scale with kinetics gene expression degradation core Hes7 across mammalian species, these key are remains unclear. In this study, we investigated if metabolic activities act as for clock, finding that they rather selective modulators. several inhibitions extended period, their effects on varied. Inhibition glycolysis decelerated protein production delay but did not influence intron delay. Electron transport chain inhibition without influencing other two processes. Combinations distinct exhibited synergistic effects. By contrast, temperature changes affected all three simultaneously. These results highlight processes, hinting scaled species may be achieved through combinations
Язык: Английский
Процитировано
2
Frontiers in Cell and Developmental Biology, Год журнала: 2024, Номер 12
Опубликована: Июнь 24, 2024
Pigs ( Sus scrofa ) are widely acknowledged as an important large mammalian animal model due to their similarity human physiology, genetics, and immunology. Leveraging the full potential of this presents significant opportunities for major advancements in fields comparative biology, disease modeling, regenerative medicine. Thus, derivation pluripotent stem cells from species can offer new tools modeling serve a stepping stone test future autologous or allogeneic cell-based therapies. Over past few decades, great progress has been made establishing porcine (pPSCs), including embryonic (pESCs) derived pre- peri-implantation embryos, induced (piPSCs) using variety cellular reprogramming strategies. However, stabilization pPSCs was not straightforward directly applying culture conditions developed optimized murine primate PSCs. Therefore, it historically challenging establish stable pPSC lines that could pass stringent pluripotency tests. Here, we review recent advances establishment We focus on evolving methods eventually led pESCs transgene-free piPSCs, well current challenges rapidly advancing field.
Язык: Английский
Процитировано
2