Discover Oncology, Год журнала: 2024, Номер 15(1)
Опубликована: Сен. 30, 2024
Язык: Английский
Investigative Ophthalmology & Visual Science, Год журнала: 2025, Номер 66(2), С. 64 - 64
Опубликована: Фев. 25, 2025
Purpose: Persistent epithelial alterations such as delayed wound healing are a key feature of diabetic corneal disease. Previously, we reported that epigenetic changes in the cornea led to suppression Wnt-5a, and addition Wnt-5a accelerated healing. In this study, set determine which Wnt receptor(s) mediated induced stimulation Methods: Human limbal cells (LECs) were isolated from postmortem non-diabetic donor eyes for single-cell RNA sequencing (scRNA-seq) DNA methylation analysis. These analyses validated by qRT-PCR, western blot, or immunostaining tissue sections. Cultured primary LECs transfected with small interfering (siRNA) specific receptors evaluate their role scratch presence absence 200 ng/mL Wnt-5a. Results: Single-cell analysis revealed differential gene expression receptors, ROR2, MCAM, FZD5, FZD6, FZD7. arrays showed hypomethylation ROR2 promoter versus 41.3% (**P < 0.01) resulting increased protein expression. Non-diabetic siRNA knockdown but not FZD7, RYK significantly decreased approximately 50% (*P 0.05) control siRNA. LECs, inhibited 40% FZD5 partially blocked could be restored Conclusions: seems mediate mainly through receptor tyrosine kinase like orphan 2 Frizzled-5 serving possible co-receptor smaller effect.
Язык: Английский
Процитировано
1
eLife, Год журнала: 2024, Номер 13
Опубликована: Май 23, 2024
The receptor tyrosine kinase ROR2 mediates noncanonical WNT5A signaling to orchestrate tissue morphogenetic processes, and dysfunction of the pathway causes Robinow syndrome, brachydactyly B, metastatic diseases. domain(s) mechanisms required for function, however, remain unclear. We solved crystal structure extracellular cysteine-rich (CRD) Kringle (Kr) domains found that, unlike other CRDs, CRD lacks signature hydrophobic pocket that binds lipids/lipid-modified proteins, such as WNTs, suggesting a novel mechanism ligand reception. Functionally, we showed CRD, but not domains, is minimally sufficient promote signaling, mutations in adjacent Kr impair secretion function. Moreover, using function-activating -perturbing antibodies against Frizzled (FZ) family WNT receptors, demonstrate involvement FZ WNT5A-ROR signaling. Thus, acts via its potentiate function super-complex includes transduce signals.
Язык: Английский
Процитировано
6
Current topics in developmental biology/Current Topics in Developmental Biology, Год журнала: 2023, Номер unknown, С. 195 - 227
Опубликована: Янв. 1, 2023
Язык: Английский
Процитировано
9
Cells, Год журнала: 2024, Номер 13(22), С. 1870 - 1870
Опубликована: Ноя. 11, 2024
Dishevelled (DVL) proteins precisely control Wnt signaling pathways with many effectors. While substantial research has advanced our understanding of DVL's role in pathways, key questions regarding its regulatory mechanisms and interactions remain unresolved. Herein, we present the recent advances perspectives on how DVL regulates signaling. The experimentally determined conserved domain structures conjunction AlphaFold-predicted are used to understand regulation. We also summarize various diseases provide insights into further directions for DVL-mediated mechanisms. These findings underscore importance as a pharmaceutical target or biological marker diseases, offering exciting potential future biomedical applications.
Язык: Английский
Процитировано
1
bioRxiv (Cold Spring Harbor Laboratory), Год журнала: 2024, Номер unknown
Опубликована: Июнь 30, 2024
Abstract Mouse embryonic fibroblasts (MEFs) derived from genetically modified mice are a valuable resource for studying gene function and regulation. The MEF system can also be combined with rescue studies to characterize the of mutant genes/proteins, such as disease-causing variants. However, primary MEFs undergo senescence soon after isolation passaging, making long-term genetic manipulations difficult. Previously described methods immortalization often inefficient or alter physiological properties cells. Here, we describe an optimized protocol immortalizing via CRISPR-mediated deletion Tp53 gene. This method is highly efficient consistently generates immortalized MEFs, iMEFs, within 14 days. Importantly, iMEFs closely resemble parent cell populations, individual cloned expanded subsequent manipulation characterization. We envision that this adopted immortalize other mouse types. Key Features CRISPR-based knockout enables in under 2 weeks. Immortalization requires Neon electroporator another comparable electroporation transfect cells CRISPR constructs.
Язык: Английский
Процитировано
0
Discover Oncology, Год журнала: 2024, Номер 15(1)
Опубликована: Сен. 30, 2024
Язык: Английский
Процитировано
0