Crk mediates Csk-Hippo signaling independently of Yap tyrosine phosphorylation to induce cell extrusion DOI

Abdul Hannan,

Qian Wang,

Yihua Wu

и другие.

bioRxiv (Cold Spring Harbor Laboratory), Год журнала: 2024, Номер unknown

Опубликована: Июль 1, 2024

Abstract Src family kinases (SFKs), including Src, Fyn and Yes, play important roles in development cancer. Despite being first discovered as the Y es- a ssociated p rotein, regulation of Yap by SFKs remains poorly understood. Here, through single-cell analysis genetic lineage tracing, we show that pan-epithelial ablation C-terminal kinase (Csk) lacrimal gland unleashes broad signaling but specifically causes extrusion apoptosis acinar progenitors at time when they are shielded myoepithelial cells from basement membrane. Csk mutants can be phenocopied constitutively active rescued deleting or Taz , indicating significant functional overlap between signaling. Although Src-induced tyrosine phosphorylation has long been believed to regulate activity, find mutating these residues both fails perturb mouse alleviate phenotype. In contrast, loses Hippo signaling-dependent serine translocates into nucleus mutants. Further chemical genetics studies demonstrate acute inhibition enhances Crk/CrkL Rac1 whereas removing Crk / CrkL Rap1 ameliorates mutant These results controls Hippo-Yap Crk/CrkL-Rac/Rap axis promote cell extrusion.

Язык: Английский

Exploring Immune Cell Diversity in the Lacrimal Glands of Healthy Mice: A Single-Cell RNA-Sequencing Atlas DOI Open Access

Qiwei Fan,

Ruyu Yan,

Yan Li

и другие.

International Journal of Molecular Sciences, Год журнала: 2024, Номер 25(2), С. 1208 - 1208

Опубликована: Янв. 19, 2024

The lacrimal gland is responsible for maintaining the health of ocular surface through production tears. However, our understanding immune system within currently limited. Therefore, in this study, we utilized single-cell RNA sequencing and bioinformatic analysis to identify analyze cells molecules present glands normal mice. A total 34,891 were obtained from mice classified into 18 distinct cell clusters using Seurat clustering. Within these populations, 26 different subpopulations identified, including T cells, innate lymphocytes, macrophages, mast dendritic B cells. Network revealed complex cell-cell interactions between with particularly significant observed among plasma Interestingly, found be main source ligands Thy1 signaling pathway, while M2 macrophages identified as primary target pathway. Moreover, some validated immunohistological techniques. Collectively, findings highlight abundance provide valuable insights complexity its relevance associated diseases.

Язык: Английский

Процитировано

6
Advances in lacrimal gland organoid development: Techniques and therapeutic applications DOI
Ahmed Abdal Dayem, Soo Bin Jang,

Nahee Lim

и другие.

Biomedicine & Pharmacotherapy, Год журнала: 2025, Номер 183, С. 117870 - 117870

Опубликована: Янв. 26, 2025

Язык: Английский

Процитировано

0
Serum and tear autoantibodies from NOD and NOR mice as potential diagnostic indicators of local and systemic inflammation in Sjögren’s disease DOI Creative Commons
Shruti Singh Kakan,

S. Abdelhamid,

Yaping Ju

и другие.

Frontiers in Immunology, Год журнала: 2025, Номер 15

Опубликована: Янв. 28, 2025

Background Sjögren’s Disease (SjD) is an autoimmune disease characterized by lymphocytic infiltration of salivary and lacrimal glands (LG). The LG produces the protein-rich aqueous component tears, SjD-associated dacryoadenitis (AD) may thus alter tear autoantibody composition. Methods presence tertiary lymphoid structures (TLS) in from two murine models AD, male non-obese diabetic (NOD) insulitis resistant (NOR) mice, were evaluated using immunofluorescence. IgG IgA reactivity serum tears these probed three studies against a panel 80-120 autoantigens microarrays relative to healthy BALB/c mice. Sources Ig investigated scRNA-Seq (GSE132420). Data analyzed R package Limma Seurat. Results Analysis immunofluorescence sections both SjD showed TLS. Only one was significantly elevated across all studies. Three autoantibodies but not Conversely, six thirteen (6 sharing same autoantigen) models. Igha Ighg2b expressing cells identified plasma cell cluster NOD.H2b LG. Conclusion NOD NOR mice with AD have distinct profiles serum. Tear isotype greater diversity than autoantibodies. TLS observed are likely source

Язык: Английский

Процитировано

0
Single-Cell RNA Sequencing Revealed Functional Conjunctival Keratinocytes Loss via TGF-β-Wnt/β-Catenin Signaling in Sjögren’s Syndrome Related Dry Eye DOI Creative Commons
Bowen Wang, Lihong Liang, Hao Zeng

и другие.

Investigative Ophthalmology & Visual Science, Год журнала: 2025, Номер 66(4), С. 43 - 43

Опубликована: Апрель 16, 2025

The role of the conjunctiva in pathophysiology Sjögren's syndrome (SS) related dry eye disease (DED) remains obscure especially view functional conjunctival epithelia. In order to illustrate effects SS, we investigated interactions between parenchymal cells and immune with single-cell RNA sequencing technique. Freshly collected from a canonical SS model was prepared for 10 × Genomics T cell receptor (TCR) sequencing. Conjunctiva Western blot, immunofluorescence, multiplex immunohistochemical (mIHC), flow cytometry. Phenol red thread test, lissamine staining, qRT-PCR were applied evaluate signs DED. DED phenotype validated model. Loss water-secreting keratinocyte projected scRNA-seq data proved by mIHC test mice. proportion Lgr4+ basal epithelial poor ability differentiate into mature increased, Wnt/β-catenin signaling upregulated it under regulation TGF-β derived macrophages. Such macrophages promoted angiogenesis through secretion VEGFA activate endothelial cells. Immuno-fibroblasts had an increased population, which implicated specifically activated chemotaxis. conjunctiva, TGF-β-Wnt/β-catenin axis downregulated formation keratinocytes accompanied infiltration pro-angiogenetic pro-fibrotic macrophage pro-inflammatory cell.

Язык: Английский

Процитировано

0
Quantitative Changes in the Proteome of Chronically Inflamed Lacrimal Glands From a Sjögren's Disease Animal Model DOI Creative Commons

Danny Toribio,

Junji Morokuma,

Dante Pellino

и другие.

Investigative Ophthalmology & Visual Science, Год журнала: 2025, Номер 66(4), С. 44 - 44

Опубликована: Апрель 17, 2025

The lacrimal gland (LG) is the major source of aqueous tears, and insufficient LG secretion leads to aqueous-deficient dry eye (ADDE) disease. To provide a foundational description LG's protein expression patterns, we prepared extracts LGs from wild-type an ADDE mouse model analyzed proteome by quantitative mass spectrometry. were isolated model, male non-obese diabetic (NOD) mice control BALB/c (n = 6 each). Protein samples in urea-based lysis buffer concentrations determined BCA method. equivalent 200 µg tryptically digested nanoflow liquid chromatography tandem spectrometry (LC-MS/MS). Proteins identified quantified using PEAKS X bioinformatics suite. Downstream differential analysis was performed MS-DAP R package. Selected significantly differentially expressed detected proteins subjected spatial immunohistochemistry. Cumulatively, LC-MS/MS-based proteomics analyses murine total 31,932 peptide sequences resulting 2617 identifications at 1% false discovery rate level. Principal component (PCA) hierarchical cluster revealed separation NOD samples. Overall, diversity consistently higher After applying global filter criteria peptide-to-protein rollup, 1750 remaining MSqRob algorithm, which 580 with statistically significant differences. Data are available via ProteomeXchange identifier PXD060937. At cellular level, up- downregulation select confirmed Our data suggest that chronic inflammation alterations proteome. Ongoing studies aim identify potentially unique, inflammation-induced could be amenable pharmacological modulation.

Язык: Английский

Процитировано

0
Single-Nuclei Characterization of Lacrimal Gland in Scopolamine-Induced Dry Eye Disease DOI Creative Commons
Yang Tang, Shengqian Dou, Chao Wei

и другие.

Investigative Ophthalmology & Visual Science, Год журнала: 2024, Номер 65(4), С. 46 - 46

Опубликована: Апрель 30, 2024

The lacrimal gland (LG) is the main organ responsible for tear secretion and an important pathogenic site dry eye disease (DED). This study aimed to comprehensively characterize LG cellular heterogeneity under normal DED conditions using single-nucleus RNA sequencing (snRNA-seq).

Язык: Английский

Процитировано

3
Analysis of the heterogeneity and complexity of murine extraorbital lacrimal gland via single-cell RNA sequencing DOI

Duliurui Huang,

Xinwei Jiao,

Shenzhen Huang

и другие.

The Ocular Surface, Год журнала: 2024, Номер 34, С. 60 - 95

Опубликована: Июнь 28, 2024

Язык: Английский

Процитировано

3
Profiles, Distribution, and Functions of Gamma Delta T Cells in Ocular Surface Homeostasis and Diseases DOI Creative Commons
Zhengze Sun,

Haolan Ji,

Yifan Zhou

и другие.

Frontiers in Bioscience-Landmark, Год журнала: 2024, Номер 29(4), С. 146 - 146

Опубликована: Апрель 11, 2024

The ocular surface microenvironment, containing the cornea, conjunctiva, and lacrimal gland, constitutes mucosal frontline of eye houses a myriad immune cells. As part unconventional T cells, gamma delta (γδ) cells differ in development functions from canonical alpha beta (αβ) They are predominantly situated sites throughout body, including tissues. Recent research has elucidated that γδ serve as primary interleukin-17A (IL-17A) source conjunctiva. play pivotal role preserving homeostasis exhibit both protective pathogenic roles diseases. This review delves into general profiles their distribution tissues, consolidates current insights different conditions dry disease, infectious keratitis, corneal wound healing, anterior chamber-associated deviation, allergic conjunctival diabetic disease. aim is to provide systemic perspective on microenvironment outline potential directions for future studies.

Язык: Английский

Процитировано

1
Crk mediates Csk-Hippo signaling independently of Yap tyrosine phosphorylation to induce cell extrusion DOI

Abdul Hannan,

Qian Wang,

Yihua Wu

и другие.

bioRxiv (Cold Spring Harbor Laboratory), Год журнала: 2024, Номер unknown

Опубликована: Июль 1, 2024

Abstract Src family kinases (SFKs), including Src, Fyn and Yes, play important roles in development cancer. Despite being first discovered as the Y es- a ssociated p rotein, regulation of Yap by SFKs remains poorly understood. Here, through single-cell analysis genetic lineage tracing, we show that pan-epithelial ablation C-terminal kinase (Csk) lacrimal gland unleashes broad signaling but specifically causes extrusion apoptosis acinar progenitors at time when they are shielded myoepithelial cells from basement membrane. Csk mutants can be phenocopied constitutively active rescued deleting or Taz , indicating significant functional overlap between signaling. Although Src-induced tyrosine phosphorylation has long been believed to regulate activity, find mutating these residues both fails perturb mouse alleviate phenotype. In contrast, loses Hippo signaling-dependent serine translocates into nucleus mutants. Further chemical genetics studies demonstrate acute inhibition enhances Crk/CrkL Rac1 whereas removing Crk / CrkL Rap1 ameliorates mutant These results controls Hippo-Yap Crk/CrkL-Rac/Rap axis promote cell extrusion.

Язык: Английский

Процитировано

0