Copine 3 “CPNE3” is a novel regulator for insulin secretion and glucose uptake in pancreatic β-cells

El‐Huneidi, Waseem; Anjum, Shabana; Mohammed, Abdul Khader; Unnikannan, Hema; Saeed, Rania; Bajbouj, Khuloud; Abu‐Gharbieh, Eman; Taneera, Jalal

doi:10.1038/s41598-021-00255-0

Cited by 15 publications

(13 citation statements)

References 43 publications

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“…A fluorescently labeled deoxyglucose analog, 2-[ N -(7-nitrobenz-2-oxa-1,3-diazol-4-yl) amino]- 2-deoxy-glucose (2-NBDG) (Invitrogen #N13195, Waltham, MA, USA), was utilized as a probe for the detection of glucose uptake in cultured cells, as previously described [ 28 ]. Briefly, after 48 h transfection, cells were incubated with the 2-NBDG reagent (1 h).…”

Section: Methodsmentioning

confidence: 99%

“…Cell viability assay was conducted using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) colorimetric assay (Sigma–Aldrich, St. Louis, MO, USA) as previously described [ 28 ]. MTT solution (10 μL) was added to transfected cells 48 h post-transfection and incubated at 37 °C for 3–4 h. The absorbance was read as 570 nm using a microplate reader (crocodile mini work-station; Crocodile Control Software) and the percentage of cell viability was calculated.…”

Section: Methodsmentioning

confidence: 99%

“…After incubation, ROS-Glo detection reagent (100 μL) was added to all wells and cells were incubated for another 20 min (RT). The relative luminescence was then detected using a plate reader [ 28 ].…”

Section: Methodsmentioning

confidence: 99%

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Expression Silencing of Mitogen-Activated Protein Kinase 8 Interacting Protein-1 Conferred Its Role in Pancreatic β-Cell Physiology and Insulin Secretion

et al. 2023

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Mitogen-activated protein kinase 8 interacting protein-1 (MAPK8IP1) gene has been recognized as a susceptibility gene for diabetes. However, its action in the physiology of pancreatic β-cells is not fully understood. Herein, bioinformatics and genetic analyses on the publicly available database were performed to map the expression of the MAPK8IP1 gene in human pancreatic islets and to explore whether this gene contains any genetic variants associated with type 2 diabetes (T2D). Moreover, a series of functional experiments were executed in a rat insulinoma cell line (INS-1 832/13) to investigate the role of the Mapk8ip1 gene in β-cell function. Metabolic engineering using RNA-sequencing (RNA-seq) data confirmed higher expression levels of MAPK8IP1 in human islets compared to other metabolic tissues. Additionally, comparable expression of MAPK8IP1 expression was detected in sorted human endocrine cells. However, β-cells exhibited higher expression of MAPK8IP1 than ductal and PSC cells. Notably, MAPK8IP1 expression was reduced in diabetic islets, and the expression was positively correlated with insulin and the β-cell transcription factor PDX1 and MAFA. Using the TIGER portal, we found that one genetic variant, “rs7115753,” in the proximity of MAPK8IP1, passes the genome-wide significance for the association with T2D. Expression silencing of Mapk8ip1 by small interfering RNA (siRNA) in INS-1 cells reduced insulin secretion, glucose uptake rate, and reactive oxygen species (ROS) production. In contrast, insulin content, cell viability, and apoptosis without cytokines were unaffected. However, silencing of Mapk8ip1 reduced cytokines-induced apoptosis and downregulated the expression of several pancreatic β-cell functional markers including, Ins1, Ins2, Pdx1, MafA, Glut2, Gck, Insr, Vamp2, Syt5, and Cacna1a at mRNA and/or protein levels. Finally, we reported that siRNA silencing of Pdx1 resulted in the downregulation of MAPK8IP1 expression in INS-1 cells. In conclusion, our findings confirmed that MAPK8IP1 is an important component of pancreatic β-cell physiology and insulin secretion.

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

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Expression Silencing of Mitogen-Activated Protein Kinase 8 Interacting Protein-1 Conferred Its Role in Pancreatic β-Cell Physiology and Insulin Secretion

et al. 2023

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“…ROS measurements was performed by the H 2 O 2 assay, as previously described [36]. In brief, 2 × 10 4 of cells/well in a total volume of 80 µL were seeded in a 96-wells plate.…”

Section: Intracellular Reactive Oxygen Species (Ros)mentioning

confidence: 99%

“…Western blot expression analysis was performed as previously described [36]. Briefly, transfected cells were collected in ice-cold PBS.…”

Section: Western Blot Analysismentioning

confidence: 99%

EXOC6 (Exocyst Complex Component 6) Is Associated with the Risk of Type 2 Diabetes and Pancreatic β-Cell Dysfunction

Sulaiman

Hachim

Khalique

et al. 2022

Biology

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EXOC6 and EXOC6B (EXOC6/6B) components of the exocyst complex are involved in the secretory granule docking. Recently, EXOC6/6B were anticipated as a molecular link between dysfunctional pancreatic islets and ciliated lung epithelium, making diabetic patients more prone to severe SARS-CoV-2 complications. However, the exact role of EXOC6/6B in pancreatic β-cell function and risk of T2D is not fully understood. Herein, microarray and RNA-sequencing (RNA-seq) expression data demonstrated the expression of EXOC6/6B in human pancreatic islets. Expression of EXOC6/6B was not affected by diabetes status. Exploration of the using the translational human pancreatic islet genotype tissue-expression resource portal (TIGER) revealed three genetic variants (rs947591, rs2488071 and rs2488073) in the EXOC6 gene that were associated (p < 2.5 × 10−20) with the risk of T2D. Exoc6/6b silencing in rat pancreatic β-cells (INS1-832/13) impaired insulin secretion, insulin content, exocytosis machinery and glucose uptake without cytotoxic effect. A significant decrease in the expression Ins1, Ins1, Pdx1, Glut2 and Vamp2 was observed in Exoc6/6b-silenced cells at the mRNA and protein levels. However, NeuroD1, Gck and InsR were not influenced compared to the negative control. In conclusion, our data propose that EXOC6/6B are crucial regulators for insulin secretion and exocytosis machinery in β-cells. This study identified several genetic variants in EXOC6 associated with the risk of T2D. Therefore, EXOC6/6B could provide a new potential target for therapy development or early biomarkers for T2D.

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YAP1-CPNE3 positive feedback pathway promotes gastric cancer cell progression

Li,

Zhong,

Shi

et al. 2024

Cell. Mol. Life Sci.

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Hippo-Yes-associated protein 1 (YAP1) plays an important role in gastric cancer (GC) progression; however, its regulatory network remains unclear. In this study, we identified Copine III (CPNE3) was identified as a novel direct target gene regulated by the YAP1/TEADs transcription factor complex. The downregulation of CPNE3 inhibited proliferation and invasion, and increased the chemosensitivity of GC cells, whereas the overexpression of CPNE3 had the opposite biological effects. Mechanistically, CPNE3 binds to the YAP1 protein in the cytoplasm, inhibiting YAP1 ubiquitination and degradation mediated by the E3 ubiquitination ligase β-transducin repeat-containing protein (β-TRCP). Thereby activating the transcription of YAP1 downstream target genes, which creates a positive feedback cycle to facilitate GC progression. Immunohistochemical analysis demonstrated significant upregulation of CPNE3 in GC tissues. Survival and Cox regression analyses indicated that high CPNE3 expression was an independent prognostic marker for GC. This study elucidated the pivotal involvement of an aberrantly activated CPNE3/YAP1 positive feedback loop in the malignant progression of GC, thereby uncovering novel prognostic factors and therapeutic targets in GC.

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Copine 3 “CPNE3” is a novel regulator for insulin secretion and glucose uptake in pancreatic β-cells

Cited by 15 publications

References 43 publications

Expression Silencing of Mitogen-Activated Protein Kinase 8 Interacting Protein-1 Conferred Its Role in Pancreatic β-Cell Physiology and Insulin Secretion

Expression Silencing of Mitogen-Activated Protein Kinase 8 Interacting Protein-1 Conferred Its Role in Pancreatic β-Cell Physiology and Insulin Secretion

EXOC6 (Exocyst Complex Component 6) Is Associated with the Risk of Type 2 Diabetes and Pancreatic β-Cell Dysfunction

YAP1-CPNE3 positive feedback pathway promotes gastric cancer cell progression

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