Anti-apoptotic activity of human matrix metalloproteinase-2 attenuates diabetes mellitus

Nishihama, Kota; Yasuma, Taro; Yano, Yutaka; Alessandro-Gabazza, Corina N. D'; Toda, Masaaki; Hinneh, Josephine A; Tonto, Prince Baffour; Takeshita, Atsuro; Totoki, Toshiaki; Mifuji-Moroka, Rumi; Kobayashi, Tetsu; Iwasa, Motoh; Takei, Yoshiyuki; Morser, John; Cann, Isaac; Gabazza, Esteban C.

doi:10.1016/j.metabol.2018.01.016

Cited by 16 publications

(23 citation statements)

References 46 publications

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“…Deregulated PI3K-Akt-IRS1 signalling is a hallmark event in beta cell dysfunction in diabetes [53,54]. PI3K-Akt signalling regulates activity of BAD, a pro-apoptotic protein, during conditions of islet stress [55,56]. We hypothesise that stress situations induce miRNAs that preserve beta cell function/ survival extracellularly in exosomes, in addition to inducing stress response and apoptosis pathways.…”

Section: Discussionmentioning

confidence: 99%

Differential expression and release of exosomal miRNAs by human islets under inflammatory and hypoxic stress

et al. 2019

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Aims/hypothesis Pancreatic islets produce non-coding microRNAs (miRNAs) that regulate islet cell function and survival. Our earlier investigations revealed that human islets undergo significant damage due to various types of stresses following transplantation and release miRNAs. Here, we sought to identify and validate exosomal miRNAs (exo-miRNAs) produced by human islets under conditions of cellular stress, preceding loss of cell function and death. We also aimed to identify islet stress signalling pathways targeted by exo-miRNAs to elucidate potential regulatory roles in islet cell stress. Methods Human islets were subjected to proinflammatory cytokine and hypoxic cell stress and miRNA from exosomes was isolated for RNA sequencing and analysis. Stress-induced exo-miRNAs were evaluated for kinetics of expression and release by intact islets for up to 48 h exposure to cytokines and hypoxia. A subset of stress-induced exo-miRNAs were assessed for recovery and detection as biomarkers of islet cell stress in a diabetic nude mouse xenotransplant model and in patients undergoing total pancreatectomy with islet auto-transplantation (TPIAT). Genes and signalling pathways targeted by stress-induced exo-miRNAs were identified by Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis and direct interactions of miRNAs with downstream signalling targets were validated in human islet cells using the miRNA Tests for Read Analysis and Prediction (MirTrap) system. Results Global exo-miRNA sequencing revealed that 879 miRNA species were released from human islets and 190 islet exo-miRNAs were differentially expressed in response to proinflammatory cytokines, hypoxia or both. Release of exo-miRNAs hsa-miR-29b-3p and hsa-miR-216a-5p was detected within 6 h of exposure to cytokines and hypoxia. The remaining subset of stressinduced exo-miRNAs, including hsa-miR-148a-3p and islet cell damage marker hsa-miR-375, showed delayed release at 24-48 h, correlating with apoptosis and cell death. Stress and damage exo-miRNAs were significantly elevated in the circulation in human-to-mouse xenotransplant models and in human transplant recipients. Elevated blood exo-miRNAs negatively correlated with post-transplant islet function based on comparisons of stress and damage exo-miRNA indices with Secretory Unit of Islet Transplant Objects (SUITO) indices. KEGG analysis and further validation of exo-miRNA targets by MirTrap analysis revealed significant enrichment of islet mRNAs involved in phosphoinositide 3-kinase/Akt and mitogen-activated protein kinase signalling pathways. Conclusions/interpretation The study identifies exo-miRNAs differentially expressed and released by islets in response to damage and stress. These exo-miRNAs could serve as potential biomarkers for assessing islet damage and predicting outcomes in islet transplantation. Notably, exo-miRNAs 29b-3p and 216a-5p could be detected in islets prior to damage-released miRNAs and indicators of cellular apoptosis and death. Thus, these stress-induced exo-miRNAs may have potential diag...

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Section: Discussionmentioning

confidence: 99%

Differential expression and release of exosomal miRNAs by human islets under inflammatory and hypoxic stress

et al. 2019

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“…33,34 Importantly, the active MMP isozyme is highly expressed in PDAC cells 35,36 and serum levels of MMP-2 have prognostic significance in pancreatic cancer patients. 39 The KRAS mutation is a critical determinant in the early stage of pancreatic ductal adenocarcinoma and is able to drive mature pancreatic cells to de-differentiate into duct-like cells and, ultimately, PDAC. 38 Taken together, these findings suggest that MMP-2 may act as a key regulator in the progress of pancreatic tumorigenesis.…”

Section: Discussionmentioning

confidence: 99%

Insulin promotes invasion and migration of KRAS^G12D mutant HPNE cells by upregulating MMP‐2 gelatinolytic activity via ERK‐ and PI3K‐dependent signalling

et al. 2019

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Objectives: Hyperinsulinemia is a risk factor for pancreatic cancer, but the function of insulin in carcinogenesis is unclear, so this study aimed to elucidate the carcinogenic effects of insulin and the synergistic effect with the KRAS mutation in the early stage of pancreatic cancer. Materials and methods:A pair of immortalized human pancreatic duct-derived cells, hTERT-HPNE E6/E7/st (HPNE) and its oncogenic KRAS G12D variant, hTERT-HPNE E6/ E7/KRAS G12D /st (HPNE-mut-KRAS), were used to investigate the effect of insulin. Cell proliferation, migration and invasion were assessed using Cell Counting Kit-8 and transwell assays, respectively. The expression of E-cadherin, N-cadherin, vimentin and matrix metalloproteinases (MMP-2, MMP-7 and MMP-9) was evaluated by Western blotting and/or qRT-PCR. The gelatinase activity of MMP-2 and MMP-9 in conditioned media was detected using gelatin zymography. The phosphorylation status of AKT, GSK3β, p38, JNK and ERK1/2 MAPK was determined by Western blotting. Results: The migration and invasion ability of HPNE cells was increased after the intro-duction of the mutated KRAS gene, together with an increased expression of MMP-2.These effects were further enhanced by the simultaneous administration of insulin.The use of MMP-2 siRNA confirmed that MMP-2 was involved in the regulation of cell invasion. Furthermore, there was a concentration-and time-dependent increase in gelatinase activity after insulin treatment, which could be reversed by an insulin receptor tyrosine kinase inhibitor (HNMPA-(AM) 3 ). In addition, insulin markedly enhanced the phosphorylation of PI3K/AKT, p38, JNK and ERK1/2 MAPK pathways, with wortmannin or LY294002 (a PI3K-specific inhibitor) and PD98059 (a MEK1-specific inhibitor) significantly inhibiting the insulin-induced increase in MMP-2 gelatinolytic activity.

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“…Interestingly, in the in vivo experiment, there was a significantly increased number of eosinophils in hMMP-2 TG mice treated with intra-tracheal eosinophils compared to their WT counterparts. MMP-2 can prolong cell survival by inhibiting apoptosis ( 21 ); therefore, it is conceivable that prolongation of eosinophil survival in an environment with high MMP-2 level explains the difference in the BALF number of eosinophils between MMP-2 TG and WT mice.…”

Section: Discussionmentioning

confidence: 99%

“…Generation and characterization of the human MMP-2 transgenic (hMMP-2 TG) mouse were previously reported ( 21 ). We developed the hMMP2-TG mice originally using inbred C57BL/6 strain, and maintained by crossing homozygotes and/or heterozygotes.…”

Section: Methodsmentioning

confidence: 99%

Role of Matrix Metalloproteinase-2 in Eosinophil-Mediated Airway Remodeling

et al. 2018

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Airway remodeling is responsible for the progressive decline of lung function in bronchial asthma. Matrix metalloproteinase-2 and fibroblast-to-myofibroblast transition are involved in tissue remodeling. Here we evaluated whether eosinophils play a role in fibroblasts-to-myofibroblasts transition and in the expression of matrix metalloproteinase-2. We co-cultured human eosinophils with human fetal lung fibroblast-1 cells, assessed the expression of remodeling-associated molecules by immunoassays and polymerase-chain reaction, and eosinophils-mediated migration of human fetal lung fibroblast-1 cells using a Boyden chamber. To clarify the participation of matrix metalloproteinase-2 in airway remodeling we administered bone marrow-derived eosinophils by intra-tracheal route to transgenic mice overexpressing the human matrix metalloproteinase-2. The expression of α-smooth muscle actin significantly increased in human fetal lung fibroblast-1 cells co-cultured with human eosinophils compared to controls. There was enhanced expression of matrix metalloproteinase-2 during fibroblast-to-myofibroblast transition. An inhibitor of matrix metalloproteinases blocked eosinophils-associated fibroblast-to-myofibroblast transition and increased migration of fibroblasts. The human matrix metalloproteinase-2 transgenic mice receiving adoptive transfer of mouse eosinophils exhibited increased inflammation and advanced airway remodeling compared to wild type mice. This study demonstrated that eosinophils induce fibroblast-to-myofibroblast transition, secretion of matrix metalloproteinase-2, accelerated migration of fibroblasts, and promote matrix metalloproteinase-2-related airway remodeling. These findings provide a novel mechanistic pathway for eosinophil-associated airway remodeling in bronchial asthma.

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Anti-apoptotic activity of human matrix metalloproteinase-2 attenuates diabetes mellitus

Cited by 16 publications

References 46 publications

Differential expression and release of exosomal miRNAs by human islets under inflammatory and hypoxic stress

Differential expression and release of exosomal miRNAs by human islets under inflammatory and hypoxic stress

Insulin promotes invasion and migration of KRAS^G12D mutant HPNE cells by upregulating MMP‐2 gelatinolytic activity via ERK‐ and PI3K‐dependent signalling

Role of Matrix Metalloproteinase-2 in Eosinophil-Mediated Airway Remodeling

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Anti-apoptotic activity of human matrix metalloproteinase-2 attenuates diabetes mellitus

Cited by 16 publications

References 46 publications

Differential expression and release of exosomal miRNAs by human islets under inflammatory and hypoxic stress

Differential expression and release of exosomal miRNAs by human islets under inflammatory and hypoxic stress

Insulin promotes invasion and migration of KRASG12D mutant HPNE cells by upregulating MMP‐2 gelatinolytic activity via ERK‐ and PI3K‐dependent signalling

Role of Matrix Metalloproteinase-2 in Eosinophil-Mediated Airway Remodeling

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Insulin promotes invasion and migration of KRAS^G12D mutant HPNE cells by upregulating MMP‐2 gelatinolytic activity via ERK‐ and PI3K‐dependent signalling