glioblastoma-relapse-post-resection-model-for-therapeutic-hydrogel

Glioblastoma Relapse Post-Resection Model for Therapeutic Hydrogel Investigations

Tumor recurrence is an important factor indicative of a poor prognosis in glioblastoma (GBM). Many studies are attempting to identify effective ...

Nanjing Medical University

Mesenchymal stem cells (MSCs) may be obtained from umbilical cord as an abundant and noninvasive source. However, the immunomodulatory properties of umbilical cord-MSCs (UC-MSCs) were poorly studied. In this study, we aimed to investigate the effects of UC-MSCs on B-cell proliferation and differentiation. UC-MSCs were found to suppress the proliferation of B cells isolated from murine spleen. Moreover, UC-MSCs markedly suppressed B-cell differentiation as shown by the decreased number of CD138+cells and reduced levels of IgM and IgG production in coculture. As revealed by transwell experiments, soluble factors produced by UC-MSCs might be involved in mediating B-cell suppression. The Blimp-1 mRNA expression was suppressed whereas the PAX-5 mRNA expression was induced in coculture. Finally, UC-MSCs modified the phosphorylation pattern of Akt and p38 pathways, which were involved in B-cell proliferation and differentiation. These results may further support the potential therapeutic use of UC-MSCs in treating autoimmune disorders.

Umbilical cord mesenchymal stem cells suppress B-cell proliferation and differentiation.

Chemokines and chemokine receptors cooperate to promote immune cell recruitment to the central nervous system (CNS). In this study, we investigated the roles of CXCR2 and CXCL1 in leukocyte recruitment to the CNS using a murine model of neuroinflammation.

CXCR2 is essential for cerebral endothelial activation and leukocyte recruitment during neuroinflammation.

Infection in male and female genital tracts can lead to infertility. The underlying mechanisms of this process remain unclear. Toll-like receptors (TLRs) recognize conserved structures and respond to pathogens by initiating signals that activate inflammatory gene transcription. Here, we demonstrate that TLR activation in sperm reduces sperm motility via signalling through myeloid differentiation factor 88 (MyD88), phosphatidylinositol 3-kinase (PI3K), and glycogen synthase kinase (GSK)-3α. Upon TLR activation, phosphorylated forms of PI3K and GSK3α were detected in the mitochondria, and the mitochondrial membrane potential was impaired in sperm. In addition, mitochondrial ATP levels were decreased after TLR agonist stimulation. Furthermore, blocking PI3K or GSK3α activation abrogated these effects and reversed the TLR-induced reduction in sperm motility. These results identify a previously unrecognized TLR signalling pathway that leads to dysfunctional sperm mitochondria, which reduce sperm motility. Our study reveals a novel mechanism by which pathogenic infection affects sperm motility and possibly leads to infertility.

TLR signalling affects sperm mitochondrial function and motility via phosphatidylinositol 3-kinase and glycogen synthase kinase-3α.

The complement system is becoming increasingly recognized as a key participant in many neurodegenerative diseases of the brain. Complement-deficient animals exhibit reduced neuroinflammation.

Complement component C3a plays a critical role in endothelial activation and leukocyte recruitment into the brain.

Bisphenol A (BPA), an environmental endocrine-disrupting organic chemical, has been positively associated with the rate of implantation failure of in vitro fertilization. However, the underlying mechanisms remain unclear. To reveal the impact and the underlying mechanism of BPA on the crosstalk between trophoblast and decidual stromal cells (DSCs), we determined whether BPA was able to affect trophoblast invasion in vitro. We found that BPA significantly inhibited CXCL8 expression in DSCs, which hindered trophoblast invasion, and activated the phosphorylation of ERK in DSCs. U0126, an inhibitor of ERK activation, remarkably rescued trophoblast invasion and the inhibition of CXCL8 expression caused by BPA treatment. Moreover, the nuclear estrogen receptor antagonist ICI 182,780 and transmembrane G protein-coupled receptor GPR30 (membrane estrogen receptor) antagonist G15 significantly blocked the phosphorylation of ERK and reversed the reduction of trophoblast invasion. In brief, BPA activated ERK through nuclear and membrane estrogen receptors and inhibited CXCL8 expression in DSCs, thereby affecting their regulation of trophoblast invasion.

Bisphenol A affects trophoblast invasion by inhibiting CXCL8 expression in decidual stromal cells.

Fingolimod (FTY720), an immunomodulator, is approved as an oral treatment for patients with relapsing forms of multiple sclerosis. Its effects are largely attributed to its mechanism of selectively retaining lymphocytes in the lymph nodes to reduce autoreactive T-cell recruitment in the CNS. In this study, we investigated the therapeutic effect of FTY720 on an animal model of CNS inflammation induced by intracerebral ventricle LPS injection. We found that FTY720 treatment significantly prevented LPS-induced neutrophil recruitment in the CNS by inhibiting leukocyte recruitment in cerebral microvessels. Furthermore, FTY720 also inhibited the expressions of adhesion molecules on the cerebral endothelium, but did not affect the expression levels of pro-inflammatory cytokines (TNF-α and IL-6) and chemokines (CXCL1 and CXCL2) in the CNS parenchyma. The inhibition of endothelial activation was accompanied by reduced phosphorylation of signaling molecules, including serine/threonine-specific protein kinase (Akt), STAT6, and nuclear factor-κB. This FTY720-attenuated inhibition of leukocyte recruitment and endothelial activation was reversed by blocking the functions of sphingosine kinase 2 or sphingosine-1-phosphate receptor 1. Our study demonstrated, for the first time, that FTY720 directly inhibits the phosphorylation of multiple signaling molecules in endothelial cells, thereby effectively blocking leukocyte recruitment in the CNS.

Fingolimod targets cerebral endothelial activation to block leukocyte recruitment in the central nervous system.

Neuropsychiatric (NP) involvement in systemic lupus erythematosus (SLE) severely impacts patients' quality of life and leads to a poor prognosis. The current therapeutic protocol, corticosteroid administration, can also induce neuropsychiatric disorders. FTY720 is an immunomodulator that selectively confines lymphocytes in lymph nodes and reduces autoreactive T cell recruitment to the central nervous system (CNS). This study aimed to identify a novel therapeutic strategy for NPSLE. B6.MRL-lpr mice were treated with oral administration of FTY720 (2 mg/kg) three times per week for 12 weeks, to evaluate its efficacy in a model of NPSLE. FTY720 significantly attenuated the impulsive and depression-like behavior of B6.MRL-lpr mice. Neuronal damage was reduced in the cortex, hippocampus, and amygdala of the FTY720-treated B6.MRL-lpr mice, as well as in TNF-α-treated HT22 cells. Additionally, FTY720 downregulated levels of inflammatory cytokines, and reduced the infiltration of T cells and neutrophils in the brain parenchyma. FTY720 also acted directly on cerebral endothelial cells and reduced the permeability of the blood-brain barrier (BBB) in B6.MRL-lpr mice, as evidenced by reduced central IgG and albumin levels. Finally, FTY720 significantly inhibited activation of PI3K/Akt/GSK3β/p65 signaling, which further reduced the expression levels of adhesion molecules in bEND.3 cells treated with B6.MRL-lpr mouse serum. Collectively, our data indicate that oral administration of FTY720 at an early stage has beneficial effects in NPSLE-model B6.MRL-lpr mice, suggesting that it may represent an effective new therapeutic strategy for NPSLE.

FTY720 attenuates behavioral deficits in a murine model of systemic lupus erythematosus.

Interleukin-33 (IL-33) is increasingly being recognized as a key immunomodulatory cytokine in many neurological diseases.

IL-33/ST2 plays a critical role in endothelial cell activation and microglia-mediated neuroinflammation modulation.

Sphingosine kinases (Sphks) are the rate-limiting enzymes in the conversion of sphingosine to biologically active sphingosine-1-phosphate. The present study aimed to determine the role of Sphk2 and its downstream targets in renal fibroblast activation and interstitial fibrosis. In the kidney interstitium of patients with renal fibrosis, Sphk2-expressing cells (mainly interstitial fibroblasts) were significantly elevated and highly correlated with disease progression in patients. In a murine model of renal interstitial fibrosis, Sphk2 was upregulated in the kidney of wild-type mice in response to disease progression. Importantly, Sphk2-knockout (KO) mice exhibited significantly lower levels of extracellular matrix (ECM) production and a suppressed inflammatory response in the kidney tissues, compared to those in their wild-type counterparts, whereas the expression of TGF-β1 was unaffected. TGF-β1 effectively upregulated Sphk2 expression in the renal interstitial fibroblast line, NRK-49F, independent of canonical Smad signaling activation. Furthermore, siRNA-mediated Sphk2 knockdown or suppression of Sphk2 activity by ABC294640 exposure effectively attenuated AKT and STAT3 activation and ECM production, but had no effects on Smad2 and Smad3 activation. Sphk2 phosphorylated Fyn to activate downstream STAT3 and AKT, thereby promoting ECM synthesis. Therefore, our findings indicate that targeting Sphk2-Fyn-STAT3/AKT signaling pathway may be a novel therapeutic approach for renal fibrosis.

Sphingosine kinase 2 cooperating with Fyn promotes kidney fibroblast activation and fibrosis via STAT3 and AKT.

Parkinson's disease (PD) is characterized by the degeneration of dopaminergic neurons and excessive microglial activation in the substantia nigra pars compacta (SNpc). In the present study, we aimed to demonstrate the therapeutic effectiveness of the potent sphingosine-1-phosphate receptor antagonist fingolimod (FTY720) in an animal model of PD induced by 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP), and to identify the potential mechanisms underlying these therapeutic effects. C57BL/6J mice were orally administered FTY720 before subcutaneous injection of MPTP. Open-field and rotarod tests were performed to determine the therapeutic effect of FTY720. The damage to dopaminergic neurons and the production of monoamine neurotransmitters were assessed using immunohistochemistry, high-performance liquid chromatography, and flow cytometry. Immunofluorescence (CD68- positive) and enzyme-linked immunosorbent assay were used to analyze the activation of microglia, and the levels of activated signaling molecules were measured using Western blotting. Our findings indicated that FTY720 significantly attenuated MPTP-induced behavioral deficits, reduced the loss of dopaminergic neurons, and increased dopamine release. FTY720 directly inhibited MPTP-induced microglial activation in the SNpc, suppressed the production of interleukin (IL)-6, IL-1β, and tumor necrosis factor-α in BV-2 microglial cells treated with 1-methyl-4-phenylpyridinium (MPP), and subsequently decreased apoptosis in SH-SY5Y neuroblastoma cells. Moreover, in MPP-treated BV-2 cells and primary microglia, FTY720 treatment significantly attenuated the increases in the phosphorylation of PI3K/AKT/GSK-3β, reduced ROS generation and p65 activation, and also inhibited the activation of NLRP3 inflammasome and caspase-1. In conclusion, FTY720 may reduce PD progression by inhibiting NLRP3 inflammasome activation via its effects on ROS generation and p65 activation in microglia. These findings provide novel insights into the mechanisms underlying the therapeutic effects of FTY720, suggesting its potential as a novel therapeutic strategy against PD. Graphical Abstract FTY720 may reduce ROS production by inhibiting the PI3K/AKT/GSK-3β signaling pathway, while at the same time reducing p65 phosphorylation, thus decreasing NLRP3 inflammasome activation through these two pathways, ultimately reducing microglia activation-induced neuronal damage.

FTY720 Inhibits MPP-Induced Microglial Activation by Affecting NLRP3 Inflammasome Activation.

Systemic lupus erythematosus (SLE) is a chronic autoimmune disease and hypocomplementemia signifies disease activity. Several studies have shown that calcium may help maintain optimum function of immune system and metabolism in SLE. The aim of our study was to analyze the relationship between total serum calcium level and SLE activity. A total of 66 patients with SLE and 214 healthy controls were included in this study. Our results showed lower serum levels of calcium ( < .001), complement C3 ( < .001), complement C4 ( < .001), and albumin ( < .001) in patients with SLE. A negative correlation was found between serum calcium level and systemic lupus erythematosus disease activity index (SLEDAI) rating ( = -0.394,  = .001). Additionally, serum level of calcium was positively correlated with serum complement C3 level ( = 0.366,  = .003) in patients with SLE, while no such correlation was found between serum calcium level and complement C4 ( = -0.190,  = .126). Likewise, patients with SLE with normal serum calcium level showed higher complement C3 level ( < .01) than that of patients with low serum calcium level. Overall, the results displayed that patients with SLE have lower serum calcium level compared to healthy controls, and the serum calcium level is positively correlated with SLEDAI rating and serum complement C3 level in patients with SLE. In conclusion, the total serum calcium level is negatively correlated with SLE disease activity.

Total Serum Calcium Level Is Negatively Correlated With Systemic Lupus Erythematosus Activity.

Iron is one of the most important trace elements in the body, and its homeostasis is essential to the normal function of the immune system. Complement component C3, which is the converging of three main pathways of complement system activation, plays a key role in the innate immunity. However, the relationship between iron homeostasis and complement C3 remains unknown. The aim of our study was to analyze the relationship between serum iron and ferritin level and complement C3 and C4. A total of 590 healthy individuals were recruited in our study. Higher serum complement C3 level (p < 0.001) was found in individuals with higher serum ferritin level (> 104.0 μg/L). Moreover, serum iron level and serum ferritin level were positively correlated with complement C3 (r = 0.133, p = 0.001; r = 0.221, p < 0.001) and complement C4 (r = 0.117, p = 0.004; r = 0.123, p = 0.003). The linear regression analysis displayed that both serum iron level and serum ferritin level were linearly correlated with serum complement C3 level (adjusted beta: 2.382, 95% CI: 0.841-3.923; adjusted beta: 42.911, 95% CI: 29.070-56.751). To explore the relationship between iron homeostasis and complement C3 further, the serum samples from C3 mice and the wild-type (WT) control mice were obtained. Significantly lower serum iron level and higher ferritin level were found in C3 mice than those in WT mice (p < 0.001; p < 0.001), indicating that complement C3 might influence iron distribution and utilization. Overall, these data suggested that serum iron and ferritin levels were correlated with complement C3. The deficiency of complement C3 may disrupt the regular iron metabolism in the body.

Serum Iron and Ferritin Levels Are Correlated with Complement C3.

Environmental pollutants, such as bisphenol A (BPA) have recently been implicated in the development of adverse birth outcomes. However, the underlying teratogenic mechanisms remain unclear. We investigated the effects of BPA on the migration and invasion of human primary extravillous trophoblast HTR-8/SVneo cells. Our results indicated that BPA reduced cell migration and invasion. Moreover, it altered the ratio of matrix metalloproteinases (MMPs) and tissue inhibitors of MMPs (TIMPs) by downregulating MMP-2 and MMP-9, and upregulating TIMP-1 and TIMP-2. Furthermore, BPA suppressed integrin β1, integrin α5, and vimentin. Interestingly, BPA-induced invasion was partially restored by G15, a membrane G-protein-coupled estrogen receptor 30 antagonist. We further revealed that 42 proteins were differentially expressed by mass spectrometry analysis, which could be divided into three categories based on gene ontology including biological process, cellular component, and molecular function. These results suggest that BPA reduces HTR-8/SVneo cell migration and invasion by downregulating MMP-2 and MMP-9, up-regulating TIMP-1 and TIMP-2, and suppressing adhesion molecules.

Exposure to environmental bisphenol A inhibits HTR-8/SVneo cell migration and invasion.

Immune-checkpoint inhibitors (ICIs) remain ineffective in a large group of non-small cell lung cancer (NSCLC) patients. Mucosal-associated invariant T (MAIT) cells, a population of unconventional innate-like T lymphocytes abundant in the human body, play important roles in human malignancies. Little is known about the immune characteristics of MAIT cells in NSCLC and correlation with prognosis and response rate of ICIs treatment.

Article	Year
Umbilical cord mesenchymal stem cells suppress B-cell proliferation and differentiation. Cellular immunology\| PubMed ID: 22414555	2012
CXCR2 is essential for cerebral endothelial activation and leukocyte recruitment during neuroinflammation. Journal of neuroinflammation\| PubMed ID: 25990934	2015
TLR signalling affects sperm mitochondrial function and motility via phosphatidylinositol 3-kinase and glycogen synthase kinase-3α. Cellular signalling\| PubMed ID: 26658093	2016
Complement component C3a plays a critical role in endothelial activation and leukocyte recruitment into the brain. Journal of neuroinflammation\| PubMed ID: 26822321	2016
Bisphenol A affects trophoblast invasion by inhibiting CXCL8 expression in decidual stromal cells. Molecular and cellular endocrinology\| PubMed ID: 28736254	2018
Fingolimod targets cerebral endothelial activation to block leukocyte recruitment in the central nervous system. Journal of leukocyte biology\| PubMed ID: 29345065	2018
FTY720 attenuates behavioral deficits in a murine model of systemic lupus erythematosus. Brain, behavior, and immunity\| PubMed ID: 29548997	2018
IL-33/ST2 plays a critical role in endothelial cell activation and microglia-mediated neuroinflammation modulation. Journal of neuroinflammation\| PubMed ID: 29728120	2018
Sphingosine kinase 2 cooperating with Fyn promotes kidney fibroblast activation and fibrosis via STAT3 and AKT. Biochimica et biophysica acta. Molecular basis of disease\| PubMed ID: 30251698	2018
FTY720 Inhibits MPP-Induced Microglial Activation by Affecting NLRP3 Inflammasome Activation. Journal of neuroimmune pharmacology : the official journal of the Society on NeuroImmune Pharmacology\| PubMed ID: 31069623	2019
Total Serum Calcium Level Is Negatively Correlated With Systemic Lupus Erythematosus Activity. Dose-response : a publication of International Hormesis Society\| PubMed ID: 32435164	2020
Serum Iron and Ferritin Levels Are Correlated with Complement C3. Biological trace element research\| PubMed ID: 32935207	2021
Exposure to environmental bisphenol A inhibits HTR-8/SVneo cell migration and invasion. Journal of biomedical research\| PubMed ID: 32981897	2020
Clinicopathological and predictive value of MAIT cells in non-small cell lung cancer for immunotherapy. Journal for immunotherapy of cancer\| PubMed ID: 36657812	2023

Dongyan Shi

Biography

JoVE Journal Publications

Other Publications