Immunotherapeutic Approaches of Rheumatoid Arthritis and the Implication on Novel Interventions for Refractoriness

Rheumatoid arthritis is an autoimmune disorder involving the chronic inflammation of affected joints which lead to the distortion and eventually destruction of the articular tissues. Clinically, many therapeutic methods are being used for RA treatment. Non-steroidal anti-inflammatory drugs (NSAIDs), steroid, and disease-modifying anti-rheumatic drugs (DMARDs) are the three main categories of intervention approaches. Among which DMARDs, targeting mainly the release of pro-inflammatory cytokines, demonstrated high efficacy because of its direct drug action that alter the underlying disease mechanisms rather than simply to mediate symptoms relieve. However, the use of DMARDs also accompanying some unwanted adverse side effects, in particular, the development of refractoriness, which hampers the successful rate of treatment. In this chapter, the conventional RA drugs will be reviewed, focusing on the currently used and latest development of DMARDs. Novel methods that could improve RA pathogenesis will also be introduced. Because of the critical role of refractory RA, the progress of the disease to develop resistance to standard drug treatment will also be described. Finally, innovative RA therapeutic methods inspired by researches concerning the pathogenesis and contemporary treatments of RA will be discussed

Potential enhancement of post-stroke angiogenic response by targeting the oligomeric aggregation of p53 protein

Tumor suppressor gene p53 and its aggregate have been found to be involved in many angiogenesis-related pathways. We explored the possible p53 aggregation formation mechanisms commonly occur after ischemic stroke, such as hypoxia and the presence of reactive oxygen species (ROS). The angiogenic pathways involving p53 mainly occur in nucleus or cytoplasm, with one exception that occurs in mitochondria. Considering the high mitochondrial density in brain and endothelial cells, we proposed that the cyclophilin D (CypD)-dependent vascular endothelial cell (VECs) necrosis pathway occurring in the mitochondria is one of the major factors that affects angiogenesis. Hence, targeting p53 aggregation, a key intermediate in the pathway, could be an alternative therapeutic target for post-stroke management

PFKFB3-driven vascular smooth muscle cell glycolysis promotes vascular calcification via the altered FoxO3 and lactate production

A link between increased glycolysis and vascular calcification has recently been reported, but it remains unclear how increased glycolysis contributes to vascular calcification. We therefore investigated the role of PFKFB3, a critical enzyme of glycolysis, in vascular calcification. We found that PFKFB3 expression was upregulated in calcified mouse VSMCs and arteries. We showed that expression of miR-26a-5p and miR-26b-5p in calcified mouse arteries was significantly decreased, and a negative correlation between Pfkfb3 mRNA expression and miR-26a-5p or miR-26b-5p was seen in these samples. Overexpression of miR-26a/b-5p significantly inhibited PFKFB3 expression in VSMCs. Intriguingly, pharmacological inhibition of PFKFB3 using PFK15 or knockdown of PFKFB3 ameliorated vascular calcification in vD3 -overloaded mice in vivo or attenuated high phosphate (Pi)-induced VSMC calcification in vitro. Consistently, knockdown of PFKFB3 significantly reduced glycolysis and osteogenic transdifferentiation of VSMCs, whereas overexpression of PFKFB3 in VSMCs induced the opposite effects. RNA-seq analysis and subsequent experiments revealed that silencing of PFKFB3 inhibited FoxO3 expression in VSMCs. Silencing of FoxO3 phenocopied the effects of PFKFB3 depletion on Ocn and Opg expression but not Alpl in VSMCs. Pyruvate or lactate supplementation, the product of glycolysis, reversed the PFKFB3 depletion-mediated effects on ALP activity and OPG protein expression in VSMCs. Our results reveal that blockade of PFKFB3-mediated glycolysis inhibits vascular calcification in vitro and in vivo. Mechanistically, we show that FoxO3 and lactate production are involved in PFKFB3-driven osteogenic transdifferentiation of VSMCs. PFKFB3 may be a promising therapeutic target for the treatment of vascular calcification.</p

Sirt7 protects against vascular calcification via modulation of reactive oxygen species and senescence of vascular smooth muscle cells

Vascular calcification is frequently seen in patients with chronic kidney disease (CKD), and significantly increases cardiovascular mortality and morbidity. Sirt7, a NAD+-dependent histone deacetylases, plays a crucial role in cardiovascular disease. However, the role of Sirt7 in vascular calcification remains largely unknown. Using in vitro and in vivo models of vascular calcification, this study showed that Sirt7 expression was significantly reduced in calcified arteries from mice administered with high dose of vitamin D3 (vD3). We found that knockdown or inhibition of Sirt7 promoted vascular smooth muscle cell (VSMC), aortic ring and vascular calcification in mice, whereas overexpression of Sirt7 had opposite effects. Intriguingly, this protective effect of Sirt7 on vascular calcification is dependent on its deacetylase activity. Unexpectedly, Sirt7 did not alter the osteogenic transition of VSMCs. However, our RNA-seq and subsequent studies demonstrated that knockdown of Sirt7 in VSMCs resulted in increased intracellular reactive oxygen species (ROS) accumulation, and induced an Nrf-2 mediated oxidative stress response. Treatment with the ROS inhibitor N-acetylcysteine (NAC) significantly attenuated the inhibitory effect of Sirt7 on VSMC calcification. Furthermore, we found that knockdown of Sirt7 delayed cell cycle progression and accelerated cellular senescence of VSMCs. Taken together, our results indicate that Sirt7 regulates vascular calcification at least in part through modulation of ROS and cellular senescence of VSMCs. Sirt7 may be a potential therapeutic target for vascular calcification.</p

Neferine induces autophagy-dependent cell death in apoptosis-resistant cancers via ryanodine receptor and Ca 2+ -dependent mechanism

From Springer Nature via Jisc Publications RouterHistory: received 2019-06-28, collection 2019-12, accepted 2019-12-16, registration 2019-12-17, online 2019-12-27, pub-electronic 2019-12-27Publication status: PublishedAbstract: Resistance of cancer cells to chemotherapy is a significant clinical concern and mechanisms regulating cell death in cancer therapy, including apoptosis, autophagy or necrosis, have been extensively investigated over the last decade. Accordingly, the identification of medicinal compounds against chemoresistant cancer cells via new mechanism of action is highly desired. Autophagy is important in inducing cell death or survival in cancer therapy. Recently, novel autophagy activators isolated from natural products were shown to induce autophagic cell death in apoptosis-resistant cancer cells in a calcium-dependent manner. Therefore, enhancement of autophagy may serve as additional therapeutic strategy against these resistant cancers. By computational docking analysis, biochemical assays, and advanced live-cell imaging, we identified that neferine, a natural alkaloid from Nelumbo nucifera, induces autophagy by activating the ryanodine receptor and calcium release. With well-known apoptotic agents, such as staurosporine, taxol, doxorubicin, cisplatin and etoposide, utilized as controls, neferine was shown to induce autophagic cell death in a panel of cancer cells, including apoptosis-defective and -resistant cancer cells or isogenic cancer cells, via calcium mobilization through the activation of ryanodine receptor and Ulk-1-PERK and AMPK-mTOR signaling cascades. Taken together, this study provides insights into the cytotoxic mechanism of neferine-induced autophagy through ryanodine receptor activation in resistant cancers

Modifizierung der Entwicklung der Prionen-Krankheit durch die Abschaltung des Galektin-3 und durch Simvastatin-Therapie

Galectin-3 is highly overexpressed in prion-infected brain tissue. Immunofluorescence double-labelling identified microglia as the major cell type expressing galectin-3. Ablation of galectin-3 did not affect PrPSc- deposition and development of gliosis. However, galectin-3−/−-mice showed prolonged survival times upon intracerebral and peripheral scrapie infections. Moreover, protein levels of the lysosomal activation marker LAMP-2 were markedly reduced in prion-infected galectin-3−/−-mice suggesting a role of galectin-3 in regulation of lysosomal functions. Lower mRNA levels of Beclin-1 and Atg5 in prion-infected wild-type and galectin-3−/−-mice point towards a possible impairment of autophagy. However, LC3-II/LC3-I ratios remained unchanged in uninfected and scrapie-infected WT- or galectin-3-/--mice indicating no general increase or decrease of autophagosome numbers. The results point towards a detrimental role of galectin-3 in prion infections of the CNS and suggest that endo-/lysosomal dysfunction in combination with reduced autophagy may contribute to disease development. The second part of this study addressed the potential of the cholesterol-lowering drug simvastatin for therapeutic intervention in a murine prion disease model. Statins have been reported to inhibit prion replication in cell cultures and to modulate inflammatory reactions. Groups of mice were intracerebrally infected with two doses of scrapie strain 139A. Simvastatin-treatment commenced 100 days postinfection. The treatment did not affect deposition of misfolded prion protein PrPSc. However, expression of marker proteins for glia activation like major histocompatibility complex class II and galectin-3 was found to be affected. Analysis of brain cholesterol synthesis and metabolism revealed a mild reduction in cholesterol precursor levels, whereas levels of cholesterol and cholesterol metabolites were unchanged. Simvastatin-treatment significantly delayed disease progression and prolonged survival times in established prion infection of the CNS (p<0003). Overall, these results suggests that the therapeutic benefit of simvastatin observed in our murine prion model is not due to the cholesterol-lowering effect of this drug. Taken together, the findings from both studies highlight the significant role of prion-induced inflammatory responses in the CNS in disease development and encourage use of immunomodulatory/anti-inflammatory drugs for therapeutic intervention in prion diseases.In Prion-infiziertem Hirngewebe zeigt Galektin-3 eine starke Überexpression. Mit Hilfe von Doppelfärbungen wurden Mikrogliazellen immunhistochemisch als Galektin-3 exprimierender Zelltyp identifiziert. Galektin-3-/--Mäuse zeigten nach intrazerebraler und peripherer Scrapie-Infektion eine verlängerte Überlebenszeit. Die Abwesenheit von Galektin-3 beeinflusste jedoch weder PrPSc-Ablagerungen noch die Entwicklung der Gliose. LAMP-2, ein Biomarker der lysosomalen Aktivität, zeigte hingegen eine deutlich verringerte Proteinexpression in Galektin-3-/--Mäusen, was auf einen möglichen Einfluss von Galektin-3 auf lysosomale Funktionen hinweist. Verminderte mRNA-Level von Beclin-1 und Atg5 in Prion-infizierten Wildtyp- und Galektin-3-/--Mäusen deuten auf eine Beeinträchtigung des Autophagie-Systems hin. Die Untersuchung der LC3-II/LC3-I Mengenverhältnisse ergab jedoch keinen Hinweis auf eine generelle Ab- oder Zunahme an Autosomen. Die Ergebnisse zeigen die schädliche Rolle des Galektin-3 bei Prion-Infektionen des ZNS, wobei eine endo-/lysosomale Dysfunktion in Kombination mit einer reduzierten Autophagie zur Krankheitsentwicklung beiträgt. Der zweite Teil dieser Arbeit beschäftigte sich mit dem therapeutischen Potential des Cholesterin-senkenden Wirkstoffs Simvastatin im murinen Prionmodell. Für Statine ist die Modulation von inflammatorischen Reaktionen und die Inhibition der Prion-Replikation in der Zellkultur beschrieben. Versuchsgruppen von Mäusen wurden mit zwei verschiedenen Infektionsdosen des Scrapie-Stammes 139A intrazerebral infiziert und 100 Tage nach Infektion mit Simvastatin behandelt. Die Behandlung hatte keinen Einfluss auf die Ablagerung des umgefalteten Prion-Proteins PrPSc, jedoch war die Expression der Gliaaktivierungsmarker MHC II (major histocompatibility complex class II) und Galektin-3 verändert. Analysen von Cholesterinsynthese und –metabolismus im Gehirn zeigten leicht reduzierte Konzentrationen von Cholesterinvorläufern aber unveränderte Konzentrationen von Cholesterin und Cholesterinmetaboliten. Die Simvastatinbehandlung führte zu einer Verzögerung des Krankheitsverlaufs und einer signifikant verlängerten Überlebenszeit bei bestehender Prion-Infektion des ZNS (p<0,01). Insgesamt deuten die Ergebnisse darauf hin, dass der therapeutische Effekt von Simvastatin in unserem murinen Prion-Modell wahrscheinlich nicht auf dessen Cholesterin-senkende Wirkung zurückzuführen ist. Zusammengefasst weisen die Ergebnisse beider Teile dieser Arbeit auf die Bedeutung der Prion-induzierten inflammatorischen Reaktion im ZNS in der Pathogenese von Prion-Krankheiten hin und lassen den Einsatz von immunmodulatorischen/anti-inflammatorischen Wirkstoffen zur therapeutischen Intervention als sinnvoll erscheinen

Additional file 1 of Modulation of alveolar macrophage and mitochondrial fitness by medicinal plant-derived nanovesicles to mitigate acute lung injury and viral pneumonia

Supplementary Material

Neferine induces autophagy-dependent cell death in apoptosis-resistant cancers via ryanodine receptor and Ca

From PubMed via Jisc Publications RouterHistory: received 2019-06-28, accepted 2019-12-16Publication status: epublishResistance of cancer cells to chemotherapy is a significant clinical concern and mechanisms regulating cell death in cancer therapy, including apoptosis, autophagy or necrosis, have been extensively investigated over the last decade. Accordingly, the identification of medicinal compounds against chemoresistant cancer cells via new mechanism of action is highly desired. Autophagy is important in inducing cell death or survival in cancer therapy. Recently, novel autophagy activators isolated from natural products were shown to induce autophagic cell death in apoptosis-resistant cancer cells in a calcium-dependent manner. Therefore, enhancement of autophagy may serve as additional therapeutic strategy against these resistant cancers. By computational docking analysis, biochemical assays, and advanced live-cell imaging, we identified that neferine, a natural alkaloid from Nelumbo nucifera, induces autophagy by activating the ryanodine receptor and calcium release. With well-known apoptotic agents, such as staurosporine, taxol, doxorubicin, cisplatin and etoposide, utilized as controls, neferine was shown to induce autophagic cell death in a panel of cancer cells, including apoptosis-defective and -resistant cancer cells or isogenic cancer cells, via calcium mobilization through the activation of ryanodine receptor and Ulk-1-PERK and AMPK-mTOR signaling cascades. Taken together, this study provides insights into the cytotoxic mechanism of neferine-induced autophagy through ryanodine receptor activation in resistant cancers