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1.
Cell Death Dis ; 13(8): 692, 2022 Aug 08.
Artigo em Inglês | MEDLINE | ID: mdl-35941108

RESUMO

Metastatic malignant melanoma is the deadliest skin cancer, and it is characterised by its high resistance to apoptosis. The main melanoma driving mutations are part of ERK pathway, with BRAF mutations being the most frequent ones, followed by NRAS, NF1 and MEK mutations. Increasing evidence shows that the MST2/Hippo pathway is also deregulated in melanoma. While mutations are rare, MST2/Hippo pathway core proteins expression levels are often dysregulated in melanoma. The expression of the tumour suppressor RASSF1A, a bona fide activator of the MST2 pathway, is silenced by promoter methylation in over half of melanomas and correlates with poor prognosis. Here, using mass spectrometry-based interaction proteomics we identified the Second Mitochondria-derived Activator of Caspases (SMAC) as a novel LATS1 interactor. We show that RASSF1A-dependent activation of the MST2 pathway promotes LATS1-SMAC interaction and negatively regulates the antiapoptotic signal mediated by the members of the IAP family. Moreover, proteomic experiments identified a common cluster of apoptotic regulators that bind to SMAC and LATS1. Mechanistic analysis shows that the LATS1-SMAC complex promotes XIAP ubiquitination and its subsequent degradation which ultimately results in apoptosis. Importantly, we show that the oncogenic BRAFV600E mutant prevents the proapoptotic signal mediated by the LATS1-SMAC complex while treatment of melanoma cell lines with BRAF inhibitors promotes the formation of this complex, indicating that inhibition of the LATS1-SMAC might be necessary for BRAFV600E-driven melanoma. Finally, we show that LATS1-SMAC interaction is regulated by the SMAC mimetic Birinapant, which requires C-IAP1 inhibition and the degradation of XIAP, suggesting that the MST2 pathway is part of the mechanism of action of Birinapant. Overall, the current work shows that SMAC-dependent apoptosis is regulated by the LATS1 tumour suppressor and supports the idea that LATS1 is a signalling hub that regulates the crosstalk between the MST2 pathway, the apoptotic network and the ERK pathway.


Assuntos
Caspases , Melanoma , Apoptose , Caspases/metabolismo , Via de Sinalização Hippo , Humanos , Melanoma/genética , Melanoma/metabolismo , Mitocôndrias/metabolismo , Proteínas Serina-Treonina Quinases/genética , Proteômica , Proteínas Proto-Oncogênicas B-raf/metabolismo
2.
Cell Death Dis ; 13(8): 693, 2022 Aug 08.
Artigo em Inglês | MEDLINE | ID: mdl-35941120

RESUMO

Renal fibrosis is a common consequence of various progressive nephropathies, including obstructive nephropathy, and ultimately leads to kidney failure. Infiltration of inflammatory cells is a prominent feature of renal injury after draining blockages from the kidney, and correlates closely with the development of renal fibrosis. However, the underlying molecular mechanism behind the promotion of renal fibrosis by inflammatory cells remains unclear. Herein, we showed that unilateral ureteral obstruction (UUO) induced Gasdermin D (GSDMD) activation in neutrophils, abundant neutrophil extracellular traps (NETs) formation and macrophage-to-myofibroblast transition (MMT) characterized by α-smooth muscle actin (α-SMA) expression in macrophages. Gsdmd deletion significantly reduced infiltration of inflammatory cells in the kidneys and inhibited NETs formation, MMT and thereby renal fibrosis. Chimera studies confirmed that Gsdmd deletion in bone marrow-derived cells, instead of renal parenchymal cells, provided protection against renal fibrosis. Further, specific deletion of Gsdmd in neutrophils instead of macrophages protected the kidney from undergoing fibrosis after UUO. Single-cell RNA sequencing identified robust crosstalk between neutrophils and macrophages. In vitro, GSDMD-dependent NETs triggered p65 translocation to the nucleus, which boosted the production of inflammatory cytokines and α-SMA expression in macrophages by activating TGF-ß1/Smad pathway. In addition, we demonstrated that caspase-11, that could cleave GSDMD, was required for NETs formation and renal fibrosis after UUO. Collectively, our findings demonstrate that caspase-11/GSDMD-dependent NETs promote renal fibrosis by facilitating inflammation and MMT, therefore highlighting the role and mechanisms of NETs in renal fibrosis.


Assuntos
Armadilhas Extracelulares , Nefropatias , Obstrução Ureteral , Caspases/metabolismo , Armadilhas Extracelulares/metabolismo , Fibrose , Humanos , Peptídeos e Proteínas de Sinalização Intracelular/genética , Peptídeos e Proteínas de Sinalização Intracelular/metabolismo , Rim/patologia , Nefropatias/patologia , Macrófagos/metabolismo , Miofibroblastos/metabolismo , Proteínas de Ligação a Fosfato/metabolismo , Proteínas Citotóxicas Formadoras de Poros , Transdução de Sinais , Obstrução Ureteral/genética
3.
Commun Biol ; 5(1): 797, 2022 Aug 08.
Artigo em Inglês | MEDLINE | ID: mdl-35941180

RESUMO

While major changes in cellular morphology during apoptosis have been well described, the subcellular changes in nuclear architecture involved in this process remain poorly understood. Imaging of nucleosomes in cortical neurons in vitro before and during apoptosis revealed that chromatin compaction precedes the activation of caspase-3 and nucleus shrinkage. While this early chromatin compaction remained unaffected by pharmacological blockade of the final execution of apoptosis through caspase-3 inhibition, interfering with the chromatin dynamics by modulation of actomyosin activity prevented apoptosis, but resulted in necrotic-like cell death instead. With super-resolution imaging at different phases of apoptosis in vitro and in vivo, we demonstrate that chromatin compaction occurs progressively and can be classified into five stages. In conclusion, we show that compaction of chromatin in the neuronal nucleus precedes apoptosis execution. These early changes in chromatin structure critically affect apoptotic cell death and are not part of the final execution of the apoptotic process in developing cortical neurons.


Assuntos
Caspases , Cromatina , Apoptose/fisiologia , Caspase 3 , Caspases/metabolismo , Neurônios/fisiologia
4.
Oncoimmunology ; 11(1): 2104551, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-35936984

RESUMO

Our previous study showed that one of the schweinfurthin compounds, 5'-methoxyschweinfurthin G (MeSG), not only enhances the anti-tumor effect of anti-PD1 antibody in the B16F10 murine melanoma model, but also provokes durable, protective anti-tumor immunity. Here we further investigated the mechanisms by which MeSG treatment induces immunogenic cell death (ICD). MeSG induced significant cell surface calreticulin (CRT) exposure in a time and concentration dependent manner as well as increased phagocytosis of tumor cells by dendritic cells in vitro. Interestingly, this CRT exposure differs from the canonical pathway in several aspects. MeSG does not cause ER stress and does not require PERK to induce CRT exposure. Caspase inhibitors partially rescue cells from MeSG-induced apoptosis, but fail to reduce CRT exposure. MeSG does not cause ERp57 exposure and the absence of ERp57 expression does not reduce CRT exposure. Finally, an intact ER to Golgi transport system is required for this phenomenon. These results lend support to the development of the schweinfurthin family as drugs to enhance clinical response to immunotherapy and highlight the need for additional research on the mechanisms of ICD induction.


Assuntos
Morte Celular Imunogênica , Neoplasias , Animais , Apoptose , Caspases/farmacologia , Morte Celular , Camundongos
5.
Front Immunol ; 13: 898819, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-35928825

RESUMO

Molecular mechanisms underlying auto-antibody-induced acantholysis in pemphigus vulgaris are subject of current research to date. To decipher the discrepancy between ubiquitous antibody binding to the epidermal desmosomes, but discontinuous disease manifestation, we were able to identify Ultraviolet A (UVA) as a cofactor for acantholysis. UVA induces interleukin (IL)-1 secretion in keratinocytes, mirroring innate immune system activation. In an in vitro keratinocyte dissociation assay increased fragmentation was observed when UVA was added to anti-Desmoglein 3 Immunoglobulins (anti-Dsg3 IgG). These results were confirmed in skin explants where UVA enhanced anti-Dsg3-mediated loss of epidermal adhesion. The UVA-mediated effect was blocked in vitro by the pan-caspase-inhibitor zVAD-fmk. Thus, we introduce UVA as a caspase-dependent exogenous cofactor for acantholysis which suggests that local innate immune responses largely contribute to overt clinical blister formation upon autoantibody binding to epidermal cells in pemphigus vulgaris.


Assuntos
Pênfigo , Acantólise/metabolismo , Caspases , Humanos , Imunidade Inata , Imunoglobulina G
6.
J Vis Exp ; (185)2022 Jul 21.
Artigo em Inglês | MEDLINE | ID: mdl-35938816

RESUMO

Caspases, a family of cysteine proteases, orchestrate programmed cell death in response to various stimuli, including microbial infections. Initially described to occur by apoptosis, programmed cell death is now known to encompass three interconnected pathways: pyroptosis, apoptosis, and necroptosis, together coined as one process, PANoptosis. Influence A virus (IAV) infection induces PANoptosis in mammalian cells by inducing the activation of different caspases, which, in turn, cleave various host as well as viral proteins, leading to processes like the activation of the host innate antiviral response or the degradation of antagonistic host proteins. In this regard, caspase 3-mediated cleavage of host cortactin, histone deacetylase 4 (HDAC4), and histone deacetylase 6 (HDAC6) has been discovered in both animal and human epithelial cells in response to the IAV infection. To demonstrate this, inhibitors, RNA interference, and site-directed mutagenesis were employed, and, subsequently, the cleavage or resistance to cleavage and the recovery of cortactin, HDAC4, and HDAC6 polypeptides were measured by western blotting. These methods, in conjunction with RT-qPCR, form a simple yet effective strategy to identify the host as well as viral proteins undergoing caspase-mediated cleavage during an infection of IAV or other human and animal viruses. The present protocol elaborates the representative results of this strategy, and the ways to make it more effective are also discussed.


Assuntos
Vírus da Influenza A , Influenza Humana , Animais , Caspases/genética , Caspases/metabolismo , Cortactina/metabolismo , Interações Hospedeiro-Patógeno , Humanos , Vírus da Influenza A/fisiologia , Mamíferos/metabolismo , Proteínas Virais/genética , Proteínas Virais/metabolismo
7.
J Vis Exp ; (185)2022 Jul 25.
Artigo em Inglês | MEDLINE | ID: mdl-35938825

RESUMO

The family of caspases is known to mediate many cellular pathways beyond cell death, including cell differentiation, axonal pathfinding, and proliferation. Since the identification of the family of cell death proteases, there has been a search for tools to identify and expand the function of specific family members in development, health, and disease states. However, many of the currently commercially available caspase tools that are widely used are not specific for the targeted caspase. In this report, we delineate the approach we have used to identify, validate, and target caspase-9 in the nervous system using a novel inhibitor and genetic approaches with immunohistochemical read-outs. Specifically, we used the retinal neuronal tissue as a model to identify and validate the presence and function of caspases. This approach enables the interrogation of cell-type specific apoptotic and non-apoptotic caspase-9 functions and can be applied to other complex tissues and caspases of interest. Understanding the functions of caspases can help to expand current knowledge in cell biology, and can also be advantageous to identify potential therapeutic targets due to their involvement in disease.


Assuntos
Caspases , Retina , Apoptose , Caspase 3/metabolismo , Caspase 9/metabolismo , Caspases/metabolismo , Diferenciação Celular , Sistema Nervoso , Retina/metabolismo
8.
BMC Neurosci ; 23(1): 50, 2022 08 09.
Artigo em Inglês | MEDLINE | ID: mdl-35945502

RESUMO

BACKGROUND: Evidences indicate that inflammasome compounds participate in amyotrophic lateral sclerosis (ALS), a fatal progressive motoneuron degenerative disease. Researchers have observed the expressions of nucleotide oligomerization domain (NOD)-like receptor protein 3 (NLRP3) related inflammasome components in specific regions of the central nervous system in different ALS models, but the cellular spatiotemporal evolution of this canonical inflammasome pathway and pyroptosis during ALS progression are unclear. METHODS: The spinal cords of hSOD1G93A mice (ALS mice) and age-matched littermates (CON mice) were dissected at pre-symptomatic stage (60 d), early- symptomatic stage (95 d), symptomatic stage (108 d) and late-symptomatic stage (122 d) of the disease. By using Nissl staining, double immunofluorescence labelling, qRT-PCR or western blot, we detected morphology change and the expression, cellular location of GSDMD, NLRP3, caspase-1 and IL-1ß in the ventral horn of lumbar spinal cords over the course of disease. RESULTS: Neural morphology changes and GSDMD+/NeuN+ double positive cells were observed in ventral horn from ALS mice even at 60 d of age, even though there were no changes of GSDMD mRNA and protein expressions at this stage compared with CON mice. With disease progression, compared with age-matched CON mice, increased expressions of GSDMD, NLRP3, activated caspase-1 and IL-1ß were detected. Double immunofluorescence labeling revealed that NLRP3, caspase-1, IL-1ß positive signals mainly localized in ventral horn neurons at pre- and early-symptomatic stages. From symptomatic stage to late-symptomatic stage, robust positive signals were co-expressed in reactive astrocytes and microglia. CONCLUSIONS: Early activation of the canonical NLRP3 inflammasome induced pyroptosis in ventral horn neurons, which may participate in motor neuron degeneration and initiate neuroinflammatory processes during ALS progression.


Assuntos
Esclerose Amiotrófica Lateral , Inflamassomos , Esclerose Amiotrófica Lateral/genética , Animais , Caspases , Modelos Animais de Doenças , Inflamassomos/metabolismo , Camundongos , Proteína 3 que Contém Domínio de Pirina da Família NLR/metabolismo , Piroptose , Superóxido Dismutase , Superóxido Dismutase-1/genética
10.
Front Immunol ; 13: 951984, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-35911774

RESUMO

Senecavirus A (SVA), also known as Seneca Valley virus, is a recently discovered picornavirus that can cause swine vesicular disease, posing a great threat to the global swine industry. It can replicate efficiently in cells, but the molecular mechanism remains poorly understood. This study determined the host's differentially expressed proteins (DEPs) during SVA infection using dimethyl labeling based on quantitative proteomics. Among the DE proteins, DDX21, a member of the DEAD (Asp-Glu-Ala-Asp)-box RNA helicase (DDX) family, was downregulated and demonstrated inhibiting SVA replication by overexpression and knockdown experiment. To antagonize this antiviral effect of DDX21, SVA infection induces the degradation of DDX21 by 2B and 3C proteins. The Co-IP results showed that 2B and 3C did not interact with DDX21, suggesting that the degradation of DDX21 did not depend on their interaction. Moreover, the 3C protein protease activity was necessary for the degradation of DDX21. Furthermore, our study revealed that the degradation of DDX21 by 2B and 3C proteins of SVA was achieved through the caspase pathway. These findings suggest that DDX21 was an effective antiviral factor for suppressing SVA infection and that SVA antagonized its antiviral effect by degrading DDX21, which will be useful to guide further studies into the mechanism of mutual regulation between SVA and the host.


Assuntos
Antivirais , Picornaviridae , Animais , Antivirais/farmacologia , Caspases , Picornaviridae/genética , Suínos , Proteínas Virais/metabolismo
11.
Biomolecules ; 12(7)2022 07 21.
Artigo em Inglês | MEDLINE | ID: mdl-35883566

RESUMO

Pyroptosis is a process of programmed cell death mediated by gasdermin (GSDM) found in recent years. In the process of pyroptosis, caspase-1 or caspase-11/4/5 is activated, which cleaves gasdermin D and separates its N-terminal pore-forming domain (PFD). The oligomers of PFD bind to the cell membrane and form macropores on the membrane, resulting in cell swelling and membrane rupture. Increasing evidence indicates that pyroptosis is involved in many diseases, including ischemia reperfusion injury. Autophagy is a highly conserved catabolic process in eukaryotic cells. It plays an important role in the survival and maintenance of cells by degrading organelles, proteins, and macromolecules in the cytoplasm and recycling degradation products. Increasing evidence shows that dysfunctional autophagy participates in many diseases. Recently, autophagy and pyroptosis have been reported to play a vital role in the process of ischemia/reperfusion injury, but the related mechanisms are not completely clear. Therefore, this article reviews the role of autophagy and pyroptosis in ischemia-reperfusion injury and analyzes the related mechanisms to provide a basis for future research.


Assuntos
Piroptose , Traumatismo por Reperfusão , Autofagia , Caspase 1/metabolismo , Caspases/metabolismo , Humanos , Piroptose/fisiologia , Traumatismo por Reperfusão/metabolismo
12.
Proc Natl Acad Sci U S A ; 119(29): e2200206119, 2022 Jul 19.
Artigo em Inglês | MEDLINE | ID: mdl-35858339

RESUMO

Human papillomaviruses (HPVs) infect the basal proliferating cells of the stratified epithelium, but the productive phase of the life cycle (consisting of viral genome amplification, late gene expression, and virion assembly) is restricted to the highly differentiated suprabasal cells. While much is known regarding the mechanisms that HPVs use to block activation of an innate immune response in undifferentiated cells, little is known concerning how HPV prevents an interferon (IFN) response upon differentiation. Here, we demonstrate that high-risk HPVs hijack a natural function of apoptotic caspases to suppress an IFN response in differentiating epithelial cells. We show that caspase inhibition results in the secretion of type I and type III IFNs that can act in a paracrine manner to induce expression of interferon-stimulated genes (ISGs) and block productive replication of HPV31. Importantly, we demonstrate that the expression of IFNs is triggered by the melanoma differentiation-associated gene 5 (MDA5)-mitochondrial antiviral-signaling protein (MAVS)-TBK1 (TANK-binding kinase 1) pathway, signifying a response to double-stranded RNA (dsRNA). Additionally, we identify a role for MDA5 and MAVS in restricting productive viral replication during the normal HPV life cycle. This study identifies a mechanism by which HPV reprograms the cellular environment of differentiating cells through caspase activation, co-opting a nondeath function of proteins normally involved in apoptosis to block antiviral signaling and promote viral replication.


Assuntos
Caspases , Papillomavirus Humano 31 , Helicase IFIH1 Induzida por Interferon , Interferons , Infecções por Papillomavirus , Replicação Viral , Caspases/metabolismo , Papillomavirus Humano 31/fisiologia , Humanos , Helicase IFIH1 Induzida por Interferon/metabolismo , Interferons/metabolismo , Infecções por Papillomavirus/imunologia , Infecções por Papillomavirus/virologia
13.
Zhen Ci Yan Jiu ; 47(7): 565-72, 2022 Jul 25.
Artigo em Chinês | MEDLINE | ID: mdl-35880271

RESUMO

OBJECTIVE: To observe the effect of electroacupuncture (EA) on the expressions of tight junction related proteins Claudin-5, ZO-1 in the colon and hippocampus, Toll-like receptor 4/nuclear factor-kappa B/NOD-like receptor protein 3 (TLR4/NF-κB/NLRP3) pathway in the hippocampus of APP/PS1 mice, so as to explore its mechanisms underlying improvement of cognitive impairment. METHODS: Eighteen 5-month-old male APP/PS1 mice were equally randomized into model and EA groups,and nine 5-month-old male C57BL/6 mice were used as the normal control. EA(2 Hz, 1 mA) was applied to "Baihui" (GV20), "Dachangshu" (BL25) and "Zusanli" (ST36) for 15 min, once daily, 5 days a week for 5 weeks. The Morris water maze swimming test was used to evaluate the mice's cognitive impairment. Nissl staining was used to observe the pathological morphology of hippocampus. The expression of amyloid ß-peptide (Aß) in brain tissue was detect by immunohistochemistry; the contents of lipopolysaccharide (LPS) in colon, serum and hippocampus were detected by ELISA; the expression levels of Claudin-5, ZO-1 in colon and hippocampus, and TLR4/NF-κB/NLRP3 pathway related proteins in hippocampus were detected by Western blot. RESULTS: Compared with the normal group, the escape latency of the mice in the model group was prolonged from the 3rd day (P<0.05, P<0.01), the number of crossing the platform and the percentage of target quadrant residence time were significantly decreased (P<0.01), and the contents of LPS in colon, serum and hippocampus were significantly increased (P<0.01), the expression levels of TLR4, NF-κB p65, NLRP3, Caspase-1, interleukin (IL)-1ß and tumor necrosis factor (TNF)-α in hippocampus and Aß in brain tissue were significantly increased (P<0.01), while the expression levels of Claudin-5, ZO-1 in colon and hippocampus were significantly decreased (P<0.01). Compared with the model group, the escape latency of mice in the EA group was shortened from the 4th day (P<0.05, P<0.01), the number of crossing the platform and the percentage of target quadrant residence time were increased (P<0.01, P<0.05), and the contents of LPS in serum and hippocampus were decreased (P<0.05), and the expression levels of TLR4, NF-κB p65, Caspase-1, NLRP3, IL-1ß, TNF-α in hippocampus and Aß in brain tissue were significantly decreased (P<0.05, P<0.01), while the expression levels of Claudin-5, ZO-1 in colon and hippocampus were significantly increased (P<0.05, P<0.01). Outcomes of Nissl staining showed dispersed arrangement of neurons with nuclear pyknosis or hyperchromasia in the hippocampus, and a decreased number of cell layers in the model group, which was relatively milder in the EA group. CONCLUSION: EA may improve the cognitive impairment of APP/PS1 mice by up-regulating the expression of Claudin-5 and ZO-1, reducing the transposition of gut-derived LPS to the central nervous system, inhibiting the over-activation of TLR4/NF-κB/NLRP3 pathway, and alleviating the inflammatory reaction of the central nervous system.


Assuntos
Doença de Alzheimer , Disfunção Cognitiva , Eletroacupuntura , Doença de Alzheimer/genética , Doença de Alzheimer/terapia , Peptídeos beta-Amiloides , Animais , Caspases , Claudina-5 , Disfunção Cognitiva/genética , Disfunção Cognitiva/terapia , Hipocampo , Lipopolissacarídeos , Masculino , Camundongos , Camundongos Endogâmicos C57BL , NF-kappa B/genética , Proteína 3 que Contém Domínio de Pirina da Família NLR/genética , Receptor 4 Toll-Like/genética
14.
Oxid Med Cell Longev ; 2022: 3345637, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-35847583

RESUMO

Caspases are an evolutionarily conserved family of proteases responsible for mediating and initiating cell death signals. In the past, the dysregulated activation of caspases was reported to play diverse but equally essential roles in neurodegenerative diseases, such as brain injury and neuroinflammatory diseases. A subarachnoid hemorrhage (SAH) is a traumatic event that is either immediately lethal or induces a high risk of stroke and neurological deficits. Currently, the prognosis of SAH after treatment is not ideal. Early brain injury (EBI) is considered one of the main factors contributing to the poor prognosis of SAH. The mechanisms of EBI are complex and associated with oxidative stress, neuroinflammation, blood-brain barrier disruption, and cell death. Based on mounting evidence, caspases are involved in neuronal apoptosis or death, endothelial cell apoptosis, and increased inflammatory cytokine-induced by apoptosis, pyroptosis, and necroptosis in the initial stages after SAH. Caspases can simultaneously mediate multiple death modes and regulate each other. Caspase inhibitors (including XIAP, VX-765, and Z-VAD-FMK) play an essential role in ameliorating EBI after SAH. In this review, we explore the related pathways mediated by caspases and their reciprocal regulation patterns after SAH. Furthermore, we focus on the extensive crosstalk of caspases as a potential area of research on therapeutic strategies for treating EBI after SAH.


Assuntos
Lesões Encefálicas , Hemorragia Subaracnóidea , Animais , Apoptose , Lesões Encefálicas/tratamento farmacológico , Caspases/metabolismo , Humanos , Estresse Oxidativo , Ratos , Ratos Sprague-Dawley , Hemorragia Subaracnóidea/metabolismo
15.
J Eur Acad Dermatol Venereol ; 36(8): 1157-1161, 2022 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-35848143

RESUMO

Pustules are among the most common lesions produced in human skin. Infections by pathogens and drug-induced reactions are frequent causes of pustule formation. In recent years immune-mediated pustular diseases have drawn attention. It is proposed to classify pustular diseases according to the initiating events and sites: purely epidermal pustules, follicular pustules or pustules noted in autoinflammatory syndromes. The unifying pathology in all of the three categories is a microinvasion of activated neutrophils into epidermal or adnexal epithelia. Formation of pustules involves established IL-17 / IL-23, IL-36 / IL-36RN driven pathology, or IL-1 /caspase-activated autoinflammation. Pathophysiology demonstrates an intriguing synergy of keratinocytes with neutrophils. This is called keratinocyte-myeloid synergy (KMS). Non-infectious pustules are formed by IFNα controlling the production of chemoattractants (IL-8, LTB4) or induced by IL-1-regulated inflammasomes and caspase/ IFNß-induced chemotaxins. The presence of physical barriers, for example, cornified cell layers (str. corneum), is instrumental in establishing chemotactic gradients and blocking migrating neutrophils. In follicular KMS-driven pustular disorders, in contrast to epidermal pustules, neutrophil-mediated toxicity propagates lasting and expanding ulcerating diseases with increased levels of circulating immunoglobulin A (IgA). Complexed IgA is suggested to propagate ongoing pustular diseases. These are prerequisites essential for developing pustules in burdensome human skin diseases.


Assuntos
Exantema , Psoríase , Caspases , Humanos , Imunoglobulina A , Interleucina-1 , Queratinócitos/patologia , Neutrófilos/patologia , Psoríase/patologia
17.
ACS Appl Bio Mater ; 5(7): 3241-3256, 2022 Jul 18.
Artigo em Inglês | MEDLINE | ID: mdl-35786838

RESUMO

The organometallic compounds are prospective candidates in the row of developing metallochemotherapeutics with the aim of overcoming the limitations of platinum drugs. In order to explore the anticancer properties of organometallic compounds with the natural medicines, two Ru(II)-p-cymene complexes containing the natural products, viz., 6-gingerol (6G) and benzylated-6-gingerdione (B-6GD) have been synthesized and characterized well. The phenolic group of the Ru(6G) complex facilitates its higher cell-free antioxidant activity than its analogue complex. Also, the same complex shows higher cytotoxicity toward A549 lung and HeLa-S3 cervical cancer cells than the Ru(B-6GD) complex but lower cytotoxicity toward A2058 metastatic melanoma cancer cells. Both complexes are shown to easily accumulate in melanoma cancer cells, and their degree of cytotoxicity in the same cells is found to be positively correlated with cell uptake. The cytotoxicity of complexes arises from their intracellular activity, mainly due to the induction of singlet oxygen production in cancer cells. The subcellular fractionation study shows that mitochondria and ER-Golgi membranes might be their predominant targets. Also, the mechanistic investigation revealed that Ru(B-6GD) induces caspase-dependent non-apoptotic cell death whereas Ru(6G) can induce caspase-independent non-apoptotic cell death. Furthermore, both complexes are found to moderately alter the adhesion properties of cancer cells, which is beneficial for antimetastatic treatment. Despite the potential pharmacological activity, Ru(6G) is encapsulated into polymer-supported liposomes to reduce its toxicity and further improve its anticancer potency. The π-conjugated yne-ene chain of polydiacetylene aids in the development of a stable nanoformulation, which achieved a slow release of the complex. Most importantly, the cancer cell uptake of the liposome-encapsulated Ru(6G) complex is 20 times enhanced and the total ROS formation in cancer cells is significantly increased compared to the non-encapsulated complex. However, the nanoformulation does not alter the antimetastatic potency of the encapsulated complex.


Assuntos
Antineoplásicos , Produtos Biológicos , Gengibre , Melanoma , Compostos Organometálicos , Rutênio , Antineoplásicos/farmacologia , Produtos Biológicos/farmacologia , Caspases/metabolismo , Linhagem Celular Tumoral , Proliferação de Células , Cimenos , Gengibre/metabolismo , Humanos , Lipossomos/farmacologia , Estrutura Molecular , Compostos Organometálicos/farmacologia , Estudos Prospectivos , Rutênio/farmacologia
18.
Int J Mol Sci ; 23(13)2022 Jun 23.
Artigo em Inglês | MEDLINE | ID: mdl-35805980

RESUMO

Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) is a cytotoxic cytokine that induces cancer cell death by binding to TRAIL receptors. Because of its selective cytotoxicity toward cancer cells, TRAIL therapeutics, such as recombinant TRAIL and agonistic antibodies targeting TRAIL receptors, have garnered attention as promising cancer treatment agents. However, many cancer cells acquire resistance to TRAIL-induced cell death. To overcome this issue, we searched for agents to sensitize cancer cells to TRAIL-induced cell death by screening a small-molecule chemical library consisting of diverse compounds. We identified a cardiac glycoside, proscillaridin A, as the most effective TRAIL sensitizer in colon cancer cells. Proscillaridin A synergistically enhanced TRAIL-induced cell death in TRAIL-sensitive and -resistant colon cancer cells. Additionally, proscillaridin A enhanced cell death in cells treated with TRAIL and TRAIL sensitizer, the second mitochondria-derived activator of caspase mimetic. Proscillaridin A upregulated TRAIL receptor expression, while downregulating the levels of the anti-cell death molecules, cellular FADD-like IL-1ß converting enzyme-like inhibitor protein and Mcl1, in a cell type-dependent manner. Furthermore, proscillaridin A enhanced TRAIL-induced cell death partly via O-glycosylation. Taken together, our findings suggest that proscillaridin A is a promising agent that enhances the anti-cancer efficacy of TRAIL therapeutics.


Assuntos
Protocolos de Quimioterapia Combinada Antineoplásica , Neoplasias do Colo , Proscilaridina , Ligante Indutor de Apoptose Relacionado a TNF , Protocolos de Quimioterapia Combinada Antineoplásica/farmacologia , Apoptose/efeitos dos fármacos , Caspases/metabolismo , Linhagem Celular Tumoral , Neoplasias do Colo/tratamento farmacológico , Neoplasias do Colo/metabolismo , Neoplasias do Colo/patologia , Sinergismo Farmacológico , Humanos , Proscilaridina/administração & dosagem , Proscilaridina/farmacologia , Receptores do Ligante Indutor de Apoptose Relacionado a TNF/metabolismo , Ligante Indutor de Apoptose Relacionado a TNF/administração & dosagem , Ligante Indutor de Apoptose Relacionado a TNF/farmacologia
19.
Dis Markers ; 2022: 9696303, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-35845136

RESUMO

Objective: To determine the expression profiling and mechanism of thioredoxin-interacting protein (TXNIP)/nucleotide-binding domain-like receptor protein 3 (NLRP3) inflammasome pathway in sciatic nerve (SN) of type 2 diabetes mellitus (T2DM) rats. Methods: Ten out of the 35 healthy SD rats (specific pathogen free) purchased were randomized into the control group, while the others were established a T2DM model by feeding a high-fat and high-sugar diet plus laparoscopic injection of 1% streptozotocin (STZ). The successfully modeled rats were subgrouped into two arms: a DM group with 10 rats and a resveratrol- (RES-) treated DM intervention group with 11 rats. Normal saline to control and DM groups. Alterations in fasting blood glucose (FBG) and body weight (BW) at different time points after administration were observed. Sciatic nerve conduction velocity (SNCV) and mechanical pain threshold (MPT) were measured. TXNIP, NLRP3, caspase-1, and interleukin- (IL-) 1ß levels in rat SN tissue were determined. Results: DM group rats showed higher FBG and lower BW than control rats at different time points (P < 0.05). The FBG of DM intervention group at 2, 4, and 6 weeks after administration was lower, and the BW at 4 and 6 weeks after dosing was higher than DM group. Higher MPT and SNCV were determined in DM intervention group versus DM group (P < 0.05). DM group rats had disordered, swollen, and dissolved SN myelin sheath structure; TXNIP inhibition led to a small amount of nerve myelin fragments and mild pathological changes. Lower TXNIP, NLRP3, caspase-1, and IL-1ß protein levels were found in DM intervention group versus DM group (P < 0.05). Conclusion: The pathogenesis of peripheral neuropathy in T2DM rats may be linked to TXNIP/NLRP3 inflammasome pathway activation, indicating the potential of this pathway as a therapeutic target for diabetic peripheral neuropathy (DPN).


Assuntos
Diabetes Mellitus Experimental , Diabetes Mellitus Tipo 2 , Animais , Caspases , Proteínas de Ciclo Celular , Diabetes Mellitus Experimental/tratamento farmacológico , Diabetes Mellitus Experimental/metabolismo , Diabetes Mellitus Experimental/patologia , Inflamassomos/metabolismo , Inflamassomos/uso terapêutico , Interleucina-1beta/genética , Interleucina-1beta/metabolismo , Proteína 3 que Contém Domínio de Pirina da Família NLR/metabolismo , Ratos , Ratos Sprague-Dawley , Nervo Isquiático/metabolismo , Nervo Isquiático/patologia
20.
Ren Fail ; 44(1): 1169-1181, 2022 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-35837696

RESUMO

Acute kidney injury (AKI) is a serious condition with high mortality. The most common cause is kidney ischemia/reperfusion (IR) injury, which is thought to be closely related to pyroptosis. Disulfiram is a well-known alcohol abuse drug, and recent studies have shown its ability to mitigate pyroptosis in mouse macrophages. This study investigated whether disulfiram could improve IR-induced AKI and elucidated the possible molecular mechanism. We generated an IR model in mouse kidneys and a hypoxia/reoxygenation (HR) injury model with murine tubular epithelial cells (MTECs). The results showed that IR caused renal dysfunction in mice and triggered pyroptosis in renal tubular epithelial cells, and disulfiram improved renal impairment after IR. The expression of proteins associated with the classical pyroptosis pathway (Nucleotide-binding oligomeric domain (NOD)-like receptor protein 3 (NLRP3), apoptosis-related specific protein (ASC), caspase-1, N-GSDMD) and nonclassical pyroptosis pathway (caspase-11, N-GSDMD) were upregulated after IR. Disulfiram blocked the upregulation of nonclassical but not all classical pyroptosis pathway proteins (NLRP3 and ASC), suggesting that disulfiram might reduce pyroptosis by inhibiting the caspase-11-GSDMD pathway. In vitro, HR increased intracellular ROS levels, the positive rate of PI staining and LDH levels in MTECs, all of which were reversed by disulfiram pretreatment. Furthermore, we performed a computer simulation of the TIR domain of TLR4 using homology modeling and identified a small molecular binding energy between disulfiram and the TIR domain. We concluded that disulfiram might inhibit pyroptosis by antagonizing TLR4 and inhibiting the caspase-11-GSDMD pathway.


Assuntos
Injúria Renal Aguda , Traumatismo por Reperfusão , Injúria Renal Aguda/tratamento farmacológico , Injúria Renal Aguda/etiologia , Injúria Renal Aguda/prevenção & controle , Animais , Caspases/metabolismo , Simulação por Computador , Dissulfiram/farmacologia , Isquemia , Rim/metabolismo , Camundongos , Proteína 3 que Contém Domínio de Pirina da Família NLR/metabolismo , Reperfusão , Traumatismo por Reperfusão/complicações , Traumatismo por Reperfusão/tratamento farmacológico , Traumatismo por Reperfusão/metabolismo , Receptor 4 Toll-Like
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