• Humans
• Cell Transformation, Neoplastic/metabolism
• MicroRNAs/genetics
• MicroRNAs/metabolism
• Mutation
• Neoplasms/pathology
• Proto-Oncogene Proteins p21(ras)/genetics
• Proto-Oncogene Proteins p21(ras)/metabolism

• R01 HD098131 NICHD NIH HHS
• R01 HD099252 NICHD NIH HHS
• T32 HL092332 NHLBI NIH HHS
• Eunice Kennedy Shriver National Institute of Child Health and Human Development (NICHD)
• Cancer Prevention and Research Institute of Texas (CPRIT)
• Andrew McDonough B+ Foundation (AMBF)
• V Foundation for Cancer Research (VFCR)
• National Heart, Lung, and Blood Institute (NHLBI)

• Cancer therapy
• cancer
• metastasis
• microRNA

• Humans
• MicroRNAs/genetics
• Neoplasms/genetics
• Neoplasms/therapy
• Neoplasms/pathology
• RNA, Small Untranslated
• Gene Expression Regulation, Neoplastic

• Ago2
• angiogenesis
• diabetes
• erectile dysfunction
• reactive oxygen species

• FAK
• cancer progression
• clinical significance
• molecular mechanisms
• prognosis

• Hepatocellular carcinoma
• RNA binding protein
• Therapeutic targets

• Biophysics
• Calcium
• Humans
• Mechanotransduction, Cellular
• MicroRNAs/genetics
• Neoplasms/genetics
• Tumor Microenvironment

• R35 GM128753 NIGMS NIH HHS

• combination therapy
• drug resistance
• focal adhesion kinase
• metastasis
• tumor microenvironment

• Argonaute-2
• Brain endothelial cells
• Glia
• Lipopolysaccharide
• Neuroprotection
• Secretome

• Animals
• Argonaute Proteins/genetics
• Argonaute Proteins/metabolism
• Argonaute Proteins/pharmacology
• Astrocytes/drug effects
• Astrocytes/metabolism
• Brain/drug effects
• Brain/metabolism
• Endothelial Cells/drug effects
• Endothelial Cells/metabolism
• Gene Silencing
• Inflammation/metabolism
• Lipopolysaccharides/pharmacology
• Mice
• Microglia/drug effects
• Microglia/metabolism

• UID/Multi/00709/2013 Fundação para a Ciência e a Tecnologia
• IF/00178/2015/CP1300/CT0001 Fundação para a Ciência e a Tecnologia
• SFRH/BD/137440/2018 Fundação para a Ciência e a Tecnologia
• POCI-01-0145-FEDER-022122 PPBI-Portuguese Platform of BioImaging
• FCT L’Oréal-UNESCO Portugal for Women in Science

• AGO2
• miRNAs
• polymorphisms
• renal cell carcinoma

• AGO2
• KRAS
• nonsmall cell lung cancer

• Animals
• Argonaute Proteins/physiology
• Carcinoma, Non-Small-Cell Lung/etiology
• Carcinoma, Non-Small-Cell Lung/genetics
• Disease Models, Animal
• Disease Progression
• Lung Neoplasms/etiology
• Lung Neoplasms/genetics
• MAP Kinase Signaling System
• Mice
• Mice, Inbred C57BL
• Proto-Oncogene Proteins p21(ras)/physiology
• Signal Transduction/physiology

• P50 CA186786 NCI NIH HHS
• R35 CA231996 NCI NIH HHS
• U01 CA214170 NCI NIH HHS

The dysregulation of RNA interference (RNAi) has often been observed in cancers, where the main focus of research has been on the small RNA molecules directing RNAi. In this review, we focus on the activity of Argonaute proteins, central components of RNAi, in tumorigenesis, and also highlight their potential applications in grading tumors and anti-cancer therapies.

Argonaute proteins (AGOs) play crucial roles in RNA-induced silencing complex (RISC) formation and activity. AGOs loaded with small RNA molecules (miRNA or siRNA) either catalyze endoribonucleolytic cleavage of target RNAs or recruit factors responsible for translational silencing and target destabilization. miRNAs are well characterized and broadly studied in tumorigenesis; nevertheless, the functions of the AGOs in cancers have lagged behind. Here, we discuss the current state of knowledge on the role of AGOs in tumorigenesis, highlighting canonical and non-canonical functions of AGOs in cancer cells, as well as the biomarker potential of AGO expression in different of tumor types. Furthermore, we point to the possible application of the AGOs in development of novel therapeutic approaches.

• Argonaute
• biomarker
• cancer
• miRNA
• therapeutics
• tumorigenesis

Hepatocellular carcinoma (HCC) is a leading cause of cancer-related morbidity and mortality globally. Despite the remarkable improvements in comprehensive HCC treatment, the underlying mechanistic details of HCC remain elusive. We screened HCC patients for differentially expressed genes (DEGs) using the Gene Expression Omnibus (GSE113850) and The Cancer Genome Atlas (TCGA) datasets. LINC01554 expression in 40 paired samples was determined by quantitative reverse transcription polymerase chain reaction (RT-qPCR), and its clinical significance was assessed. LINC01554 was found to have a gain-of-function role in HCC in vitro. Additionally, the bioinformatics analysis of the genes co-expressed with LINC01554 was performed using the Co-LncRNA website, and potential molecular mechanisms were investigated using the Gene Ontology and Kyoto Encyclopaedia of Genes and Genomes resources and validated by in vitro experiments. A total of 229 DEGs were identified from the GSE113850 dataset. Among the identified DEGs, three long non-coding RNAs (lncRNAs) (DIO3OS, LINC01554, and LINC01093) with |logFC| ≥2 and P<0.05 were screened. A total of 148 lncRNAs with |logFC| ≥1 and P<0.05 were identified from TCGA dataset. Low LINC01554 expression levels were significantly correlated with overall survival, pathological stage, hepatitis B infection, tumour size, portal vein tumour thrombus, and TNM stage. Using gain-of-function assays, we further showed that LINC01554 inhibited the proliferation, migration, and invasion of the HCCLM9 and SK-Hep1 cells and promoted G0/G1 arrest, but it did not significantly affect apoptosis. Western blotting revealed that LINC01554 overexpression resulted in increased ZO-1 and E-cadherin expression levels, but decreased N-cadherin and vimentin expression levels. Moreover, LINC01554 overexpression inhibited Akt, p-Akt, β-catenin, and p-Gsk3β expression. Our results showed that LINC01554 repressed HCC cell invasiveness and epithelial-to-mesenchymal transition partly by inhibiting Wnt and PI3K-Akt signalling in vitro. Taken together, our findings provide new insights into the molecular mechanisms underlying HCC tumourigenesis and implicate LINC01554 as a potential target for HCC therapy.

• FAK inhibitors
• Focal adhesion kinase
• ovarian cancer
• platinum compounds
• resistance
• taxanes.

Regulation of gene expression has emerged as a fundamental element of transcript homeostasis. Key effectors in this process are the Argonautes (AGOs), highly specialized RNA-binding proteins (RBPs) that form complexes, such as the RNA-Induced Silencing Complex (RISC). AGOs dictate post-transcriptional gene-silencing by directly loading small RNAs and repressing their mRNA targets through small RNA-sequence complementarity. The four human highly-conserved family-members (AGO1, AGO2, AGO3, and AGO4) demonstrate multi-faceted and versatile roles in transcriptome’s stability, plasticity, and functionality. The post-translational modifications of AGOs in critical amino acid residues, the nucleotide polymorphisms and mutations, and the deregulation of expression and interactions are tightly associated with aberrant activities, which are observed in a wide spectrum of pathologies. Through constantly accumulating information, the AGOs’ fundamental engagement in multiple human diseases has recently emerged. The present review examines new insights into AGO-driven pathology and AGO-deregulation patterns in a variety of diseases such as in viral infections and propagations, autoimmune diseases, cancers, metabolic deficiencies, neuronal disorders, and human infertility. Altogether, AGO seems to be a crucial contributor to pathogenesis and its targeting may serve as a novel and powerful therapeutic tool for the successful management of diverse human diseases in the clinic.

• RNA binding proteins
• argonaute proteins
• autoimmune disease
• cancer
• human disease
• infertility
• metabolic disorders
• neuronal deficiencies
• post-transcriptional regulation
• viral infection

• Inflammation
• Oncogenes
• Regulation
• Tumor suppressor genes
• miRNA

AGO2 (Argonaute RISC Catalytic Component 2) plays an important role in small RNA-guided gene silencing processes. It has been implied in tumorigenesis of different types of tumors. In this study, we found that AGO2 expression was remarkably increased in human hepatocellular carcinoma (HCC) tissues when compared with adjacent noncancerous tissues. High expression of AGO2 was associated with poor prognosis in HCC patients. The CRISPR/Cas9-mediated knockout of AGO2 in SMMC-7721 cells inhibited cell proliferation and induced significant G1 phase arrest of cell cycle. Inhibition of cell migration was also observed in SMMC-7721 AGO2^−/− cells. In vivo experiments showed that tumors grew slower in nude mice transplanted with AGO2^−/− cells than in SMMC-7721 cell-derived xenograft mice. Microarray analysis and western blot analysis revealed that AGO2 depletion decreased expression of Survivin, Vimentin, and Snail. Overexpression of AGO2 in SMMC-7721 and Huh-7 cells could reverse the knockout-induced inhibition effects on either cell behaviors or expression of Survivin, Vimentin, and Snail Therefore, our data demonstrated that AGO2 might facilitate HCC tumorigenesis and metastasis through modulating expression of Survivin, Vimentin, and Snail.

The most common malignant tumor of the digestive system is HCC. However, the mechanism and pathogenesis of HCC occurrence and progress are still unknown. LncRNA is closely related to the occurrence and progress of HCC. It is important to investigate the effect and role of lncRNA in HCC.

LncRNA microarray assay was used to screen the differential expression profile of lncRNA. SNHG11, miR-184 and GO2 expression was analyzed by RT-PCR. The ability of SNHG11 to serve as a sponge for miRNA and the fact that miR-184 directly targets mRNA were revealed by dual luciferase assay and RIP. Apoptosis and autophagy related proteins were detected by Western blot. Cell proliferation, invasion, migration, and apoptosis were detected by CCK-8 assay, wound healing assay, transwell assay, and flow cytometry.

LncRNA microarray assay and RT-PCR results revealed that the expression of SNHG11 was increased in HCC tumor tissues and also upregulated in HCC cells. SNHG11 had a connection with poor survival rate in HCC. In addition, dual luciferase assay and RIP results revealed that SNHG11 serves as a sponge for miR-184 and miR-184 directly targets AGO2. Pearson correlation analysis showed that SNHG11 with miR-184 and miR-184 with AGO2 were negative correlations, and SNHG11 with AGO2 was a positive correlation. Cell function assay and Western blot showed SNHG4/miR-184/AGO2 regulatory loop was critical for HCC cell proliferation, migration, apoptosis, and autophagy.

Our study demonstrated that the expression of SNHG11 is higher in HCC; moreover, SNHG11 promotes proliferation, migration, apoptosis, and autophagy by regulating AGO2 via miR-184 in HCC. Our verification of the role of SNHG11 may provide a novel biomarker for the diagnosis, therapy, and prognosis of HCC.

• LncRNA SNHG11
• apoptosis
• biomarker
• hepatocellular cancer

• focal adhesion kinase
• small-cell lung cancer
• targeted therapy

• - Fonds Spécial de Recherche (FSR)
• - Fondation Willy and Marcy De Vooght
• 1.E.108.19F Fonds pour la Formation à la Recherche dans l'Industrie et dans l'Agriculture
• 7.4624.15 (Télévie) Fonds De La Recherche Scientifique - FNRS
• FMG-2011-BR-02, FMG-2013-BR-02, FMG-2014-BR-01, FMG-2015-BR-02, FMG-2016-BR-02, FMG-2017-BR-04, and FMG-2018-BR-01 Fondation Mont-Godinne
• - Secteurs des Sciences de la Santé

• Ago2
• Amplification
• Argonaute 2
• Biomarker
• Breast
• Cancer
• Cytoplasmic
• DFS
• Disease
• EIF2C2
• Expression
• Gene
• IHC
• Intensity
• Midbody
• Pattern
• RFS
• Relapse
• Staining
• Survival
• Tumour
• mRNA
• miRNA
• microRNA

Insulin signaling is mediated by a highly integrated network that controls glucose metabolism, protein synthesis, cell growth, and differentiation. Our previous work indicates that the insulin receptor tyrosine kinase substrate (IRTKS), also known as BAI1-associated protein 2-like 1 (BAIAP2L1), is a novel regulator of insulin network, but the mechanism has not been fully studied. In this work we reveal that IRTKS co-localizes with Src homology (SH2) containing inositol polyphosphate 5-phosphatase-2 (SHIP2), and the SH3 domain of IRTKS directly binds to SHIP2’s catalytic domain INPP5c. IRTKS suppresses SHIP2 phosphatase to convert phosphatidylinositol 3,4,5-triphosphate (PI(3,4,5)P₃, PIP3) to phosphatidylinositol (3,4) bisphosphate (PI(3,4)P₂). IRTKS-knockout significantly increases PI(3,4)P₂ level and decreases cellular PI(3,4,5)P₃ content. Interestingly, the interaction between IRTKS and SHIP2 is dynamically regulated by insulin, which feeds back and affects the tyrosine phosphorylation of IRTKS. Furthermore, IRTKS overexpression elevates PIP3, activates the AKT–mTOR signaling pathway, and increases cell proliferation. Thereby, IRTKS not only associates with insulin receptors to activate PI3K but also interacts with SHIP2 to suppress its activity, leading to PIP3 accumulation and the activation of the AKT–mTOR signaling pathway to modulate cell proliferation.

• IRTKS
• PI(3,4,5)P3
• SHIP2
• insulin pathway

• AGO2
• DICER
• acute lymphoblastic leukaemia
• microRNA biogenesis

• FIP200
• LKB1
• PI3K
• PTEN
• anoikis
• chaperon
• dimerization
• miRNA
• motility
• regulation

• extracellular vesicles
• hepatocellular carcinoma
• cancer-associated fibroblasts
• hepatic stellate cells

• Animals
• Carcinoma, Hepatocellular/drug therapy
• Carcinoma, Hepatocellular/pathology
• Cell Line, Tumor
• Cell Proliferation/genetics
• Exosomes
• Extracellular Vesicles
• Female
• Gene Expression Regulation, Neoplastic/drug effects
• Humans
• Liver Neoplasms/drug therapy
• Liver Neoplasms/pathology
• Mice
• MicroRNAs/pharmacology
• Reverse Transcriptase Polymerase Chain Reaction

• K08 DK090154 NIDDK NIH HHS
• R01 CA083650 NCI NIH HHS
• R01 CA190040 NCI NIH HHS
• R01 EB017742 NIBIB NIH HHS

• 3T3 protocol
• Ago2
• Mouse embryonic fibroblast

• Adenocarcinoma/drug therapy
• Adenocarcinoma/enzymology
• Adenocarcinoma/genetics
• Adenocarcinoma/pathology
• Adenocarcinoma of Lung
• Adenosine Deaminase/genetics
• Adenosine Deaminase/metabolism
• Cell Line
• Cell Movement
• Enzyme Stability
• Focal Adhesion Kinase 1/antagonists & inhibitors
• Focal Adhesion Kinase 1/genetics
• Focal Adhesion Kinase 1/metabolism
• Humans
• Lung Neoplasms/drug therapy
• Lung Neoplasms/enzymology
• Lung Neoplasms/genetics
• Lung Neoplasms/pathology
• Neoplasm Proteins/antagonists & inhibitors
• Neoplasm Proteins/genetics
• Neoplasm Proteins/metabolism
• RNA-Binding Proteins/genetics
• RNA-Binding Proteins/metabolism

• U54 CA137788 NCI NIH HHS
• R01 CA132580 NCI NIH HHS
• P30 CA008748 NCI NIH HHS
• R01 CA192399 NCI NIH HHS
• R01 CA136705 NCI NIH HHS

GADD45A (growth arrest and DNA damage inducible alpha), a stress response gene induced by genotoxic and nongenotoxic stresses, is implicated in various key processes, including the control of cell cycle checkpoints and DNA repair. The expression of GADD45A is directly regulated by numerous transcription factors, with p53 being the most representative. Moreover, post-transcriptional regulation also plays a role in GADD45A expression. However, little is known about the regulatory effects of microRNAs (miRNAs) on GADD45A expression. As a potential tumour suppressor, miR-138 has pleiotropic biological functions in various cancers. We have previously reported p53-mediated activation of miR-138 in human non-small-cell lung cancer (NSCLC) cells. In this study, we found that miR-138 specifically targeted AGO2, which affects the stability and maturation of miR-130b. Decreased expression of miR-130b promoted the expression of GADD45A and resulted in the G2/M phase arrest and proliferation inhibition in human NSCLC cells. Our results suggested that p53 could alternatively upregulate GADD45A in human NSCLC cells through a post-transcriptional pathway in which miR-138 is involved.

• AGO2
• FAK/PI3K/AKT signaling pathway
• cell proliferation
• hypopharyngeal cancer

Viral infection triggers a series of signaling cascades and host innate immune responses, including interferon (IFN) production, which depends on coordinated activity of multiple transcription factors. IFN regulatory factor 3 (IRF3) and transcriptional coactivator CREB binding protein (CBP) and/or p300 are core factors that participate in transcriptional complex formation in the nucleus. In general, cells balance the production of IFNs through suppressive and stimulative mechanisms, but viral infections can disrupt such equilibrium. This study determined that H5N1 viral infection reduced the distribution of human argonaute 2 (AGO2) in A549 cell nucleus. AGO2 did not block phosphorylation, nuclear translocation, and DNA binding ability of IRF3 but inhibited its association with CBP. Therefore, this newly revealed mechanism shows that cellular response leads to transfer of AGO2 from cell nucleus and promotes IFN-β expression to increase host survival during viral infection.

• AGO2
• CBP/p300
• IRF3
• negative regulation
• type I interferon

• cancer stem cells
• focal adhesion kinase
• hepatocellular carcinoma
• metastasis
• proliferation

• Carcinoma, Hepatocellular/enzymology
• Carcinoma, Hepatocellular/genetics
• Carcinoma, Hepatocellular/pathology
• Disease Progression
• Focal Adhesion Protein-Tyrosine Kinases/genetics
• Focal Adhesion Protein-Tyrosine Kinases/metabolism
• Gene Expression Regulation, Enzymologic
• Gene Expression Regulation, Neoplastic
• Humans
• Liver Neoplasms/enzymology
• Liver Neoplasms/genetics
• Liver Neoplasms/pathology
• Models, Genetic
• Neoplasm Invasiveness
• Neoplastic Stem Cells/metabolism

• MMP2/9
• diindolylmethane (DIM)
• focal adhesion kinase (FAK)
• hepatocellular carcinoma
• pTEN

MicroRNAs (miRs) are small non-coding RNAs that regulate most cellular protein networks by targeting mRNAs for translational inhibition or degradation. Dicer, a type III endoribonuclease, is a critical component in microRNA biogenesis and is required for mature microRNA production. Abnormal Dicer expression occurs in numerous cancer types and correlates with poor patient prognosis. For example, increased Dicer expression in melanoma is associated with more aggressive tumors (higher tumor mitotic index and depth of invasion) and poor patient prognosis. However, the role that Dicer plays in melanoma development and immune evasion remains unclear. Here, we report on a newly discovered relationship between Dicer expression and tumor immunogenicity. To investigate Dicer's role in regulating melanoma immunogenicity, Dicer knockdown studies were performed. We found that B16F0-Dicer deficient cells exhibited decreased tumor growth compared to control cells and were capable of inducing anti-tumor immunity. The decrease in tumor growth was abrogated in immunodeficient NSG mice and was shown to be dependent upon CD8⁺ T cells. Dicer knockdown also induced a more responsive immune gene profile in melanoma cells. Further studies demonstrated that CD8⁺ T cells preferentially killed Dicer knockdown tumor cells compared to control cells. Taken together, we present evidence which links Dicer expression to tumor immunogenicity in melanoma.

• CD8+ T cells
• Dicer
• immunogenicity
• melanoma
• microRNA

• Cell migration
• Cell proliferation
• Hepatocellular carcinoma
• XPO5

• Cell migration
• Cell proliferation
• Hepatocellular carcinoma
• XPO5

Argonaute 2 (AGO2), a central component of RNA-induced silencing complex, plays critical roles in cancer. We examined whether the single nucleotide polymorphisms (SNPs) of AGO2 were related to the risk of nasopharyngeal carcinoma (NPC).

Twenty-five tag SNPs within AGO2 were genotyped in Guangxi population consisting of 855 NPC patients and 1036 controls. The SNPs significantly associated with NPC were further replicated in Guangdong population consisting of 996 NPC patients and 972 controls. Functional experiments were conducted to examine the biologic roles of AGO2 in NPC.

A significantly increased risk of advanced lymph node metastasis of NPC was identified for the AGO2 rs3928672 GA + AA genotype compared with GG genotype in both the Guangxi and Guangdong populations (combined odd ratio = 2.08, 95 % confidence interval = 1.44-3.01, P = 8.60 × 10⁻⁵). Moreover, the AGO2 protein expression levels of rs3928672 GA + AA genotype carriers were higher than the GG genotype carriers in the NPC tissues (P = 0.041), and AGO2 was significantly over-expressed in NPC tissues compared with non-cancerous nasopharyngeal tissues (P = 0.011). In addition, AGO2 knockdown reduced cell proliferation, induced apoptosis, and inhibited migration of NPC cells. Furthermore, gene expression microarray showed that genes altered following AGO2 knockdown were clustered in tumorigenesis and metastasis relevant pathways.

Our findings suggest that the genetic polymorphism in AGO2 may be a risk factor for the advanced lymph node metastasis of NPC in Chinese populations, and AGO2 acts as an oncogene in the development of NPC.

The online version of this article (doi:10.1186/s12885-015-1895-4) contains supplementary material, which is available to authorized users.

• HCC
• LNX
• PDZRN4
• overexpression
• proliferation

AGO2 (Argonaute 2, EIF2C2) is the only member in AGO family with catalytic activity and of extreme importance during small RNAs guided gene silencing processes. The structural investigations have provided insights into details and functional mechanisms of the four major domains within AGO2. As a multifunction player, AGO2 has been revealed involved in tumorgenesis through miRNAs-dependent or independent ways. And nowadays, AGO2 has also been more importantly found ectopically over-expressed in carcinomas and closely associated with aspects of cancers in means of interacting with well-known tumor factors. Here, we provide a review on structural insights, functional mechanisms, novel roles and relationship with carcinomas of AGO2.

• Argonaute-2
• angiogenesis
• carcinomas
• microRNAs

Hepatocellular carcinoma (HCC) is one of the leading causes of cancer-related deaths globally. The identity and role of cell surface molecules driving complex biological events leading to HCC progression are poorly understood, hence representing major lacunae in HCC therapies. Here, combining SILAC quantitative proteomics and biochemical approaches, we uncover a critical oncogenic role of Agrin, which is overexpressed and secreted in HCC. Agrin enhances cellular proliferation, migration and oncogenic signalling. Mechanistically, Agrin’s extracellular matrix sensor activity provides oncogenic cues to regulate Arp2/3-dependent ruffling, invadopodia formation and epithelial–mesenchymal transition through sustained focal adhesion integrity that drives liver tumorigenesis. Furthermore, Agrin signalling through Lrp4-muscle-specific tyrosine kinase (MuSK) forms a critical oncogenic axis. Importantly, antibodies targeting Agrin reduced oncogenic signalling and tumour growth in vivo. Together, we demonstrate that Agrin is frequently upregulated and important for oncogenic property of HCC, and is an attractive target for antibody therapy.

The proteoglycan Agrin is known to be expressed in neurons and muscle and to bind ECM protein laminin. Here the authors report that Agrin promotes hepatocellular carcinoma by stimulating proliferation, decreasing focal adhesion, increasing invasiveness and promoting an epithelial-to-mesenchymal transition.

• focal adhesion kinase
• tumor growth
• metastasis
• tumor microenvironment
• fak inhibitors

• Animals
• Focal Adhesion Kinase 1/antagonists & inhibitors
• Focal Adhesion Kinase 1/metabolism
• Humans
• Molecular Targeted Therapy/methods
• Neoplasm Invasiveness
• Neoplasms/metabolism
• Neoplasms/pathology
• Protein Kinase Inhibitors/pharmacology
• Signal Transduction
• Small Molecule Libraries/pharmacology
• Stromal Cells/enzymology
• Tumor Microenvironment

• T32 CA121938 NCI NIH HHS
• CA180769 NCI NIH HHS
• R01 CA102310 NCI NIH HHS
• CA102310 NCI NIH HHS
• R01 CA180769 NCI NIH HHS
• T32‑CA121938 NCI NIH HHS

• calpain small subunit 1
• focal adhesion kinase
• hepatocellular carcinoma
• metastasis
• prognosis