Many Labs 2: Investigating Variation in Replicability Across Samples and Settings

We conducted preregistered replications of 28 classic and contemporary published findings, with protocols that were peer reviewed in advance, to examine variation in effect magnitudes across samples and settings. Each protocol was administered to approximately half of 125 samples that comprised 15,305 participants from 36 countries and territories. Using the conventional criterion of statistical significance (p < .05), we found that 15 (54%) of the replications provided evidence of a statistically significant effect in the same direction as the original finding. With a strict significance criterion (p < .0001), 14 (50%) of the replications still provided such evidence, a reflection of the extremely highpowered design. Seven (25%) of the replications yielded effect sizes larger than the original ones, and 21 (75%) yielded effect sizes smaller than the original ones. The median comparable Cohen’s ds were 0.60 for the original findings and 0.15 for the replications. The effect sizes were small (< 0.20) in 16 of the replications (57%), and 9 effects (32%) were in the direction opposite the direction of the original effect. Across settings, the Q statistic indicated significant heterogeneity in 11 (39%) of the replication effects, and most of those were among the findings with the largest overall effect sizes; only 1 effect that was near zero in the aggregate showed significant heterogeneity according to this measure. Only 1 effect had a tau value greater than .20, an indication of moderate heterogeneity. Eight others had tau values near or slightly above .10, an indication of slight heterogeneity. Moderation tests indicated that very little heterogeneity was attributable to the order in which the tasks were performed or whether the tasks were administered in lab versus online. Exploratory comparisons revealed little heterogeneity between Western, educated, industrialized, rich, and democratic (WEIRD) cultures and less WEIRD cultures (i.e., cultures with relatively high and low WEIRDness scores, respectively). Cumulatively, variability in the observed effect sizes was attributable more to the effect being studied than to the sample or setting in which it was studied.UCR::Vicerrectoría de Investigación::Unidades de Investigación::Ciencias Sociales::Instituto de Investigaciones Psicológicas (IIP

Dysregulation of integrin-linked kinase (ILK) signaling during colorectal carcinogenesis : modulation of signaling pathways by non-steroidal anti-inflammatory drugs (NSAID)

One of the most common events involved in the development of human colon cancer is the mutation of the adenomatous polyposis coli (APC) gene. This protein through its interactions with Axin and GSK3β, serves to regulate the cytosolic levels of β-catenin. Mutation of the APC gene, which impairs complex formation, results in the stabilization of β-catenin. Stabilization is believed to coincide with the translocation of β-catenin to the nucleus, where it up-regulates a number of genes implicated in oncogenesis. Interestingly, stable over-expression of the integrin-linked kinase (ILK) in rat intestinal epithelial cells has been demonstrated to modulate p-catenin sub-cellular localization and function. However, the significance of this finding in human colorectal carcinogenesis is unclear. To determine if ILK signaling was disrupted in colorectal carcinogenesis, this signaling pathway was characterized during various stages of development beginning with the earliest lesion, the adenomatous polyp. The results from these studies demonstrated that ILK was significantly overexpressed and exhibited an increased phosphotransferase activity in polyps resected from patients diagnosed with familial adenomatous polyposis. Changes in ILK activity reflected changes on downstream targets, predominantly GSK3β. In addition to this, dramatic increases in ILK immunoreactivity were observed in all abnormal crypts from sporadic polyps, when compared with the normal appearing crypts, within the same resected specimens. To delineate whether these changes in ILK signaling could be generalized for colon cancer, this signaling nexus was also investigated in both primary lesions as well as secondary deposits within regional lymph nodes. The results from these studies demonstrate that ILK was significantly hyperexpressed in malignant acini from either the primary or secondary site in relation to the normal crypts within the same lesion. Furthermore, over-expression of the ILK protein coincided with an increase in the MBP phosphotransferase activity of the immunoprecipitated ILK in colon cancer in approximately 63% of the primary lesions examined. In addition to this, the data indicated that there was a direct correlation between the protein expression of ILK and the protein levels of Lef-1 in the cases of colon cancer that were analyzed. As aspirin and sulindac have been demonstrated to elicit chemopreventative effects in colon cancer, I tested whether non-steroidal antiinflammatory agents targeted the ILK signaling nexus in vivo. Both of these drugs inhibited the serum-induced activation of ILK and PKB, modulated serine-9 phosphorylation on GSK3β, and down-regulated Tcf-4 transcriptional activity. In addition to this, sulilndac was shown to also inhibit another protein kinase that is known to influence p-catenin, protein kinase CK2. Furthermore, the data demonstrated that over-expression of ILK, PKB or CK2 in a cell culture system, inhibited NSAID mediated apoptosis. In conclusion, dysregulation of the ILK signaling nexus appears to be an early event during the development of colon cancer and it is possible that selective inhibition of this kinase might be an important chemopreventative/chemotherapeutic strategy in the colon.Medicine, Faculty ofMedicine, Department ofExperimental Medicine, Division ofGraduat

A prospective cohort study of 14-3-3 eta in ACPA and/or RF-positive patients with arthralgia

14-3-3η (eta) is a novel serum/plasma protein biomarker involved in the upregulation of inflammatory and joint damage factors. We analysed the association of 14-3-3η with the development of clinically apparent arthritis in a cohort of subjects with arthralgia and positivity for at least one serologic marker: rheumatoid factor (RF) or anti-citrullinated protein antibody (ACPA). Measurement of 14-3-3η in plasma collected on entry into the cohort. For this study, 144 subjects with a minimum of 2.5 (median and maximum 5) years of follow-up were available. The relationship between presence and levels of 14-3-3η and development of arthritis was investigated. Arthritis occurred in 43 (30 %) of the 144 subjects after a median of 15 months. 14-3-3η was detectable up to 5 years before onset of clinical arthritis and was present significantly more often (36 % versus 14 %; relative risk 2.5, 95 % confidence interval 1.2-5.6; p = 0.02) and at significantly higher levels (median 0.95 versus 0.28 ng/ml; p = 0.02) in subjects developing arthritis compared with those who did not. 14-3-3η levels/positivity and ACPA, but not RF, were univariately associated with the development of arthritis while generalized linear model analysis with RF and ACPA as obligatory factors could not return an incremental benefit with 14-3-3η. 14-3-3η was detectable prior to the onset of arthritis and was associated with arthritis development in arthralgia subjects pre-selected for positivity of RF or ACPA. Its power to predict onset of arthritis independent of ACPA and RF requires a new study in which patients are not pre-selected based on ACPA and/or RF seropositivit

14-3-3η Promotes Invadosome Formation via the FOXO3–Snail Axis in Rheumatoid Arthritis Fibroblast-like Synoviocytes

Erosive destruction of joint structures is a critical event in the progression of rheumatoid arthritis (RA), in which fibroblast-like synoviocytes (FLS) are the primary effectors. We previously reported that the ability of RA FLS to degrade extracellular matrix (ECM) components depends on the formation of actin-rich membrane protrusions, called invadosomes, through processes that remain elusive. 14-3-3η belongs to a family of scaffolding proteins involved in a wide range of cellular functions, and its expression is closely related to joint damage and disease activity in RA patients. In this study, we sought to assess the role of 14-3-3η in joint damage by examining its contribution to the invadosome formation phenotype of FLS. Using human primary FLS, we show that 14-3-3η expression is closely associated with their ability to form invadosomes. Furthermore, knockdown of 14-3-3η using shRNAs decreases the level of invadosome formation in RA FLS, whereas addition of the recombinant protein to FLS from healthy individuals promotes their formation. Mechanistic studies suggest that 14-3-3η regulates invadosome formation by increasing Snail expression, a mechanism that involves nuclear exclusion of the transcription repressor FOXO3. Our results implicate the 14-3-3η–FOXO3–Snail axis in promoting the aggressive ECM-degrading phenotype of RA FLS, and suggest a role for this scaffolding protein in cartilage degradation

14-3-3η is a novel mediator associated with the pathogenesis of rheumatoid arthritis and joint damage

Introduction: The aim of this study was to investigate whether 14-3-3η, a specific isoform of a family of proteins regulating processes such as cellular signalling, activates cell-signalling pathways and induces factors known to contribute to the pathophysiology of rheumatoid arthritis (RA). We also investigated whether 14-3-3η is associated with more severe disease in both early and established RA. Methods We investigated the effect of 14-3-3η on the activation of RA-relevant signalling cascades and induction of proinflammatory mediators that contribute to the joint damage process. 14-3-3η titres from 33 patients with early RA (mean RA duration = 1.8 months) and from 40 patients with established RA were measured in serum drawn at the 3-year time point of the Behandel Strategieën study. The relationship between 14-3-3η titres and standard clinical variables was investigated by correlation analysis. The association with radiographic damage and radiographic progression over at least a 2-year period was investigated using univariate and multivariate regression analyses. Results 14-3-3η activated selected members of the mitogen-activated protein kinase (MAPK) family, mainly extracellular regulated kinase 1/2 and c-Jun kinase, but not p38MAPK. Activation by 14-3-3η, using levels spanning the concentration range found in RA patient serum, resulted in the induction of inflammatory transcripts such as interleukin 1 (IL-1) and IL-6 and factors linked to the joint damage process, such as receptor activator of nuclear factor κB ligand and matrix metalloproteinase 1. Serum 14-3-3η correlated significantly with rheumatoid factor (RF) (r = 0.43) and anticitrullinated protein antibodies (ACPAs) (r = 0.31) in the early RA cohort, but not with C-reactive protein (CRP) or the Disease Activity Score in 28 joints in either cohort. Serum 14-3-3η concentrations were significantly higher in patients with radiographically assessed joint damage and in those who had radiographic progression. By multivariate analysis, we show that 14-3-3η complemented markers such as CRP, RF and ACPA in informing RA radiographic status and/or progression. Conclusions Extracellular 14-3-3η activates key signalling cascades and induces factors associated with the pathogenesis of RA at concentrations found in patients with RA, and its expression is higher in patients with radiographic damage and RA progression.Medicine, Faculty ofSurgery, Department ofNon UBCReviewedFacult