Wnt signaling induces transcription, spatial proximity, and translocation of fusion gene partners in human hematopoietic cells.

Chromosomal translocations are frequently associated with a wide variety of cancers, particularly hematologic malignancies. A recurrent chromosomal abnormality in acute myeloid leukemia is the reciprocal translocation t(8;21) that fuses RUNX1 and ETO genes. We report here that Wnt/ß-catenin signaling increases the expression of ETO and RUNX1 genes in human hematopoietic progenitors. We found that ß-catenin is rapidly recruited into RNA polymerase II transcription factories (RNAPII-Ser5) and that ETO and RUNX1 genes are brought into close spatial proximity upon Wnt3a induction. Notably, long-term treatment of cells with Wnt3a induces the generation a frequent RUNX1-ETO translocation event. Thus, Wnt/ß-catenin signaling induces transcription and translocation of RUNX1 and ETO fusion gene partners, opening a novel window to understand the onset/development of leukemia.

Alternative RUNX1 Promoter Regulation by Wnt/ß-Catenin Signaling in Leukemia Cells and Human Hematopoietic Progenitors.

Two distantly located promoter regions regulate the dynamic expression of RUNX genes during development: distal P1 and proximal P2 promoters. We have recently described that ß-catenin increases total Runx1 mRNA levels in human CD34(+) hematopoietic progenitors and enhances spatial proximity with its translocation partner ETO. Here, we report that induction of Wnt/ß-catenin signaling in HL60 and Jurkat leukemia-derived cell lines and CD34(+) progenitors selectively activate the production of the longer distal P1-Runx1 mRNA isoform. Gain- and loss-of-function experiments revealed that the differential increase in P1-Runx1 expression is accomplished through a minimal ß-catenin responsive region that includes a highly conserved TCF/LEF-binding element, located -20/-16 bp upstream of the canonical distal P1-Runx1 transcription start site. We conclude that the distal P1-Runx1 promoter is a direct transcriptional target of Wnt/ß-catenin signaling that may be important in normal hematopoiesis or its transition into malignant stem cells during the onset or progression of leukemia.

Early Transcriptional Changes Induced by Wnt/ß-Catenin Signaling in Hippocampal Neurons.

Wnt/ß-catenin signaling modulates brain development and function and its deregulation underlies pathological changes occurring in neurodegenerative and neurodevelopmental disorders. Since one of the main effects of Wnt/ß-catenin signaling is the modulation of target genes, in the present work we examined global transcriptional changes induced by short-term Wnt3a treatment (4 h) in primary cultures of rat hippocampal neurons. RNAseq experiments allowed the identification of 170 differentially expressed genes, including known Wnt/ß-catenin target genes such as Notum, Axin2, and Lef1, as well as novel potential candidates Fam84a, Stk32a, and Itga9. Main biological processes enriched with differentially expressed genes included neural precursor (GO:0061364, p-adjusted = 2.5 × 10-7), forebrain development (GO:0030900, p-adjusted = 7.3 × 10-7), and stem cell differentiation (GO:0048863 p-adjusted = 7.3 × 10-7). Likewise, following activation of the signaling cascade, the expression of a significant number of genes with transcription factor activity (GO:0043565, p-adjusted = 4.1 × 10-6) was induced. We also studied molecular networks enriched upon Wnt3a activation and detected three highly significant expression modules involved in glycerolipid metabolic process (GO:0046486, p-adjusted = 4.5 × 10-19), learning or memory (GO:0007611, p-adjusted = 4.0 × 10-5), and neurotransmitter secretion (GO:0007269, p-adjusted = 5.3 × 10-12). Our results indicate that Wnt/ß-catenin mediated transcription controls multiple biological processes related to neuronal structure and activity that are affected in synaptic dysfunction disorders.

Health status of Pomatoschistus microps populations in relation to pollution and natural stressors: implications for ecological risk assessment.

Effects induced on wild populations by recurrent environmental contamination may difficult the ecological risk assessment of punctual pollution events such as oil spills. Here, the issue was addressed by comparing the health status of Pomatoschistus microps populations from four NW Iberian estuaries, using an integrated chemical-biological monitoring. Despite high seasonal variability, the parameters measured discriminated estuaries with different contamination levels and associated biological effects with chemical and abiotic stress. The decreased health status of fish from polluted sites strengthens the need of considering pollution-induced background effects and seasonal variability when assessing impacts and risks of oil and other chemical spills.

Retrospective qualitative analysis of ecological networks under environmental perturbation: a copper-polluted intertidal community as a case study.

The coast of Chañaral Bay in northern Chile has been affected by copper mine wastes for decades. This sustained perturbation has disrupted the intertidal community in several ways, but the mechanisms behind the observed shifts in local biodiversity remain poorly understood. Our main goal was to identify the species (lumped into trophic groups) belonging to the Chañaral intertidal community that, being directly affected by copper pollution, contributed primarily to the generation of the observed changes in community structure. These groups of species were called initiators. We applied a qualitative modelling approach based only on the sign and direction of effects among species, and present a formula for predicting changes in equilibrium abundances considering stress on multiple variables simultaneously. We then applied this technique retrospectively to identify the most likely set of initiators. Our analyses allowed identification of a unique set of four initiators in the studied intertidal system (a group of algae, sessile invertebrates, a group of herbivores and starfish), which were hypothesized to be the primary drivers of the observed changes in community structure. In addition, a hypothesis was derived about how the perturbation affected these initiators. The hypothesis is that pollution affected negatively the population growth rate of both algae and sessile invertebrates and suppressed the interaction between herbivores and starfish. Our analytic approach, focused on identifying initiators, constitutes an advance towards understanding the mechanisms underlying human-driven ecosystem disruption and permits identifying species that may serve as a focal point for community management and restoration.

IgG targeting distinct seasonal coronavirus- conserved SARS-CoV-2 spike subdomains correlates with differential COVID-19 disease outcomes.

Despite SARS-CoV-2 being a "novel" virus, early detection of anti-spike IgG in severe COVID-19 patients may be caused by the amplification of humoral memory responses against seasonal coronaviruses. Here, we examine this phenomenon by characterizing anti-spike IgG responses in non-hospitalized convalescent individuals across a spectrum of COVID-19 severity. We observe that disease severity positively correlates with anti-spike IgG levels, IgG cross-reactivity against other betacoronaviruses (ß-CoVs), and FcγR activation. Analysis of IgG targeting ß-CoV-conserved and non-conserved immunodominant epitopes within the SARS-CoV-2 spike protein revealed epitope-specific relationships: IgG targeting the conserved heptad repeat (HR) 2 region significantly correlates with milder disease, while targeting the conserved S2'FP region correlates with more severe disease. Furthermore, a lower HR2-to-S2'FP IgG-binding ratio correlates with greater disease severity, with ICU-hospitalized COVID-19 patients showing the lowest HR2/S2'FP ratios. These findings suggest that HR2/S2'FP IgG profiles may predict disease severity and offer insight into protective versus deleterious humoral recall responses.

Sustained Antibody-Dependent NK Cell Functions in Mild COVID-19 Outpatients During Convalescence.

The coronavirus disease 2019 (COVID19) pandemic has left researchers scrambling to identify the humoral immune correlates of protection from COVID-19. To date, the antibody mediated correlates of virus neutralization have been extensively studied. However, the extent that non-neutralizing functions contribute to anti-viral responses are ill defined. In this study, we profiled the anti-spike antibody subtype/subclass responses, along with neutralization and antibody-dependent natural killer cell functions in 83 blood samples collected between 4 and 201 days post-symptoms onset from a cohort of COVID-19 outpatients. We observed heterogeneous humoral responses against the acute respiratory syndrome coronavirus 2 (SARS-CoV-2) spike protein. Overall, anti-spike profiles were characterized by a rapid rise of IgA and sustained IgG titers. In addition, strong antibody-mediated natural killer effector responses correlated with milder disease and being female. While higher neutralization profiles were observed in males along with increased severity. These results give an insight into the underlying function of antibodies beyond neutralization and suggest that antibody-mediated natural killer cell activity is a key function of the humoral response against the SARS-CoV-2 spike protein.

Wnt/ß-catenin signaling stimulates the expression and synaptic clustering of the autism-associated Neuroligin 3 gene.

Synaptic abnormalities have been described in individuals with autism spectrum disorders (ASD). The cell-adhesion molecule Neuroligin-3 (Nlgn3) has an essential role in the function and maturation of synapses and NLGN3 ASD-associated mutations disrupt hippocampal and cortical function. Here we show that Wnt/ß-catenin signaling increases Nlgn3 mRNA and protein levels in HT22 mouse hippocampal cells and primary cultures of rat hippocampal neurons. We characterized the activity of mouse and rat Nlgn3 promoter constructs containing conserved putative T-cell factor/lymphoid enhancing factor (TCF/LEF)-binding elements (TBE) and found that their activity is significantly augmented in Wnt/ß-catenin cell reporter assays. Chromatin immunoprecipitation (ChIP) assays and site-directed mutagenesis experiments revealed that endogenous ß-catenin binds to novel TBE consensus sequences in the Nlgn3 promoter. Moreover, activation of the signaling cascade increased Nlgn3 clustering and co- localization with the scaffold PSD-95 protein in dendritic processes of primary neurons. Our results directly link Wnt/ß-catenin signaling to the transcription of the Nlgn3 gene and support a functional role for the signaling pathway in the dysregulation of excitatory/inhibitory neuronal activity, as is observed in animal models of ASD.

Effects of copper on early developmental stages of Lessonia nigrescens Bory (Phaeophyceae).

Copper effects on the early developmental gametophytic and sporophytic stages of the kelp Lessonia nigrescens were tested in gradients of increasing concentrations of ASV-labile copper. The results demonstrated a high sensitivity to copper of all life-history stages of the alga, where even the lowest tested concentration affected spore release as well as their subsequent settlement. More significant, concentrations higher than 7.87 microg L(-1) totally interrupted the development of the spores after they settle. This effect led to a failure in the formation of male and female gametophytes and, as a consequence, to a complete disruption of the normal life cycle of the kelp. Thus, we suggest that the absence of L. nigrescens from copper-enriched environments results from the high sensitivity of its early life cycle stages, which limits growth and maturation of the gametophytic microscopic phase and, as a consequence, prevents development of the macroscopic sporophytic phase.

Micro-evolution due to pollution: possible consequences for ecosystem responses to toxic stress.

Polluting events can change community structure and ecosystem functioning. Selection of genetically inherited tolerance on exposed populations, here referred as micro-evolution due to pollution, has been recognized as one of the causes of these changes. However, there is a gap between studies addressing this process and those assessing effects at higher levels of biological organization. In this review we attempt to address these evolutionary considerations into the ecological risk assessment (ERA) of polluting events and to trigger the discussion about the consequences of this process for the ecosystem response to toxic stress. We provide clear evidence that pollution drives micro-evolutionary processes in several species. When this process occurs, populations inhabiting environments that become polluted may persist. However, due to the existence of ecological costs derived from the loss of genetic variability, negative pleiotropy with fitness traits and/or from physiological alterations, micro-evolution due to pollution may alter different properties of the affected populations. Despite the existence of empirical evidence showing that safety margins currently applied in the ERA process may account for pollution-induced genetic changes in tolerance, information regarding long-term ecological consequences at higher levels of biological organization due to ecological costs is not explicitly considered in these procedures. In relation to this, we present four testable hypotheses considering that micro-evolution due to pollution acts upon the variability of functional response traits of the exposed populations and generates changes on their functional effect traits, therefore, modifying the way species exploit their ecological niches and participate in the overall ecosystem functioning.

Synergistic roles of climate warming and human occupation in Patagonian megafaunal extinctions during the Last Deglaciation.

The causes of Late Pleistocene megafaunal extinctions (60,000 to 11,650 years ago, hereafter 60 to 11.65 ka) remain contentious, with major phases coinciding with both human arrival and climate change around the world. The Americas provide a unique opportunity to disentangle these factors as human colonization took place over a narrow time frame (~15 to 14.6 ka) but during contrasting temperature trends across each continent. Unfortunately, limited data sets in South America have so far precluded detailed comparison. We analyze genetic and radiocarbon data from 89 and 71 Patagonian megafaunal bones, respectively, more than doubling the high-quality Pleistocene megafaunal radiocarbon data sets from the region. We identify a narrow megafaunal extinction phase 12,280 ± 110 years ago, some 1 to 3 thousand years after initial human presence in the area. Although humans arrived immediately prior to a cold phase, the Antarctic Cold Reversal stadial, megafaunal extinctions did not occur until the stadial finished and the subsequent warming phase commenced some 1 to 3 thousand years later. The increased resolution provided by the Patagonian material reveals that the sequence of climate and extinction events in North and South America were temporally inverted, but in both cases, megafaunal extinctions did not occur until human presence and climate warming coincided. Overall, metapopulation processes involving subpopulation connectivity on a continental scale appear to have been critical for megafaunal species survival of both climate change and human impacts.

Wnt/ß-catenin signaling in Alzheimer's disease.

Alzheimer's disease is a neurodegenerative disorder that causes a progressive decline of mental and cognitive processes such as memory, judgment and reasoning. We proposed earlier that a sustained loss of function of Wnt/ß- catenin signaling components underlies the onset and progression of the disease. Here, we discuss recent data on the involvement of Wnt/b-catenin signaling on amyloid precursor protein (APP) processing, Aß peptide neurotoxicity, τ phosphorylation, and modulation of Apolipoprotein E function in the brain. We conclude that several components of the cascade are actively engaged in the events leading to AD neuropathology and propose that compounds that mimic activation of this signaling cascade, such as lithium, should be considered for therapeutic intervention in Alzheimer's patients. In summary, data accumulated during the past decade confirm some important predictions of our hypothesis where components of this signaling cascade are actively engaged in the events leading to AD neuropathology and that compounds that mimic activation of this signaling cascade, such as lithium, should be considered for therapeutic intervention in Alzheimer's patients.

A novel functional low-density lipoprotein receptor-related protein 6 gene alternative splice variant is associated with Alzheimer's disease.

We previously found that single nucleotide polymorphisms in the low-density lipoprotein receptor-related protein 6 (LRP6) gene are associated with Alzheimer's disease (AD). Here, we studied the posttranscriptional metabolism of the LRP6 message scanning sequentially the 23 LRP6 exons in human tissues and found a novel LRP6 isoform that completely skips exon 3 (LRP6Δ3) in all tissues examined and was also conserved in mice. Expression levels of the LRP6 isoforms were determined in 47 cortical brain messenger (m)RNA samples including 22 AD cases, 11 control subjects, and 14 individuals with other neurological disorders. LRP6Δ3 mRNA levels were significantly augmented in AD brains compared with controls (1.6-fold; p = 0.037) or other pathological samples (2-fold; p = 0.007). Functional analysis in Wnt/ß-catenin signaling assays revealed that skipping of exon 3 reduced significantly the signaling activity of the LRP6 coreceptor. We conclude that the LRP6Δ3 isoform is a novel splice variant, which shows diminished Wnt/ß-catenin signaling activity and might have a functional role in individuals with AD.

Individual, population and community level effects of subtle anthropogenic contamination in estuarine meiobenthos.

The study presented here searched for the level of taxonomic resolution required to detect the effects of low-level chronic pollution on estuarine meiobenthic communities. Meiofauna from two sites, with special attention to harpacticoid copepods, was analysed at different taxonomic levels of aggregation using uni- and multivariate methods. Adaptation processes that could buffer biodiversity disruptions were also considered through the analysis of fitness-related and tolerance traits in the harpacticoid copepod Paronychocamptus nanus. Results showed that uni- and multivariate analyses could be inadequate when assessing subtle anthropogenic contamination. Instead, the assessment of inter-population differences in tolerance to the main source of stress rises as a required procedure if potential effects of this type of contamination are being investigated. Specifically, a 96 h acute toxicity test performed with populations from the affected site appears as a faster and reliable general tool to assess impacts of low-level chronic pollution in estuaries.

Cadmium--copper antagonism in seaweeds inhabiting coastal areas affected by copper mine waste disposals.

Cadmium and copper accumulation by macroalgae was studied in a coastal area exposed to upwelling events and high levels of Cu, the latter resulting from mine disposals. Eight species were studied, and all had very high concentrations of Cd outside of the Cu-contaminated area. Cu in algal tissues was much higher in contaminated than in reference sites. High Cu appeared to suppress Cd bioaccumulation; Cd in algal tissues was much lower in the Cu-contaminated area than in the reference sites. Transplant experiments with Lessonia nigrescens revealed a depuration of Cd in individuals transplanted to areas with high Cu. However, Cd depuration occurs more slowly than Cu uptake. These differences suggest that while Cd and Cu are linked mechanistically, itis nota simple substitution. Overall, the work confirms that macroalgae are useful indicators of metal contamination and may be used as in situ biomonitors for labile forms of metals, like free Cu2+. However, antagonistic relationships between metals must be clearly understood in order to properly interpret their concentrations in macroalgae.

Effects of cypermethrin on marine plankton communities: a simulated field study using mesocosms.

In earlier single-species toxicity tests we showed the negative effects on the calanoid copepod Acartia tonsa upon exposure to cypermethrin, a pesticide used in treatment for sea lice in salmon farming. In the present study we assessed effects at a higher level of biological organization and under a more realistic exposure scenario using mesocosms. The results showed that simulated field studies (SFSs) could be conducted with the mesocosms designed here. When cypermethrin was applied inside these mesocosms, its concentration decreased exponentially following a first-order kinetics model. The pesticide immediately reduced zooplankton density and biodiversity not only directly, by killing copepods, but also indirectly, by increasing the numbers of rotifers. Zooplankton density recovered after treatment, but zooplankton biodiversity remained altered. In an open environment, however, the rapid dissipation of the pesticide, coupled with population processes of compensation, migration, and immigration, may lead to recovery of the affected zooplankton communities.