EMID2 is a novel biotherapeutic for aggressive cancers identified by in vivo screening.

BACKGROUND: New drugs to tackle the next pathway or mutation fueling cancer are constantly proposed, but 97% of them are doomed to fail in clinical trials, largely because they are identified by cellular or in silico screens that cannot predict their in vivo effect. METHODS: We screened an Adeno-Associated Vector secretome library (> 1000 clones) directly in vivo in a mouse model of cancer and validated the therapeutic effect of the first hit, EMID2, in both orthotopic and genetic models of lung and pancreatic cancer. RESULTS: EMID2 overexpression inhibited both tumor growth and metastatic dissemination, consistent with prolonged survival of patients with high levels of EMID2 expression in the most aggressive human cancers. Mechanistically, EMID2 inhibited TGFß maturation and activation of cancer-associated fibroblasts, resulting in more elastic ECM and reduced levels of YAP in the nuclei of cancer cells. CONCLUSION: This is the first in vivo screening, precisely designed to identify proteins able to interfere with cancer cell invasiveness. EMID2 was selected as the most potent protein, in line with the emerging relevance of the tumor extracellular matrix in controlling cancer cell invasiveness and dissemination, which kills most of cancer patients.

A ligand-insensitive UNC5B splicing isoform regulates angiogenesis by promoting apoptosis.

The Netrin-1 receptor UNC5B is an axon guidance regulator that is also expressed in endothelial cells (ECs), where it finely controls developmental and tumor angiogenesis. In the absence of Netrin-1, UNC5B induces apoptosis that is blocked upon Netrin-1 binding. Here, we identify an UNC5B splicing isoform (called UNC5B-Δ8) expressed exclusively by ECs and generated through exon skipping by NOVA2, an alternative splicing factor regulating vascular development. We show that UNC5B-Δ8 is a constitutively pro-apoptotic splicing isoform insensitive to Netrin-1 and required for specific blood vessel development in an apoptosis-dependent manner. Like NOVA2, UNC5B-Δ8 is aberrantly expressed in colon cancer vasculature where its expression correlates with tumor angiogenesis and poor patient outcome. Collectively, our data identify a mechanism controlling UNC5B's necessary apoptotic function in ECs and suggest that the NOVA2/UNC5B circuit represents a post-transcriptional pathway regulating angiogenesis.

Genetic lineage tracing reveals poor angiogenic potential of cardiac endothelial cells.

AIMS: Cardiac ischaemia does not elicit an efficient angiogenic response. Indeed, lack of surgical revascularization upon myocardial infarction results in cardiomyocyte death, scarring, and loss of contractile function. Clinical trials aimed at inducing therapeutic revascularization through the delivery of pro-angiogenic molecules after cardiac ischaemia have invariably failed, suggesting that endothelial cells in the heart cannot mount an efficient angiogenic response. To understand why the heart is a poorly angiogenic environment, here we compare the angiogenic response of the cardiac and skeletal muscle using a lineage tracing approach to genetically label sprouting endothelial cells. METHODS AND RESULTS: We observed that overexpression of the vascular endothelial growth factor in the skeletal muscle potently stimulated angiogenesis, resulting in the formation of a massive number of new capillaries and arterioles. In contrast, response to the same dose of the same factor in the heart was blunted and consisted in a modest increase in the number of new arterioles. By using Apelin-CreER mice to genetically label sprouting endothelial cells we observed that different pro-angiogenic stimuli activated Apelin expression in both muscle types to a similar extent, however, only in the skeletal muscle, these cells were able to sprout, form elongated vascular tubes activating Notch signalling, and became incorporated into arteries. In the heart, Apelin-positive cells transiently persisted and failed to give rise to new vessels. When we implanted cancer cells in different organs, the abortive angiogenic response in the heart resulted in a reduced expansion of the tumour mass. CONCLUSION: Our genetic lineage tracing indicates that cardiac endothelial cells activate Apelin expression in response to pro-angiogenic stimuli but, different from those of the skeletal muscle, fail to proliferate and form mature and structured vessels. The poor angiogenic potential of the heart is associated with reduced tumour angiogenesis and growth of cancer cells.

Alternative splicing in endothelial cells: novel therapeutic opportunities in cancer angiogenesis.

Alternative splicing (AS) is a pervasive molecular process generating multiple protein isoforms, from a single gene. It plays fundamental roles during development, differentiation and maintenance of tissue homeostasis, while aberrant AS is considered a hallmark of multiple diseases, including cancer. Cancer-restricted AS isoforms represent either predictive biomarkers for diagnosis/prognosis or targets for anti-cancer therapies. Here, we discuss the contribution of AS regulation in cancer angiogenesis, a complex process supporting disease development and progression. We consider AS programs acting in a specific and non-redundant manner to influence morphological and functional changes involved in cancer angiogenesis. In particular, we describe relevant AS variants or splicing regulators controlling either secreted or membrane-bound angiogenic factors, which may represent attractive targets for therapeutic interventions in human cancer.

Interactive effect of 5-HTTLPR genotype and age on sources of cortical rhythms in healthy women.

This study was aimed to localize the effects of 5-HTTLPR (serotonin-transporter-linked polymorphic region) on the age differences of spontaneous EEG activity in women using neuroimaging analysis sLORETA (Standardized Low Resolution brain Electromagnetic Tomography). DNA samples extracted from cheek swabs and resting-state EEG recorded at 60 standard leads were collected from young (YW, N=86, 18-35years) and older (OW, N=45; 55-80years) healthy women. We have shown that advanced age was associated with increased posterior EEG desynchronization in S'/S'. S' (LG allele was grouped with S alleles owing to its functional equivalence and this group was labeled as S') genotype carriers denoted by decrease of delta - beta1 current source density, and to a lesser extent in L/L homozygotes denoted by decrease in delta activity. In heterozygotes OW, as compared with heterozygotes YW, higher source density estimates of beta1 in frontal and temporal cortex were observed. Age differences were more pronounced in the right hemisphere in S'/S' and L/L carriers and in the left hemisphere in heterozygotes. We also found that in OW, current source density estimates of theta, alpha1, alpha2, alpha3 and beta1 sources in the right occipital lobe were higher in S'/L than in S'/S' carriers. These results may have implications for understanding 5-HTT-dependent variation in the effect of aging on brain activity.

Age-related differences in electroencephalogram connectivity and network topology.

To better understand age-related differences in brain function and behavior, connectivity between brain regions was estimated from electroencephalogram source time series in eyes closed versus eyes open resting condition. In beta band, decrease of connectivity upon eyes opening was more pronounced in younger than in older participants. The extent of this decrease was associated with reaction time in attention tasks, and this relationship was fully mediated by participants' age, implying that physiological processes, which lead to age-related slowing, include changes in beta reactivity. Graph-theoretical analysis showed a decrease of modularity and clustering in beta and gamma band networks in older adults, implying that age makes brain networks more random. The overall number of nodes identified as hubs in posterior cortical regions decreased in older participants. At the same time, increase of connectedness of anterior nodes, probably reflecting compensatory activation of the anterior attentional system, was observed in beta-band network of older adults. These findings show that normal aging mostly affects interactions in beta band, which are probably involved in attentional processes.

EEG correlates of spontaneous self-referential thoughts: a cross-cultural study.

The default mode network (DMN) has been mostly investigated using positron emission tomography and functional magnetic resonance imaging (fMRI) and has received mixed support in electroencephalographic (EEG) studies. In this study, after sLORETA transformation of EEG data, we applied group spatial independent component analysis which is routinely used in fMRI research. In three large and diverse samples coming from two different cultures (Russian and Taiwanese), spontaneous EEG data and retrospective questionnaire measures of subject's state, thoughts, and feelings during the EEG registration were collected. Regression analyses showed that appearance of spontaneous self-referential thoughts was best predicted by enhanced alpha activity within the DMN. Diminished theta and delta activity in the superior frontal gyrus and enhanced beta activity in the postcentral gyrus added to the prediction. The enhanced alpha activity prevailed in the posterior DMN hub in Russian, but in the anterior DMN hub in Taiwanese participants. Possible cross-cultural differences in personality and attitudes underlying this difference are discussed.

Association of verbal and figural creative achievement with polymorphism in the human serotonin transporter gene.

The purpose of this study was to examine the potential association between the S (short) and L (long) alleles of the 5-HTTLPR polymorphism of the serotonin transporter (5-HTT) gene and verbal and figural creative ability. Sixty-two unrelated Caucasian university students (29 men and 33 women) participated in the experiment. The results showed a significant association between verbal and figural creativity scores and the 5-HTTLPR polymorphism. The subjects with S/S and L/S genotypes demonstrated higher verbal creativity scores in comparison with the L/L genotype carriers. The carriers of S/S genotype demonstrated also higher figural creativity scores in comparison with the carries of L/S and L/L genotypes. Thus, it is the first report on a significant association between the 5-HTTLPR polymorphism and creative achievements. As the 5-HTTLPR polymorphism is associated with genetically defined alteration in the brain serotonergic neurotransmission our result provides an evidence of the involvement of the central serotonin system in creativity regulation.

EEG mapping in seasonal affective disorder.

Given that the nature of hemispheric dysfunction is different in heterogeneous disorders, in the present investigation EEG power mapping was applied to establish neurophysiological profiles that might potentially discriminate patients with seasonal affective disorder (SAD) among other affective disorders. The baseline resting EEG activity was recorded from 31 depressed SAD patients and 30 controls. Power in the delta, theta-1, theta-2, alpha, beta-1 and beta-2 frequency bands was extracted by Fourier transformation. Patients were found to have a lower delta (in central, parietal, occipital, temporal, posterior-temporal areas), theta-1 (in central and parietal), theta-2 (in anterior-frontal, parietal, occipital) and alpha activity (in anterior-frontal, midfrontal, central, parietal and occipital areas) than controls. SAD subjects showed, compared to controls, an asymmetrical distribution of delta, theta-1, theta-2 and alpha activity in parietal and temporal regions due to an increase of EEG power over the right electrode sites, and beta activity in the lateral frontal region due to an increase of beta power over the right electrode site. It is assumed that differential hemispheric contributions of EEG spectra may discriminate between the varieties of depression or different depressive states.