Sphingosine kinases regulate ER contacts with late endocytic organelles and cholesterol trafficking.

Membrane contact sites (MCS), close membrane apposition between organelles, are platforms for interorganellar transfer of lipids including cholesterol, regulation of lipid homeostasis, and co-ordination of endocytic trafficking. Sphingosine kinases (SphKs), two isoenzymes that phosphorylate sphingosine to the bioactive sphingosine-1-phosphate (S1P), have been implicated in endocytic trafficking. However, the physiological functions of SphKs in regulation of membrane dynamics, lipid trafficking and MCS are not known. Here, we report that deletion of SphKs decreased S1P with concomitant increases in its precursors sphingosine and ceramide, and markedly reduced endoplasmic reticulum (ER) contacts with late endocytic organelles. Expression of enzymatically active SphK1, but not catalytically inactive, rescued the deficit of these MCS. Although free cholesterol accumulated in late endocytic organelles in SphK null cells, surprisingly however, cholesterol transport to the ER was not reduced. Importantly, deletion of SphKs promoted recruitment of the ER-resident cholesterol transfer protein Aster-B (also called GRAMD1B) to the plasma membrane (PM), consistent with higher accessible cholesterol and ceramide at the PM, to facilitate cholesterol transfer from the PM to the ER. In addition, ceramide enhanced in vitro binding of the Aster-B GRAM domain to phosphatidylserine and cholesterol liposomes. Our study revealed a previously unknown role for SphKs and sphingolipid metabolites in governing diverse MCS between the ER network and late endocytic organelles versus the PM to control the movement of cholesterol between distinct cell membranes.

Neutrophilia in severe asthma is reduced in Ormdl3 overexpressing mice.

Genome-wide association studies have linked the ORM (yeast)-like protein isoform 3 (ORMDL3) to asthma severity. Although ORMDL3 is a member of a family that negatively regulates serine palmitoyltransferase (SPT) and thus biosynthesis of sphingolipids, it is still unclear whether ORMDL3 and altered sphingolipid synthesis are causally related to non-Th2 severe asthma associated with a predominant neutrophil inflammation and high interleukin-17 (IL-17) levels. Here, we examined the effects of ORMDL3 overexpression in a preclinical mouse model of allergic lung inflammation that is predominantly neutrophilic and recapitulates many of the clinical features of severe human asthma. ORMDL3 overexpression reduced lung and circulating levels of dihydrosphingosine, the product of SPT. However, the most prominent effect on sphingolipid levels was reduction of circulating S1P. The LPS/OVA challenge increased markers of Th17 inflammation with a predominant infiltration of neutrophils into the lung. A significant decrease of neutrophil infiltration was observed in the Ormdl3 transgenic mice challenged with LPS/OVA compared to the wild type and concomitant decrease in IL-17, that plays a key role in the pathogenesis of neutrophilic asthma. LPS decreased survival of murine neutrophils, which was prevented by co-treatment with S1P. Moreover, S1P potentiated LPS-induced chemotaxis of neutrophil, suggesting that S1P can regulate neutrophil survival and recruitment following LPS airway inflammation. Our findings reveal a novel connection between ORMDL3 overexpression, circulating levels of S1P, IL-17 and neutrophil recruitment into the lung, and questions the potential involvement of ORMDL3 in the pathology, leading to development of severe neutrophilic asthma.

A new preclinical model of western diet-induced progression of non-alcoholic steatohepatitis to hepatocellular carcinoma.

Non-alcoholic steatohepatitis (NASH) results from the accumulation of excessive liver lipids leading to hepatocellular injury, inflammation, and fibrosis that greatly increase the risk for hepatocellular carcinoma (HCC). Despite the well-characterized clinical and histological pathology for NASH-driven HCC in humans, its etiology remains unclear and there is a deficiency in pre-clinical models that recapitulate the progression of the human disease. Therefore, we developed a new mouse model amenable to genetic manipulations and gene targeting that mimics the gradual NASH to HCC progression observed in humans. C57BL/6NJ mice were fed a Western high-fat diet and sugar water (HFD/SW) and monitored for effects on metabolism, liver histology, tumor development, and liver transcriptome for up to 54 weeks. Chronic HFD/SW feeding led to significantly increased weight gain, serum and liver lipid levels, liver injury, and glucose intolerance. Hepatic pathology progressed and mice developed hepatocellular ballooning, inflammation, and worse fibrosis was apparent at 16 weeks, greatly increased through 32 weeks, and remained elevated at 54 weeks. Importantly, hepatocellular cancer spontaneously developed in 75% of mice on HFD/SW, half of which were HCC, whereas none of the mice on the chow diet developed HCC. Chronic HFD/SW induced molecular markers of de novo lipogenesis, endoplasmic reticulum stress, inflammation, and accumulation of p62, all of which also participate in the human pathology. Moreover, transcriptome analysis revealed activation of HCC-related genes and signatures associated with poor prognosis of human HCC. Overall, we have identified a new preclinical model that recapitulates known hallmarks of NASH-driven HCC that can be utilized for future molecular mechanistic studies of this disease.

mTORC1 and SGLT2 Inhibitors-A Therapeutic Perspective for Diabetic Cardiomyopathy.

Diabetic cardiomyopathy is a critical diabetes-mediated co-morbidity characterized by cardiac dysfunction and heart failure, without predisposing hypertensive or atherosclerotic conditions. Metabolic insulin resistance, promoting hyperglycemia and hyperlipidemia, is the primary cause of diabetes-related disorders, but ambiguous tissue-specific insulin sensitivity has shed light on the importance of identifying a unified target paradigm for both the glycemic and non-glycemic context of type 2 diabetes (T2D). Several studies have indicated hyperactivation of the mammalian target of rapamycin (mTOR), specifically complex 1 (mTORC1), as a critical mediator of T2D pathophysiology by promoting insulin resistance, hyperlipidemia, inflammation, vasoconstriction, and stress. Moreover, mTORC1 inhibitors like rapamycin and their analogs have shown significant benefits in diabetes and related cardiac dysfunction. Recently, FDA-approved anti-hyperglycemic sodium-glucose co-transporter 2 inhibitors (SGLT2is) have gained therapeutic popularity for T2D and diabetic cardiomyopathy, even acknowledging the absence of SGLT2 channels in the heart. Recent studies have proposed SGLT2-independent drug mechanisms to ascertain their cardioprotective benefits by regulating sodium homeostasis and mimicking energy deprivation. In this review, we systematically discuss the role of mTORC1 as a unified, eminent target to treat T2D-mediated cardiac dysfunction and scrutinize whether SGLT2is can target mTORC1 signaling to benefit patients with diabetic cardiomyopathy. Further studies are warranted to establish the underlying cardioprotective mechanisms of SGLT2is under diabetic conditions, with selective inhibition of cardiac mTORC1 but the concomitant activation of mTORC2 (mTOR complex 2) signaling.

CRISPR/Cas9 deletion of ORMDLs reveals complexity in sphingolipid metabolism.

The serine palmitoyltransferase (SPT) complex catalyzes the rate-limiting step in the de novo biosynthesis of ceramides, the precursors of sphingolipids. The mammalian ORMDL isoforms (ORMDL1-3) are negative regulators of SPT. However, the roles of individual ORMDL isoforms are unclear. Using siRNA against individual ORMDLs, only single siORMDL3 had modest effects on dihydroceramide and ceramide levels, whereas downregulation of all three ORMDLs induced more pronounced increases. With the CRISPR/Cas9-based genome-editing strategy, we established stable single ORMDL3 KO (ORMDL3-KO) and ORMDL1/2/3 triple-KO (ORMDL-TKO) cell lines to further understand the roles of ORMDL proteins in sphingolipid biosynthesis. While ORMDL3-KO modestly increased dihydroceramide and ceramide levels, ORMDL-TKO cells had dramatic increases in the accumulation of these sphingolipid precursors. SPT activity was increased only in ORMDL-TKO cells. In addition, ORMDL-TKO but not ORMDL3-KO dramatically increased levels of galactosylceramides, glucosylceramides, and lactosylceramides, the elevated N-acyl chain distributions of which broadly correlated with the increases in ceramide species. Surprisingly, although C16:0 is the major sphingomyelin species, it was only increased in ORMDL3-KO, whereas all other N-acyl chain sphingomyelin species were significantly increased in ORMDL-TKO cells. Analysis of sphingoid bases revealed that although sphingosine was only increased 2-fold in ORMDL-TKO cells, levels of dihydrosphingosine, dihydrosphingosine-1-phosphate, and sphingosine-1-phosphate were hugely increased in ORMDL-TKO cells and not in ORMDL3-KO cells. Thus, ORMDL proteins may have a complex, multifaceted role in the biosynthesis and regulation of cellular sphingolipids.

Repression of human and mouse brain inflammaging transcriptome by broad gene-body histone hyperacetylation.

Brain "inflammaging," a low-grade and chronic inflammation, is a major hallmark for aging-related neurodegenerative diseases. Here, by profiling H3K27ac and gene expression patterns in human and mouse brains, we found that age-related up-regulated (Age-Up) and down-regulated (Age-Down) genes have distinct H3K27ac patterns. Although both groups show promoter H3K27ac, the Age-Up genes, enriched for inflammation-related functions, are additionally marked by broad H3K27ac distribution over their gene bodies, which is progressively reduced during aging. Age-related gene expression changes can be predicted by gene-body H3K27ac level. Contrary to the presumed transcription activation function of promoter H3K27ac, we found that broad gene-body hyper H3K27ac suppresses overexpression of inflammaging genes. Altogether, our findings revealed opposite regulations by H3K27ac of Age-Up and Age-Down genes and a mode of broad gene-body H3K27ac in repressing transcription.

History of Psychology Publish and Perish: Psychology's Most Prolific Authors Are Not Always the Ones We Remember.

What is the relationship between being highly prolific in the realm of publication and being remembered as a great psychologist of the past? In this study, the PsycINFO database was used to identify the historical figures who wrote the most journal articles during the half-century from 1890 to 1939. Although a number of the 10 most prolific authors are widely remembered for their influence on the discipline today-E. L. Thorndike, Karl Pearson, E. B. Titchener, Henri Pi6ron-the majority are mostly forgotten. The data were also separated into the 5 distinct decades. Once again, a mixture of eminent and obscure individuals made appearances. Most striking, perhaps, was the great increase in articles published over the course of the half-century-approximately doubling each decade-and the enormous turnover in who was most prolific, decade over decade. In total, 100 distinct individuals appeared across just 5 lists of about 25 names each.

The Evolution of the American Journal of Psychology, 1904-1918: A Network Investigation.

In an earlier article, we used digital historical methods to examine the first 14 volumes (1887-1903) of The American Journal of Psychology (AJP) by creating networks of the vocabularies used in every substantive article the journal published in those years. These networks showed us the major research groups that had been gathered together by the journal's founder-editor, G. Stanley Hall, and how the intellectual composition of the journal (and the discipline) changed over that 17-year period. In the present article we extended that study forward, creating networks for the next 15 volumes (1904-1918), broken into time blocks of 5 years each. Our findings, in brief, were that vision research (especially color) continued to play as consistent and prominent role in the journal as it had since the start; that memory, association, and some higher mental processes were active areas of research throughout; that Hall continued to have an on-again, off-again relationship with theoretical and philosophical psychology; that intelligence-measuring methods, psychoanalysis, and psychophysics were periodically popular enough that they each generated a cluster in 1 of the 3 time blocks; and that Titchener's participation in the editing of AJP led to its becoming the major outlet for his Structuralist research program (an absence that we noted in our earlier studies of the journal Psychological Review).

BRIDGE OVER TROUBLED WATERS? THE MOST "CENTRAL" MEMBERS OF PSYCHOLOGY AND PHILOSOPHY ASSOCIATIONS CA. 1900.

There are many different ways to assess the significance of historical figures. Often we look at the influence of their writings, or at the important offices they held with disciplinary institutions such as universities, journals, and scholarly societies. In this study, however, we took a novel approach: we took the complete memberships, ca. 1900, of four organizations-the American Psychological Association, the Western Philosophical Association, the American Philosophical Association, and the Southern Society for Philosophy and Psychology-and visualized them as a network. We then identified individuals who "bridged" between two or more of these groups and considered what might be termed their "centrality" to the psychological-philosophical community of their time. First, we examined these figures qualitatively, briefly describing their lives and careers. Then we approached the problem mathematically, considering several alternative technical realizations of "centrality" and then explaining our reasons for choosing eigenvector centrality as the best for our purposes. We found a great deal of overlap among the results of the qualitative and quantitative approaches, but also some telling differences. J. Mark Baldwin, Edward Buchner, Christine Ladd Franklin, and Frank Thilly consistently emerged as highly central figures. Some more marginal figures such as Max Meyer, and Frederick J. E. Woodbridge, Edward A. Pace, Edward H. Griffin played interesting roles as well.

On the Meanings of Self-Regulation: Digital Humanities in Service of Conceptual Clarity.

Self-regulation is of interest both to psychologists and to teachers. But what the word means is unclear. To define it precisely, two studies examined the American Psychological Association's system of controlled vocabulary-specifically, the 447 associated terms it presents-and used techniques from the Digital Humanities to identify 88 closely related concepts and six broad conceptual clusters. The resulting analyses show how similar ideas are interrelated: self-control, self-management, self-observation, learning, social behavior, and the personality constructs related to self-monitoring. A full-color network map locates these concepts and clusters relative to each other. It also highlights some of the interests of different audiences, which can be described heuristically using two axes that have been labeled abstract versus practical and self-oriented versus other-oriented.

Midlife gene expressions identify modulators of aging through dietary interventions.

Dietary interventions are effective ways to extend or shorten lifespan. By examining midlife hepatic gene expressions in mice under different dietary conditions, which resulted in different lifespans and aging-related phenotypes, we were able to identify genes and pathways that modulate the aging process. We found that pathways transcriptionally correlated with diet-modulated lifespan and physiological changes were enriched for lifespan-modifying genes. Intriguingly, mitochondrial gene expression correlated with lifespan and anticorrelated with aging-related pathological changes, whereas peroxisomal gene expression showed an opposite trend. Both organelles produce reactive oxygen species, a proposed causative factor of aging. This finding implicates a contribution of peroxisome to aging. Consistent with this hypothesis, lowering the expression levels of peroxisome proliferation genes decreased the cellular peroxide levels and extended the lifespan of Drosophila melanogaster and Caenorhabditis elegans. These findings show that transcriptional changes resulting from dietary interventions can effectively reflect causal factors in aging and identify previously unknown or under-appreciated longevity pathways, such as the peroxisome pathway.

The Evolution of The American Journal of Psychology 1, 1887-1903: A Network Investigation.

The American Journal of Psychology (AJP) was the first academic journal in the United States dedicated to the "new" scientific form of the discipline. But where did the journal's founding owner/editor, G. Stanley Hall, find the "psychologists" he needed to fill the pages of such a venture 1887, when he was still virtually the only professor of psychology in the country? To investigate this question we used the substantive vocabularies of every full article published in AJP's first 14 volumes to generate networks of verbally similar articles. These networks reveal the variety of research communities that Hall drew on to launch and support the journal. Three separate networks, corresponding to 3 successive time blocks, show how Hall's constellation of participating research communities changed over AJP's first 17 years. Many of these communities started with rather nebulous boundaries but soon began to differentiate into groups of more distinct specialties. Some topics declined over time, but new ones regularly appeared to replace them. We sketch a quasievolutionary model to describe the intellectual ecology of AJP's early years.

Searching for the structure of early American psychology: Networking Psychological Review, 1909-1923.

This study continues a previous investigation of the intellectual structure of early American psychology by presenting and analyzing 3 networks that collectively include every substantive article published in Psychological Review during the 15-year period from 1909 to 1923. The networks were laid out such that articles (represented by the network's nodes) that possessed strongly correlated vocabularies were positioned closer to each other spatially than articles with weakly correlated vocabularies. We identified distinct research communities within the networks by locating and interpreting the clusters of lexically similar articles. We found that the Psychological Review was in some turmoil during this period compared with its first 15 years attributable, first, to Baldwin's unexpected departure in 1910; second, to the pressures placed on the discipline by United States entry into World War I; and, third, to the emergence of specialty psychology journals catering to research communities that had once published in the Review. The journal emerged from these challenges, however, with a better-defined mission: to serve as the chief repository of theoretical psychology in the United States.

Searching for the structure of early American psychology: Networking Psychological Review, 1894-1908.

This study investigated the intellectual structure of early American psychology by generating 3 networks that collectively included every substantive article published in Psychological Review during the 15-year period from the journal's start in 1894 until 1908. The networks were laid out so that articles with strongly correlated vocabularies were positioned close to each other spatially. Then, we identified distinct research communities by locating and interpreting article clusters within the networks. We found that, from the first 5-year time block to the second, psychological specialties rapidly differentiated themselves from each other. Between the second and third 5-year time blocks, however, the number of specialties shrunk. We discuss the degree to which this shift may have been attributable either to a change in the journal's editorship in 1904, or to a broader crisis of confidence, beginning that same year, in the use of "consciousness" as the discipline's defining concept.

Systematic prediction of pharmacodynamic drug-drug interactions through protein-protein-interaction network.

Identifying drug-drug interactions (DDIs) is a major challenge in drug development. Previous attempts have established formal approaches for pharmacokinetic (PK) DDIs, but there is not a feasible solution for pharmacodynamic (PD) DDIs because the endpoint is often a serious adverse event rather than a measurable change in drug concentration. Here, we developed a metric "S-score" that measures the strength of network connection between drug targets to predict PD DDIs. Utilizing known PD DDIs as golden standard positives (GSPs), we observed a significant correlation between S-score and the likelihood a PD DDI occurs. Our prediction was robust and surpassed existing methods as validated by two independent GSPs. Analysis of clinical side effect data suggested that the drugs having predicted DDIs have similar side effects. We further incorporated this clinical side effects evidence with S-score to increase the prediction specificity and sensitivity through a Bayesian probabilistic model. We have predicted 9,626 potential PD DDIs at the accuracy of 82% and the recall of 62%. Importantly, our algorithm provided opportunities for better understanding the potential molecular mechanisms or physiological effects underlying DDIs, as illustrated by the case studies.

Beyond the schools of psychology 2: a digital analysis of psychological review, 1904-1923.

In order to better understand the broader trends and points of contention in early American psychology, it is conventional to organize the relevant material in terms of "schools" of psychology-structuralism, functionalism, etc. Although not without value, this scheme marginalizes many otherwise significant figures, and tends to exclude a large number of secondary, but interesting, individuals. In an effort to address these problems, we grouped all the articles that appeared in the second and third decades of Psychological Review into five-year blocks, and then cluster analyzed each block by the articles' verbal similarity to each other. This resulted in a number of significant intellectual "genres" of psychology that are ignored by the usual "schools" taxonomy. It also made "visible" a number of figures who are typically downplayed or ignored in conventional histories of the discipline, and it provide us with an intellectual context in which to understand their contributions.

Overexpression of ORMDL3 confers sexual dimorphism in diet-induced non-alcoholic steatohepatitis.

OBJECTIVE: The bioactive sphingolipid metabolites ceramide and sphingosine-1-phosphate (S1P) accumulate with overnutrition and have been implicated in non-alcoholic steatohepatitis (NASH) development. ORMDL3, a negative regulator of the rate-limiting step in ceramide biosynthesis, has been identified as an obesity-related gene. Therefore, we assessed the role of ORMDL3 in diet-induced obesity and development of NASH. METHODS: Globally overexpressing Ormdl3-Flag transgenic mice (ORMDL3TG) were fed a western high-fat, carbohydrate and cholesterol enriched diet, with high fructose-glucose drinking water. Physiological, biochemical and sphingolipidomic analyses were employed to measure the effect of ORMDL3 overexpression on NASH development. RESULTS: ORMDL3TG male but not female mice fed a western high-fat diet and sugar water had exacerbated adipocyte hypertrophy together with increased severity of white adipose inflammation and fibrosis. Hepatic steatosis, dyslipidemia, impaired glucose homeostasis, hyperinsulinemia, and insulin resistance were significantly more severe only in obese ORMDL3TG male mice that accompanied dramatic liver fibrosis, inflammation, and formation of hepatic crown-like structures, which are unique features of human and murine NASH. Obesogenic diet induces ORMDL expression in male mice but reduces it in females. Mechanistically, overexpression of Ormdl3 lowered the levels of S1P and ceramides only in obese female mice and antithetically increased them in tissues of obese males. ORMDL3TG male mice exhibited a much greater induction of the UPR, propagating ER stress that contributed to their early development of NASH. CONCLUSIONS: This study uncovered a previously unrecognized role for ORMDL3 in sexual dimorphism important for the development and progression of NASH.

Sphingosine Kinase 2 and p62 Regulation are Determinants of Sexual Dimorphism in Hepatocellular Carcinoma.

OBJECTIVE: Hepatocellular carcinoma (HCC) is the third leading cause of cancer mortality, and its incidence is increasing due to endemic obesity. HCC is sexually dimorphic in both humans and rodents with higher incidence in males, although the mechanisms contributing to these correlations remain unclear. Here, we examined the role of sphingosine kinase 2 (SphK2), the enzyme that regulates the balance of bioactive sphingolipid metabolites, sphingosine-1-phosphate (S1P) and ceramide, in gender specific MASH-driven HCC. METHODS: Male and female mice were fed a high fat diet with sugar water, a clinically relevant model that recapitulates MASH-driven HCC in humans followed by physiological, biochemical cellular and molecular analyses. In addition, correlations with increased risk of HCC recurrence were determined in patients. RESULTS: Here, we report that deletion of SphK2 protects both male and female mice from Western diet-induced weight gain and metabolic dysfunction without affecting hepatic lipid accumulation or fibrosis. However, SphK2 deficiency decreases chronic diet-induced hepatocyte proliferation in males but increases it in females. Remarkably, SphK2 deficiency reverses the sexual dimorphism of HCC, as SphK2-/- male mice are protected whereas the females develop liver cancer. Only in male mice, chronic western diet induced accumulation of the autophagy receptor p62 and its downstream mediators, the antioxidant response target NQO1, and the oncogene c-Myc. SphK2 deletion repressed these known drivers of HCC development. Moreover, high p62 expression correlates with poor survival in male HCC patients but not in females. In hepatocytes, lipotoxicity-induced p62 accumulation is regulated by sex hormones and prevented by SphK2 deletion. Importantly, high SphK2 expression in male but not female HCC patients is associated with a more aggressive HCC differentiation status and increased risk of cancer recurrence. CONCLUSIONS: This work identifies SphK2 as a potential regulator of HCC sexual dimorphism and suggests SphK2 inhibitors now in clinical trials could have opposing, gender-specific effects in patients.

An exploratory digital analysis of the early years of G. Stanley Hall's American Journal of Psychology and Pedagogical Seminary.

In this article, we present the results of an exploratory digital analysis of the contents of the two journals founded in the late 19th century by American psychologist G. Stanley Hall. Using the methods of the increasingly popular digital humanities, some key attributes of the American Journal of Psychology (AJP) and the Pedagogical Seminary (PS) are identified. Our analysis reaffirms some of Hall's explicit aims for the two periodicals, while also revealing a number of other features of the journals, as well as of the people who published within their pages, the methodologies they employed, and the institutions at which they worked. Notably, despite Hall's intent that his psychological journal be strictly an outlet for scientific research, the journal-like its sister pedagogically focused publication-included an array of methodologically diverse research. The multiplicity of research styles that characterize the content of Hall's journals in their initial years is, in part, a consequence of individual researchers at times crossing methodological lines and producing a diverse body of research. Along with such variety within each periodical, it is evident that the line between content appropriate to one periodical rather than the other was fluid rather than absolute. The full results of this digitally informed analysis of Hall's two journals suggest a number of novel avenues for future research and demonstrate the utility of digital methods as applied to the history of psychology. (PsycINFO Database Record (c) 2013 APA, all rights reserved).

Beyond the schools of psychology 1: a digital analysis of Psychological Review, 1894-1903.

Traditionally, American psychology at the turn of the twentieth century has been framed as a competition among a number of "schools": structuralism, functionalism, behaviorism, etc. But this is only one way in which the "structure" of the discipline can be conceived. Most psychologists did not belong to a particular school, but they still worked within loose intellectual communities, and so their work was part of an implicit psychological "genre," if not a formalized "school." In this study, we began the process of discovering the underlying genres of American psychology at the turn of the twentieth century by taking the complete corpus of articles from the journal Psychological Review during the first decade of its publication and conducting a statistical analysis of the vocabularies they employed to see what clusters of articles naturally emerged. Although the traditional functionalist school was among the clusters we found, we also found distinct research traditions around the topics of color vision, spatial vision, philosophy/metatheory, and emotion. In addition, momentary clusters corresponding to important debates (e.g., the variability hypothesis) appeared during certain years, but not others.