A function of spalt major as a sequence-specific DNA binding transcription factor mediates repression of knirps in the Drosophila wing imaginal disc.

The Spalt transcriptional regulators participate in a variety of cell fate decisions during multicellular development. Vertebrate Spalt proteins have been mostly associated to the organization of heterochromatic regions, but they also contribute regulatory functions through binding to A/T rich motives present in their target genes. The developmental processes in which the Drosophila spalt genes participate are well known through genetic analysis, but the mechanism by which the Spalt proteins regulate transcription are still unknown. Furthermore, despite the prominent changes in gene expression associated to mutations in the spalt genes, the specific DNA sequences they bind are unknow. Here, we analyze a DNA fragment present in the regulatory region of the knirps gene. Spalt proteins are candidate repressors of knirps expression during the formation of the venation pattern in the wing disc, and we identified a minimal conserved 30bp sequence that binds to Spalt major both in vivo and in vitro. This sequence mediates transcriptional repression in the central region of the wing blade, constituting the first confirmed case of a direct regulatory interaction between Spalt major and its target DNA in Drosophila. Interestingly, we also find similar sequences in a set of eight novel candidate Spalt target genes, pointing to a common mechanism of transcriptional repression mediated by Spalt proteins.

Functional requirements of protein kinases and phosphatases in the development of the Drosophila melanogaster wing.

Protein kinases and phosphatases constitute a large family of conserved enzymes that control a variety of biological processes by regulating the phosphorylation state of target proteins. They play fundamental regulatory roles during cell cycle progression and signaling, among other key aspects of multicellular development. The complement of protein kinases and phosphatases includes approximately 326 members in Drosophila, and they have been the subject of several functional screens searching for novel components of signaling pathways and regulators of cell division and survival. These approaches have been carried out mostly in cell cultures using RNA interference to evaluate the contribution of each protein in different functional assays and have contributed significantly to assign specific roles to the corresponding genes. In this work, we describe the results of an evaluation of the Drosophila complement of kinases and phosphatases using the wing as a system to identify their functional requirements in vivo. We also describe the results of several modifying screens aiming to identify among the set of protein kinases and phosphatases additional components or regulators of the activities of the epidermal growth factor and insulin receptors signaling pathways.

Genome-wide phenotypic RNAi screen in the Drosophila wing: phenotypic description of functional classes.

The Drosophila genome contains approximately 14,000 protein-coding genes encoding all the necessary information to sustain cellular physiology, tissue organization, organism development, and behavior. In this manuscript, we describe in some detail the phenotypes in the adult fly wing generated after knockdown of approximately 80% of Drosophila genes. We combined this phenotypic description with a comprehensive molecular classification of the Drosophila proteins into classes that summarize the main expected or known biochemical/functional aspect of each protein. This information, combined with mRNA expression levels and in situ expression patterns, provides a simplified atlas of the Drosophila genome, from housekeeping proteins to the components of the signaling pathways directing wing development, that might help to further understand the contribution of each gene group to wing formation.

Genome-wide phenotypic RNAi screen in the Drosophila wing: global parameters.

We have screened a collection of UAS-RNAi lines targeting 10,920 Drosophila protein-coding genes for phenotypes in the adult wing. We identified 3653 genes (33%) whose knockdown causes either larval/pupal lethality or a mutant phenotype affecting the formation of a normal wing. The most frequent phenotypes consist of changes in wing size, vein differentiation, and patterning, defects in the wing margin and in the apposition of the dorsal and ventral wing surfaces. We also defined 16 functional categories encompassing the most relevant aspect of each protein function and assigned each Drosophila gene to one of these functional groups. This allowed us to identify which mutant phenotypes are enriched within each functional group. Finally, we used previously published gene expression datasets to determine which genes are or are not expressed in the wing disc. Integrating expression, phenotypic and molecular information offers considerable precision to identify the relevant genes affecting wing formation and the biological processes regulated by them.

Association Between Dairy Product Consumption and Colorectal Cancer Risk in Adults: A Systematic Review and Meta-Analysis of Epidemiologic Studies.

Dairy product consumption may decrease colorectal cancer (CRC) risk, but very few studies have evaluated the association between different types of dairy products and CRC location. The aim of this systematic review and meta-analysis was to examine the associations between dairy product consumption and CRC incidence. Summary RRs and ORs with 95% CIs were estimated. A total of 15 cohort studies and 14 case-control studies comprising a total of >22,000 cases were included in the quantitative synthesis. The cohort studies showed a consistent significant decrease in CRC risk associated with higher consumption of total dairy products (RR: 0.80; 95% CI: 0.70, 0.91) and total milk (RR: 0.82; 95% CI: 0.76, 0.88) compared with the CRC risk associated with lower consumption. These studies also showed a significant protective association between low-fat milk consumption and CRC (RR: 0.76; 95% CI: 0.66, 0.88), but only for colon cancer (RR: 0.73; 95% CI: 0.61, 0.87). Cheese consumption was inversely associated with the risk of CRC (RR: 0.85; 95% CI: 0.76, 0.96) and proximal colon cancer (RR: 0.74; 95% CI: 0.60, 0.91). No significant associations with CRC were found for the consumption of low-fat dairy products, whole milk, fermented dairy products, or cultured milk. Most of these associations were not supported by the case-control studies. In conclusion, high consumption of total dairy products and total milk was associated with a lower risk of developing CRC at any anatomic location, including the proximal and distal colon and the rectum. Low-fat milk consumption was associated with a lower risk of CRC, but this association was restricted to colon cancer. Cheese consumption was associated with the prevention of CRC, specifically proximal colon cancer. Further studies on larger samples and with longer follow-up periods, along with appropriately designed and executed clinical trials, are warranted to determine whether dairy product consumption affects CRC development.

Association of lifestyle factors and inflammation with sarcopenic obesity: data from the PREDIMED-Plus trial.

BACKGROUND: Sarcopenia is a progressive age-related skeletal muscle disorder associated with increased likelihood of adverse outcomes. Muscle wasting is often accompanied by an increase in body fat, leading to 'sarcopenic obesity'. The aim of the present study was to analyse the association of lifestyle variables such as diet, dietary components, physical activity (PA), body composition, and inflammatory markers, with the risk of sarcopenic obesity. METHODS: A cross-sectional analysis based on baseline data from the PREDIMED-Plus study was performed. A total of 1535 participants (48% women) with overweight/obesity (body mass index: 32.5 ± 3.3 kg/m2 ; age: 65.2 ± 4.9 years old) and metabolic syndrome were categorized according to sex-specific tertiles (T) of the sarcopenic index (SI) as assessed by dual-energy X-ray absorptiometry scanning. Anthropometrical measurements, biochemical markers, dietary intake, and PA information were collected. Linear regression analyses were carried out to evaluate the association between variables. RESULTS: Subjects in the first SI tertile were older, less physically active, showed higher frequency of abdominal obesity and diabetes, and consumed higher saturated fat and less vitamin C than subjects from the other two tertiles (all P < 0.05). Multiple adjusted linear regression models evidenced significant positive associations across tertiles of SI with adherence to the Mediterranean dietary score (P-trend < 0.05), PA (P-trend < 0.0001), and the 30 s chair stand test (P-trend < 0.0001), whereas significant negative associations were found with an inadequate vitamin C consumption (P-trend < 0.05), visceral fat and leucocyte count (all P-trend < 0.0001), and some white cell subtypes (neutrophils and monocytes), neutrophil-to-lymphocyte ratio, and platelet count (all P-trend < 0.05). When models were additionally adjusted by potential mediators (inflammatory markers, diabetes, and waist circumference), no relevant changes were observed, only dietary variables lost significance. CONCLUSIONS: Diet and PA are important regulatory mediators of systemic inflammation, which is directly involved in the sarcopenic process. A healthy dietary pattern combined with exercise is a promising strategy to limit age-related sarcopenia.

Fatty Acids Composition of Blood Cell Membranes and Peripheral Inflammation in the PREDIMED Study: A Cross-Sectional Analysis.

There is limited evidence from epidemiological studies for the inflammatory or anti-inflammatory properties of fatty acids in blood cell membranes. Therefore, this study examined associations between baseline (n = 282) and 1-year (n = 143) changes in the levels of fatty acids in blood cell membranes with circulating inflammatory markers in older adults at high cardiovascular risk. The data for this cross-sectional analysis was obtained from a case-control study within the PREDIMED study. Linear regression with elastic net penalty was applied to test associations between measured fatty acids and inflammatory markers. Several fatty acids were associated with interferon-γ (IFNγ) and interleukins (ILs) IL-6, IL-8, and IL-10 at baseline and additionally also with IL-1b at 1 year. Omega-6 fatty acids were consistently positively associated with pro-inflammatory IL-6 and IL-8 at baseline. Omega-3 fatty acids including C20:5n3 and C18:3n3 were negatively associated with IFN-γ at 1 year. It is interesting to note that the cis and trans forms of C16:1n7 at 1 year were oppositely associated with the inflammatory markers. C16:1n7trans was negatively associated with IFN-γ, IL-6, IL-8, IL-10, and IL-1b, whereas C16:1n7cis was positively associated with IL-1b. This study adds to the growing body of evidence suggesting potential differences in inflammatory or anti-inflammatory properties of fatty acids in blood cell membranes.

Lifestyle factors and visceral adipose tissue: Results from the PREDIMED-PLUS study.

BACKGROUND: Visceral adipose tissue (VAT) is a strong predictor of cardiometabolic health, and lifestyle factors may have a positive influence on VAT depot. This study aimed to assess the cross-sectional associations between baseline levels of physical activity (PA), sedentary behaviours (SB) and adherence to the Mediterranean diet (MedDiet) with VAT depot in older individuals with overweight/obesity and metabolic syndrome. METHODS: Baseline data of the PREDIMED-Plus study including a sample of 1,231 Caucasian men and women aged 55-75 years were used. Levels of leisure-time PA (total, light, and moderate-to-vigorous, in METs·min/day) and SB (total and TV-viewing, in h/day) were evaluated using validated questionnaires. Adherence to the MedDiet was evaluated using a 17-item energy-restricted MedDiet (erMedDiet) screener. The chair-stand test was used to estimate the muscle strength. VAT depot was assessed with DXA-CoreScan. Multivariable adjusted linear regression models were used to evaluate the association between lifestyle factors and VAT. For the statistics we had used multiadjusted linear regression models. RESULTS: Total leisure-time PA (100 METs·min/day: ß -24.3g, -36.7;-11.9g), moderate-to-vigorous PA (ß -27.8g, 95% CI -40.8;-14.8g), chair-stand test (repeat: ß -11.5g, 95% CI -20.1;-2.93g) were inversely associated, and total SB (h/day: ß 38.2g, 95% CI 14.7;61.7) positively associated with VAT. Light PA, TV-viewing time and adherence to an erMedDiet were not significantly associated with VAT. CONCLUSIONS: In older adults with overweigh/obesity and metabolic syndrome, greater PA, muscle strength, and lower total SB were associated with less VAT depot. In this study, adherence to an erMedDiet was not associated with lower VAT.

Leisure-time physical activity at moderate and high intensity is associated with parameters of body composition, muscle strength and sarcopenia in aged adults with obesity and metabolic syndrome from the PREDIMED-Plus study.

AIMS: We aimed to examine the associations of leisure-time physical activity (PA) and sedentary behavior (SB) with the prevalence of sarcopenia, body composition and muscle strength among older adults having overweight/obesity and metabolic syndrome, from the PREDIMED-Plus trial. METHODS: Cross-sectional baseline analysis including 1539 men and women (65 ± 5 y). Sarcopenia was defined as low muscle mass (according to FNIH cut-offs) plus low muscle strength (lowest sex-specific tertile for 30-s chair-stand test). We applied multivariable-adjusted Cox regression with robust variance and constant time (given the cross-sectional design) for the associations of self-reported leisure-time PA and SB with sarcopenia; and multivariable-linear regression for the associations with dual-energy X-ray absorptiometry (DXA)-derived bone mass, fat mass, lean mass and lower-limb muscle strength. RESULTS: Inverse associations were observed between sarcopenia and each hourly increment in total [prevalence ratio 0.81 (95% confidence interval, 0.70, 0.93)], moderate [0.80 (0.66, 0.97)], vigorous [0.51 (0.32, 0.84)], and moderate-vigorous PA (MVPA) [0.74 (0.62, 0.89)]. Incrementing 1-h/day total-PA and MVPA was inversely associated with body-mass-index, waist circumference (WC), fat mass, and positively associated with bone mass and lower-limb muscle strength (all P <.05). One h/day increase in total SB, screen-based SB and TV-viewing was positively associated with body-mass-index, WC and fat mass. Light-PA was not significantly associated with any outcome. CONCLUSIONS: Total-PA and PA at moderate and high intensities may protect against the prevalence of sarcopenia, have a beneficial role on body composition and prevent loss of muscle strength. SB, particularly TV-viewing, may have detrimental effects on body composition in older adults at high cardiovascular risk.

The Effect of Nutrients and Dietary Supplements on Sperm Quality Parameters: A Systematic Review and Meta-Analysis of Randomized Clinical Trials.

Infertility, which affects ∼15% of the world's population, is a global public health issue recognized by the WHO. Therefore, it is of major clinical and public health importance to investigate whether modifiable lifestyle factors-such as stress, drug use, smoking, alcohol intake, and diet-may influence human fertility. A systematic review and meta-analysis of randomized clinical trials (RCTs) from the MEDLINE-PubMed database was conducted to assess the effect of nutrients, dietary supplements, or food on sperm quality parameters. In total, 28 articles were included for qualitative analysis and 15 for quantitative meta-analysis. Total sperm concentrations [expressed as mean differences (MDs); 95% CIs, in spermatozoa (spz)/mL] were increased by selenium (3.91 × 106 spz/mL; 3.08, 4.73 spz/mL), zinc (1.48 × 106 spz/mL; 0.69, 2.27 spz/mL), omega-3 (n-3) fatty acids (10.98 × 106 spz/mL; 10.25, 11.72 spz/mL), and coenzyme Q10 (CoQ10) (5.93 × 106 spz/mL; 5.36, 6.51 spz/mL). Sperm counts were increased by ω-3 fatty acids (18.70 × 106 spz/mL; 16.89, 20.51 spz/mL) and CoQ10 supplementation (10.15 × 106 spz/mL; 8.34, 11.97 spz/mL). Sperm total motility was increased by selenium (3.30%; 2.95%, 3.65%), zinc (7.03%; 6.03%, 8.03%), ω-3 fatty acids (7.55%; 7.09%, 8.01%), CoQ10 (5.30%; 4.98%, 5.62%), and carnitines (7.84%; 6.54%, 9.13%), whereas sperm progressive motility was increased only after supplementation with carnitines (7.45%; 6.24%, 8.67%). Finally, sperm morphology was enhanced by selenium (1.87%; 1.50%, 2.24%), ω-3 fatty acid (0.91%; 0.69%, 1.13%), CoQ10 (1.06%; 0.72%, 1.41%), and carnitine (4.91%; 3.68%, 6.15%) supplementation. This meta-analysis of RCTs suggests that some dietary supplements could beneficially modulate sperm quality parameters and affect male fertility. However, results must be cautiously interpreted due to the limited sample size of the meta-analyzed studies and the considerable observed interstudy heterogeneity.The present study and the corresponding search protocol were registered at the PROSPERO registry at http://www.crd.york.ac.uk/PROSPERO as CRD42017058380.

Cross-sectional associations of objectively-measured sleep characteristics with obesity and type 2 diabetes in the PREDIMED-Plus trial.

Study Objectives: To examine independent and combined associations of sleep duration and sleep variability with body composition, obesity and type 2 diabetes (T2D) in elders at high cardiovascular risk. Methods: Cross-sectional analysis of 1986 community-dwelling elders with overweight/obesity and metabolic syndrome from PREDIMED-Plus trial. Associations of accelerometry-derived sleep duration and sleep variability with body mass index (BMI), waist circumference (WC) and body composition were assessed fitting multivariable-adjusted linear regression models. Prevalence ratios (PR) and 95% confidence intervals (CI) for obesity and T2D were obtained using multivariable-adjusted Cox regression with constant time. "Bad sleepers" (age-specific non-recommended sleep duration plus sleep variability above the median) and "good sleepers" (age-specific recommended sleep duration plus sleep variability below the median) were characterized by combining sleep duration and sleep variability, and their associations with these outcomes were examined. Results: One hour/night increment in sleep duration was inversely associated with BMI (ß -0.38 kg/m2 [95% CI -0.54, -0.23]), WC (ß -0.86 cm [95% CI -1.25, -0.47]), obesity (PR 0.96 [95% CI 0.93, 0.98]), T2D (PR 0.93 [95% CI 0.88, 0.98]) and other DXA-derived adiposity-related measurements (android fat and trunk fat, all p < .05). Each 1-hour increment in sleep variability was positively associated with T2D (PR 1.14 [95% CI 1.01, 1.28]). Compared with "good sleepers," "bad sleepers" were positively associated with obesity (PR 1.12 [95% CI 1.01, 1.24]) and T2D (PR 1.62 [95% CI 1.28, 2.06]). Conclusions: This study revealed cross-sectional associations of sleep duration with adiposity parameters and obesity. Sleep duration and sleep variability were associated with T2D. Considering simultaneously sleep duration and sleep variability could have additional value, particularly for T2D, as they may act synergistically.

Higher dietary glycemic index and glycemic load values increase the risk of osteoporotic fracture in the PREvención con DIeta MEDiterránea (PREDIMED)-Reus trial.

Background: High glucose and insulin concentrations seem to have a negative impact on bone health. However, the relation between the dietary glycemic index (DGI) and the dietary glycemic load (DGL), which has proved to be effective at modulating blood glucose concentrations after carbohydrate consumption, has yet to be explored in relation to bone health. Objective: The aim of the study was to examine the associations between the DGI or DGL and the risk of osteoporotic-related fractures in an elderly Mediterranean population. Design: The study was conducted in 870 subjects aged 55-80 y at high cardiovascular disease risk participating in the PREvención con DIeta MEDiterránea (PREDIMED)-Reus study. The DGI and DGL were estimated from validated food-frequency questionnaires with the use of the international glycemic index and glycemic load values, with glucose as reference. Data on osteoporotic fractures were acquired from a systematic review of medical records. We used Cox proportional hazard models to assess the risk of osteoporotic fracture according to tertiles of average DGI and DGL. Results: A total of 114 new cases of osteoporotic-related fractures were documented after a mean follow-up of 8.9 y. Participants in the highest tertile of DGI and DGL had a significantly higher risk of osteoporotic fractures than those in the lowest tertile after adjusting for potential confounders (HR: 1.80; 95% CI: 1.03, 3.15 and HR: 3.20; 95% CI: 1.25, 8.18, respectively). Conclusions: A high DGI and DGL are associated with a higher risk of osteoporosis-related fractures in an elderly Mediterranean population at high cardiovascular disease risk. This trial was registered at isrctn.com as ISRCTN35739639.

Dietary Magnesium and Cardiovascular Disease: A Review with Emphasis in Epidemiological Studies.

Magnesium (Mg) is an essential dietary element for humans involved in key biological processes. A growing body of evidence from epidemiological studies, randomized controlled trials (RCTs) and meta-analyses have indicated inverse associations between Mg intake and cardiovascular diseases (CVD). The present review aims to summarize recent scientific evidence on the topic, with a focus on data from epidemiological studies assessing the associations between Mg intake and major cardiovascular (CV) risk factors and CVD. We also aimed to review current literature on circulating Mg and CVD, as well as potential biological processes underlying these observations. We concluded that high Mg intake is associated with lower risk of major CV risk factors (mainly metabolic syndrome, diabetes and hypertension), stroke and total CVD. Higher levels of circulating Mg are associated with lower risk of CVD, mainly ischemic heart disease and coronary heart disease. Further, RCTs and prospective studies would help to clarify whether Mg intake and Mg circulating levels may also protect against other CVDs and CVD death.

Legume consumption is inversely associated with type 2 diabetes incidence in adults: A prospective assessment from the PREDIMED study.

BACKGROUND & AIMS: Legumes, a low-energy, nutrient-dense and low glycemic index food, have shown beneficial effects on glycemic control and adiposity. As such, legumes are widely recommended in diabetic diets, even though there is little evidence that their consumption protects against type 2 diabetes. Therefore the aim of the present study was to examine the associations between consumption of total legumes and specific subtypes, and type 2 diabetes risk. We also investigated the effect of theoretically substituting legumes for other protein- or carbohydrate-rich foods. METHODS: Prospective assessment of 3349 participants in the PREvención con DIeta MEDiterránea (PREDIMED) study without type 2 diabetes at baseline. Dietary information was assessed at baseline and yearly during follow-up. We used Cox regression models to estimate hazard ratios (HRs) and 95% confidence intervals (95% CIs) for type-2 diabetes incidence according to quartiles of cumulative average consumption of total legumes, lentils, chickpeas, dry beans and fresh peas. RESULTS: During a median follow-up of 4.3 years, 266 new cases of type 2 diabetes occurred. Individuals in the highest quartile of total legume and lentil consumption had a lower risk of diabetes than those in the lowest quartile (HR: 0.65; 95% CI: 0.43, 0.96; P-trend = 0.04; and HR: 0.67; 95% CI: 0.46-0.98; P-trend = 0.05, respectively). A borderline significant association was also observed for chickpeas consumption (HR 0.68; 95% CI: 0.46, 1.00; P-trend = 0.06). Substitutions of half a serving/day of legumes for similar servings of eggs, bread, rice or baked potato was associated with lower risk of diabetes incidence. CONCLUSIONS: A frequent consumption of legumes, particularly lentils, in the context of a Mediterranean diet, may provide benefits on type 2 diabetes prevention in older adults at high cardiovascular risk. TRIAL REGISTRATION: The trial is registered at http://www.controlled-trials.com (ISRCTN35739639). Registration date: 5th October 2005.

Leisure-time physical activity, sedentary behaviors, sleep, and cardiometabolic risk factors at baseline in the PREDIMED-PLUS intervention trial: A cross-sectional analysis.

Limited data exists on the interrelationships between physical activity (PA), sedentary behaviors and sleep concerning cardiometabolic risk factors in aged adults at high cardiovascular disease risk. Our aim was to examine independent and joint associations between time spent in leisure-time PA, sedentary behaviors and sleep on the prevalence of obesity, type 2 diabetes (T2D) and components of the metabolic syndrome (MetS) in Mediterranean individuals at high cardiovascular risk. Cross-sectional analyses were performed on baseline data from 5776 Spanish adults (aged 55-75y in men; 60-75y in women) with overweight/obesity and MetS, from October 2013 to October 2016, in the PREDIMED-PLUS trial. Employing multivariable-adjusted Cox regression with robust variance and constant time (given the cross-sectional design), higher prevalence of obesity, T2D and abdominal obesity as component of the MetS were associated with greater time in TV-viewing (Relative Risk, RR: 1.02, 95%CI: 1.01, 1.03; RR:1.04, 95%CI: 1.02, 1.06 and RR: 1.01 95%CI: 1.00, 1.02; respectively, all P < .01). Conversely, greater time in moderate-vigorous PA (MVPA) was associated with lower prevalence of obesity, T2D, abdominal obesity and low HDL-cholesterol (RR: 0.95, 95%CI: 0.93, 0.97; RR: 0.94, 95%CI: 0.89, 0.99; RR: 0.97, 95%CI: 0.96, 0.98; and RR: 0.95, 95%CI: 0.91, 0.99, respectively, all P < .05). For these outcomes, theoretically substituting 1-h/day of MVPA for 1-h/day TV-viewing was also significantly associated with lower prevalence (RR 0.91 to 0.97, all P < .05). Similar lower RR in these outcomes was observed when substituting 1-h/day of MVPA for 1-h/day of sleeping. Longer time watching TV and not meeting MVPA recommendations were jointly associated with higher RR of the prevalence of obesity and T2D. We concluded that, in senior individuals at high cardiovascular risk, greater time spent on MVPA and fewer on sedentary behaviors was inversely associated with prevalence of obesity, T2D, and some of the components of MetS.

A Search for Genes Mediating the Growth-Promoting Function of TGFß in the Drosophila melanogaster Wing Disc.

Transforming Growth Factor ß (TGFß) signaling has a complex influence on cell proliferation, acting to stop cell division in differentiating cells, but also promoting cell division in immature cells. The activity of the pathway in Drosophila is mostly required to stimulate the proliferation of neural and epithelial tissues. Most interestingly, this function is not absolutely required for cell division, but it is needed for these tissues to reach their correct size. It is not known how TGFß signaling promotes cell division in imaginal discs, or what the interactions between TGFß activity and other signaling pathways regulating cell proliferation are. In this work, we have explored the disc autonomous function of TGFß that promotes wing imaginal disc growth. We have studied the genetic interactions between TGFß signaling and other pathways regulating wing disc growth, such as the Insulin and Hippo/Salvador/Warts pathways, as well as cell cycle regulators. We have also identified a collection of TGFß candidate target genes affecting imaginal growth using expression profiles. These candidates correspond to genes participating in the regulation of a variety of biochemical processes, including different aspects of cell metabolism, suggesting that TGFß could affect cell proliferation by regulating the metabolic fitness of imaginal cells.

Stepwise reprogramming of liver cells to a pancreas progenitor state by the transcriptional regulator Tgif2.

The development of a successful lineage reprogramming strategy of liver to pancreas holds promises for the treatment and potential cure of diabetes. The liver is an ideal tissue source for generating pancreatic cells, because of its close developmental origin with the pancreas and its regenerative ability. Yet, the molecular bases of hepatic and pancreatic cellular plasticity are still poorly understood. Here, we report that the TALE homeoprotein TGIF2 acts as a developmental regulator of the pancreas versus liver fate decision and is sufficient to elicit liver-to-pancreas fate conversion both ex vivo and in vivo. Hepatocytes expressing Tgif2 undergo extensive transcriptional remodelling, which represses the original hepatic identity and, over time, induces a pancreatic progenitor-like phenotype. Consistently, in vivo forced expression of Tgif2 activates pancreatic progenitor genes in adult mouse hepatocytes. This study uncovers the reprogramming activity of TGIF2 and suggests a stepwise reprogramming paradigm, whereby a 'lineage-restricted' dedifferentiation step precedes the identity switch.

The Spalt Transcription Factors Generate the Transcriptional Landscape of the Drosophila melanogaster Wing Pouch Central Region.

The Drosophila genes spalt major (salm) and spalt-related (salr) encode Zn-finger transcription factors regulated by the Decapentaplegic (Dpp) signalling pathway in the wing imaginal disc. The function of these genes is required for cell survival and proliferation in the central region of the wing disc, and also for vein patterning in the lateral regions. The identification of direct Salm and Salr target genes, and the analysis of their functions, are critical steps towards understanding the genetic control of growth and patterning of the Drosophila wing imaginal disc by the Dpp pathway. To identify candidate Salm/Salr target genes, we have compared the expression profile of salm/salr knockdown wing discs with control discs in microarray experiments. We studied by in situ hybridization the expression pattern of the genes whose mRNA levels varied significantly, and uncovered a complex transcription landscape regulated by the Spalt proteins in the wing disc. Interestingly, candidate Salm/Salr targets include genes which expression is turned off and genes which expression is positively regulated by Salm/Salr. Furthermore, loss-of-function phenotypic analysis of these genes indicates, for a fraction of them, a requirement for wing growth and patterning. The identification and analysis of candidate Salm/Salr target genes opens a new avenue to reconstruct the genetic structure of the wing, linking the activity of the Dpp pathway to the development of this epithelial tissue.

Glimpse into Hox and tale regulation of cell differentiation and reprogramming.

During embryonic development, cells become gradually restricted in their developmental potential and start elaborating lineage-specific transcriptional networks to ultimately acquire a unique differentiated state. Hox genes play a central role in specifying regional identities, thereby providing the cell with critical information on positional value along its differentiation path. The exquisite DNA-binding specificity of the Hox proteins is frequently dependent upon their interaction with members of the TALE family of homeodomain proteins. In addition to their function as Hox-cofactors, TALE homeoproteins control multiple crucial developmental processes through Hox-independent mechanisms. Here, we will review recent findings on the function of both Hox and TALE proteins in cell differentiation, referring mostly to vertebrate species. In addition, we will discuss the direct implications of this knowledge on cell plasticity and cell reprogramming.

Mutually exclusive signaling signatures define the hepatic and pancreatic progenitor cell lineage divergence.

Understanding how distinct cell types arise from multipotent progenitor cells is a major quest in stem cell biology. The liver and pancreas share many aspects of their early development and possibly originate from a common progenitor. However, how liver and pancreas cells diverge from a common endoderm progenitor population and adopt specific fates remains elusive. Using RNA sequencing (RNA-seq), we defined the molecular identity of liver and pancreas progenitors that were isolated from the mouse embryo at two time points, spanning the period when the lineage decision is made. The integration of temporal and spatial gene expression profiles unveiled mutually exclusive signaling signatures in hepatic and pancreatic progenitors. Importantly, we identified the noncanonical Wnt pathway as a potential developmental regulator of this fate decision and capable of inducing the pancreas program in endoderm and liver cells. Our study offers an unprecedented view of gene expression programs in liver and pancreas progenitors and forms the basis for formulating lineage-reprogramming strategies to convert adult hepatic cells into pancreatic cells.