In silico whole-genome screening for cancer-related single-nucleotide polymorphisms located in human mRNA untranslated regions

BACKGROUND: A promising application of the huge amounts of genetic data currently available lies in developing a better understanding of complex diseases, such as cancer. Analysis of publicly available databases can help identify potential candidates for genes or mutations specifically related to the cancer phenotype. In spite of their huge potential to affect gene function, no systematic attention has been paid so far to the changes that occur in untranslated regions of mRNA. RESULTS: In this study, we used Expressed Sequence Tag (EST) databases as a source for cancer-related sequence polymorphism discovery at the whole-genome level. Using a novel computational procedure, we focused on the identification of untranslated region (UTR)-localized non-coding Single Nucleotide Polymorphisms (UTR-SNPs) significantly associated with the tumoral state. To explore possible relationships between genetic mutation and phenotypic variation, bioinformatic tools were used to predict the potential impact of cancer-associated UTR-SNPs on mRNA secondary structure and UTR regulatory elements. We provide a comprehensive and unbiased description of cancer-associated UTR-SNPs that may be useful to define genotypic markers or to propose polymorphisms that can act to alter gene expression levels. Our results suggest that a fraction of cancer-associated UTR-SNPs may have functional consequences on mRNA stability and/or expression. CONCLUSION: We have undertaken a comprehensive effort to identify cancer-associated polymorphisms in untranslated regions of mRNA and to characterize putative functional UTR-SNPs. Alteration of translational control can change the expression of genes in tumor cells, causing an increase or decrease in the concentration of specific proteins. Through the description of testable candidates and the experimental validation of a number of UTR-SNPs discovered on the secreted protein acidic and rich in cysteine (SPARC) gene, this report illustrates the utility of a cross-talk between in silico transcriptomics and cancer genetics

Sphingosine-1-phosphate activates chemokine-promoted myeloma cell adhesion and migration involving α4β1 integrin function

51 p.-7 fig.-1 tab.-2 fig.supl.Myeloma cell adhesion dependent on α4β1 integrin is crucial for the progression of multiple myeloma (MM). The α4β1-dependent myeloma cell adhesion is up-regulated by the chemokine CXCL12, and pharmacological blockade of the CXCL12 receptor CXCR4 leads to defective myeloma cell homing to bone marrow (BM). Sphingosine-1-phosphate (S1P) regulates immune cell trafficking upon binding to G-protein-coupled receptors. Here we show that myeloma cells express S1P1, a receptor for S1P. We found that S1P up-regulated the α4β1-mediated myeloma cell adhesion and transendothelial migration stimulated by CXCL12. S1P promoted generation of high-affinity α4β1 that efficiently bound the α4β1 ligand VCAM-1, a finding that was associated with S1P-triggered increase in talin-β1 integrin association. Furthermore, S1P cooperated with CXCL12 for enhancement of α4β1-dependent adhesion strengthening and spreading. CXCL12 and S1P activated the DOCK2-Rac1 pathway, which was required for stimulation of myeloma cell adhesion involving α4β1. Moreover, in vivo analyses indicated that S1P contributes to optimizing the interactions of MM cells with the BM microvasculture and for their lodging inside the bone marrow. The regulation of α4β1-dependent adhesion and migration of myeloma cells by CXCL12-S1P combined activities might have important consequences for myeloma disease progressionThis study was supported by the Ministerio de Ciencia e Innovación (Grant Nos SAF2011-24022 to JT, SAF2009-07035 to AGP, SAF2009-11037 to AH, RD06/0020/0011 to JT and AGP, RD06/0020/0006 to NCG and MG and PI081825 to MG), the Comunidad de Madrid (Grant No. P2010/BMD-2314 to AGP, JT and AH) and the Fundación de Investigación Médica Mutua Madrileña (to AGP and MG). AH was also funded by the FP7-People-2009-RG (Grant No. 246655), a Ramón y Cajal Fellowship (Grant No. RYC-2007-00697) and the Pro-CNIC Foundation.Peer reviewe

Epidemiología de las lesiones endometriales en mujeres obesas mayores de 50 años / Epidemiology of the endometrial lesions in obese women older than 50 years old

Se realizó una investigación retrospectiva y analítica, con el objetivo de identificar los factores de riesgo asociados a las lesiones endometriales en las mujeres obesas mayores de 50 años en el municipio Pinar del Río. El universo de estudio estuvo constituido por la totalidad de las mujeres residentes en el área de salud Hermanos Cruz. Se seleccionó un grupo de estudio conformado por las mujeres obesas mayores de 50 años y un grupo de control conformado por las mujeres mayores de 50 años no obesas. Para obtener los datos, se utilizó una encuesta-formulario, donde se incluyeron variables generales de la mujer, antecedentes personales y familiares e historia ginecobstétrica, la cual fue aplicada a ambos grupos, con previo consentimiento. Se realizó un estudio ultrasonográfico a las mujeres incluidas en la investigación y uno histológico del endometrio cuando el examen ultrasonográfico resultó anormal (mayor de 4 mm en posmenopáusicas y de 14 mm en premenopáusicas.). Se confeccionó una base de datos con las variables estudiadas. Los resultados obtenidos se agruparon en cuadros (simples y de contingencia). Para su procesamiento estadístico se utilizaron la frecuencia absoluta y porcentual, el estadígrafo Ji cuadrado, pruebas de probabilidad de Fisher, los odds ratio (crudo y ponderado) y Z. La obesidad estuvo asociada con mayor probabilidad de tener un resultado ultrasonográfico anormal de la medida del grosor endometrial y de lesiones premalignas y cáncer endometrial, principalmente cuando existieron antecedentes familiares de cáncer de mama o colon y antecedentes personales de menarquía precoz, paridad y sangrado uterino anormal. Palabras clave: adenocarcinoma, neoplasias, epidemiología,hiperplasia endometrial,biopsia. ABSTRACT A retrospective and analytic research was carried out with the purpose of identifying risk factors associated with endometrial lesions in obese women older than 50 years old in Pinar del Rio municipality. The universe was comprised of the total of women living at "Hermanos Cruz" health sector. A group of obese women older than 50 years old and a control group of non-obese women older than 50 years old were chosen. To collect data, a survey-form was used; including general variables of women, personal, familial, gynecological and obstetric records. The survey was applied for both groups having a previous consent. A sonographic study was conducted to women participating in the research and performing an endometrial-histological examination when the Sonography showed abnormalities (greater 4mm in postmenopausal and 14mm in premenopausal women). A database using the variables studied was created; and the results were grouped in charts (simple and of contingency). In the statistical process absolute frequency and percentage, chi square, Fisher's probability test, odds ratio (raw and adjusted), and Z test were used. Obesity was associated with a major probability of having an abnormal sonographic result of the endometrial thickness, premalignant lesions and endometrial cancer; mainly when familial history of breast or colon cancers existed as well as personal records of early menarche, parity and abnormal uterine bleeding. Key words: Adenocarcinoma, Endometrial neoplasm, epidemiology, Endometrial hyperplasia, biopsy

Initiatives to Counter Fake News in Selected Countries: Argentina, Brazil, Canada, China, Egypt, France, Germany, Israel, Japan, Kenya, Malaysia, Nicaragua, Russia, Sweden, United Kingdom

Comparative Summary by Peter Roudik, Director of Legal Research This report examines the legal approaches of fifteen countries, representing all regions of the world, to the emerging problem of manipulation with “fake news” using mass and social media, especially the impact of fake news on ongoing political processes and elections, and the legislative measures undertaken to counteract the dissemination of false information. Fake news as a phenomenon is not new and has been known since ancient times, but the present-day proliferation of digital and social media platforms, which allow for much broader distribution of information to a global audience, makes the need to counter fake news much more acute. With the exception of Japan, which appears to be the only country in this study where fake news scandals are limited to newspapers and tweeted messages that have no outside influence, a fact explained by the difficulty of the Japanese language for foreigners, the widespread distribution of false information and its impact on decision making and democratic processes is becoming a challenge worldwide. In 2017, a parliamentary committee in Egypt identified the dissemination of 53,000 false rumors over a period of two months. In Germany, 59% of survey participants stated that they had encountered fake news, and in some segments of the population this number was up to almost 80%. In Kenya, a country where 90% of the population has access to high-speed internet, 90% of surveyed users said that they received false or inaccurate information regarding the recent elections through social media. The countries included in this study are addressing the fake news problem through one or more of the following four approaches: • In the absence of legislation that expressly addresses the objectivity of news posted on social media, some of the surveyed countries apply relevant provisions of existing civil, criminal, administrative, and other laws regulating the media, elections, and anti-defamation (Canada, Japan, Nicaragua, Sweden, and the United Kingdom), even though these laws, enacted in the pre-internet era, do not always reflect current technological and telecommunications developments. • Others are choosing to enact new and more focused legislation that imposes sanctions on social media networks that spread false news, usually imposing fines and ordering the removal of information identified as false (China, Egypt, France, Germany, Israel, Malaysia, and Russia). In Malaysia and Egypt these provisions apply extraterritorially. • Another option reflected in the country surveys is to engage election authorities and digital platforms to secure a well-informed electorate, either by identifying and blocking fake news, providing fact-checking resources for the general public, or through the mass publication of “real” news during election season and beyond (Argentina, the UK, China, and Malaysia). Argentina, for example, is considering legislation that would create a Commission for the Verification of Fake News within the National Election Chamber. During national election campaigns, the Commission would recognize, label, and prevent the distribution of news considered “of doubtful credibility.” Both the UK and China have programs in place to systematically rebut fake news by publishing reliable information, while Malaysia provides a fact-checking portal. • Some of the countries are also addressing the issue in a more general way by educating citizens about the dangers of fake news (Sweden and Kenya). Sweden starts at a young age, having enlisted a famous cartoon character to teach children about the dangers of fake news through a cartoon strip that illustrates what happens to the bear’s super-strength when false rumors are circulated about him. The US Embassy in Kenya launched a media literacy campaign in 2018, initially aimed at the Kenya chapter of the Young African Leaders Initiative, with the specific goal of stopping the dissemination of fake news. Among the countries surveyed, there is no common position regarding the definition of “fake news” and its scope. The UK government attempts to avoid use of the term altogether, instead using the words “disinformation” and “misinformation.” Countries with established anti-fake news laws have more elaborate terminology. Malaysian legislation defines fake news as “any news, information, data and reports, which is or are wholly or partly false, whether in the form of features, visuals or audio recordings or in any other form capable of suggesting words or ideas.” Russia passed a law penalizing the publication of fake news in March 2019, defining the term as “socially-significant false information distributed under the guise of truthful messages if they create a threat of endangering people’s lives, health, or property; create possibilities for mass violations of public order or public security; or may hinder the work of transportation and social infrastructure, credit institutions, lines of communications, industry, and energy enterprises.” China has made it a crime to “fabricate false information on [a] dangerous situation, epidemic, disaster or alert and disseminate such information via [an] information network or any other media while clearly knowing that it is fabricated, thereby seriously disturbing public order.” Relying on the 1881 Freedom of the Press Law, France has made it illegal to “disturb public peace through the publication, dissemination, or reproduction of fake news in bad faith.” The bad-faith publication, dissemination, or reproduction of forged or altered items, or items falsely attributed to third parties, is also prohibited. Broad definitions are usually found in the laws of those countries that are rated low in indices related to freedom of speech, and such laws are often viewed by human rights organizations as government attempts to further restrict free speech and stifle opposition. The new Malaysian government tried unsuccessfully to repeal a 2018 act under which the government is required to “take measures to remove” the publication of recognized false information and imprison the publisher for up to six years. In Canada and Kenya courts have found anti-fake news provisions unconstitutional as a violation of freedom of expression and have thus suspended the implementation of such provisions. Following the events of the United States 2016 election campaign, several countries introduced legal mechanisms aimed at protecting the integrity of the democratic process, although depending on country specifics these laws apply to varied actors. In Sweden, the focus was on self-regulation by professional organizations of journalists and other media providers and strengthening ethics rules. This solution followed the European Union’s approach where an EU-wide voluntary Code of Practice on Disinformation has been introduced. However, in view of the inadequacy of voluntary measures taken by social media platforms, Germany enacted the Network Enforcement Act in 2017. While this Act does not create new obligations for social media, it The Law Library of Congress 3 Initiatives to Counter Fake News: Comparative Summary imposes heavy fines for noncompliance with existing legislation and creates rules for the investigation and removal of illegal content hosted by networks with a very large number of registered users. French law also provides for special preventive measures that need to be implemented by operators of large-scale online platforms. Russian law distinguishes between news published by online media, news aggregators, and individual social network users. There are specific rules for the removal of information and the liability of authors, publishers, and internet providers depending on the type of the online platform. Some countries are also taking steps to prevent foreign influence in their national elections. Interesting examples include an Israeli bill targeting foreign propaganda that, if passed, would allow the head of the Central Election Commission, who serves as a Supreme Court justice, to issue injunctions preventing the receipt of prohibited donations, monetary or otherwise, under current law. Under French law, a judge may order any measures necessary to stop the online dissemination of misleading information during the three months preceding an election. During the same period, foreign television broadcasts may be suspended if they deliver false information. The governments of several countries included in this report recognize that a substantive response to disinformation could be an effective way to tackle fake news. The British government’s position is that it is more important to inform citizens of the facts than to simply rebut false information. For this purpose, a Rapid Response Unit within the executive branch monitors news and engages with the public online. In China, a government online platform called “Refuting Rumors” was launched to broadcast “real” news sourced from government agencies and state-owned media. A similar web portal allowing the public to check the authenticity of news found online has been established in Malaysia. In addition, Kenya and Sweden have general education campaigns aimed at young people in place to counter the fake news trend, as noted above. The individual country surveys that follow analyze current and proposed initiatives to limit the spread of false information undertaken at the national level, each country’s challenges associated with these efforts, and efforts undertaken by national governments to secure the validity and accuracy of legal information

Protein Translation Inhibition is Involved in the Activity of the Pan-PIM Kinase Inhibitor PIM447 in Combination with Pomalidomide-Dexamethasone in Multiple Myeloma

Background: Proviral Insertion site for Moloney murine leukemia virus (PIM) kinases are overexpressed in hematologic malignancies, including multiple myeloma. Previous preclinical data from our group demonstrated the anti-myeloma effect of the pan-PIM kinase inhibitor PIM447. Methods: Based on those data, we evaluate here, by in vitro and in vivo studies, the activity of the triple combination of PIM447 + pomalidomide + dexamethasone (PIM-Pd) in multiple myeloma. Results: Our results show that the PIM-Pd combination exerts a potent anti-myeloma effect in vitro and in vivo, where it markedly delays tumor growth and prolongs survival of treated mice. Mechanism of action studies performed in vitro and on mice tumor samples suggest that the combination PIM-Pd inhibits protein translation processes through the convergent inhibition of c-Myc and mTORC1, which subsequently disrupts the function of eIF4E. Interestingly the MM pro-survival factor IRF4 is also downregulated after PIM-Pd treatment. As a whole, all these molecular changes would promote cell cycle arrest and deregulation of metabolic pathways, including glycolysis and lipid biosynthesis, leading to inhibition of myeloma cell proliferation. Conclusions: Altogether, our data support the clinical evaluation of the triple combination PIM-Pd for the treatment of patients with multiple myeloma.This work was supported by funding from Spanish FIS (PI15/00067, PI15/02156 and PI18/01600) and FEDER, AECC (GCB120981SAN), Junta de Castilla y León, Consejería de Sanidad (GRS 862/A/13 and BIO/SA05/14), Fundación Memoria de D. Samuel Solórzano Barruso of the University of Salamanca (FS/22-2015), Fundación Ramón Areces (FRA16/003), Sociedad Española de Hematología y Hemoterapia and Centro en Red de Medicina Regenerativa y Terapia Celular de Castilla y León. E.M.O. was supported by an Inplant grant from IDIVAL. T.P. is supported by a grant from AECC (INVES18043PAÍN)

Stroma-Mediated Resistance to S63845 and Venetoclax through MCL-1 and BCL-2 Expression Changes Induced by miR-193b-3p and miR-21-5p Dysregulation in Multiple Myeloma.

BH3-mimetics targeting anti-apoptotic proteins such as MCL-1 (S63845) or BCL-2 (venetoclax) are currently being evaluated as effective therapies for the treatment of multiple myeloma (MM). Interleukin 6, produced by mesenchymal stromal cells (MSCs), has been shown to modify the expression of anti-apoptotic proteins and their interaction with the pro-apoptotic BIM protein in MM cells. In this study, we assess the efficacy of S63845 and venetoclax in MM cells in direct co-culture with MSCs derived from MM patients (pMSCs) to identify additional mechanisms involved in the stroma-induced resistance to these agents. MicroRNAs miR-193b-3p and miR-21-5p emerged among the top deregulated miRNAs in myeloma cells when directly co-cultured with pMSCs, and we show their contribution to changes in MCL-1 and BCL-2 protein expression and in the activity of S63845 and venetoclax. Additionally, direct contact with pMSCs under S63845 and/or venetoclax treatment modifies myeloma cell dependence on different BCL-2 family anti-apoptotic proteins in relation to BIM, making myeloma cells more dependent on the non-targeted anti-apoptotic protein or BCL-XL. Finally, we show a potent effect of the combination of S63845 and venetoclax even in the presence of pMSCs, which supports this combinatorial approach for the treatment of MM

Expression of p53 protein isoforms predicts survival in patients with multiple myeloma

Loss and/or mutation of the TP53 gene are associated with short survival in multiple myeloma, but the p53 landscape goes far beyond. At least 12 p53 protein isoforms have been identified as a result of a combination of alternative splicing, alternative promoters and/or alternative transcription site starts, which are grouped as α, β, γ, from transactivation domain (TA), long, and short isoforms. Nowadays, there are no studies evaluating the expression of p53 isoforms and its clinical relevance in multiple myeloma (MM). We used capillary nanoimmunoassay to quantify the expression of p53 protein isoforms in CD138‐purified samples from 156 patients with newly diagnosed MM who were treated as part of the PETHEMA/GEM2012 clinical trial and investigated their prognostic impact. Quantitative real‐time polymerase chain reaction was used to corroborate the results at RNA levels. Low and high levels of expression of short and TAp53β/γ isoforms, respectively, were associated with adverse prognosis in MM patients. Multivariate Cox models identified high levels of TAp53β/γ (hazard ratio [HR], 4.49; p < .001) and high‐risk cytogenetics (HR, 2.69; p < .001) as independent prognostic factors associated with shorter time to progression. The current cytogenetic‐risk classification was notably improved when expression levels of p53 protein isoforms were incorporated, whereby high‐risk MM expressing high levels of short isoforms had significantly longer survival than high‐risk patients with low levels of these isoforms. This is the first study that demonstrates the prognostic value of p53 isoforms in MM patients, providing new insights on the role of p53 protein dysregulation in MM biology

Preclinical evaluation of the simultaneous inhibition of MCL-1 and BCL-2 with the combination of S63845 and venetoclax in multiple myeloma

This work was supported by the Spanish ISCIII-FIS and FEDER Funds (PI 15/00067 and PI 15/02156) and the Regional Health Council of Castilla y León (GRS 1604/A/17). EMA was supported by a grant from the Regional Education Council of Castilla y León co-financed by the European Social Fund