Emerging concepts in drug discovery for cancer therapy.

These are exciting times to be involved in biomedical research. The transition from a scientific hypothesis to a testable therapy is now faster than ever, and new technologies that facilitate drug development are constantly emerging. The rapid development of COVID-19 vaccines provides a clear example of the current pace of drug discovery and its clinical implementation. Importantly, the wide availability of technologies that speed up drug development has also reached oncology, and investigators can now consider several independent strategies when looking to develop a new therapy. This thematic issue provides an overview of recent advances in cancer drug discovery, in areas such as fragment-based drug development, targeting the DNA damage response or the MYC oncogene, senolytic therapies and computational tools that facilitate drug discovery and the selection of treatments in personalized medicine. We are confident that reading these reviews will help those interested in the development of cancer therapies get a broader and updated view of available opportunities and challenges.Research in OF laboratory is funded by grants from the Spanish Ministry of Science, Innovation and Universities (PID2021-128722OB-I00, co-financed with European FEDER funds), the Spanish Association Against Cancer (AECC; PROYE20101FERN), Cancerfonden (180640) and the Swedish Research Council (538-2014-31).S

Efectos del programa de inducción docente, en el desempeño de los docentes noveles de las regiones de Lima Provincias y Ancash, 2016

La presente investigación titulada: Efectos del programa de inducción docente, en el desempeño de los docentes noveles de las regiones de Lima Provincias y Ancash, 2016; tuvo como objetivo determinar los efectos de la aplicación del programa de inducción docente, en el desempeño de los docentes noveles de las regiones de Lima Provincias y Ancash, en el año 2016. Se utilizó el diseño pre–experimental, la población fue de 90 docentes noveles nombrados en el año 2016 en las II.EE. del Estado, de las regiones de Lima Provincias y Ancash. El tipo de muestreo asumida fue la muestra censal. El instrumento que se utilizó fue la ficha de observación del desempeño del docente novel, elaborado por el Ministerio de Educación (2016) y validado para efecto de este estudio. La conclusión general obtenida es que la aplicación del programa de inducción docente mejora significativamente el desempeño de los docentes noveles de las regiones Lima Provincias y Ancash, en el año 2016.This research entitled: Effects of the teacher induction program on the performance of new teachers in the regions of Lima Provinces and Ancash, 2016; aimed to determine the effects of the application of the teacher induction program on the performance of new teachers in the regions of Lima Provinces and Ancash, in 2016. The pre-experimental design was used, the population was 90 teachers novices appointed in 2016 in the II.EE. of the State, of the regions of Lima Provinces and Ancash. The type of sampling assumed was the census sample. The instrument used was the new teacher's performance observation sheet, prepared by the Ministry of Education (2016) and validated for the purposes of this study. The general conclusion obtained is that the application of the teacher induction program significantly improves the performance of novice teachers in the Lima Provinces and Ancash regions, in 2016

Civil aspects of the judicial sale of real unmovable property in Italy and Spain

La tesi di dottorato ha come oggetto di studio gli aspetti sostanziali della vendita giudiziale da un punto di vista comparatistico tra Spagna e Italia. Si tratta di un tema che non è stato analizzato profondamente dalle più recenti dottrine spagnola e italiana, che è caratterizzato da una notevole complessità tecnica visto il suo forte carattere interdisciplinare (in particolare, processuale-sostanziale) e la cui vigenza attuale è stata intensificata dalla grande quantità di processi di esecuzione che in questo periodo di instabilità economica sono una constante nei Tribunali spagnoli e italiani. Il fatto che costituisca un’istituzione giuridica inquadrata nell’ambito del processo di espropriazione forzata fa sì che questa possieda un accentuato carattere processuale e, infatti, è il legislatore processuale che si occupa per la maggior parte del suo regime giuridico. Tuttavia, al di là di questa sua natura evidentemente processuale, essa produce una serie di effetti sostanziali di enorme importanza, che sono proprio l’oggetto della tesi di dottorato, nello specifico ambito dei beni immobili (considerando la grande importanza economica e pratica dei diritti di questa natura nella nostra realtà socio-economica).The PhD thesis seeks the study of the civil aspects of the judicial sale from a comparative point of view between Italy and Spain. It is a subject which hasn't been deeply analyzed by the recent Italian and Spanish doctrine, and which is characterized by a considerable technical complexity due to its interdisciplinary nature (mainly, procedural and civil). It is also a current issue in this period of economic crisis in which foreclosure processes are a day by day reality of our Courts. The fact of being an institution framed in the foreclosure process outlines its procedural nature, but that cannot hide the fact of producing important civil effects of great importance which are precisely the aim of the PhD thesis

ATM regulates ATR chromatin loading in response to DNA double-strand breaks

DNA double-strand breaks (DSBs) are among the most deleterious lesions that can challenge genomic integrity. Concomitant to the repair of the breaks, a rapid signaling cascade must be coordinated at the lesion site that leads to the activation of cell cycle checkpoints and/or apoptosis. In this context, ataxia telangiectasia mutated (ATM) and ATM and Rad-3–related (ATR) protein kinases are the earliest signaling molecules that are known to initiate the transduction cascade at damage sites. The current model places ATM and ATR in separate molecular routes that orchestrate distinct pathways of the checkpoint responses. Whereas ATM signals DSBs arising from ionizing radiation (IR) through a Chk2-dependent pathway, ATR is activated in a variety of replication-linked DSBs and leads to activation of the checkpoints in a Chk1 kinase–dependent manner. However, activation of the G2/M checkpoint in response to IR escapes this accepted paradigm because it is dependent on both ATM and ATR but independent of Chk2. Our data provides an explanation for this observation and places ATM activity upstream of ATR recruitment to IR-damaged chromatin. These data provide experimental evidence of an active cross talk between ATM and ATR signaling pathways in response to DNA damage

A Genome-wide CRISPR Screen Identifies CDC25A as a Determinant of Sensitivity to ATR Inhibitors

One recurring theme in drug development is to exploit synthetic lethal properties as means to preferentially damage the DNA of cancer cells. We and others have previously developed inhibitors of the ATR kinase, shown to be particularly genotoxic for cells expressing certain oncogenes. In contrast, the mechanisms of resistance to ATR inhibitors remain unexplored. We report here on a genome-wide CRISPR-Cas9 screen that identified CDC25A as a major determinant of sensitivity to ATR inhibition. CDC25A-deficient cells resist high doses of ATR inhibitors, which we show is due to their failure to prematurely enter mitosis in response to the drugs. Forcing mitotic entry with WEE1 inhibitors restores the toxicity of ATR inhibitors in CDC25A-deficient cells. With ATR inhibitors now entering the clinic, our work provides a better understanding of the mechanisms by which these compounds kill cells and reveals genetic interactions that could be used for their rational use.We thank the laboratories of Feng Zhang and Kosuke Yusa for sharing all CRISPR-related plasmids used here through Addgene (plasmids 42230, 50946, and 50947) and Edna Fonseca for her comments on the manuscript. Research was funded by Fundacion Botin, Banco Santander, through its Santander Universities Global Division and by grants from the Spanish Ministry of Economy and Competitiveness (MINECO) (SAF2011-23753 and SAF2014-57791-REDC), Fundacio La Marato de TV3, the Howard Hughes Medical Institute, and the European Research Council (ERC-617840) to O.F.-C.; by a PhD fellowship from La Caixa Foundation to C.M.-R.; by grants from MINECO to S.R. (RYC2011-09242 and SAF2013-49147P, this last project co-financed with European FEDER funds); and by a grant from MINECO (SAF2013-44866-R) to S.O.S

A Chemical Screen Identifies Compounds Capable of Selecting for Haploidy in Mammalian Cells

The recent availability of somatic haploid cell lines has provided a unique tool for genetic studies in mammals. However, the percentage of haploid cells rapidly decreases in these cell lines, which we recently showed is due to their overgrowth by diploid cells present in the cultures. Based on this property, we have now performed a phenotypic chemical screen in human haploid HAP1 cells aiming to identify compounds that facilitate the maintenance of haploid cells. Our top hit was 10-Deacetyl-baccatin-III (DAB), a chemical precursor in the synthesis of Taxol, which selects for haploid cells in HAP1 and mouse haploid embryonic stem cultures. Interestingly, DAB also enriches for diploid cells in mixed cultures of diploid and tetraploid cells, including in the colon cancer cell line DLD-1, revealing a general strategy for selecting cells with lower ploidy in mixed populations of mammalian cells.We would like to thank the members of the O.F.-C. laboratory and MonicaAlvarez-Fernandez for insightful comments and the Transgenic Mice, FlowCytometry, and Confocal Microscopy Units from the CNIO for their technicalhelp. T.O. was funded by a PhD fellowship from Boehringer IngelheimFonds. Research was funded by Fundacion Botı n, Banco Santander throughits Santander Universities Global Division, and by grants from MINECO(SAF2014-57791-REDC and SAF2014-59498-R to O.F.-C., SAF-2013-44866-R to S.O., and SAF2013-49147-P and SAF2016-80874-P to S.R.; pro-jects that were co-financed with ERDF-EU funds) and the EuropeanResearch Council (ERC-617840). Research at the G.d.C. laboratory is fundedby the AECC Scientific Foundation (LABAE16017DECA).S

NSMCE2 suppresses cancer and aging in mice independently of its SUMO ligase activity.

The SMC5/6 complex is the least understood of SMC complexes. In yeast, smc5/6 mutants phenocopy mutations in sgs1, the BLM ortholog that is deficient in Bloom's syndrome (BS). We here show that NSMCE2 (Mms21, in Saccharomyces cerevisiae), an essential SUMO ligase of the SMC5/6 complex, suppresses cancer and aging in mice. Surprisingly, a mutation that compromises NSMCE2-dependent SUMOylation does not have a detectable impact on murine lifespan. In contrast, NSMCE2 deletion in adult mice leads to pathologies resembling those found in patients of BS. Moreover, and whereas NSMCE2 deletion does not have a detectable impact on DNA replication, NSMCE2-deficient cells also present the cellular hallmarks of BS such as increased recombination rates and an accumulation of micronuclei. Despite the similarities, NSMCE2 and BLM foci do not colocalize and concomitant deletion of Blm and Nsmce2 in B lymphocytes further increases recombination rates and is synthetic lethal due to severe chromosome mis-segregation. Our work reveals that SUMO- and BLM-independent activities of NSMCE2 limit recombination and facilitate segregation; functions of the SMC5/6 complex that are necessary to prevent cancer and aging in mice.The authors want to thank Jordi Torres and Mark O'Driscoll for comments on the manuscript. Work in OF laboratory related to this project was supported by Fundacion Botin, by Banco Santander through its Santander Universities Global Division and by grants from MINECO (SAF2011-23753 and SAF2014-57791-REDC), Howard Hughes Medical Institute, and the European Research Council (ERC-617840). Work in JM laboratory was funded by a grant from MINECO (BFU2013-49153P).S

POLD3 Is Haploinsufficient for DNA Replication in Mice

The Pold3 gene encodes a subunit of the Polδ DNA polymerase complex. Pold3 orthologs are not essential in Saccharomyces cerevisiae or chicken DT40 cells, but the Schizosaccharomyces pombe ortholog is essential. POLD3 also has a specialized role in the repair of broken replication forks, suggesting that POLD3 activity could be particularly relevant for cancer cells enduring high levels of DNA replication stress. We report here that POLD3 is essential for mouse development and is also required for viability in adult animals. Strikingly, even Pold3(+/-) mice were born at sub-Mendelian ratios, and, of those born, some presented hydrocephaly and had a reduced lifespan. In cells, POLD3 deficiency led to replication stress and cell death, which were aggravated by the expression of activated oncogenes. Finally, we show that Pold3 deletion destabilizes all members of the Polδ complex, explaining its major role in DNA replication and the severe impact of its deficiency.Research was funded by Fundacion Botin, Banco Santander, through its Santander Universities Global Division, and by grants from the Spanish Ministry of Economy and Competitiveness (MINECO) (SAF2014-59498-R; SAF2014-57791-REDC), Fundacio La Marato de TV3, the Howard Hughes Medical Institute, and the European Research Council (ERC-617840) to O.F.-C.; by a Marie Curie International Outgoing Fellowshp (IOF) from the FP7 Marie Curie Actions and a grant from MINECO (BFU2014-55168-JIN) that was co-funded by European Regional Development Funds (FEDER) to E.L.; by a grant from MINECO (BFU2013-49153) to J.M.; and by the European Commission (ERC grant ONIDDAC) to T.D.H.S

Efficacy of ATR inhibitors as single agents in Ewing sarcoma

Ewing sarcomas (ES) are pediatric bone tumors that arise from a driver translocation, most frequently EWS/FLI1. Current ES treatment involves DNA damaging agents, yet the basis for the sensitivity to these therapies remains unknown. Oncogene-induced replication stress (RS) is a known source of endogenous DNA damage in cancer, which is suppressed by ATR and CHK1 kinases. We here show that ES suffer from high endogenous levels of RS, rendering them particularly dependent on the ATR pathway. Accordingly, two independent ATR inhibitors show in vitro toxicity in ES cell lines as well as in vivo efficacy in ES xenografts as single agents. Expression of EWS/FLI1 or EWS/ERG oncogenic translocations sensitizes non-ES cells to ATR inhibitors. Our data shed light onto the sensitivity of ES to genotoxic agents, and identify ATR inhibitors as a potential therapy for Ewing Sarcomas.We would want to thank Enrique de Alava for providing ES lines. Work in O.F. laboratory was supported by Fundación Botín, by Banco Santander through its Santander Universities Global Division and by grants from MINECO (SAF2014-57791-REDC and SAF2014-59498-R), Fundació La Marato de TV3, Howard Hughes Medical Institute and the European Research Council (ERC-617840). The A.N. laboratory was supported by the Intramural Research Program of the NIH, the National Cancer Institute, the Center for Cancer Research, an Ellison Medical Foundation Senior Scholar in Aging, and the Alex Lemonade Stand Foundation Award. J.A. laboratory is supported by Asociación Pablo Ugarte, ASION-La Hucha de Tomás, Fundación La Sonrisa de Alex and Instituto de Salud Carlos III (PI12/00816 and Spanish Cancer Network RTICC RD12/0036/0027). A.L. laboratory was supported by the Danish National Research Foundation (DNRF115), Danish Council for Independent Research (Sapere Aude, DFF-Starting Grant 2014) and Danish Cancer Society (KBVU-2014).S

On the conservation of the slow conformational dynamics within the amino acid kinase family: NAGK the paradigm

N-Acetyl-L-Glutamate Kinase (NAGK) is the structural paradigm for examining the catalytic mechanisms and dynamics of amino acid kinase family members. Given that the slow conformational dynamics of the NAGK (at the microseconds time scale or slower) may be rate-limiting, it is of importance to assess the mechanisms of the most cooperative modes of motion intrinsically accessible to this enzyme. Here, we present the results from normal mode analysis using an elastic network model representation, which shows that the conformational mechanisms for substrate binding by NAGK strongly correlate with the intrinsic dynamics of the enzyme in the unbound form. We further analyzed the potential mechanisms of allosteric signalling within NAGK using a Markov model for network communication. Comparative analysis of the dynamics of family members strongly suggests that the low-frequency modes of motion and the associated intramolecular couplings that establish signal transduction are highly conserved among family members, in support of the paradigm sequence→structure→dynamics→function © 2010 Marcos et al