The Evidence REVEAL Study: Exploring the Use of Real-World Evidence and Complex Clinical Trial Design by the European Pharmaceutical Industry.

The rapid evolution of science and technology allows innovative approaches to generate new types of evidence about the effectiveness of medical product development so as to speed up patients' access to better diagnostics and treatment. Our study explored how two emerging approaches, the use of real-world evidence (RWE) and complex clinical trial (CCT) design, are currently being used by the pharmaceutical industry to support premarketing authorization of medical product development and reviewed the international landscape for regulatory acceptance of such novel approaches. Combining evidence from a literature review, company survey, and interviews with international regulators and experts, we found that 80% of Europe-based pharmaceutical companies have used RWE and 50% have used CCTs, in some capacity. Further, we present case examples of how companies are using these approaches and how international regulators are preparing for such developments. To conclude, we provide a set of recommendations for European industry and regulators to consider so that these novel approaches achieve their full potential within the EU regulatory system.

Homologous recombination repair intermediates promote efficient de novo telomere addition at DNA double-strand breaks.

The healing of broken chromosomes by de novo telomere addition, while a normal developmental process in some organisms, has the potential to cause extensive loss of heterozygosity, genetic disease, or cell death. However, it is unclear how de novo telomere addition (dnTA) is regulated at DNA double-strand breaks (DSBs). Here, using a non-essential minichromosome in fission yeast, we identify roles for the HR factors Rqh1 helicase, in concert with Rad55, in suppressing dnTA at or near a DSB. We find the frequency of dnTA in rqh1Δ rad55Δ cells is reduced following loss of Exo1, Swi5 or Rad51. Strikingly, in the absence of the distal homologous chromosome arm dnTA is further increased, with nearly half of the breaks being healed in rqh1Δ rad55Δ or rqh1Δ exo1Δ cells. These findings provide new insights into the genetic context of highly efficient dnTA within HR intermediates, and how such events are normally suppressed to maintain genome stability.

Evidence on mechanisms and tools for use of health information for decision-making

The World Health Assembly in 2005 urged Member States to establish or strengthen knowledge transfer mechanisms to support evidence-informed health policies and health care delivery. The European Health Information Initiative was set up to strengthen the use of evidence, information and research for policy-making in the WHO European Region. While good-quality health information is a key component for decision-making, it needs to be packaged and communicated in an effective way to policy-makers, the end-users. This report describes tools and mechanisms that can help to increase the use of health information in policy development. Packaging tools include synthesis methods, such as policy briefs, and visualization methods. Application tools include surveillance data and modelling/simulation to explore the behaviour and performance of processes and interventions. Dissemination and communication tools include health information-sharing platforms, newsletters and person-to-person communications. Finally, linkage and exchange tools such as knowledge networks facilitate the dissemination and refining of health information, thus increasing the chance of its translation into policy.

Evidence on mechanisms and tools for use of health information for decision-making

The World Health Assembly in 2005 urged Member States to establish or strengthen knowledge transfer mechanisms to support evidence-informed health policies and health care delivery. The European Health Information Initiative was set up to strengthen the use of evidence, information and research for policy-making in the WHO European Region. While good-quality health information is a key component for decision-making, it needs to be packaged and communicated in an effective way to policy-makers, the end-users. This report describes tools and mechanisms that can help to increase the use of health information in policy development. Packaging tools include synthesis methods, such as policy briefs, and visualization methods. Application tools include surveillance data and modelling/simulation to explore the behaviour and performance of processes and interventions. Dissemination and communication tools include health information-sharing platforms, newsletters and person-to-person communications. Finally, linkage and exchange tools such as knowledge networks facilitate the dissemination and refining of health information, thus increasing the chance of its translation into policy.

Фактические данные о механизмах и инструментах использования информации здравоохранения для принятия решений

В 2005 г. Всемирная ассамблея здравоохранения призвала государства-члены создать новые или укрепить имеющиеся механизмы для передачи знаний в поддержку проведения научно обоснованной политики и организации медицинской помощи. Для того чтобы поддержать использование фактических данных, информации и научных исследований при выработке политики, в Европейском регионе ВОЗ была основана и введена в действие Европейская инициатива в области информации здравоохранения. Качественная информация здравоохранения является ключевым компонентом в процессе принятия решений, и ее необходимо эффективно «упаковывать» и доносить до руководителей, определяющих политику, которые являются конечными пользователями этой информации. В настоящем докладе описаны инструменты и механизмы, призванные содействовать более активному использованию информации здравоохранения в процессе выработки политики. Так, к инструментам «упаковки» информации относятся методы синтеза (например, составление аналитических записок по вопросам политики) и визуализации. К инструментам применения относятся данные эпиднадзора, а также имитационное моделирование, позволяющее изучать ход и результаты осуществления процессов и вмешательств. Инструменты распространения и коммуникации – это платформы для обмена информацией здравоохранения, новостные бюллетени, а также личное общение. Наконец, инструменты выстраивания связей и обмена, такие как неформальные сети по обмену знаниями, содействуют распространению и уточнению информации здравоохранения, тем самым увеличивая шансы на ее претворение в конкретные меры политики.

Tel1ATM dictates the replication timing of short yeast telomeres.

Telomerase action is temporally linked to DNA replication. Although yeast telomeres are normally late replicating, telomere shortening leads to early firing of subtelomeric DNA replication origins. We show that double-strand breaks flanked by short telomeric arrays cause origin firing early in S phase at late-replicating loci and that this effect on origin firing time is dependent on the Tel1(ATM) checkpoint kinase. The effect of Tel1(ATM) on telomere replication timing extends to endogenous telomeres and is stronger than that elicited by Rif1 loss. These results establish that Tel1(ATM) specifies not only the extent but also the timing of telomerase recruitment.

Tpz1TPP1 SUMOylation reveals evolutionary conservation of SUMO-dependent Stn1 telomere association.

Elongation of the telomeric overhang by telomerase is counteracted by synthesis of the complementary strand by the CST complex, CTC1(Cdc13)/Stn1/Ten1. Interaction of budding yeast Stn1 with overhang-binding Cdc13 is increased by Cdc13 SUMOylation. Human and fission yeast CST instead interact with overhang-binding TPP1/POT1. We show that the fission yeast TPP1 ortholog, Tpz1, is SUMOylated. Tpz1 SUMOylation restricts telomere elongation and promotes Stn1/Ten1 telomere association, and a SUMO-Tpz1 fusion protein has increased affinity for Stn1. Our data suggest that SUMO inhibits telomerase through stimulation of Stn1/Ten1 action by Tpz1, highlighting the evolutionary conservation of the regulation of CST function by SUMOylation.

Protein phosphatase 1 recruitment by Rif1 regulates DNA replication origin firing by counteracting DDK activity.

The firing of eukaryotic origins of DNA replication requires CDK and DDK kinase activities. DDK, in particular, is involved in setting the temporal program of origin activation, a conserved feature of eukaryotes. Rif1, originally identified as a telomeric protein, was recently implicated in specifying replication timing in yeast and mammals. We show that this function of Rif1 depends on its interaction with PP1 phosphatases. Mutations of two PP1 docking motifs in Rif1 lead to early replication of telomeres in budding yeast and misregulation of origin firing in fission yeast. Several lines of evidence indicate that Rif1/PP1 counteract DDK activity on the replicative MCM helicase. Our data suggest that the PP1/Rif1 interaction is downregulated by the phosphorylation of Rif1, most likely by CDK/DDK. These findings elucidate the mechanism of action of Rif1 in the control of DNA replication and demonstrate a role of PP1 phosphatases in the regulation of origin firing.

Failed gene conversion leads to extensive end processing and chromosomal rearrangements in fission yeast.

Loss of heterozygosity (LOH), a causal event in cancer and human genetic diseases, frequently encompasses multiple genetic loci and whole chromosome arms. However, the mechanisms by which such extensive LOH arises, and how it is suppressed in normal cells is poorly understood. We have developed a genetic system to investigate the mechanisms of DNA double-strand break (DSB)-induced extensive LOH, and its suppression, using a non-essential minichromosome, Ch(16), in fission yeast. We find extensive LOH to arise from a new break-induced mechanism of isochromosome formation. Our data support a model in which Rqh1 and Exo1-dependent end processing from an unrepaired DSB leads to removal of the broken chromosome arm and to break-induced replication of the intact arm from the centromere, a considerable distance from the initial lesion. This process also promotes genome-wide copy number variation. A genetic screen revealed Rhp51, Rhp55, Rhp57 and the MRN complex to suppress both isochromosome formation and chromosome loss, in accordance with these events resulting from extensive end processing associated with failed homologous recombination repair.

Break-induced loss of heterozygosity in fission yeast: dual roles for homologous recombination in promoting translocations and preventing de novo telomere addition.

Loss of heterozygosity (LOH), a causal event in tumorigenesis, frequently encompasses multiple genetic loci and whole chromosome arms. However, the mechanisms leading to such extensive LOH are poorly understood. We investigated the mechanisms of DNA double-strand break (DSB)-induced extensive LOH by screening for auxotrophic marker loss approximately 25 kb distal to an HO endonuclease break site within a nonessential minichromosome in Schizosaccharomyces pombe. Extensive break-induced LOH was infrequent, resulting from large translocations through both allelic crossovers and break-induced replication. These events required the homologous recombination (HR) genes rad32(+), rad50(+), nbs1(+), rhp51(+), rad22(+), rhp55(+), rhp54(+), and mus81(+). Surprisingly, LOH was still observed in HR mutants, which resulted predominantly from de novo telomere addition at the break site. De novo telomere addition was most frequently observed in rad22Delta and rhp55Delta backgrounds, which disrupt HR following end resection. Further, levels of de novo telomere addition, while increased in ku70Delta rhp55Delta strains, were reduced in exo1Delta rhp55Delta and an rhp55Delta strain overexpressing rhp51. These findings support a model in which HR prevents de novo telomere addition at DSBs by competing for resected ends. Together, these results suggest that the mechanisms of break-induced LOH may be predicted from the functional status of the HR machinery.

HacA-independent induction of chaperone-encoding gene bipA in Aspergillus niger strains overproducing membrane proteins.

Transcription of two unfolded protein response genes, hacA and bipA, was examined in Aspergillus niger strains overproducing membrane proteins. Despite elevated bipA mRNA levels, no 5'-truncated hacA transcript was detected, raising the possibility of a hacA-independent induction of bipA mRNA under the stress of membrane protein overproduction in A. niger.