A 3′UTR polymorphism modulates mRNA stability of the oncogene and drug target Polo-like Kinase 1

BACKGROUND: The Polo-like Kinase 1 (PLK1) protein regulates cell cycle progression and is overexpressed in many malignant tissues. Overexpression is associated with poor prognosis in several cancer entities, whereby expression of PLK1 shows high inter-individual variability. Although PLK1 is extensively studied, not much is known about the genetic variability of the PLK1 gene. The function of PLK1 and the expression of the corresponding gene could be influenced by genomic variations. Hence, we investigated the gene for functional polymorphisms. Such polymorphisms could be useful to investigate whether PLK1 alters the risk for and the course of cancer and they could have an impact on the response to PLK1 inhibitors. METHODS: The coding region, the 5′ and 3′UTRs and the regulatory regions of PLK1 were systematically sequenced. We determined the allele frequencies and genotype distributions of putatively functional SNPs in 120 Caucasians and analyzed the linkage and haplotype structure using Haploview. The functional analysis included electrophoretic mobility shift assay (EMSA) for detected variants of the silencer and promoter regions and reporter assays for a 3′UTR polymorphism. RESULTS: Four putatively functional polymorphisms were detected and further analyzed, one in the silencer region (rs57973275), one in the core promoter region (rs16972787), one in intron 3 (rs40076) and one polymorphism in the 3′untranslated region (3′UTR) of PLK1 (rs27770). Alleles of rs27770 display different secondary mRNA structures and showed a distinct allele-dependent difference in mRNA stability with a significantly higher reporter activity of the A allele (p < 0.01). CONCLUSION: The present study provides evidence that at least one genomic variant of PLK1 has functional properties and influences expression of PLK1. This suggests polymorphisms of the PLK1 gene as an interesting target for further studies that might affect cancer risk, tumor progression as well as the response to PLK1 inhibitors

Ultracold chemical reactions of a single Rydberg atom in a dense gas

Within a dense environment ($\rho \approx 10^{14}\,$atoms/cm$^3$) at ultracold temperatures ($T < 1\,\mu{}\text{K}$), a single atom excited to a Rydberg state acts as a reaction center for surrounding neutral atoms. At these temperatures almost all neutral atoms within the Rydberg orbit are bound to the Rydberg core and interact with the Rydberg atom. We have studied the reaction rate and products for $nS$ $^{87}$Rb Rydberg states and we mainly observe a state change of the Rydberg electron to a high orbital angular momentum $l$, with the released energy being converted into kinetic energy of the Rydberg atom. Unexpectedly, the measurements show a threshold behavior at $n\approx 100$ for the inelastic collision time leading to increased lifetimes of the Rydberg state independent of the densities investigated. Even at very high densities ($\rho\approx4.8\times 10^{14}\,\text{cm}^{-3}$), the lifetime of a Rydberg atom exceeds $10\,\mu\text{s}$ at $n > 140$ compared to $1\,\mu\text{s}$ at $n=90$. In addition, a second observed reaction mechanism, namely Rb$_2^+$ molecule formation, was studied. Both reaction products are equally probable for $n=40$ but the fraction of Rb$_2^+$ created drops to below 10$\,$% for $n\ge90$.Comment: 13 pages, 13 figure

Impact of Protein Intake during Weight Loss on Preservation of Fat-Free Mass, Resting Energy Expenditure, and Physical Function in Overweight Postmenopausal Women: A Randomized Controlled Trial

Introduction: Weight loss in old age increases the risk of sarcopenia caused by the age-related reduction of fat-free mass (FFM). Due to the strong correlation between FFM and resting energy expenditure (REE), the maintenance of this must also be considered. Besides, the physical function (PF) must be maintained. Objective: The impact of protein intake on changes in FFM, REE, and PF during weight loss in overweight postmenopausal women was investigated. Methods: Fifty-four postmenopausal women (BMI 30.9 ± 3.4; age 59 ± 7 years) were randomized into 2 groups receiving energy-restricted diets with either 0.8 g (normal protein; NP) or 1.5 g protein/kg body weight (high protein; HP) for 12 weeks, followed by a 6-month follow-up phase with an ad libitum food intake. FFM, REE, and PF (strength, endurance, and balance) were measured at baseline, after weight loss, and after follow-up. Results: Forty-six women completed the weight loss intervention and 29 were followed up. The weight loss was –4.6 ± 3.6 kg (HP) and –5.2 ± 3.4 kg (NP; both p &#x3c; 0.001) and the weight regain during follow-up was 1.3 ± 2.8 kg (HP; p = 0.03) and 0.4 ± 2.5 kg (NP; p = 0.39), with no differences between groups. Similar decreases in FFM (–0.9 ± 1.1 [HP] vs. –1.0 ± 1.3 kg [NP]) and REE (–862 ± 569 [HP] vs. –1,000 ± 561 kJ [NP]; both p &#x3c; 0.001) were observed in both groups. During follow-up, no changes in FFM were detected in either group, whereas in the NP group the REE increased again (+138 ± 296; p = 0.02). The main determinants of FFM loss were the energy deficit and the speed of weight loss. In the NP group, the Short Physical Performance Battery score improved with weight loss (+0.6 ± 0.8; p &#x3c; 0.001) and handgrip strength decreased (–1.7 ± 3.4 kg; p &#x3c; 0.001), whereas no changes were observed in the HP group. Conclusions: An HP weight-loss diet without exercise had no impact on preservation of FFM and REE but may help to maintain muscle strength in postmenopausal women

Identification of tumor-specific Salmonella Typhimurium promoters and their regulatory logic

Conventional cancer therapies are often limited in effectiveness and exhibit strong side effects. Therefore, alternative therapeutic strategies are demanded. The employment of tumor-colonizing bacteria that exert anticancer effects is such a novel approach that attracts increasing attention. For instance, Salmonella enterica serovar Typhimurium has been used in many animal tumor models as well as in first clinical studies. These bacteria exhibit inherent tumoricidal effects. In addition, they can be used to deliver therapeutic agents. However, bacterial expression has to be restricted to the tumor to prevent toxic substances from harming healthy tissue. Therefore, we screened an S. Typhimurium promoter-trap library to identify promoters that exclusively drive gene expression in the cancerous tissue. Twelve elements could be detected that show reporter gene expression in tumors but not in spleen and liver. In addition, a DNA motif was identified that appears to be necessary for tumor specificity. Now, such tumor-specific promoters can be used to safely express therapeutic proteins by tumor-colonizing S. Typhimurium directly in the neoplasia

Tumor Invasion of Salmonella enterica Serovar Typhimurium Is Accompanied by Strong Hemorrhage Promoted by TNF-α

BACKGROUND:Several facultative anaerobic bacteria with potential therapeutic abilities are known to preferentially colonize solid tumors after systemic administration. How they efficiently find and invade the tumors is still unclear. However, this is an important issue to be clarified when bacteria should be tailored for application in cancer therapy. METHODOLOGY/PRINCIPAL FINDINGS:We describe the initial events of colonization of an ectopic transplantable tumor by Salmonella enterica serovar Typhimurium. Initially, after intravenous administration, bacteria were found in blood, spleen, and liver. Low numbers were also detected in tumors associated with blood vessels as could be observed by immunohistochemistry. A rapid increase of TNF-alpha in blood was observed at that time, in addition to other pro-inflammatory cytokines. This induced a tremendous influx of blood into the tumors by vascular disruption that could be visualized in H&E stainings and quantified by hemoglobin measurements of tumor homogenate. Most likely, together with the blood, bacteria were flushed into the tumor. In addition, blood influx was followed by necrosis formation, bacterial growth, and infiltration of neutrophilic granulocytes. Depletion of TNF-alpha retarded blood influx and delayed bacterial tumor-colonization. CONCLUSION:Our findings emphasize similarities between Gram-negative tumor-colonizing bacteria and tumor vascular disrupting agents and show the involvement of TNF-alpha in the initial phase of tumor-colonization by bacteria

Einblicke in die bakterielle Tumorbesiedlung

Several facultative anaerobic bacterial strains are known to preferentially accumulate and proliferate within solid tumors when given systemically. Their ability to naturally target to and accumulate in solid tumors suggests their use as transport vehicles that carry therapeutic molecules directly into solid tumors. Although facultative anaerobic bacteria have already been tested in experimental anticancer therapies for several decades, only little is known about the mechanisms that are involved in bacterial colonization of solid tumors and about bacteria-host interactions inside the tumor. The present work deals with several aspects of bacterial tumor colonization and the impacts of bacterial settlement on infiltration and composition of cells of the immune system, on interactions of bacteria with such cells and on influences of these interactions and of the tumor microenvironment on the bacteria and their behavior. The findings of this work do not only shed light on the mechanism of bacterial tumor colonization and bacteria-host interactions inside solid tumors but also explain restrictions of bacteria-mediated gene transfer into solid tumors. Moreover, bacterial dissemination and bacterial accumulation inside solid tumors could be improved by depleting host neutrophils. Further extensions of these findings may help to improve bacterial tumor therapies and may give rise to robust clinically relevant use of bacteria in anticancer therapies.Verschiedene fakultativ anaerobe Bakterien reichern sich nach systemischer Verabreichung bevorzugt in soliden Tumoren an und proliferieren dort. Die Fähigkeit der Bakterien, gezielt Tumore zu besiedeln und sich dort zu akkumulieren, macht sie zu idealen Vektoren für die Tumortherapie. So könnten diese Bakterien beispielsweise als Transporter verwendet werden, die therapeutische Moleküle direkt in den Tumor transportieren. Obwohl fakultativ anaerobe Bakterien bereits seit mehreren Jahrzehnten experimentell für einen Einsatz in der Krebstherapie gestestet werden, ist nur wenig über die Interaktionen zwischen Bakterien und Wirtszellen in Tumor bekannt, oder über die Mechanismen, die bei der bakteriellen Tumorbesiedlung eine Rolle spielen. Die vorliegende Arbeit behandelt unterschiedliche Aspekte der bakteriellen Tumorbesiedlung und deren Auswirkung auf Infiltration und Zusammensetzung der Immunzellen im Tumor, auf Interaktionen der Bakterien mit diesen Zellen sowie den Einfluss dieser Interaktionen und des Tumormicroenvironments auf die Bakterien und auf deren Verhalten. Die vorliegende Arbeit gibt nicht nur neue Einblicke in die bakterielle Tumorbesiedlung, sie zeigt zudem Gründe auf, die erklären, warum bakterienvermittelter Gentransfer in solide Tumore nur bedingt möglich ist. Weiterhin konnte durch die Depletion von neutrophilen Granulozyten die Verteilung der Bakterien im Tumor verbessert und die bakterielle Besiedlung der Tumore verstärkt werden. Auf dieser Basis könnten bakterielle Tumortherapien in Zukunft verbessert werden und so eine klinisch relevante Nutzung der Bakterien in der Krebstherapie ermöglichen