Despite advances in therapy, specific subtypes Acute Myeloid Leukaemia (AML) remains largely incurable. The first-in-class RNA Polymerase I (Pol I) inhibitor, CX-5461, has demonstrated promising activity in both haematological malignancies and solid tumours by selectively inhibiting ribosome biogenesis to induce nucleolar stress, a critical vulnerability in rapidly proliferating cancer cells. CX-5461 has undergone clinical trials for the treatment of both solid and haematological malignancies, and 2nd generation Pol I inhibitor PMR-116 has also entered clinical trials, underscoring the translational potential of drugs that target RNA Polymerase I. To enhance the therapeutic efficacy of Pol I inhibition and prevent the emergence of resistance, we conducted a cell line-based unbiased screen of FDA-approved drugs to identify compounds that might synergise with CX-5461. This screen revealed that the pan-CDK inhibitors Dinaciclib and Flavopiridol enhance nucleolar stress pathway (NSP) activation and are strong candidates for combinatorial therapy with CX-5461. Further analysis showed that the combination of CX-5461 and Dinaciclib acts synergistically across a genetically diverse panel of human AML cell lines. This synergy is dependent on an intact NSP, with both agents independently stabilising p53, but with distinct phenotypic outcomes: Dinaciclib induces rapid apoptosis, whereas CX-5461 primarily enforces cell cycle arrest. This functional complementarity results in efficient tumour cell clearance and likely accounts for the delayed onset of therapy resistance. Importantly, combination treatment with CX-5461 and Dinaciclib significantly improved survival in murine models of AML and reduced colony formation in primary human AML samples. Together these findings provide preclinical evidence for a novel combination treatment strategy that leverages nucleolar stress and cell cycle control to enhance treatment outcomes in AML, paving the way for clinical translation of Pol I-CDK co-targeting therapies. ### Competing Interest Statement The authors have declared no competing interest. National Health and Medical Research Council, https://ror.org/011kf5r70, 1158732, 2002741, 1116999, 2009504 Australian Government−s National Collaborative Research Infrastructure Strategy (NCRIS) Coordination for the Improvement of Higher Education Personnel (CAPES) foundation, Brazil

The properties of superheavy elements probe extremes of physics and chemistry. They are synthesised at accelerator laboratories using nuclear fusion, where two atomic nuclei collide, stick together (capture), then with low probability evolve to a compact superheavy nucleus. The fundamental microscopic mechanisms controlling fusion are not fully understood, limiting predictive capability. Even capture, considered to be the simplest stage of fusion, is not matched by models. Here we show that collisions of 40Ca with 208Pb, experience an 'explosion' of mass and charge transfers between the nuclei before capture, with unexpectedly high probability and complexity. Ninety different partitions of the protons and neutrons between the projectile-like and target-like nuclei are observed. Since each is expected to have a different probability of fusion, the early stages of collisions may be crucial in superheavy element synthesis. Our interpretation challenges the current view of fusion, explains both the successes and failures of current capture models, and provides a framework for improved models.

Background: To understand superheavy element synthesis reactions, quantifying the role of quantum shells in quasifission dynamics is important. In reactions with actinide nuclides, a wide peak in the binary quasifission mass yield is seen, centered close to the 208Pb mass. It is generally attributed to the 208Pb spherical closed shells causing a valley in the potential-energy surface, attracting flux to these mass splits. However, an early experiment studying 48Ca, 50Ti+238U reactions showed strong evidence that sequential fission plays an important role in generating the observed peak. These conflicting interpretations have not been resolved up to now.Purpose: This work aims to measure quasifission mass spectra for reactions with nuclei lighter than 208Pb, having negligible sequential fission, to search for systematic features correlated with the proton shells known to affect low-energy fission mass distributions of the same actinide elements.Methods: Systematic measurements have been made at energies near and below the capture barriers (where quasifission is most prominent) of mass-angle distributions for fission following collisions of 48Ti projectiles with even-even nuclides from 154Sm to 200Hg. Mean excitation energies above the ground-states ranged from 51 to 33 MeV, respectively.Results: With increasing compound nucleus atomic number ZCN, a rapid transition occurs from fission having characteristics of fusion-fission to fast quasifission. The heaviest reactions form 240Cf, 244Fm, and 248No. Low -energy fission of neighboring isotopes is mass asymmetric, correlated with proton number Z = 56. However, peak quasifission yields are at mass-symmetry for all reactions. There appears to be a very small (P-3%) systematic excess of yield correlated with Z = 56, however this is at the limit of sensitivity of the experiment.Conclusions: No significant (>3%) systematic features are seen in the quasifission mass spectra that can be unambiguously identified as resulting from shells. This small influence may result from attenuation of shell effects due to the excitation energy introduced, even in these near-barrier reactions giving low excitation energies typical of superheavy element synthesis reactions.

Background: At energies above the capture barrier, coupled-channels (CC) calculations with a standard nuclear potential diffuseness (0.65 fm) increasingly overestimate experimental capture cross section as the charge product (${Z}_{1}{Z}_{2}$) of the colliding nuclei increases. It has been suggested this may be linked to energy dissipation outside the capture barrier.Purpose: We investigate quantitatively the role of energy dissipation in suppressing capture in heavy ion fusion reactions.Method: The yields of sequential fission, including that resulting from deep inelastic collisions, and of fission following capture were determined simultaneously for collisions of $^{18}\mathrm{O}, ^{30}\mathrm{Si}, ^{34}\mathrm{S}$, and $^{40}\mathrm{Ca}+^{232}\mathrm{Th}$ at a range of energies around the respective capture barriers.Results: The ratio of experimental to CC capture cross sections was found to decrease with increasing ${Z}_{1}{Z}_{2}$. Conversely, the ratio of sequential fission to capture-fission increased with increasing ${Z}_{1}{Z}_{2}$. The sum of sequential and capture fission agrees quite well with the CC cross sections.Conclusions: The experimental capture fission and sequential fission cross sections, and their comparison with CC calculations, give a consistent picture that the increase in density overlap at the capture barrier with increasing ${Z}_{1}{Z}_{2}$ of the colliding nuclei is correlated with increasing energy dissipative processes. These compete increasingly strongly with capture as the ${Z}_{1}{Z}_{2}$ of the reaction increases. For the $^{40}\mathrm{Ca}$ reaction, the total fission yield exceeds expectations from capture model calculations, indicating that deep inelastic processes occur both from trajectories that would have led to capture and also from more peripheral trajectories.

The nucleolar surveillance pathway (NSP) monitors nucleolar fidelity and responds to nucleolar stresses (i.e., inactivation of ribosome biogenesis) by mediating the inhibitory binding of ribosomal proteins (RPs) to mouse double minute 2 homolog (MDM2), a nuclear-localised E3 ubiquitin ligase, which results in p53 accumulation. Inappropriate activation of the NSP has been implicated in the pathogenesis of collection of human diseases termed “ribosomopathies”, while drugs that selectively activate the NSP are now in trials for cancer. Despite the clinical significance, the precise molecular mechanism(s) regulating the NSP remain poorly understood. Using genome-wide loss of function screens, we demonstrate the ribosome biogenesis (RiBi) axis as the most potent class of genes whose disruption stabilises p53. Furthermore, we identified a novel suite of genes critical for the NSP, including a novel mammalian protein implicated in 5S ribonucleoprotein particle (5S-RNP) biogenesis, HEATR3. By selectively disabling the NSP, we unexpectedly demonstrate that a functional NSP is required for the ability of all nuclear acting stresses tested, including DNA damage, to robustly induce p53 accumulation. Together, our data demonstrates that the NSP has evolved as the dominant central integrator of stresses that regulate nuclear p53 abundance, thus ensuring RiBi is hardwired to cellular proliferative capacity.

Psi, the sole FUSE Binding Protein (FUBP) family single stranded DNA/RNA binding protein in Drosophila , is essential for proper cell and tissue growth, however its mechanism of function remains unclear. Here we use Targeted DamID combined with RNA-sequencing to generate the first genome-wide binding and expression profiles for Psi. Surprisingly, we demonstrate Psi drives growth in the Drosophila wing through transcriptional repression of key developmental pathways (e.g. Wnt, Notch and TGFβ). Thus, Psi patterns tissue growth by directly repressing transcription of developmental growth suppressors. Analysis of direct Psi targets identified novel growth inhibitors, including Tolkin (Zinc metallopeptidase implicated in TGFβ signalling), Ephexin (Rho-GEF) and emp (CD36 scavenger receptor-related protein). Their depletion not only suppressed impaired growth associated with Psi knockdown, but alone was sufficient to drive wing overgrowth. Thus, Psi drives wing growth twofold, through direct activation of Myc and through transcriptional repression of growth inhibitors comprising core developmental pathways.

A search for production of the superheavy elements with atomic numbers 119 and 120 was performed in the Ti-50 + Bk-249 and Ti-50 + Cf-249 fusion-evaporation reactions, respectively, at the gas-filled recoil separator TASCA at GSI Darmstadt, Germany. Over four months of irradiation, the Bk-249 target partially decayed into Cf-249, which allowed for a simultaneous search for both elements. Neither was detected at cross-section sensitivity levels of 65 and 200 fb for the Ti-50 + Bk-249 and Ti-50 + Cf-249 reactions, respectively, at a midtarget beam energy of E-lab = 281.5 MeV. The nonobservation of elements 119 and 120 is discussed within the concept of fusion-evaporation reactions including various theoretical predictions on the fission-barrier heights of superheavy nuclei in the region of the island of stability.

J. Khuyagbaatar,1,2,* A. Yakushev,2 Ch. E. Düllmann,1,2,3 D. Ackermann,2,† L.-L. Andersson,1 M. Asai,4 M. Block,2 R. A. Boll,5 H. Brand,2 D. M. Cox,6,‡ M. Dasgupta,7 X. Derkx,1,3 A. Di Nitto,3 K. Eberhardt,1,3 J. Even,1,§ M. Evers,7 C. Fahlander,8 U. Forsberg,8 J. M. Gates,9 N. Gharibyan,10 P. Golubev,8 K. E. Gregorich,9 J. H. Hamilton,11 W. Hartmann,2 R.-D. Herzberg,6 F. P. Heßberger,1,2 D. J. Hinde,7 J. Hoffmann,2 R. Hollinger,2 A. Hübner,2 E. Jäger,2 B. Kindler,2 J. V. Kratz,3 J. Krier,2 N. Kurz,2 M. Laatiaoui,2 S. Lahiri,12 R. Lang,2 B. Lommel,2 M. Maiti,12,‖ K. Miernik,5 S. Minami,2 A. Mistry,6,¶ C. Mokry,1,3 H. Nitsche,9,** J. P. Omtvedt,13 G. K. Pang,9 P. Papadakis,6,14 D. Renisch,3 J. Roberto,5 D. Rudolph,8 J. Runke,2 K. P. Rykaczewski,5 L. G. Sarmiento,8 M. Schädel,2,4 B. Schausten,2 A. Semchenkov,13 D. A. Shaughnessy,10 P. Steinegger,15,16 J. Steiner,2 E. E. Tereshatov,10,†† P. Thörle-Pospiech,1,3 K. Tinschert,2 T. Torres De Heidenreich,2 N. Trautmann,3 A. Türler,15,16 J. Uusitalo,14 D. E. Ward,8 M. Wegrzecki,17 N. Wiehl,1,3 S. M. Van Cleve,5 and V. Yakusheva1 1Helmholtz Institute Mainz, 55099 Mainz, Germany 2GSI Helmholtzzentrum für Schwerionenforschung, 64291 Darmstadt, Germany 3Johannes Gutenberg-Universität Mainz, 55099 Mainz, Germany 4Advanced Science Research Center, Japan Atomic Energy Agency, Tokai, Ibaraki 319-1195, Japan 5Oak Ridge National Laboratory, Oak Ridge, Tennessee 37831, USA 6University of Liverpool, Liverpool L69 7ZE, United Kingdom 7The Australian National University, Canberra, ACT 0200, Australia 8Lund University, 22100 Lund, Sweden 9Lawrence Berkeley National Laboratory, Berkeley, California 94720, USA 10Lawrence Livermore National Laboratory, Livermore, California 94551, USA 11Vanderbilt University, Nashville, Tennessee 37235, USA 12Saha Institute of Nuclear Physics, Kolkata 700064, India 13University of Oslo, 0315 Oslo, Norway 14University of Jyväskylä, 40351 Jyväskylä, Finland 15Paul Scherrer Institute, 5232 Villigen, Switzerland 16University of Bern, 3012 Bern, Switzerland 17The Institute of Electron Technology, 02-668 Warsaw, Poland

The three-dimensional organization of the genome contributes to its maintenance and regulation. While chromosomal regions associate with nucleolar ribosomal RNA genes (rDNA), the biological significance of rDNA-genome interactions and whether they are dynamically regulated during disease remain unclear. rDNA chromatin exists in multiple inactive and active states and their transition is regulated by the RNA polymerase I transcription factor UBTF. Here, using a MYC-driven lymphoma model, we demonstrate that during malignant progression the rDNA chromatin converts to the open state, which is required for tumor cell survival. Moreover, this rDNA transition co-occurs with a reorganization of rDNA-genome contacts which correlate with gene expression changes at associated loci, impacting gene ontologies including B-cell differentiation, cell growth and metabolism. We propose that UBTF-mediated conversion to open rDNA chromatin during malignant transformation contributes to the regulation of specific gene pathways that regulate growth and differentiation through reformed long-range physical interactions with the rDNA.

distribution of leukocyte lineages in the BM of CHIP carriers.In patients harboring TET2 mutations.Surprisingly, we did not determine any alterations in HSPCs in patients with a DNMT3A mutation, which contrasts the results in knock-out mouse models, showing an increased stem cell self-renewal.Whether non-mutated HSPCs are also affected in individuals with TET2 mutations in a paracrine, cell-extrinsic fashion, caused by an inflammatory milieu due to altered cytokine production, requires further investigation.

Small RNA pathway components and small RNA profiles of flowering plant egg cells are largely unexplored, mainly because they are not easily accessible but deeply buried inside the ovary. We describe here the utilization of proliferating callus tissue that adopted transcriptome features of Arabidopsis egg cell as a tool to explore small RNA pathway components and small RNA profiles in egg cells. We furthermore complement our studies with mRNA-Seq data from isolated Arabidopsis egg cells and provide data validation by promoter-reporter studies and whole mount in situ hybridization. Sequencing of small RNA libraries demonstrate the predominance of TE-derived siRNAs in the egg cell-related callus. TE-features and expression profiles suggest post-transcriptional silencing of activated Gypsy-like LTR retrotransposons, whereas the majority of class II DNA transposons belonging to Copia, CACTA, hAT-like and Mutator superfamilies are subjected to transcriptional silencing. Small RNA-seq furthermore led to the identification of differentially expressed known and novel miRNAs whose expression in the egg cell was verified by small RNA whole mount in situ hybridization. Both the strong expression of miRNAs in the egg-cell-adjoining synergids and the secretion of miRNAs into the micropyle suggest hitherto undescribed roles for these accessory cells in intercellular communication with the egg cell and the arriving pollen tube. In conclusion, our datasets provide valuable and comprehensive resources to study small RNA pathways and small-RNA-mediated epigenetic reprogramming during egg cell differentiation and the onset of plant embryogenesis.

Khuyagbaatar, J.; Yakushev, A.; Düllmann, Ch E.; Ackermann, D.; Andersson, L. L.; Asai, M.; Block, M.; Boll, R. A.; Brand, H.; Cox, D. M.; Dasgupta, M.; Derkx, X.; Di Nitto, A.; Eberhardt, K.; Even, J.; Evers, M.; Fahlander, C.; Forsberg, U.; Gates, J. M.; Gharibyan, N.; Golubev, P.; Gregorich, K. E.; Hamilton, J. H.; Hartmann, W.; Herzberg, R. D.; Heßberger, F. P.; Hinde, D. J.; Hoffmann, J.; Hollinger, R.; Hübner, A.; Jäger, E.; Kindler, B.; Kratz, J. V.; Krier, J.; Kurz, N.; Laatiaoui, M.; Lahiri, S.; Lang, R.; Lommel, B.; Maiti, M.; Miernik, K.; Minami, S.; Mistry, A.; Mokry, C.; Nitsche, H.; Omtvedt, J. P.; Pang, G. K.; Papadakis, P.; Renisch, D.; Roberto, J.

The heaviest currently known nuclei, which have up to 118 protons, have been produced in Ca-48 induced reactions with actinide targets. Among them, the element tennessine (Ts), which has 117 protons, has been synthesized by fusing Ca-48 with the radioactive target Bk-249, which has a half-life of 327 d. The experiment was performed at the gas-filled recoil separator TASCA. Two long and two short a decay chains were observed. The long chains were attributed to the decay of Ts-294. The possible origin of the short-decay chains is discussed in comparison with the known experimental data. They are found to fit with the decay chain patterns attributed to Ts-293. The present experimental results confirm the previous findings at the Dubna Gas-Filled Recoil Separator on the decay chains originating from the nuclei assigned to Ts.

Background: The formation of superheavy elements (SHEs) by fusion of two massive nuclei is severely inhibited by the competing quasifission process. Lowexcitation energies favor SHE survival against fusion-fission competition. In "cold" fusion with spherical target nuclei near Pb-208, SHE yields are largest at beam energies significantly below the average capture barrier. In "hot" fusion with statically deformed actinide nuclei, this is not the case. Here the elongated deformation-aligned configurations in sub-barrier capture reactions inhibits fusion (formation of a compact compound nucleus), instead favoring rapid reseparation through quasifission. Purpose: To determine the probabilities of fast and slow quasifission in reactions with prolate statically deformed actinide nuclei, through measurement and quantitative analysis of the dependence of quasifission characteristics at beam energies spanning the average capture barrier energy. Methods: The Australian National University Heavy Ion Accelerator Facility and CUBE fission spectrometer have been used to measure fission and quasifission mass and angle distributions for reactions with projectiles from C to S, bombarding Th and U target nuclei. Results: Mass-asymmetric quasifission occurring on a fast time scale, associated with collisions with the tips of the prolate actinide nuclei, shows a rapid increase in probability with increasing projectile charge, the transition being centered around projectile atomic number ZP = 14. For mass-symmetric fission events, deviations of angular anisotropies from expectations for fusion fission, indicating a component of slower quasifission, suggest a similar transition, but centered around ZP similar to 8. Conclusions: Collisions with the tips of statically deformed prolate actinide nuclei show evidence for two distinct quasifission processes of different time scales. Their probabilities both increase rapidly with the projectile charge. The probability of fusion can be severely suppressed by these two quasifission processes, since the sub-barrier heavy element yield is likely to be determined by the product of the probabilities of surviving each quasifission process.

The nucleolus is a dynamic subnuclear compartment that has a number of different functions, but its primary role is to coordinate the production and assembly of ribosomes. For well over 100 years, pathologists have used changes in nucleolar number and size to stage diseases such as cancer. New information about the nucleolus' broader role within the cell is leading to the development of drugs which directly target its structure as therapies for disease. Traditionally, it has been difficult to develop high-throughput image analysis pipelines to measure nucleolar changes due to the broad range of morphologies observed. In this study, we describe a simple high-content image analysis algorithm using Harmony software (PerkinElmer), with a PhenoLOGIC™ machine-learning component, that can measure and classify three different nucleolar morphologies based on nucleolin and fibrillarin staining ("normal," "peri-nucleolar rings" and "dispersed"). We have utilized this algorithm to determine the changes in these classes of nucleolar morphologies over time with drugs known to alter nucleolar structure. This approach could be further adapted to include other parameters required for the identification of new therapies that directly target the nucleolus.

Received 21 June 2017DOI:https://doi.org/10.1103/PhysRevC.96.029901©2017 American Physical SocietyPhysics Subject Headings (PhySH)Research AreasFissionLow & intermediate energy heavy-ion reactionsNuclear Physics

miRNAs play critical roles in heart disease. In addition to differential miRNA expression, miRNA-mediated control is also affected by variable miRNA processing or alternative 3′-end cleavage and polyadenylation (APA) of their mRNA targets. To what extent these phenomena play a role in the heart remains unclear. We sought to explore miRNA processing and mRNA APA in cardiomyocytes, and whether these change during cardiac hypertrophy. Thoracic aortic constriction (TAC) was performed to induce hypertrophy in C57BL/6J mice. RNA extracted from cardiomyocytes of sham-treated, pre-hypertrophic (2days post-TAC), and hypertrophic (7days post-TAC) mice was subjected to small RNA- and poly(A)-test sequencing (PAT-Seq). Differential expression analysis matched expectations; nevertheless we identified ~400 mRNAs and hundreds of noncoding RNA loci as altered with hypertrophy for the first time. Although multiple processing variants were observed for many miRNAs, there was little change in their relative proportions during hypertrophy. PAT-Seq mapped ~48,000 mRNA 3′-ends, identifying novel 3′ untranslated regions (3′UTRs) for over 7000 genes. Importantly, hypertrophy was associated with marked changes in APA with a net shift from distal to more proximal mRNA 3′-ends, which is predicted to decrease overall miRNA repression strength. We independently validated several examples of 3′UTR proportion change and showed that alternative 3′UTRs associate with differences in mRNA translation. Our work suggests that APA contributes to altered gene expression with the development of cardiomyocyte hypertrophy and provides a rich resource for a systems-level understanding of miRNA-mediated regulation in physiological and pathological states of the heart.

To discover new regulatory pathways in B lymphoma cells, we performed a combined analysis of experimental, clinical and global gene expression data. We identified a specific cluster of genes that was coherently expressed in primary lymphoma samples and suppressed by activation of the B cell receptor (BCR) through αIgM treatment of lymphoma cells in vitro. This gene cluster, which we called BCR.1, includes numerous cell cycle regulators. A reduced expression of BCR.1 genes after BCR activation was observed in different cell lines and also in CD10+ germinal center B cells. We found that BCR activation led to a delayed entry to and progression of mitosis and defects in metaphase. Cytogenetic changes were detected upon long-term αIgM treatment. Furthermore, an inverse correlation of BCR.1 genes with c-Myc co-regulated genes in distinct groups of lymphoma patients was observed. Finally, we showed that the BCR.1 index discriminates activated B cell-like and germinal centre B cell-like diffuse large B cell lymphoma supporting the functional relevance of this new regulatory circuit and the power of guided clustering for biomarker discovery.

Background: The presence of neutron transfer channels with positive Q values can enhance sub-barrier fusion cross sections. Recent measurements of the fusion excitation functions for Ni-58+Sn-132,Sn-124 found that the fusion enhancement due to the influence of neutron transfer is smaller than that in Ca-40+Sn-132,Sn-124 although the Q values for multineutron transfer are comparable.Purpose: To investigate the differences observed between the fusion of Sn + Ni and Sn + Ca.Methods: Fusion excitation functions for Ti-46,Ti-50+Sn-124 have been measured at energies near the Coulomb barrier.Results: A comparison of the barrier distributions for Ti-46+Sn-124 and Ca-40+Sn-124 shows that the Ca-40+Sn-124 system has a barrier strength resulting from the coupling to the very collective octupole state in Ca-40 at an energy significantly lower than the uncoupled barrier.Conclusions: The large sub-barrier fusion enhancement in Ca-40 induced reactions is attributed to both couplings to neutron transfer and inelastic excitation, with the octupole vibration of Ca-40 playing a major role.

Background: Fission fragments from heavy ion collisions with actinide nuclei show mass-asymmetric and mass-symmetric components. The relative probabilities of these two components vary rapidly with beam energy with respect to the capture barrier, indicating a strong dependence on the alignment of the deformed nucleus with the partner in the collisions.Purpose: To study the characteristics of the mass-asymmetric quasifission component by reproducing the experimental mass-angle distributions to investigate mass evolution and sticking times.Methods: Fission fragment mass-angle distributions were measured for the S-34 + Th-232 reaction. Simulations to match the measurements were made by using a classical phenomenological approach. Mass ratio distributions and angular distributions of the mass-asymmetric quasifission component were simultaneously fit to constrain the free parameters used in the simulation.Results: The mass-asymmetric quasifission component-predominantly originating from tip (axial) collisions with the prolate deformed Th-232-is found to be peaked near A = 200 at all energies and center-of-mass angles. A Monte Carlo model using the standard mass equilibration time constant of 5.2 x 10(-21) s predicts more symmetric mass splits. Three different hypotheses assuming (i) a mass halt at A = 200, (ii) a slower mass equilibration time, or (iii) a Fermi-type mass drift function reproduced the main experimental features.Conclusions: In tip collisions for the S-34 + Th-232 reaction, mass-asymmetric fission with A similar to 200 is the dominant outcome. The average sticking time is found to be similar to 7 x 10(-21) s, independent of the scenario used for mass evolution.