Modulation of PKM alternative splicing by PTBP1 promotes gemcitabine resistance in pancreatic cancer cells

Pancreatic ductal adenocarcinoma (PDAC) is an aggressive and incurable disease. Poor prognosis is due to multiple reasons, including acquisition of resistance to gemcitabine, the first-line chemotherapeutic approach. Thus, there is a strong need for novel therapies, targeting more directly the molecular aberrations of this disease. We found that chronic exposure of PDAC cells to gemcitabine selected a subpopulation of cells that are drug-resistant (DR-PDAC cells). Importantly, alternative splicing (AS) of the pyruvate kinase gene (PKM) was differentially modulated in DR-PDAC cells, resulting in promotion of the cancer-related PKM2 isoform, whose high expression also correlated with shorter recurrence-free survival in PDAC patients. Switching PKM splicing by antisense oligonucleotides to favor the alternative PKM1 variant rescued sensitivity of DR-PDAC cells to gemcitabine and cisplatin, suggesting that PKM2 expression is required to withstand drug-induced genotoxic stress. Mechanistically, upregulation of the polypyrimidine-tract binding protein (PTBP1), a key modulator of PKM splicing, correlated with PKM2 expression in DR-PDAC cell lines. PTBP1 was recruited more efficiently to PKM pre-mRNA in DR- than in parental PDAC cells. Accordingly, knockdown of PTBP1 in DR-PDAC cells reduced its recruitment to the PKM pre-mRNA, promoted splicing of the PKM1 variant and abolished drug resistance. Thus, chronic exposure to gemcitabine leads to upregulation of PTBP1 and modulation of PKM AS in PDAC cells, conferring resistance to the drug. These findings point to PKM2 and PTBP1 as new potential therapeutic targets to improve response of PDAC to chemotherapy.Oncogene advance online publication, 3 August 2015; doi:10.1038/onc.2015.270

Effect of disorder on the vortex-lattice melting transition

We use a three dimensional stacked triangular network of Josephson junctions as a model for the study of vortex structure in the mixed state of high Tc superconductors. We show that the addition of disorder destroys the first order melting transition occurring for clean samples. The melting transition splits in two different (continuous) transitions, ocurring at temperatures Ti and Tp (>Ti). At Ti the perpendicular-to-field superconductivity is lost, and at Tp the parallel-to-field superconductivity is lost. These results agree well with recent experiments in YBaCuO.Comment: 4 pages + 2 figure

A new structural class of serine protease inhibitors revealed by the structure of the hirustasin–kallikrein complex

AbstractBackground: Hirustasin belongs to a class of serine protease inhibitors characterized by a well conserved pattern of cysteine residues. Unlike the closely related inhibitors, antistasin/ghilanten and guamerin, which are selective for coagulation factor Xa or neutrophil elastase, hirustasin binds specifically to tissue kallikrein. The conservation of the pattern of cysteine residues and the significant sequence homology suggest that these related inhibitors possess a similar three-dimensional structure to hirustasin.Results: The crystal structure of the complex between tissue kallikrein and hirustasin was analyzed at 2.4 Å resolution. Hirustasin folds into a brick-like structure that is dominated by five disulfide bridges and is sparse in secondary structural elements. The cysteine residues are connected in an abab cdecde pattern that causes the polypeptide chain to fold into two similar motifs. As a hydrophobic core is absent from hirustasin the disulfide bridges maintain the tertiary structure and present the primary binding loop to the active site of the protease. The general structural topography and disulfide connectivity of hirustasin has not previously been described.Conclusions: The crystal structure of the kallikrein–hirustasin complex reveals that hirustasin differs from other serine protease inhibitors in its conformation and its disulfide bond connectivity, making it the prototype for a new class of inhibitor. The disulfide pattern shows that the structure consists of two domains, but only the C-terminal domain interacts with the protease. The disulfide pattern of the N-terminal domain is related to the pattern found in other proteins. Kallikrein recognizes hirustasin by the formation of an antiparallel β sheet between the protease and the inhibitor. The P1 arginine binds in a deep negatively charged pocket of the enzyme. An additional pocket at the periphery of the active site accommodates the sidechain of the P4 valine

First-Order Melting of a Moving Vortex Lattice: Effects of Disorder

We study the melting of a moving vortex lattice through numerical simulations with the current driven 3D XY model with disorder. We find that there is a first-order phase transition even for large disorder when the corresponding equilibrium transition is continuous. The low temperature phase is an anisotropic moving glass.Comment: Important changes from original version. Finite size analysis of results has been added. Figure 2 has been changed. There is a new additional Figure. To be published in Physical Review Letter

Plasticity and memory effects in the vortex solid phase of twinned YBa2Cu3O7 single crystals

We report on marked memory effects in the vortex system of twinned YBa2Cu3O7 single crystals observed in ac susceptibility measurements. We show that the vortex system can be trapped in different metastable states with variable degree of order arising in response to different system histories. The pressure exerted by the oscillating ac field assists the vortex system in ordering, locally reducing the critical current density in the penetrated outer zone of the sample. The robustness of the ordered and disordered states together with the spatial profile of the critical current density lead to the observed memory effects

Direct observation of the washboard noise of a driven vortex lattice in a high-temperature superconductor, Bi2Sr2CaCu2Oy

We studied the conduction noise spectrum in the vortex state of a high-temperature superconductor, Bi2Sr2CaCu2Oy, subject to a uniform driving force. Two characteristic features, a broadband noise (BBN) and a narrow-band noise (NBN), were observed in the vortex-solid phase. The origin of the large BBN was determined to be plastic motion of the vortices, whereas the NBN was found to originate from the washboard modulation of the translational velocity of the driven vortices. We believe this to be the first observation ofComment: 4 pages, 4 figures, to appear in Phys. Rev. Let

Hall Anomaly and Vortex-Lattice Melting in Superconducting Single Crystal YBa2Cu3O7-d

Sub-nanovolt resolution longitudinal and Hall voltages are measured in an ultra pure YBa2Cu3O7-d single crystal. The Hall anomaly and the first-order vortex-lattice melting transition are observed simultaneously. Changes in the dynamic behavior of the vortex solid and liquid are correlated with features of the Hall conductivity sxy. With the magnetic field oriented at an angle from the twin-boundaries, the Hall conductivity sharply decreases toward large negative values at the vortex-lattice melting transition.Comment: 6 pages, 2 figures included, Postscript, to appear in Phys. Rev. Let

Melting and Dimensionality of the Vortex Lattice in Underdoped YBa2Cu3O6.60

Muon spin rotation measurements of the magnetic field distribution in the vortex state of the oxygen deficient high-Tc superconductor YBa{2}Cu{3}O{6.60} reveal a vortex-lattice melting transition at much lower temperature than that in the fully oxygenated material. The transition is best described by a model in which adjacent layers of ``pancake'' vortices decouple in the liquid phase. Evidence is also found for a pinning-induced crossover from a solid 3D to quasi-2D vortex lattice, similar to that observed in the highly anisotropic superconductor Bi{2+x}Sr{2-x}CaCu{2}O{8+y}.Comment: 8 pages, 4 figures, 5 postscript file

Flux-line entanglement as the mechanism of melting transition in high-temperature superconductors in a magnetic field

The mechanism of the flux-line-lattice (FLL) melting in anisotropic high-T_c superconductors in ${\bf B}\parallel {\bf \hat{c}}$ is clarified by Monte Carlo simulations of the 3D frustrated XY model. The percentage of entangled flux lines abruptly changes at the melting temperature T_m, while no sharp change can be found in the number and size distribution of vortex loops around T_m. Therefore, the origin of this melting transition is the entanglement of flux lines. Scaling behaviors of physical quantities are consistent with the above mechanism of the FLL melting. The Lindemann number is also evaluated without any phenomenological arguments.Comment: 10 pages, 5 Postscript figures, RevTeX; changed content and figures, Phys. Rev. B Rapid Commun. in pres

Is there a Phase Transition to the Flux Lattice State?

The sharp drops in the resistance and magnetization which are usually attributed to a phase transition from the vortex liquid state to a crystal state are explained instead as a crossover between three and two dimensional behavior, which occurs when the phase coherence length in the liquid becomes comparable to the sample thickness. Estimates of the width of the crossover region and the phase coherence length scales are in agreement with experiment.Comment: 4 pages, RevTe