デュシェンヌ型筋ジストロフィー患者由来iPS細胞を用いた初期病態再現

京都大学0048新制・課程博士博士(医学)甲第19229号医博第4028号新制||医||1011(附属図書館)32228京都大学大学院医学研究科医学専攻(主査)教授 髙橋 良輔, 教授 妻木 範行, 教授 井上 治久学位規則第4条第1項該当Doctor of Medical ScienceKyoto UniversityDFA

A Zebrafish Chemical Suppressor Screening Identifies Small Molecule Inhibitors of the Wnt/β-catenin Pathway

SummaryGenetic screening for suppressor mutants has been successfully used to identify important signaling regulators. Using an analogy to genetic suppressor screening, we developed a chemical suppressor screening method to identify inhibitors of the Wnt/β-catenin signaling pathway. We used zebrafish embryos in which chemically induced β-catenin accumulation led to an “eyeless” phenotype and conducted a pilot screening for compounds that restored eye development. This approach allowed us to identify geranylgeranyltransferase inhibitor 286 (GGTI-286), a geranylgeranyltransferase (GGTase) inhibitor. Our follow-up studies showed that GGTI-286 reduces nuclear localization of β-catenin and transcription dependent on β-catenin/T cell factor in mammalian cells. In addition to pharmacological inhibition, GGTase gene knockdown also attenuates the nuclear function of β-catenin. Overall, we validate our chemical suppressor screening as a method for identifying Wnt/β-catenin pathway inhibitors and implicate GGTase as a potential therapeutic target for Wnt-activated cancers

Orexin receptor-1 in the locus coeruleus plays an important role in cue-dependent fear memory consolidation

The noradrenergic (NA) projections arising from the locus ceruleus (LC) to the amygdala and bed nucleus of the stria terminalis have been implicated in the formation of emotional memory. Since NA neurons in the LC (LC-NA neurons) abundantly express orexin receptor-1 (OX1R) and receive prominent innervation by orexin-producing neurons, we hypothesized that an OX1R-mediated pathway is involved in the physiological fear learning process via regulation of LC-NA neurons. To evaluate this hypothesis, we examined the phenotype of Ox1r-/- mice in the classic cued and contextual fear-conditioning test. We found that Ox1r-/- mice showed impaired freezing responses in both cued and contextual fear-conditioning paradigms. In contrast, Ox2r-/- mice showed normal freezing behavior in the cued fear-conditioning test, while they exhibited shorter freezing time in the contextual fear-conditioning test. Double immunolabeling of Fos and tyrosine hydroxylase showed that double-positive LC-NA neurons after test sessions of both cued and contextual stimuli were significantly fewer in Ox1r-/- mice. AAV-mediated expression of OX1R in LC-NA neurons in Ox1r-/- mice restored the freezing behavior to the auditory cue to a comparable level to that in wild-type mice in the test session. Decreased freezing time during the contextual fear test was not affected by restoring OX1R expression in LC-NA neurons. These observations support the hypothesis that the orexin system modulates the formation and expression of fear memory via OX1R in multiple pathways. Especially, OX1R in LC-NA neurons plays an important role in cue-dependent fear memory formation and/or retrieval. © 2013 the authors

Defect of Interferon γ Leads to Impaired Wound Healing through Prolonged Neutrophilic Inflammatory Response and Enhanced MMP-2 Activation.

Interferon (IFN)-γ is mainly secreted by CD4+ T helper 1 (Th1), natural killer (NK) and NKT cells after skin injury. Although IFN-γ is well known regarding its inhibitory effects on collagen synthesis by fibroblasts in vitro, information is limited regarding its role in wound healing in vivo. In the present study, we analyzed how the defect of IFN-γ affects wound healing. Full-thickness wounds were created on the backs of wild type (WT) C57BL/6 and IFN-γ-deficient (KO) mice. We analyzed the percent wound closure, wound breaking strength, accumulation of leukocytes, and expression levels of COL1A1, COL3A1, and matrix metalloproteinases (MMPs). IFN-γKO mice exhibited significant attenuation in wound closure on Day 10 and wound breaking strength on Day 14 after wound creation, characteristics that are associated with prolonged neutrophil accumulation. Expression levels of COL1A1 and COL3A1 mRNA were lower in IFN-γKO than in WT mice, whereas expression levels of MMP-2 (gelatinase) mRNA were significantly greater in IFN-γKO than in WT mice. Moreover, under neutropenic conditions created with anti-Gr-1 monoclonal antibodies, wound closure in IFN-γKO mice was recovered through low MMP-2 expression levels. These results suggest that IFN-γ may be involved in the proliferation and maturation stages of wound healing through the regulation of neutrophilic inflammatory responses

Azimuthal anisotropy of neutral pion production in Au+Au collisions at sqrt(s_NN) = 200 GeV: Path-length dependence of jet quenching and the role of initial geometry

We have measured the azimuthal anisotropy of pi0's for 1 < pT < 18 GeV/c for Au+Au collisions at sqrt s_NN = 200 GeV. The observed anisotropy shows a gradual decrease in 3 < pT < 7 - 10 GeV/c, but remains positive beyond 10 GeV/c. The magnitude of this anisotropy is under-predicted, up to at least 10 GeV/c, by current perturbative QCD (pQCD) energy-loss model calculations. An estimate of the increase in anisotropy expected from initial-geometry modification due to gluon saturation effects and initial-geometry fluctuations is insufficient to account for this discrepancy. Calculations which implement a path length dependence steeper than what is implied by current pQCD energy-loss models, show reasonable agreement with the data.Comment: 384 authors, 6 pages text, 3 figures. Submitted to Phys. Rev. Lett. Plain text data tables for the points plotted in figures for this and previous PHENIX publications are (or will be) publicly available at http://www.phenix.bnl.gov/papers.htm

Transverse momentum and centrality dependence of dihadron correlations in Au+Au collisions at sqrt(s_NN)=200 GeV: Jet-quenching and the response of partonic matter

Azimuthal angle \Delta\phi correlations are presented for charged hadrons from dijets for 0.4 < p_T < 10 GeV/c in Au+Au collisions at sqrt(s_NN) = 200 GeV. With increasing p_T, the away-side distribution evolves from a broad to a concave shape, then to a convex shape. Comparisons to p+p data suggest that the away-side can be divided into a partially suppressed "head" region centered at Delta\phi ~ \pi, and an enhanced "shoulder" region centered at Delta\phi ~ \pi +/- 1.1. The p_T spectrum for the "head" region softens toward central collisions, consistent with the onset of jet quenching. The spectral slope for the "shoulder" region is independent of centrality and trigger p_T, which offers constraints on energy transport mechanisms and suggests that the "shoulder" region contains the medium response to energetic jets.Comment: 420 authors from 58 institutions, 6 pages, 4 figures. Submitted to Physical Review Letters. Plain text data tables for the points plotted in figures for this and previous PHENIX publications are (or will be) publicly available at http://www.phenix.bnl.gov/papers.htm

Transverse momentum dependence of J/psi polarization at midrapidity in p+p collisions at sqrt(s)=200 GeV

We report the measurement of the transverse momentum dependence of inclusive J/psi polarization in p+p collisions at sqrt(s)=200 GeV performed by the PHENIX Experiment at RHIC. The polarization is studied in the helicity, Gottfried-Jackson, and Collins-Soper frames for p_T < 5 GeV/c and |y| < 0.35. The J/psi polarization in the helicity and Gottfried-Jackson frames is consistent with zero for all transverse momenta, with a slight (1.8 sigma) trend towards longitudinal polarization for transverse momenta above 2 GeV/c. No conclusion is allowed due to the limited acceptance in the Collins-Soper frame and the uncertainties of the current data. The results are compared to observations for other collision systems and center of mass energies and to different quarkonia production models.Comment: 384 authors from 62 institutions, 11 pages, 9 figures, 2 tables. v2 is expanded version submitted to Physical Review D. Plain text data tables for the points plotted in figures are available at http://www.phenix.bnl.gov/papers.htm

Particle-species dependent modification of jet-induced correlations in Au+Au collisions at sqrt(s_NN) = 200 GeV

We report PHENIX measurements of the correlation of a trigger hadron at intermediate transverse momentum (2.5<p_{T,trig}<4 GeV/c), with associated mesons or baryons at lower p_{T,assoc}, in Au+Au collisions at sqrt(s_NN) = 200 GeV. The jet correlations for both baryons and mesons show similar shape alterations as a function of centrality, characteristic of strong modification of the away-side jet. The ratio of jet-associated baryons to mesons for this jet increases with centrality and p_{T,assoc} and, in the most central collisions, reaches a value similar to that for inclusive measurements. This trend is incompatible with in-vacuum fragmentation, but could be due to jet-like contributions from correlated soft partons which recombine upon hadronization.Comment: 344 authors, 4 pages text, RevTeX, 4 figures. Submitted to Physical Review Letters. Plain text data tables for the points plotted in figures for this and previous PHENIX publications are (or will be) publicly available at http://www.phenix.bnl.gov/papers.htm

High p_T Direct Photon and pi^0 Triggered Azimuthal Jet Correlations in sqrt(s)=200 GeV p+p Collisions

Correlations of charged hadrons of 1 < pT < 10 GeV/c with high pT direct photons and pi^ 0 mesons in the range 5 <pT < 15 GeV/c are used to study jet fragmentation in the photon+jet and di-jet channels, respectively. The magnitude of the partonic transverse momentum, kT, is obtained by comparing to a model incorporating a Gaussian kT smearing. The sensitivity of the associated charged hadron spectra to the underlying fragmentation function is tested and the data are compared to calculations using recent global fit results. The shape of the direct photon-associated hadron spectrum as well as its charge asymmetry are found to be consistent with a sample dominated by quark-gluon Compton scattering. No significant evidence of fragmentation photon correlated production is observed within experimental uncertainties.Comment: 431 authors, 18 pages, 18 figures, 4 tables, RevTeX-4. Submitted to Physical Review D. Plain text data tables for the points plotted in figures for this and previous PHENIX publications are (or will be) publicly available at http://www.phenix.bnl.gov/papers.htm