Quiescent Fraction Research Articles

THE DISTRIBUTION OF SATELLITES AROUND MASSIVE GALAXIES AT 1 <z< 3 IN ZFOURGE/CANDELS: DEPENDENCE ON STAR FORMATION ACTIVITY

We study the statistical distribution of satellites around star-forming and quiescent central galaxies at 1<z<3 using imaging from the FourStar Galaxy Evolution Survey (ZFOURGE) and the Cosmic Assembly Near-IR Deep Extragalactic Legacy Survey (CANDELS). The deep near-IR data select satellites down to $\log(M/M_\odot)>9$ at z<3. The radial satellite distribution around centrals is consistent with a projected NFW profile. Massive quiescent centrals, $\log(M/M_\odot)>10.78$, have $\sim$2 times the number of satellites compared to star-forming centrals with a significance of 2.7$\sigma$ even after accounting for differences in the centrals' stellar-mass distributions. We find no statistical difference in the satellite distributions of intermediate-mass quiescent and star-forming centrals, $10.48<\log(M/M_\odot)<10.78$. Comparing to the Guo2011 semi-analytic model, the excess number of satellites indicates that quiescent centrals have halo masses 0.3 dex larger than star-forming centrals, even when the stellar-mass distributions are fixed. We use a simple toy model that relates halo mass and quenching, which roughly reproduces the observed quenched fractions and the differences in halo mass between star-forming and quenched galaxies only if galaxies have a quenching probability that increases with halo mass from $\sim$0 for $\log(M_h/M_\odot)\sim$11 to $\sim$1 for $\log(M_h/M_\odot)\sim$13.5. A single halo-mass quenching threshold is unable to reproduce the quiescent fraction and satellite distribution of centrals. Therefore, while halo quenching may be an important mechanism, it is unlikely to be the only factor driving quenching. It remains unclear why a high fraction of centrals remain star-forming even in relatively massive halos.

Human marrow stromal cells downsize the stem cell fraction of lung cancers by fibroblast growth factor 10.

The functional interplay between cancer cells and marrow stromal cells (MSCs) has attracted a great deal of interest due to the MSC tropism for tumors but remains to be fully elucidated. In this study, we investigated human MSC-secreted paracrine factors that appear to have critical functions in cancer stem cell subpopulations. We show that MSC-conditioned medium reduced the cancer stem cell-enriched subpopulation, which was detected as a side population and quiescent (G0) cell cycle fraction in human lung cancer cells by virtue of fibroblast growth factor 10 (FGF10). This reduction of the stem cell-enriched fraction was also observed in lung cancer cells supplemented with recombinant human FGF10 protein. Moreover, supplementary FGF10 attenuated the expression of stemness genes encoding transcription factors, such as OCT3/4 and SOX2, and crippled the self-renewal capacity of lung cancer cells, as evidenced by the impaired formation of floating spheres in the suspension culture. We finally confirmed the therapeutic potential of the FGF10 treatment, which rendered lung cancer cells prone to a chemotherapeutic agent, probably due to the reduced cancer stem cell subpopulation. Collectively, these results add further clarification to the molecular mechanisms underlying MSC-mediated cancer cell kinetics, facilitating the development of future therapies.

OBSERVATIONS OF ENVIRONMENTAL QUENCHING IN GROUPS IN THE 11 GYR SINCEz= 2.5: DIFFERENT QUENCHING FOR CENTRAL AND SATELLITE GALAXIES

We present direct observational evidence for star formation quenching in galaxy groups in the redshift range 0<z<2.5. We utilize a large sample of nearly 6000 groups, selected by fixed cumulative number density from three photometric catalogs, to follow the evolving quiescent fractions of central and satellite galaxies over roughly 11 Gyr. At z~0, central galaxies in our sample range in stellar mass from Milky Way/M31 analogs (M=6.5x10^10 M\solar) to nearby massive ellipticals (M=1.5x10^11 M\solar). Satellite galaxies in the same groups reach masses as low as twice that of the Large Magellanic Cloud (M=6.5x10^9 M\solar). Using statistical background subtraction, we measure the average rest-frame colors of galaxies in our groups and calculate the evolving quiescent fractions of centrals and satellites over seven redshift bins. Our analysis shows clear evidence for star formation quenching in group halos, with a different quenching onset for centrals and their satellite galaxies. Using halo mass estimates for our central galaxies, we find that star formation shuts off in centrals when typical halo masses reach between 10^12 and 10^13 M\solar, consistent with predictions from the halo quenching model. In contrast, satellite galaxies in the same groups most likely undergo quenching by environmental processes, whose onset is delayed with respect to their central galaxy. Although star formation is suppressed in all galaxies over time, the processes that govern quenching are different for centrals and satellites. While mass plays an important role in determining the star formation activity of central galaxies, quenching in satellite galaxies is dominated by the environment in which they reside.

The pre-processing of subhaloes in SDSS groups and clusters

We investigate pre-processing using the observed quenched fraction of group and cluster galaxies in the Yang et al. (2007) SDSS-DR7 group catalogue in the redshift range of 0.01 < z < 0.045. We categorize group galaxies as virialized, infall or backsplash and we apply a combination of the Dressler-Shectman statistic and group member velocities to identify subhaloes. On average the fraction of galaxies that reside in subhaloes is a function of host halo mass, where more massive systems have a higher fraction of subhalo galaxies both in the overall galaxy and infall populations. Additionally, we find that between 2 < r_200 < 3 the quiescent fraction is higher in the subhalo population with respect to both the field and non-subhalo populations. At these large radii (2 < r_200 < 3), the majority of galaxies (~ 80 %) belong to the infall population and therefore, we attribute the enhanced quenching to infalling subhalo galaxies, indicating that pre-processing has occurred in the subhalo population. We conclude that pre-processing plays a significant role in the observed quiescent fraction, but only for the most massive (M_halo > 10^14.5 M_sun) systems in our sample.

The connection between galaxy structure and quenching efficiency

Using data from the SDSS-DR7, including structural measurements from 2D surface brightness fits with GIM2D, we show how the fraction of quiescent galaxies depends on galaxy stellar mass $M_*$, effective radius $R_e$, fraction of $r-$band light in the bulge, $B/T$, and their status as a central or satellite galaxy at $0.01<z<0.2$. For central galaxies we confirm that the quiescent fraction depends not only on stellar mass, but also on $R_e$. The dependence is particularly strong as a function of $M_*/R_e^\alpha$, with $\alpha\sim 1.5$. This appears to be driven by a simple dependence on $B/T$ over the mass range $9 < \log(M_*/M_\odot) < 11.5$, and is qualitatively similar even if galaxies with $B/T>0.3$ are excluded. For satellite galaxies, the quiescent fraction is always larger than that of central galaxies, for any combination of $M_*$, $R_e$ and $B/T$. The quenching efficiency is not constant, but reaches a maximum of $\sim 0.7$ for galaxies with $9 < \log(M_*/M_\odot) < 9.5$ and $R_e<1$ kpc. This is the same region of parameter space in which the satellite fraction itself reaches its maximum value, suggesting that the transformation from an active central galaxy to a quiescent satellite is associated with a reduction in $R_e$ due to an increase in dominance of a bulge component.

THE PAN-STARRS1 MEDIUM-DEEP SURVEY: THE ROLE OF GALAXY GROUP ENVIRONMENT IN THE STAR FORMATION RATE VERSUS STELLAR MASS RELATION AND QUIESCENT FRACTION OUT TOz∼ 0.8

Using a large sample of field and group galaxies drawn from the Pan-STARRS1 Medium-Deep Survey, we present the specific star formation rate (SSFR) - stellar mass (M*) relation, as well as the quiescent fraction versus M* relation in different environments. We confirm that the fraction of quiescent galaxies is strongly dependent on environment at a fixed stellar mass, but that the amplitude and the slope of the star-forming sequence is similar between the field and groups: in other words, the SSFR-density relation at a fixed stellar mass is primarily driven by the change in the star-forming and quiescent fractions between different environments rather than a global suppression in the star formation rate for the star-forming population. However, when we restrict our sample to the cluster-scale environments ($M>10^{14}M_{solar}$), we find a global reduction in the SSFR of the star forming sequence of $17\%$ at 4$\sigma$ confidence as opposed to its field counterpart. After removing the stellar mass dependence of the quiescent fraction seen in field galaxies, the excess in the quiescent fraction due to the environment quenching in groups and clusters is found to increase with stellar mass. We argue that these results are in favor of galaxy mergers to be the primary environment quenching mechanism operating in galaxy groups whereas strangulation is able to reproduce the observed trend in the environment quenching efficiency and stellar mass relation seen in clusters. Our results also suggest that the relative importance between mass quenching and environment quenching depends on stellar mass -- the mass quenching plays a dominant role in producing quiescent galaxies for more massive galaxies, while less massive galaxies are quenched mostly through the environmental effect, with the transition mass around $1-2\times10^{10}M_{solar}$ in the group/cluster environment. (abridged)

Star formation and environmental quenching of GEEC2 group galaxies at z ∼ 1

We present new analysis from the GEEC2 spectroscopic survey of galaxy groups at $0.8<z<1$. Our previous work revealed an intermediate population between the star-forming and quiescent sequences and a strong environmental dependence in the fraction of quiescent galaxies. Only $\sim5$ per cent of star-forming galaxies in both the group and field sample show a significant enhancement in star formation, which suggests that quenching is the primary process in the transition from the star-forming to the quiescent state. To model the environmental quenching scenario, we have tested the use of different exponential quenching timescales and delays between satellite accretion and the onset of quenching. We find that with no delay, the quenching timescale needs to be long in order to match the observed quiescent fraction, but then this model produces too many intermediate galaxies. Fixing a delay time of 3 Gyr, as suggested from the local universe, produces too few quiescent galaxies. The observed fractions are best matched with a model that includes a delay that is proportional to the dynamical time and a rapid quenching timescale ($\sim0.25$ Gyr), but this model also predicts intermediate galaxies H{\delta} strength higher than that observed. Using stellar synthesis models, we have tested other scenarios, such as the rejuvenation of star formation in early-type galaxies and a portion of quenched galaxies possessing residual star formation. If environment quenching plays a role in the GEEC2 sample, then our work suggests that only a fraction of intermediate galaxies may be undergoing this transition and that quenching occurs quite rapidly in satellite galaxies ($\lesssim0.25$ Gyr).

Do group dynamics play a role in the evolution of member galaxies?

We examine galaxy groups from the present epoch to z = 1 to explore the impact of group dynamics on galaxy evolution. We use group catalagues from the Sloan Digital Sky Survey (SDSS), the Group Environment and Evolution Collaboration (GEEC) and the high redshift GEEC2 sample to study how the observed member properties depend on galaxy stellar mass, group dynamical mass and dynamical state of the host group. We find a strong correlation between the fraction of non-star-forming (quiescent) galaxies and galaxy stellar mass, but do not detect a significant difference in the quiescent fraction with group dynamical mass, within our sample halo mass range of 10^13-10^14.5 M_sun, or with dynamical sate. However, at a redshift of approximately 0.4 we do see some evidence that the quiescent fraction in low mass galaxies (log(M_star/M_sun) < 10.5) is lower in groups with substructure. Additionally, our results show that the fraction of groups with non-Gaussian velocity distributions increases with redshift to roughly z = 0.4, while the amount of detected substructure remains constant to z = 1. Based on these results, we conclude that for massive galaxies (log(M_star/M_sun_ > 10.5), evolution is most strongly correlated to the stellar mass of a galaxy with little or no additional effect related to either the group dynamical mass or dynamical state. For low mass galaxies, we do see some evidence of a correlation between the quiescent fraction and the amount of detected substructure, highlighting the need to probe further down the stellar mass function to elucidate the role of the environment in galaxy evolution.

Visualizing the effect of tumor microenvironments on radiation-induced cell kinetics in multicellular spheroids consisting of HeLa cells

In this study, we visualized the effect of tumor microenvironments on radiation-induced tumor cell kinetics. For this purpose, we utilized a multicellular spheroid model, with a diameter of ∼500μm, consisting of HeLa cells expressing the fluorescent ubiquitination-based cell-cycle indicator (Fucci). In live spheroids, a confocal laser scanning microscope allowed us to clearly monitor cell kinetics at depths of up to 60μm. Surprisingly, a remarkable prolongation of G2 arrest was observed in the outer region of the spheroid relative to monolayer-cultured cells. Scale, an aqueous reagent that renders tissues optically transparent, allowed visualization deeper inside spheroids. About 16h after irradiation, a red fluorescent cell fraction, presumably a quiescent G0 cell fraction, became distinct from the outer fraction consisting of proliferating cells, most of which exhibited green fluorescence indicative of G2 arrest. Thereafter, the red cell fraction began to emit green fluorescence and remained in prolonged G2 arrest. Thus, for the first time, we visualized the prolongation of radiation-induced G2 arrest in spheroids and the differences in cell kinetics between the outer and inner fractions.

Galaxy evolution in groups and clusters: satellite star formation histories and quenching time-scales in a hierarchical Universe

Satellite galaxies in groups and clusters are more likely to have low star formation rates (SFR) and lie on the red-sequence than central (field) galaxies. Using galaxy group/cluster catalogs from SDSS DR7, together with a cosmological N-body simulation to track satellite orbits, we examine the star formation histories and quenching timescales of satellites of M_star > 5 x 10^9 M_sun at z=0. We first explore satellite infall histories: group preprocessing and ejected orbits are critical aspects of satellite evolution, and properly accounting for these, satellite infall typically occurred at z~0.5, or ~5 Gyr ago. To obtain accurate initial conditions for the SFRs of satellites at their time of first infall, we construct an empirical parametrization for the evolution of central galaxy SFRs and quiescent fractions. With this, we constrain the importance and efficiency of satellite quenching as a function of satellite and host halo mass, finding that satellite quenching is the dominant process for building up all quiescent galaxies at M_star < 10^10 M_sun. We then constrain satellite star formation histories, finding a 'delayed-then-rapid' quenching scenario: satellite SFRs evolve unaffected for 2-4 Gyr after infall, after which star formation quenches rapidly, with an e-folding time of < 0.8 Gyr. These quenching timescales are shorter for more massive satellites but do not depend on host halo mass: the observed increase in satellite quiescent fraction with halo mass arises simply because of satellites quenching in a lower mass group prior to infall (group preprocessing), which is responsible for up to half of quenched satellites in massive clusters. Because of the long time delay before quenching starts, satellites experience significant stellar mass growth after infall, nearly identical to central galaxies. This fact provides key physical insight into the subhalo abundance matching method.

Modified MuDPIT Separation Identified 4488 Proteins in a System-wide Analysis of Quiescence in Yeast

A modified multidimensional protein identification technology (MudPIT) separation was coupled to an LTQ Orbitrap Velos mass spectrometer and used to rapidly identify the near-complete yeast proteome from a whole cell tryptic digest. This modified online two-dimensional liquid chromatography separation consists of 39 strong cation exchange steps followed by a short 18.5 min reversed-phase (RP) gradient. A total of 4269 protein identifications were made from 4189 distinguishable protein families from yeast during log phase growth. The "Micro" MudPIT separation performed as well as a standard MudPIT separation in 40% less gradient time. The majority of the yeast proteome can now be routinely covered in less than a days' time with high reproducibility and sensitivity. The newly devised separation method was used to detect changes in protein expression during cellular quiescence in yeast. An enrichment in the GO annotations "oxidation reduction", "catabolic processing" and "cellular response to oxidative stress" was seen in the quiescent cellular fraction, consistent with their long-lived stress resistant phenotypes. Heterogeneity was observed in the stationary phase fraction with a less dense cell population showing reductions in KEGG pathway categories of "Ribosome" and "Proteasome", further defining the complex nature of yeast populations present during stationary phase growth. In total, 4488 distinguishable protein families were identified in all cellular conditions tested.

Targeting Chronic Myeloid Leukemia Stem Cells

Chronic myeloid leukemia (CML) is a clonal myeloproliferative disorder that is characterized by the presence of the fusion oncogene BCR-ABL that encodes the tyrosine kinase BCR-ABL. Constitutive expression of BCR-ABL leads to the unregulated production of mature myeloid cells in the bone marrow and their subsequent release into the blood. Untreated, CML will progress from a chronic to accelerated phase over a number of years before quickly proceeding to a terminal blast crisis phase, reminiscent of acute leukemia. The advent of tyrosine kinase inhibitors has led to much improved management of the disease, but these drugs do not provide a cure as they are unable to eradicate the most primitive, quiescent fraction of CML stem cells. This review looks at recent research into targeting CML stem cells and focuses on major signalling pathways of interest.

Thrombopoietin-Receptor Signalling Induces Proliferation of Dormant HSC.

AbstractAbstract 2343Lifelong blood production is maintained by a very rare population of self-renewing hematopoietic stem cells (HSCs) in bone marrow (BM). Proliferation, differentiation and survival of HSC toward stepwise hematopoietic cell development needs to be tightly controlled by cell intrinsic and extrinsic factors, as excess or insufficient production of mature blood cells potentially leads to neoplasia or aplasia. HSCs and progenitors (HSPCs) are equipped with cell surface receptors for different cytokines or chemokines (Kaushansky, NEJM 2006), and thus can integrate external signals, leading finally to proliferation and subsequent increase of hematopoietic cells in demand. Some of these regulatory pathways are already exploited in clinical settings: CXCR4 antagonists for HSPC mobilization, human granulocyte colony-stimulating factor (hG-CSF) for HSC mobilization and myeloid regeneration, and thrombopoietin agonists (THPO) for improving thrombocytopenia. However, despite their clinical use, little is known about the influence of these molecules on HSC.We established in vivo HSC divisional tracking with CFSE (5(6)-carboxyfluorescein diacetate N-succinimidyl ester). This allows to track single HSC division with high resolution, and subsequently to test biological activity of HSC-containing fractions (LKS) with different divisional history (Takizawa et al., JEM 2011). Using this system we evaluated the effects of systemic administration of human fms-related tyrosine kinase 3 ligand (hFlt3L), hG-CSF (Filgrastim), CXCR4 antagonist (AMD3100), and the THPO receptor (cMpl) agonist (Romiplostim) on HSC division. CFSE-labeled LKS were transferred into non-irradiated steady-state recipients. The non-dividing cell fraction was defined by the CFSE profile of CD4+CD62L+ T cells. One week after transplantation mice were injected with PBS or respective reagents daily or every other day for over one week. Three weeks after transfer, phenotypic BM analysis demonstrated that most of donor LKS had undergone several divisions while a small fraction of LKS remained undivided in PBS treated control mice (Figure 1a), containing long term self-renewing HSCs with at least 20–30% frequency (Takizawa et al., JEM 2011). Administration of hFlt3L increased the percentage of intermediate (1–5x divided) and fast cycling (>5x divided) LKS, which mainly contains CD150- Flt3+ multipotent progenitor cells (Figure 1b). Upon injections of cMpl agonist all donor LKS divided more than once, leaving no cells in quiescent fraction, with substantial expansion of CD150+ cells in the divided fraction. CXCR4 antagonist and hG-CSF administration had little effect on LKS proliferation. These data suggest that cMpl agonist drives dormant cells into proliferation, whereas hG-CSF has little effect on LKS division.To determine whether cMpl agonist increases the turnover of functionally defined, bona fide HSCs, we performed secondary transplantation of 0–1, 2–4, and ≥5x divided LKS. Twenty fast- (≥5x divided cells at 3 weeks), slow-cycling (2–4x divided) or relatively dormant LKS Flt3- cMpl+ cells (0–1x divided) were sorted from mice treated with PBS or cMpl agonist, and transplanted into lethally irradiated mice. Early results demonstrate increased percentage of secondary recipient engrafted with 2–4x divided cells from primary animals treated with cMpl agonist compared to those cells from PBS treated controlOur results thus suggest that cMpl agonists have mitogenic activity not only for megakaryocyte progenitors but also for HSCs. How far this holds true in the human species needs to be determined. However, it should be taken in consideration given clinical data on evolution of pre-existing clonal myeloid diseases under cMpl agonist treatment (Dantoni, ASH abstract 2011), and also when treatment is applied long-term to patients with primary non-clonal hematopoietic diseases as immune thrombocytopenia or aplastic anemia. [Display omitted] Disclosures:No relevant conflicts of interest to declare.

EVOLUTION OF QUIESCENT AND STAR-FORMING GALAXIES SINCEz∼ 1.5 AS A FUNCTION OF THEIR VELOCITY DISPERSIONS

We measure stellar masses and structural parameters for 5,500 quiescent and 20,000 star-forming galaxies at 0.3<z\leq1.5 in the Newfirm Medium Band Survey COSMOS and UKIDSS UDS fields. We combine these measurements to infer velocity dispersions and determine how the number density of galaxies at fixed inferred dispersion, or the Velocity Dispersion Function (VDF), evolves with time for each population. We show that the number of galaxies with high velocity dispersions appears to be surprisingly stable with time, regardless of their star formation history. Furthermore, the overall VDF for star-forming galaxies is constant with redshift, extending down to the lowest velocity dispersions probed by this study. The only galaxy population showing strong evolution are quiescent galaxies with low inferred dispersions, whose number density increases by a factor of ~4 since z=1.5. This build-up leads to an evolution in the quiescent fraction of galaxies such that the threshold dispersion above which quiescent galaxies dominate the counts moves to lower velocity dispersion with time. We show that our results are qualitatively consistent with a simple model in which star-forming galaxies quench and are added to the quiescent population. In order to compensate for the migration into the quiescent population, the velocity dispersions of star-forming galaxies must increase, with a rate that increases with dispersion.

THE PHYSICAL ORIGINS OF THE MORPHOLOGY-DENSITY RELATION: EVIDENCE FOR GAS STRIPPING FROM THE SLOAN DIGITAL SKY SURVEY

We provide a physical interpretation and explanation of the morphology-density relation for galaxies, drawing on stellar masses, star formation rates, axis ratios and group halo masses from the Sloan Digital Sky Survey. We first re-cast the classical morphology-density relation in more quantitative terms, using low star formation rate (quiescence) as a proxy for early-type morphology and dark matter halo mass from a group catalog as a proxy for environmental density: for galaxies of a given stellar mass the quiescent fraction is found to increase with increasing dark matter halo mass. Our novel result is that - at a given stellar mass - quiescent galaxies are significantly flatter in dense environments, implying a higher fraction of disk galaxies. Supposing that the denser environments differ simply by a higher incidence of quiescent disk galaxies that are structurally similar to star-forming disk galaxies of similar mass, explains simultaneously and quantitatively these quiescence -nvironment and shape-environment relations. Our findings add considerable weight to the slow removal of gas as the main physical driver of the morphology-density relation, at the expense of other explanations.

The impact of respiratory motion on tumor quantification and delineation in static PET/CT imaging

Our aim is to investigate the impact of respiratory motion on tumor quantification and delineation in static PET/CT imaging using a population of patient respiratory traces. A total of 1295 respiratory traces acquired during whole body PET/CT imaging were classified into three types according to the qualitative shape of their signal histograms. Each trace was scaled to three diaphragm motion amplitudes (6 mm, 11 mm and 16 mm) to drive a whole body PET/CT computer simulation that was validated with a physical phantom experiment. Three lung lesions and one liver lesion were simulated with diameters of 1 cm and 2 cm. PET data were reconstructed using the OS-EM algorithm with attenuation correction using CT images at the end-expiration phase and respiratory-averaged CT. The errors of the lesion maximum standardized uptake values (SUVmax) and lesion volumes between motion-free and motion-blurred PET/CT images were measured and analyzed. For respiration with 11 mm diaphragm motion and larger quiescent period fraction, respiratory motion can cause a mean lesion SUVmax underestimation of 28% and a mean lesion volume overestimation of 130% in PET/CT images with 1 cm lesions. The errors of lesion SUVmax and volume are larger for patient traces with larger motion amplitudes. Smaller lesions are more sensitive to respiratory motion than larger lesions for the same motion amplitude. Patient respiratory traces with relatively larger quiescent period fraction yield results less subject to respiratory motion than traces with long-term amplitude variability. Mismatched attenuation correction due to respiratory motion can cause SUVmax overestimation for lesions in the lower lung region close to the liver dome. Using respiratory-averaged CT for attenuation correction yields smaller mismatch errors than those using end-expiration CT. Respiratory motion can have a significant impact on static oncological PET/CT imaging where SUV and/or volume measurements are important. The impact is highly dependent upon motion amplitude, lesion location and size, attenuation map and respiratory pattern. To overcome the motion effect, motion compensation techniques may be necessary in clinical practice to improve the tumor quantification for determining the response to therapy or for radiation treatment planning.

Hypoxia-Adapted CML Cells Are More Primitive Population and Are Eradicated by Glyoxalase-1 Inhibitors.

Abstract 2166Poster Board II-143Normal hematopoietic stem cells reside in the epiphyses of the bone marrow that is a low oxygenated area and are protected against ROS-induced DNA damages. Recent study showed that the hypoxic environment plays a crucial role not only in maintaining stem cells but also in tumorigenesis. Hypoxia induces dramatical changes in cell characteristics including cell cycle quiescence, self renewal potency, and shift in energy production from an aerobic to anaerobic pathway, and it induces resistance to a variety of cell death stimuli. Chronic myelogenous leukemia (CML) is a disorder of hematopoietic stem cells caused by the constitutive activation of the Bcr-Abl tyrosine kinase. Tyrosine kinase inhibitors (TKIs) have led to marked improvement in prognosis of CML patients. However, CML cells could not be eradicated completely by TKI alone because quiescent CML stem cells are less sensitive to such molecular target drugs. Therefore, we speculate that the adaptation of leukemic cells to hypoxia in the bone marrow niche alters their characteristics contributing to minimal residual disease.We first evaluated the oxygen levels of engrafted leukemic cells by pimonidazol (hypoxia specific marker) staining.We transplanted K562 cells to the bone marrow of NOD/SCID/gcnull (NOG) mice and found that those cells engrafted and survived in the epiphysis of the bone marrow where O2 concentrations are less than 1.3%. Then, we generated two hypoxia-adapted (HA) CML subclones from K562 and KCL22 by cultivating under 1.0% O2, and were denoted as K562/HA and KCL22/HA, respectively. Both cell lines survived and proliferated continuously for years under 1.0% O2conditions, although their growth was slower than that of their parental counterparts under 20% O2conditions. Interestingly, HA-CML cells exhibited several unique characteristics compared to their parental cells. First, these HA cells showed higher transplantation efficacy in NOG mice. The transplanted HA cells grow more rapidly in vivo than the parental cells and mice transplanted with HA cells died earlier. Next, the percentage of G0 fractions in K562 and K562/HA cells were 0.87 ± 0.58 % and 4.9 ± 2.1 %, respectively, indicating that K562/HA cells included more quiescent fractions than the parental K562. Hoechst staining analysis confirmed that HA cell lines include more SP (side population) fractions than their parental cells, indicating that HA cells contains more dormant cells. We next examined the signaling pathway of HA cell lines. Despite the unchanged levels of AKT, STAT, and ERK phosphrylation, BCR-ABL phsophrylation was suppressed in HA cells. Both of HA cell lines showed higher expression of b-catenin which is considered essential for survival and self-renewality of CML stem cells. Furthermore, HA cells were less sensitive to TKIs (imatinib, dasatinib, and bafetinib) and chemotherapeutic agents (daunorubicin and busulfan). Taken together, our HA cell lines have characteristics of more primitive CML cell populations resistant to cytotoxic agents.We next examined the energy metabolites such as adenosine triphosphate (ATP), glucose consumption, and lactate production in HA cells. The amounts of ATP in K562/HA and KCL22/HA cells decreased, whereas glucose consumption and lactate production increased compared with those in their parental cell lines. These findings indicate that ATP production of HA cells depends on glycolysis. Furthermore, we found higher expression and kinase activity of Glyoxalase-1 (Glo-I). Glo-I is an enzyme that detoxifies glycolysis-specific cytocidal byproducts in glycolytis system. Glo-I inhibitors such as S-p-bromobenzylglutathione cyclopentyl diester (BBGC), 2-crotonyloxymethyl-4,5,6-trihydroxycylohex-2-enone (COTC), and methyl-gerfelin were much more cytotoxic against HA-CML cells than their parental cells in vitro. Notably, when K562/HA-transplanted mice were treated with 100 mg/kg/day BBGC for 8 days, the treated mice survived longer than the untreated mice (Figure 1). These findings suggest that Glo-1 plays an important role in primitive CML cells survival under hypoxia. [Display omitted] In conclusion, Glo-1 is a novel attractive target against hypoxia-adapted primitive CML cells in the bone marrow milieu. Investigation of hypoxia-specific pathways and roles on CML cells could develop novel therapeutic approach targeting TKIs resistance. Disclosures:No relevant conflicts of interest to declare.

Expansion of Normal and Leukemic Hematopoietic Progenitor Cells by PTH Requires bFGF Activation of c-Kit and Its Downstream JAK2/STAT5 Signaling.

Abstract 2511Poster Board II-488 Introduction:The signaling mechanisms which orchestrate hematopoietic stem and progenitor cell (HSPC) expansion, and proliferation of leukemic progenitor cells are not fully understood. In vivo parathyroid hormone (PTH) treatment was shown to expand HSPC via activation of bone forming osteoblasts (Calvi et al., Nature, 2003). PTH induced bone remodeling via activation of both osteoblasts and osteoclasts is a basic fibroblast growth factor (bFGF) dependent event (Okada et al., JBC, 2003; Hurley et al., BBRC, 2006). Different FGF receptors are expressed on various sub-types of HSPC and also on bone marrow (BM) stromal cells. This ligand, alone or in combination with other cytokines, promotes ex vivo expansion of both human and murine HSPC. Interestingly, bFGF mediated expansion of HSPC in vitro, in a culture of total BM cells, but not of purified Sca-1+/c-Kit+/Lineage− (SKL) murine HSPC (Yeoh et al., Stem Cells, 2006), implying that HSPC expansion by bFGF is also mediated indirectly via the microenvironment. KG1a is a human CD34+ progenitor AML cell line which harbors a mutation due to a chromosomal translocation, resulting in a constitutively active isoform of the FGF receptor (Gu et al., Blood, 2006). We used this cell line to elucidate the mechanism behind FGF mediated expansion of human CD34+ progenitors. The goal of this study is to reveal the roles of PTH and bFGF in regulation of normal and leukemic HSPC expansion in vivo and the downstream signaling machinery. Results:In order to understand how cytokines control HSPC proliferation and expansion in vivo, we repeatedly injected PTH to wild type (WT) and bFGF knock-out (KO) mice (5 injections per week) for 4 weeks. We observed that PTH administration to WT mice increased the levels of primitive SKL progenitors in the BM and spleen, decreased the percentage of G0 quiescent SKL cells, and upregulated c-Kit expression on the Lin−/Sca-1+ primitive progenitor population. Surprisingly, these effects were not observed in bFGF KO mice treated with PTH, revealing that PTH induced expansion of HSPC requires bFGF signaling. Repeated bFGF injections for only 1 week (total of 7 injections), significantly increased the numbers and frequency of hematopoietic progenitor cells (HPC) with only a minor effect on differentiated white blood cell numbers in the murine BM and spleen. HSPC expansion by bFGF was accompanied with splenomegaly, a phenotype resembling myelo-proliferative like diseases. In addition we found that bFGF downregulated SDF-1 and upregulated SCF expression on BM stromal cells together with upregulation of c-Kit expression on Lin−/Sca-1+ primitive progenitors. Inhibition of FGF signaling in KG1a cells using a specific FGFR inhibitor reduced STAT5 phosphorylation levels and increased the G0 quiescent cell fraction. Moreover, specific inhibitors of FGFR, JAK2 and STAT5, completely blocked human leukemic KG1a proliferation, implying for the involvement of this signaling pathway in expansion of immature human CD34+ AML cells. Conclusions:Our preliminary results reveal that bFGF is down stream of PTH induced HSPC expansion in vivo and that this ligand can expand progenitor cells in their natural BM and spleen microenvironments. bFGF rapidly mediates higher expansion of murine SKL HSPC within 1 week as compared to PTH expansion within 4 weeks. PTH and bFGF induced expansion is mediated both directly and indirectly by upregulating c-Kit receptors on HSPC and its ligand, SCF expression by stromal cells. Our in vitro experiments with malignant human KG1a cells indicate that FGF signaling activates STAT5 via JAK2 to promote expansion of both normal and leukemic progenitor cells. This activation is most probably mediated via SCF/c-Kit signaling since AML cells express high levels of these molecules (Ramenghi et al., Stem Cells, 1994) and therefore may bypass the dependency on the stromal microenvironment. Further studies will provide insights on how PTH/bFGF and their downstream mediators like c-Kit and SCF control normal and leukemic HSPC expansion and their evolving stromal microenvironments. These insights may be used for the development of drugs to inhibit expansion and survival of leukemic HSPC in patients. Moreover, they may also contribute to future development of therapeutic applications aimed at expanding normal HSPC prior to their mobilization or following clinical HSPC transplantation protocols. Disclosures:No relevant conflicts of interest to declare.

The fraction of quiescent massive galaxies in the early Universe

Aims: The aim of this work is to collect a complete, mass--selected sample of galaxies with very low specific star formation rate, for a comparison with the prediction of recent theoretical models. Method: We use the 24/K flux ratio, complemented by the SED fitting to the full 0.35-8.0 mum spectral distribution, to select quiescent galaxies from z~0.4 to z~4 in the GOODS--MUSIC sample. Our observational selection can be translated into thresholds on the specific star formation rate SFR/M_*, that can be used to compare with the theoretical predictions. Results: We find that, in the framework of the well known global decline of the quiescent fraction with redshift, a non-negligible fraction ~15-20% of massive galaxies with very low specific star formation rate exists up to z~4, including a tail of "Red&Dead" galaxies with SFR/M_*<10^{-11}/yr. Recent theoretical models vary to a large extent in the prediction of the fraction of galaxies with very low specific star formation rates, and are unable to provide a global match to our data.

High GATA-2 expression inhibits human hematopoietic stem and progenitor cell function by effects on cell cycle

AbstractEvidence suggests the transcription factor GATA-2 is a critical regulator of murine hematopoietic stem cells. Here, we explore the relation between GATA-2 and cell proliferation and show that inducing GATA-2 increases quiescence (G0 residency) of murine and human hematopoietic cells. In human cord blood, quiescent fractions (CD34+CD38−HoechstloPyronin Ylo) express more GATA-2 than cycling counterparts. Enforcing GATA-2 expression increased quiescence of cord blood cells, reducing proliferation and performance in long-term culture-initiating cell and colony-forming cell (CFC) assays. Gene expression analysis places GATA-2 upstream of the quiescence regulator MEF, but enforcing MEF expression does not prevent GATA-2–conferred quiescence, suggesting additional regulators are involved. Although known quiescence regulators p21CIP1 and p27KIP1 do not appear to be responsible, enforcing GATA-2 reduced expression of regulators of cell cycle such as CCND3, CDK4, and CDK6. Enforcing GATA-2 inhibited human hematopoiesis in vivo: cells with highest exogenous expression (GATA-2hi) failed to contribute to hematopoiesis in nonobese diabetic–severe combined immunodeficient (NOD-SCID) mice, whereas GATA-2lo cells contributed with delayed kinetics and low efficiency, with reduced expression of Ki-67. Thus, GATA-2 activity inhibits cell cycle in vitro and in vivo, highlighting GATA-2 as a molecular entry point into the transcriptional program regulating quiescence in human hematopoietic stem and progenitor cells.