Slow-Cycling Cancer Cells: A Dissertation

Tumor recurrence after chemotherapy is a major cause of patient morbidity and mortality. Recurrences are thought to be due to small subsets of stem-like cancer cells that are able to survive chemotherapy and drive tumor re-growth. A more complete understanding of stem-like cancer cell regulation is required to develop therapies to better target and eliminate these cells. Slow-cycling stem cells are integral components of adult epithelial tissues and may give rise to cancer stem cell populations that share similar characteristics. These slow-cycling adult stem cells are inherently resistant to traditional forms of chemotherapy and transference of this characteristic may help to explain therapy resistance in cancer stem cell populations. Using a novel application for the proliferation marker CFSE, we have identified populations of slow-cycling cancer cells with tumor initiating capabilities. As predicted, slow-cycling cancer cells exhibit a multi-fold increase in chemotherapy resistance and retain the ability to re-enter the cell cycle. Furthermore, we observed consistent over-expression of the CDK5 activator, p35, in slow-cycling cancer cells. Manipulation of p35 expression in cancer cells affects cell cycle distribution and survival when these cells are treated with traditional forms of chemotherapy. Additionally, we demonstrate that alterations in p35 expression affect BCL2 levels, suggesting a mechanism for the survival phenotype. Combined, our data suggest a model whereby slow-cycling stem-like cancer cells utilize the p35/CDK5 complex to slow cell cycling speed and promote resistance to chemotherapy. Future p35 targeting, in combination with traditional forms of chemotherapy, may help eliminate these cells and reduce tumor recurrence rates, increasing long-term patient survival

Branes, Rings and Matrix Models in Minimal (Super)string Theory

We study both bosonic and supersymmetric (p,q) minimal models coupled to Liouville theory using the ground ring and the various branes of the theory. From the FZZT brane partition function, there emerges a unified, geometric description of all these theories in terms of an auxiliary Riemann surface M_{p,q} and the corresponding matrix model. In terms of this geometric description, both the FZZT and ZZ branes correspond to line integrals of a certain one-form on M_{p,q}. Moreover, we argue that there are a finite number of distinct (m,n) ZZ branes, and we show that these ZZ branes are located at the singularities of M_{p,q}. Finally, we discuss the possibility that the bosonic and supersymmetric theories with (p,q) odd and relatively prime are identical, as is suggested by the unified treatment of these models.Comment: 72 pages, 3 figures, improved treatment of FZZT and ZZ branes, minor change

Loss of F-box only protein 2 (Fbxo2) disrupts levels and localization of select NMDA receptor subunits, and promotes aberrant synaptic connectivity

NMDA receptors (NMDARs) play an essential role in some forms of synaptic plasticity, learning, and memory. Therefore, these receptors are highly regulated with respect to their localization, activation, and abundance both within and on the surface of mammalian neurons. Fundamental questions remain, however, regarding how this complex regulation is achieved. Using cell-based models and F-box Only Protein 2 (Fbxo2) knock-out mice, we found that the ubiquitin ligase substrate adaptor protein Fbxo2, previously reported to facilitate the degradation of the NMDAR subunit GluN1 in vitro, also functions to regulate GluN1 and GluN2A subunit levels in the adult mouse brain. In contrast, GluN2B subunit levels are not affected by the loss of Fbxo2. The loss of Fbxo2 results in greater surface localization of GluN1 and GluN2A, together with increases in the synaptic markers PSD-95 and Vglut1. These synaptic changes do not manifest as neurophysiological differences or alterations in dendritic spine density in Fbxo2 knock-out mice, but result instead in increased axo-dendritic shaft synapses. Together, these findings suggest that Fbxo2 controls the abundance and localization of specific NMDAR subunits in the brain and may influence synapse formation and maintenance

A Note on Background (In)dependence

In general quantum systems there are two kinds of spacetime modes, those that fluctuate and those that do not. Fluctuating modes have normalizable wavefunctions. In the context of 2D gravity and ``non-critical'' string theory these are called macroscopic states. The theory is independent of the initial Euclidean background values of these modes. Non-fluctuating modes have non-normalizable wavefunctions and correspond to microscopic states. The theory depends on the background value of these non-fluctuating modes, at least to all orders in perturbation theory. They are superselection parameters and should not be minimized over. Such superselection parameters are well known in field theory. Examples in string theory include the couplings $t_k$ (including the cosmological constant) in the matrix models and the mass of the two-dimensional Euclidean black hole. We use our analysis to argue for the finiteness of the string perturbation expansion around these backgrounds.Comment: 16 page

Resistance exercise initiates mechanistic target of rapamycin (mTOR) translocation and protein complex co-localisation in human skeletal muscle

The mechanistic target of rapamycin (mTOR) is a central mediator of protein synthesis in skeletal muscle. We utilized immunofluorescence approaches to study mTOR cellular distribution and protein-protein co-localisation in human skeletal muscle in the basal state as well as immediately, 1 and 3 h after an acute bout of resistance exercise in a fed (FED; 20 g Protein/40 g carbohydrate/1 g fat) or energy-free control (CON) state. mTOR and the lysosomal protein LAMP2 were highly co-localised in basal samples. Resistance exercise resulted in rapid translocation of mTOR/LAMP2 towards the cell membrane. Concurrently, resistance exercise led to the dissociation of TSC2 from Rheb and increased in the co-localisation of mTOR and Rheb post exercise in both FED and CON. In addition, mTOR co-localised with Eukaryotic translation initiation factor 3 subunit F (eIF3F) at the cell membrane post-exercise in both groups, with the response significantly greater at 1 h of recovery in the FED compared to CON. Collectively our data demonstrate that cellular trafficking of mTOR occurs in human muscle in response to an anabolic stimulus, events that appear to be primarily influenced by muscle contraction. The translocation and association of mTOR with positive regulators (i.e. Rheb and eIF3F) is consistent with an enhanced mRNA translational capacity after resistance exercise

Authigenic carbonates from the Cascadia subduction zone and their relation to gas hydrate stability

Authigenic carbonates are intercalated with massive gas hydrates in sediments of the Cascadia margin. The deposits were recovered from the uppermost 50 cm of sediments on the southern summit of the Hydrate Ridge during the RV Sonne cruise SO110. Two carbonate lithologies that differ in chemistry, mineralogy, and fabric make up these deposits. Microcrystalline high-magnesium calcite (14 to 19 mol% MgCO3) and aragonite are present in both semiconsolidated sediments and carbonate-cemented clasts. Aragonite occurs also as a pure phase without sediment impurities. It is formed by precipitation in cavities as botryoidal and isopachous aggregates within pure white, massive gas hydrate. Variations in oxygen isotope values of the carbonates reflect the mineralogical composition and define two end members: a Mg-calcite with δ18O =4.86‰ PDB and an aragonite with δ18O =3.68‰ PDB. On the basis of the ambient bottom-water temperature and accepted equations for oxygen isotope fractionation, we show that the aragonite phase formed in equilibrium with its pore-water environment, and that the Mg-calcite appears to have precipitated from pore fluids enriched in 18O. Oxygen isotope enrichment probably originates from hydrate water released during gas-hydrate destabilization

Molecular rationale for the use of PI3K/AKT/mTOR pathway inhibitors in combination with crizotinib in ALK-mutated neuroblastoma

Mutations in the ALK tyrosine kinase receptor gene represent important therapeutic targets in neuroblastoma, yet their clinical translation has been challenging. The ALKF1174L mutation is sensitive to the ALK inhibitor crizotinib only at high doses and mediates acquired resistance to crizotinib in ALK-translocated cancers. We have shown that the combination of crizotinib and an inhibitor of downstream signaling induces a favorable response in transgenic mice bearing ALKF1174L/MYCN-positive neuroblastoma. Here, we investigated the molecular basis of this effect and assessed whether a similar strategy would be effective in ALK-mutated tumors lacking MYCN overexpression. We show that in ALK-mutated, MYCN-amplified neuroblastoma cells, crizotinib alone does not affect mTORC1 activity as indicated by persistent RPS6 phosphorylation. Combined treatment with crizotinib and an ATP-competitive mTOR inhibitor abrogated RPS6 phosphorylation, leading to reduced tumor growth and prolonged survival in ALKF1174L/MYCN-positive models compared to single agent treatment. By contrast, this combination, while inducing mTORC1 downregulation, caused reciprocal upregulation of PI3K activity in ALK-mutated cells expressing wild-type MYCN. Here, an inhibitor with potency against both mTOR and PI3K was more effective in promoting cytotoxicity when combined with crizotinib. Our findings should enable a more precise selection of molecularly targeted agents for patients with ALK-mutated tumors

D-brane Decay in Two-Dimensional String Theory

We consider unstable D0-branes of two dimensional string theory, described by the boundary state of Zamolodchikov and Zamolodchikov [hep-th/0101152] multiplied by the Neumann boundary state for the time coordinate $t$. In the dual description in terms of the $c=1$ matrix model, this D0-brane is described by a matrix eigenvalue on top of the upside down harmonic oscillator potential. As suggested by McGreevy and Verlinde [hep-th/0304224], an eigenvalue rolling down the potential describes D-brane decay. As the eigenvalue moves down the potential to the asymptotic region it can be described as a free relativistic fermion. Bosonizing this fermion we get a description of the state in terms of a coherent state of the tachyon field in the asymptotic region, up to a non-local linear field redefinition by an energy-dependent phase. This coherent state agrees with the exponential of the closed string one-point function on a disk with Sen's marginal boundary interaction for $t$ which describes D0-brane decay.Comment: 19 pages, harvmac, minor change

DNA methylation among firefighters

Firefighters are exposed to carcinogens and have elevated cancer rates. We hypothesized that occupational exposures in firefighters would lead to DNA methylation changes associated with activation of cancer pathways and increased cancer risk. To address this hypothesis, we collected peripheral blood samples from 45 incumbent and 41 new recruit nonsmoking male firefighters and analyzed the samples for DNA methylation using an Illumina Methylation EPIC 850k chip. Adjusting for age and ethnicity, we performed: 1) genome-wide differential methylation analysis; 2) genome-wide prediction for firefighter status (incumbent or new recruit) and years of service; and 3) Ingenuity Pathway Analysis (IPA). Four CpGs, including three in the YIPF6, MPST, and PCED1B genes, demonstrated above 1.5-fold statistically significant differential methylation after Bonferroni correction. Genome-wide methylation predicted with high accuracy incumbent and new recruit status as well as years of service among incumbent firefighters. Using IPA, the top pathways with more than 5 gene members annotated from differentially methylated probes included Sirtuin signaling pathway, p53 signaling, and 5' AMP-activated protein kinase (AMPK) signaling. These DNA methylation findings suggest potential cellular mechanisms associated with increased cancer risk in firefighters.US Federal Emergency Management Agency Assistance to Firefighters Grant program [EMW-2014-FP-00200]Open access journalThis item from the UA Faculty Publications collection is made available by the University of Arizona with support from the University of Arizona Libraries. If you have questions, please contact us at [email protected]

Indirect comparison of interventions using published randomised trials: systematic review of PDE-5 inhibitors for erectile dysfunction

BACKGROUND: There are no randomised and properly blinded trials directly comparing one PDE-5 inhibitor with another in a normal home setting. Valid indirect comparisons with a common comparator must examine equivalent doses, similar duration, similar populations, with the same outcomes reported in the same way. METHODS: Published randomised, double-blind trials of oral PDE-5 inhibitors for erectile dysfunction were sought from reference lists in previous reviews and electronic searching. Analyses of efficacy and harm were carried out for each treatment, and results compared where there was a common comparator and consistency of outcome reporting, using equivalent doses. RESULTS: Analysis was limited by differential reporting of outcomes. Sildenafil trials were clinically and geographically more diverse. Tadalafil and vardenafil trials tended to use enriched enrolment. Using all trials, the three interventions were similar for consistently reported efficacy outcomes. Rates of successful intercourse for sildenafil, tadalafil and vardenafil were 65%, 62%, and 59%, with placebo rates of 23–28%. The rates of improved erections were 76%, 75% and 71%, respectively, with placebo rates of 22–24%, and NNTs of 1.9 or 2.0. Reporting of withdrawals was less consistent, but all-cause withdrawals for sildenafil, tadalafil and vardenafil were 8% 13% and 20%. All three drugs were well tolerated, with headache being the most commonly reported event at 13–17%. There were few serious adverse events. CONCLUSION: There were differences between trials in outcomes reported, limiting comparisons, and the most useful outcomes were not reported. For common outcomes there was similar efficacy between PDE-5 inhibitors