The Identification of Novel Mechanisms in Neuronal Development and Degeneration

The goal of this dissertation is to further understand two key, broad processes which occur over the course of a neuron\u27s lifetime: its development and possible degeneration in disease. We identify novel components in both of these processes and attempt to understand the functional significance as well as the mechanism each component uses to exert its effects. We begin with work done focusing on how the neuron\u27s dendritic tree develops. The development of neurons has two phases: (1) a first phase relying on a genetic program and (2) a second phase that uses synaptic activity to guide the fine tuning of connections. We are primarily interested in understanding how neurons develop within the motor system using cues from synaptic activity. One type of activity-dependent development in the motor system is driven by AMPA receptors assembled with the GluA1 subunit using the scaffolding protein, SAP97. In Chapter 2, we describe our finding that the small protein, CRIPT, functions in the development of the neuron\u27s dendritic tree within the motor system. We show that CRIPT is expressed during the time in development when activity-dependent remodeling is occurring the in spinal cord. Additionally, we show that CRIPT binds to the PDZ3 domain of SAP97 using its C-terminus. Finally, we show that CRIPT is necessary for proper dendritic growth of the motor system and normal motor system responses in vivo. There are a number of diseases that specifically target motor neurons. One adult-onset disease is Amyotrophic Lateral Sclerosis (ALS). Like many neurodegenerative diseases, it is a multifactorial disease and a contributing factor is the lack of proper protein quality control and the formation of large protein aggregates within motor neurons. In Chapter 3, we focus on how endoplasmic reticulum-associated degradation (ERAD) components might affect mutant proteins in ALS and their toxicity. ERAD is responsible for maintaining the proper folding of proteins that are to be trafficked to the cell surface or secreted from the cell. Although many proteins associated with ALS are not ERAD substrates, a number of ERAD components have been found to interact with ALS-causing mutant proteins and have been found to be mutated in forms of familial ALS. We identify new modifiers within the ERAD pathway of the toxicity of two ALS-linked proteins, mutant TDP-43 and mutant SOD1. We focus on the mechanism of one suppressor of ALS-linked proteotoxicity, the loss of RAD-23. The levels of RAD23 are increased in ALS models and reducing RAD-23 levels can suppress phenotypes in ALS models, as well as suppress motor neuron toxicity of the mutant proteins. We further show that reduced RAD-23 is able to accelerate the turnover of mutant proteins associated with ALS. Finally, we show that there is a mislocalization of RAD-23 protein and increased RAD-23 protein expression in post mortem spinal cord tissue of ALS patients, suggesting that RAD-23 may be a new novel target in the treatment of ALS. Collectively, this work identifies and describes novel proteins and mechanisms involved in the development and pathobiology of motor neurons

31st Annual Meeting and Associated Programs of the Society for Immunotherapy of Cancer (SITC 2016) : part two

Background The immunological escape of tumors represents one of the main ob- stacles to the treatment of malignancies. The blockade of PD-1 or CTLA-4 receptors represented a milestone in the history of immunotherapy. However, immune checkpoint inhibitors seem to be effective in specific cohorts of patients. It has been proposed that their efficacy relies on the presence of an immunological response. Thus, we hypothesized that disruption of the PD-L1/PD-1 axis would synergize with our oncolytic vaccine platform PeptiCRAd. Methods We used murine B16OVA in vivo tumor models and flow cytometry analysis to investigate the immunological background. Results First, we found that high-burden B16OVA tumors were refractory to combination immunotherapy. However, with a more aggressive schedule, tumors with a lower burden were more susceptible to the combination of PeptiCRAd and PD-L1 blockade. The therapy signifi- cantly increased the median survival of mice (Fig. 7). Interestingly, the reduced growth of contralaterally injected B16F10 cells sug- gested the presence of a long lasting immunological memory also against non-targeted antigens. Concerning the functional state of tumor infiltrating lymphocytes (TILs), we found that all the immune therapies would enhance the percentage of activated (PD-1pos TIM- 3neg) T lymphocytes and reduce the amount of exhausted (PD-1pos TIM-3pos) cells compared to placebo. As expected, we found that PeptiCRAd monotherapy could increase the number of antigen spe- cific CD8+ T cells compared to other treatments. However, only the combination with PD-L1 blockade could significantly increase the ra- tio between activated and exhausted pentamer positive cells (p= 0.0058), suggesting that by disrupting the PD-1/PD-L1 axis we could decrease the amount of dysfunctional antigen specific T cells. We ob- served that the anatomical location deeply influenced the state of CD4+ and CD8+ T lymphocytes. In fact, TIM-3 expression was in- creased by 2 fold on TILs compared to splenic and lymphoid T cells. In the CD8+ compartment, the expression of PD-1 on the surface seemed to be restricted to the tumor micro-environment, while CD4 + T cells had a high expression of PD-1 also in lymphoid organs. Interestingly, we found that the levels of PD-1 were significantly higher on CD8+ T cells than on CD4+ T cells into the tumor micro- environment (p < 0.0001). Conclusions In conclusion, we demonstrated that the efficacy of immune check- point inhibitors might be strongly enhanced by their combination with cancer vaccines. PeptiCRAd was able to increase the number of antigen-specific T cells and PD-L1 blockade prevented their exhaus- tion, resulting in long-lasting immunological memory and increased median survival

Inclusive quarkonium production in pp collisions at s=5.02\sqrt{s} = 5.02 TeV

This article reports on the inclusive production cross section of several quarkonium states, $\mathrm{J}/\psi$, $\psi {\rm (2S)}$, $\Upsilon\rm(1S)$, $\Upsilon\rm(2S)$, and $\Upsilon\rm(3S)$, measured with the ALICE detector at the LHC, in \pp collisions at $\sqrt{s} = 5.02$ TeV. The analysis is performed in the dimuon decay channel at forward rapidity ($2.5 < y < 4$). The measured cross sections, assuming unpolarized quarkonia, are: $\sigma_{\mathrm{J}/\psi} = 5.88 \pm 0.03 \pm 0.34\ \mu$b, $\sigma_{\psi {\rm (2S)}} = 0.87 \pm 0.06 \pm 0.10\ \mu$b, $\sigma_{\Upsilon\rm(1S)} = 45.5 \pm 3.9 \pm 3.5$ nb, $\sigma_{\Upsilon\rm(2S)} = 22.4 \pm 3.2 \pm 2.7$ nb, and $\sigma_{\Upsilon\rm(3S)} = 4.9 \pm 2.2 \pm 1.0$ nb, where the first (second) uncertainty is the statistical (systematic) one. The transverse-momentum ($p_{\rm T}$) and rapidity ($y$) differential cross sections for $\mathrm{J}/\psi$, $\psi {\rm (2S)}$, $\Upsilon\rm(1S)$, and the $\psi {\rm (2S)}$-to-$\mathrm{J}/\psi$ cross section ratios are presented. For the first time, the cross sections of the three $\Upsilon$ states, as well as the $\psi {\rm (2S)}$ one as a function of $p_{\rm T}$ and $y$, are measured at $\sqrt{s} = 5.02$ TeV at forward rapidity. These measurements also significantly extend the $\mathrm{J}/\psi$$p_{\rm T}$ reach with respect to previously published results. A comparison with ALICE measurements in pp collisions at $\sqrt{s} = 2.76$, 7, 8, and 13 TeV is presented and the energy dependence of quarkonium production cross sections is discussed. Finally, the results are compared with the predictions from several production models

Neutral to charged kaon yield fluctuations in Pb – Pb collisions at sNN\sqrt{s_{\rm NN}} = 2.76 TeV

We present the first measurement of event-by-event fluctuations in the kaon sector in Pb – Pb collisions at $\sqrt{s_{\rm NN}}$ = 2.76 TeV with the ALICE detector at the LHC. The robust fluctuation correlator ν$_{dyn}$ is used to evaluate the magnitude of fluctuations of the relative yields of neutral and charged kaons, as well as the relative yields of charged kaons, as a function of collision centrality and selected kinematic ranges. While the correlator ν$_{dyn}$[K$^+$,K$^−$] exhibits a scaling approximately in inverse proportion of the charged particle multiplicity, ν$_{dyn}$[K$_S^0$,K$^\pm$] features a significant deviation from such scaling. Within uncertainties, the value of ν$_{dyn}$[K$_S^0$,K$^\pm$] is independent of the selected transverse momentum interval, while it exhibits a pseudorapidity dependence. The results are compared with HIJING, AMPT and EPOS–LHC predictions, and are further discussed in the context of the possible production of disoriented chiral condensates in central Pb – Pb collisions

First study of the two-body scattering involving charm hadrons

This article presents the first measurement of the interaction between charm hadrons and nucleons. The two-particle momentum correlations of $pD^-$ and $\bar{p}D^+$ pairs are measured by the ALICE Collaboration in high-multiplicity pp collisions at $\sqrt{s}$ = 13 TeV. The data are compatible with the Coulomb-only interaction hypothesis within (1.1–1.5)σ. The level of agreement slightly improves if an attractive nucleon $(N)\bar{D}$ strong interaction is considered, in contrast to most model predictions which suggest an overall repulsive interaction. This measurement allows for the first time an estimation of the 68% confidence level interval for the isospin I=0 inverse scattering length of the $N\bar{D}$ state $f_{0,I=0}^{-1}$∈[-0.4,0.9] fm$^{-1}$, assuming negligible interaction for the isospin I=1 channel

Measurement of the angle between jet axes in Pb−-Pb collisions at sNN=5.02\sqrt{s_{\rm NN}} = 5.02 TeV

International audienceThis letter presents the first measurement of the angle between different jet axes (denoted as ${\Delta}R$) in Pb$-$Pb collisions. The measurement is carried out in the 0$-$10% most-central events at $\sqrt{s_{\rm NN}} = 5.02$ TeV. Jets are assembled by clustering charged particles at midrapidity using the anti-$k_{\rm T}$ algorithm with resolution parameters $R=0.2$ and $0.4$ and transverse momenta in the intervals $40 < p_{\rm T}^{\rm ch jet} < 140$ GeV/$c$ and $80 < p_{\rm T}^{\rm ch jet} < 140$ GeV/$c$, respectively. Measurements at these low transverse momenta enhance the sensitivity to quark$-$gluon plasma (QGP) effects. A comparison to models implementing various mechanisms of jet energy loss in the QGP shows that the observed narrowing of the Pb$-$Pb distribution relative to pp can be explained if quark-initiated jets are more likely to emerge from the medium than gluon-initiated jets. These new measurements discard intra-jet $p_{\rm T}$ broadening as described in a model calculation with the BDMPS formalism as the main mechanism of energy loss in the QGP. The data are sensitive to the angular scale at which the QGP can resolve two independent splittings, favoring mechanisms that incorporate incoherent energy loss

First measurement of the ∣t∣|t|-dependence of incoherent J/ψ\psi photonuclear production

International audienceThe first measurement of the cross section for incoherent photonuclear production of J/$\psi$ vector meson as a function of the Mandelstam $|t|$ variable is presented. The measurement was carried out with the ALICE detector at midrapidity, $|y|<0.8$, using ultra-peripheral collisions of Pb nuclei at a centre-of-mass energy per nucleon pair $\sqrt{s_{\mathrm{NN}}} = 5.02$ TeV. This rapidity interval corresponds to a Bjorken-$x$ range $(0.3$$-$$1.4)\times 10^{-3}$. Cross sections are reported in five $|t|$ intervals in the range $0.04<|t|<1$~GeV$^2$ and compared to the predictions of different models. Models that ignore quantum fluctuations of the gluon density in the colliding hadron predict a $|t|$-dependence of the cross section much steeper than in data. The inclusion of such fluctuations in the same models provides a better description of the data

Multiplicity-dependent production of Σ(1385)±\Sigma(1385)^{\pm} and Ξ(1530)0\Xi(1530)^{0} in pp collisions at s=13\sqrt{s}=13 TeV

International audienceThe production yields of the $\Sigma(1385)^{\pm}$ and $\Xi(1530)^{0}$ resonances are measured in pp collisions at $\sqrt{s}=13$ TeV with ALICE. The measurements are performed as a function of the charged particle multiplicity $\langle \mathrm{d}N_\mathrm{ch}/\mathrm{d}\eta \rangle$, which is related to the energy density produced in the collision. The results include transverse momentum ($p_{\rm T}$) distributions, $p_{\rm T}$-integrated yields, mean transverse momenta of $\Sigma(1385)^{\pm}$ and $\Xi(1530)^{0}$, as well as ratios of the $p_{\rm T}$-integrated resonance yields relative to yields of other hadron species. The $\Sigma(1385)^{\pm}/\pi^{\pm}$ and $\Xi(1530)^{0}/\pi^{\pm}$ yield ratios are consistent with the trend of the enhancement of strangeness production from low to high multiplicity pp collisions, which was previously observed for strange and multi-strange baryons. The yield ratio between the measured resonances and the long-lived baryons with the same strangeness content exhibits a hint of a mild increasing trend at low multiplicity, despite too large uncertainties to exclude the flat behaviour. The results are compared to predictions from models such as EPOS-LHC and PYTHIA 8 with Rope shoving. The latter provides the best description of the multiplicity dependence of the $\Sigma(1385)^{\pm}$ and $\Xi(1530)^{0}$ production in pp collisions at $\sqrt{s}=13$ TeV

Charged-particle production as a function of the relative transverse activity classifier in pp, p−-Pb, and Pb−-Pb collisions at the LHC

International audienceMeasurements of charged-particle production in pp, p$-$Pb, and Pb$-$Pb collisions in the toward, away, and transverse regions with the ALICE detector are discussed. These regions are defined event-by-event relative to the azimuthal direction of the charged trigger particle, which is the reconstructed particle with the largest transverse momentum ($p_{\mathrm{T}}^{\rm trig}$) in the range $8<p_{\mathrm{T}}^{\rm trig}<15$ GeV$/c$. The toward and away regions contain the primary and recoil jets, respectively; both regions are accompanied by the underlying event (UE). In contrast, the transverse region perpendicular to the direction of the trigger particle is dominated by the so-called UE dynamics, and includes also contributions from initial- and final-state radiation. The relative transverse activity classifier, $R_{\mathrm{T}}=N_{\mathrm{ch}}^{\mathrm{T}}/\langle N_{\mathrm{ch}}^{\mathrm{T}}\rangle$, is used to group events according to their UE activity, where $N_{\mathrm{ch}}^{\mathrm{T}}$ is the charged-particle multiplicity per event in the transverse region and $\langle N_{\mathrm{ch}}^{\mathrm{T}}\rangle$ is the mean value over the whole analysed sample. The energy dependence of the $R_{\mathrm{T}}$ distributions in pp collisions at $\sqrt{s}=2.76$, 5.02, 7, and 13 TeV is reported, exploring the Koba-Nielsen-Olesen (KNO) scaling properties of the multiplicity distributions. The first measurements of charged-particle $p_{\rm T}$ spectra as a function of $R_{\mathrm{T}}$ in the three azimuthal regions in pp, p$-$Pb, and Pb$-$Pb collisions at $\sqrt{s_{\rm NN}}=5.02$ TeV are also reported. Data are compared with predictions obtained from the event generators PYTHIA 8 and EPOS LHC. This set of measurements is expected to contribute to the understanding of the origin of collective-like effects in small collision systems (pp and p$-$Pb)