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

J/ψ\psi-hadron correlations at midrapidity in pp collisions at s\sqrt{s} = 13 TeV

International audienceWe report on the measurement of inclusive, non-prompt, and prompt J/$\psi$-hadron correlations by the ALICE Collaboration at the CERN Large Hadron Collider in pp collisions at a center-of-mass energy of 13 TeV. The correlations are studied at midrapidity ($|y| < 0.9$) in the transverse momentum ranges $p_{\rm T} < 40~\text{GeV}/c$ for the J/$\psi$ and $0.15 < p_{\rm T} < 10$ GeV/$c$ and $|\eta|<0.9$ for the associated hadrons. The measurement is based on minimum bias and high multiplicity data samples corresponding to integrated luminosities of $L_{\text{int}} = 34~\text{nb}^{-1}$ and $L_{\text{int}} = 6.9~\text{pb}^{-1}$, respectively. In addition, two more data samples are employed, requiring, on top of the minimum bias condition, a threshold on the tower energy of $E = 4$ and $9~\text{GeV}$ in the ALICE electromagnetic calorimeters, which correspond to integrated luminosities of $L_{\text{int}} = 0.9~\text{pb}^{-1}$ and $L_{\text{int}} = 8.4~\text{pb}^{-1}$, respectively. The results are presented as associated hadron yields per J/$\psi$ trigger as a function of the azimuthal angle difference between the associated hadrons and J/$\psi$ mesons. The integrated near-side and away-side correlated yields are also extracted as a function of the J/$\psi$ transverse momentum. The measurements are discussed in comparison to PYTHIA calculations

First measurement of A = 4 (anti)hypernuclei at the LHC

International audienceIn this Letter, the first evidence of the ${}^4_{\bar{\Lambda}}\overline{\mathrm{He}}$ antihypernucleus is presented, along with the first measurement at the LHC of the production of (anti)hypernuclei with mass number $A=4$, specifically (anti)${}^4_{\Lambda}\mathrm{H}$ and (anti)${}^4_{\Lambda}\mathrm{He}$. In addition, the antiparticle-to-particle ratios for both hypernuclei (${}^4_{\bar{\Lambda}}\overline{\mathrm{H}}$ / ${}^4_{\Lambda}\mathrm{H}$~and ${}^4_{\bar{\Lambda}}\overline{\mathrm{He}}$ / ${}^4_{\Lambda}\mathrm{He}$) are shown, which are sensitive to the baryochemical potential of the strongly-interacting matter created in heavy-ion collisions. The results are obtained from a data sample of central Pb--Pb collisions, collected during the 2018 LHC data-taking at a center-of-mass energy per nucleon pair of $\sqrt{s_{\mathrm{NN}}} =$ 5.02 TeV. The yields measured for the average of the charge-conjugated states are found to be $[0.78 \; \pm \; 0.19 \; \mathrm{(stat.)} \; \pm \; 0.17 \; \mathrm{(syst.)}] \times 10^{-6}$ for the (anti)${}^4_{\Lambda}\mathrm{H}$ and $[1.08 \; \pm \; 0.34 \; \mathrm{(stat.)} \; \pm \; 0.20 \; \mathrm{(syst.)}] \times 10^{-6}$ for the (anti)${}^4_{\Lambda}\mathrm{He}$, and the measured antiparticle-to-particle ratios are in agreement with unity. The presence of (anti)${}^4_{\Lambda}\mathrm{H}$ and (anti)${}^4_{\Lambda}\mathrm{He}$ excited states is expected to strongly enhance the production yield of these hypernuclei. The yield values exhibit a combined deviation of 3.3$\sigma$ from the theoretical ground-state-only expectation, while the inclusion of the excited states in the calculations leads to an agreement within 0.6$\sigma$ with the present measurements. Additionally, the measured (anti)${}^4_{\Lambda}\mathrm{H}$ and (anti)${}^4_{\Lambda}\mathrm{He}$ masses are compatible with the world-average values within the uncertainties

Particle production as a function of charged-particle flattenicity in pp collisions at s\sqrt{s} = 13 TeV

International audienceThis paper reports the first measurement of the transverse momentum ($p_{\mathrm{T}}$) spectra of primary charged pions, kaons, (anti)protons, and unidentified particles as a function of the charged-particle flattenicity in pp collisions at $\sqrt{s}=13$ TeV. Flattenicity is a novel event shape observable that is measured in the pseudorapidity intervals covered by the V0 detector, $2.8<\eta<5.1$ and $-3.7<\eta<-1.7$. According to QCD-inspired phenomenological models, it shows sensitivity to multiparton interactions and is less affected by biases towards larger $p_{\mathrm{T}}$ due to local multiplicity fluctuations in the V0 acceptance than multiplicity. The analysis is performed in minimum-bias (MB) as well as in high-multiplicity events up to $p_{\mathrm{T}}=20$ GeV/$c$. The event selection requires at least one charged particle produced in the pseudorapidity interval $|\eta|<1$. The measured $p_{\mathrm{T}}$ distributions, average $p_{\mathrm{T}}$, kaon-to-pion and proton-to-pion particle ratios, presented in this paper, are compared to model calculations using PYTHIA 8 based on color strings and EPOS LHC. The modification of the $p_{\mathrm{T}}$-spectral shapes in low-flattenicity events that have large event activity with respect to those measured in MB events develops a pronounced peak at intermediate $p_{\mathrm{T}}$ ($2<p_{\mathrm{T}}<8$ GeV/$c$), and approaches the vicinity of unity at higher $p_{\mathrm{T}}$. The results are qualitatively described by PYTHIA, and they show different behavior than those measured as a function of charged-particle multiplicity based on the V0M estimator

Multimuons in cosmic-ray events as seen in ALICE at the LHC

International audienceALICE is a large experiment at the CERN Large Hadron Collider. Located 52 meters underground, its detectors are suitable to measure muons produced by cosmic-ray interactions in the atmosphere. In this paper, the studies of the cosmic muons registered by ALICE during Run 2 (2015--2018) are described. The analysis is limited to multimuon events defined as events with more than four detected muons ($N_\mu>4$) and in the zenith angle range $0^{\circ}100$) obtained with QGSJET and SIBYLL is compatible with the data, while EPOS-LHC produces a significantly lower rate ($55\%$ of the measured rate). For both QGSJET and SIBYLL, the rate is close to the data when the composition is assumed to be dominated by heavy elements, an outcome compatible with the average energy $E_\mathrm{prim} \sim 10^{17}$~eV of these events. This result places significant constraints on more exotic production mechanisms

Measurement of the inclusive isolated-photon production cross section in pp and Pb−-Pb collisions at sNN=5.02\sqrt{s_{\rm NN}}=5.02 TeV

International audienceThe ALICE Collaboration at the CERN LHC has measured the inclusive production cross section of isolated photons at midrapidity as a function of the photon transverse momentum ($p_{\rm T}^{\gamma}$), in Pb-Pb collisions in different centrality intervals, and in pp collisions, at centre-of-momentum energy per nucleon pair of $\sqrt{s_{\mathrm{NN}}}=5.02$ TeV. The photon transverse momentum range is between 10-14 and 40-140 GeV/$c$, depending on the collision system and on the Pb-Pb centrality class. The result extends to lower $p_{\rm T}^{\gamma}$ than previously published results by the ATLAS and CMS experiments at the same collision energy. The covered pseudorapidity range is $|\eta^{\gamma}| <0.67$. The isolation selection is based on a charged particle isolation momentum threshold $p_{\rm T}^{\rm iso,~ch} = 1.5$ GeV/$c$ within a cone of radii $R=0.2$ and $0.4$. The nuclear modification factor is calculated and found to be consistent with unity in all centrality classes, and also consistent with the HG-PYTHIA model, which describes the event selection and geometry biases that affect the centrality determination in peripheral Pb-Pb collisions. The measurement is compared to next-to-leading order perturbative QCD calculations and to the measurements of isolated photons and Z$^0$ bosons from the CMS experiment, which are all found to be in agreement

Particle production as a function of charged-particle flattenicity in pp collisions at s\sqrt{s} = 13 TeV

International audienceThis paper reports the first measurement of the transverse momentum ($p_{\mathrm{T}}$) spectra of primary charged pions, kaons, (anti)protons, and unidentified particles as a function of the charged-particle flattenicity in pp collisions at $\sqrt{s}=13$ TeV. Flattenicity is a novel event shape observable that is measured in the pseudorapidity intervals covered by the V0 detector, $2.8<\eta<5.1$ and $-3.7<\eta<-1.7$. According to QCD-inspired phenomenological models, it shows sensitivity to multiparton interactions and is less affected by biases towards larger $p_{\mathrm{T}}$ due to local multiplicity fluctuations in the V0 acceptance than multiplicity. The analysis is performed in minimum-bias (MB) as well as in high-multiplicity events up to $p_{\mathrm{T}}=20$ GeV/$c$. The event selection requires at least one charged particle produced in the pseudorapidity interval $|\eta|<1$. The measured $p_{\mathrm{T}}$ distributions, average $p_{\mathrm{T}}$, kaon-to-pion and proton-to-pion particle ratios, presented in this paper, are compared to model calculations using PYTHIA 8 based on color strings and EPOS LHC. The modification of the $p_{\mathrm{T}}$-spectral shapes in low-flattenicity events that have large event activity with respect to those measured in MB events develops a pronounced peak at intermediate $p_{\mathrm{T}}$ ($2<p_{\mathrm{T}}<8$ GeV/$c$), and approaches the vicinity of unity at higher $p_{\mathrm{T}}$. The results are qualitatively described by PYTHIA, and they show different behavior than those measured as a function of charged-particle multiplicity based on the V0M estimator

First measurement of Ds1(1+)(2536)+\mathrm{D_{s1}}(1^{+})(2536)^+ and Ds2∗(2+)(2573)+\mathrm{D_{s2}^{*}(2^{+})(2573)^+} production in proton--proton collisions at s=13\sqrt{s} = 13 TeV at the LHC

International audienceThe production yields of the orbitally excited charm-strange mesons $\mathrm{D_{s1}(1^{+})(2536)^+}$ and $\mathrm{D_{s2}^{*}(2^{+})(2573)^+}$ were measured for the first time in proton--proton (pp) collisions at a center-of-mass energy of $\sqrt{s} =13$ TeV with the ALICE experiment at the LHC. The $\mathrm{D_{s1}^+}$ and $\mathrm{D_{s2}^{*+}}$ mesons were measured at midrapidity ($|y|<0.5$) in minimum-bias and high-multiplicity pp collisions in the transverse-momentum interval $2 < p_{\rm T} < 24$ GeV/$c$. Their production yields relative to the $\mathrm{D_{s}^{+}}$ ground-state yield were found to be compatible between minimum-bias and high-multiplicity collisions, as well as with previous measurements in $\mathrm{e^\pm p}$ and $\mathrm{e^{+}e^{-}}$ collisions. The measured $\mathrm{D_{s1}^+/D_{s}^{+}}$ and $\mathrm{D_{s2}^{*+}/D_{s}^{+}}$ yield ratios are described by statistical hadronization models and can be used to tune the parameters governing the production of excited charm-strange hadrons in Monte Carlo generators, such as PYTHIA 8