xix, 140 p. ; ill. (some col.) A print copy of this thesis is available through the UO Libraries. Search the library catalog for the location and call number. / Using approximately 350 million à + à - pair events recorded with the BaBar detector at the Stanford Linear Accelerator Center between 1999 and 2006, a search has been made for neutrinoless, lepton-flavor violating tau decays to three lighter leptons. All six decay modes consistent with conservation of electric charge and energy have been considered. With signal selection efficiencies of 5-12%, we obtain 90% confidence level upper limits on the branching fraction B (à [arrow right] [cursive l][cursive l][cursive l]) in the range (4 - 8) à 10 -8 . / Adviser: Eric Torrence

Ce mémoire de thèse présente une étude de l’identification des candidats tau par différents algorithmes dans les collisions pp au LHC et impliquant différents processus physiques : électrofaibles, production de quarks top, SUSY et QCD. Les leptons tau sont identifiés géométriquement dans la voie de désintégration hadronique car les jets issus de ces désintégrations sont généralement plus étroits et mieux isolés que les jets QCD. Une discrimination supplémentaire est appliquée en exigeant la présence d’un des modes de désintégration hadronique dominants du tau contenant un ou trois pions chargés et jusqu’à deux pions neutres. Les jets QCD produits en très grande quantité dans les collisions proton-proton du LHC constituent une source de bruit de fond importante pour les analyses impliquant l’identification de leptons tau. L’analyse des jets QCD produits dans le LHC pour les études de mauvaise identification porte sur 36 pb−1 de données enregistrées par le détecteur CMS pendant l’année 2010. Les efficacités d’identification et les taux de mauvaise identification nous révèlent une forte dépendance en les caractéristiques de l’évènement étudié : les efficacités sont globalement plus basses dans les évènements présentant une activité hadronique importante comme les évènements SUSY et tt que dans les évènements QCD mous et électrofaibles. Les effets de l'environnement sur l’identification de tau sont paramétrés à l’aide de deux variables globales : somme(ET) et la multiplicité de hadrons chargés. Les efficacités chutent typiquement de 80% entre les multiplicités de hadrons les plus faibles et le plus fortes. / The work presented in this thesis deals with tau identified by different algorithms in pp collision at LHC and in various kind of physics processes : Electroweak, top quark production, SUSY and QCD. Tau leptons are geometrically identified in their hadronic decay mode because the jets coming from those decays are generally more collimated and isolated than QCD jets. An additional discrimination is applied in asking the presence of one of the dominant # decay modes containing one or three charged pions and up to two neutral pions. The QCD jets produced in large quantities in LHC proton-proton collisions constitute a large background source for analyses involving tau-ID. The analysis of QCD jets produced in LHC for fake-rate studies processed 36pb−1 of data registered during year 2010. Identification efficiencies and fake-rate distributions reveals a strong dependency on event type : distributions are globally lower in events having a large hadronic activity like SUSY and tt events than in electroweak and QCD events. The environment effects of the collision on the identification of tau leptons can be parameterized by two global variables : somme(ET) and charged hadron multiplicity. Efficiencies drop down by 80% between the lowest and the highest charged hadron multiplicities.

abstract: Alzheimer's Disease (AD) is the sixth leading cause of death in the United States and the most common form of dementia. Its cause remains unknown, but it is known to involve two hallmark pathologies: Amyloid Beta plaques and neurofibrillary tangles (NFTs). Several proteins have been implicated in the formation of neurofibrillary tangles, including Tau and S100B. S100B is a dimeric protein that is typically found bound to Ca(II) or Zn(II). These experiments relate to the involvement of S100B in Alzheimer's Disease-related processes and the results suggest that future research of S100B is warranted. Zn(II)-S100B was found to increase the rate at which tau assembled into paired helical filaments, as well as affect the rate at which tubulin polymerized into microtubules and the morphology of SH-SY5Y neuroblastoma cells after 72 hours of incubation. Zn(II)-S100B also increased the firing rate of hippocampal neurons after 36 hours of incubation. Together, these results suggest several possibilities: Zn(II)-S100B may be a key part of the formation of paired helical filaments (PHFs) that subsequently form NFTs. Zn(II)-S100B may also be competing with tau to bind tubulin, which could lead to an instability of microtubules and subsequent cell death. This finding aligns with the neurodegeneration that is commonly seen in AD and which could be a result of this microtubule instability. Ultimately, these results suggest that S100B is likely involved in several AD-related processes, and if the goal is to find an efficient and effective therapeutic target for AD, the relationship between S100B, particularly Zn(II)-S100B, and tau needs to be further studied. / Dissertation/Thesis / M.S. Applied Biological Sciences 2013

La pathologie Tau est une lésion commune à plus d’une vingtaine de maladies neurodégénératives, regroupées sous le terme de Tauopathies. Elle correspond à l’accumulation intracellulaire de matériel fibrillaire constitué de protéines Tau hyper- et anormalement phosphorylées. Dans la maladie d’Alzheimer (MA), démence la plus courante chez la personne âgée, la progression et la distribution topographique de ces agrégats évoluent au cours du temps et sont corrélés aux déficits cognitifs observés. A ce jour, les traitements sont principalement symptomatiques. Cependant, plusieurs stratégies thérapeutiques sont étudiées parmi lesquelles l’immunothérapie. Les travaux présentés dans cette thèse ont pour objectif d’étudier les effets de l’immunothérapie Tau, active et passive, dans un modèle transgénique murin mimant la pathologie Tau de type Alzheimer. Les souris THY-Tau22, surexpriment une isoforme de Tau humaine mutée sur deux sites et sous contrôle d’un promoteur neuronal. Ce modèle présente dès l’âge de trois mois des altérations progressives de l’apprentissage et de la mémoire en parallèle d’une accumulation de protéine Tau principalement au niveau hippocampique, sans perte neuronale majeure, ce qui lui confère les caractéristiques d’un stade précoce de MA. Au sein du modèle THY-Tau22, on retrouve la protéine Tau phosphorylée en Ser422, un épitope particulièrement pertinent pour l’immunothérapie. En effet, la pSer422 est un épitope de phosphorylation anormale unique, présent dans la plupart des Tauopathies. Dans ces travaux de thèse, nous montrons que la vaccination précoce contre la Ser422 phosphorylée de la protéine Tau peut prévenir l'altération de mémoire spatiale mesurée par le test du labyrinthe en Y. Cette diminution de l'atteinte cognitive est associée à une diminution de la phosphorylation anormale de Tau au niveau de l'hippocampe et à une réduction significative des espèces insolubles de Tau. Les résultats de cette vaccination nous ont amenés à générer un anticorps monoclonal dirigé contre la pSer422 (2H9) afin d’évaluer les effets de l’immunothérapie passive. Selon la même cinétique d’âge, nous avons injecté, chaque semaine, par voie intrapéritonéale, des souris THY-Tau22 avec 5mg/kg et 10mg/kg de 2H9 ou une solution saline. Cette approche prévient l’apparition de déficits de mémoire spatiale mesurée par les tests du labyrinthe en Y et de la piscine de Morris. Les analyses immunohistochimiques révèlent également une réduction des protéines Tau anormalement phosphorylées au niveau de l’hippocampe. Afin d’étudier les mécanismes sous-jacents à l’immunothérapie anti Tau, nous avons injecté des anticorps anti-phosphoTau par stéréotaxie au niveau de l’hippocampe de souris THY-Tau22. Nous montrons, qu’une fois au sein du cerveau, ils sont capables d’entrer dans les neurones contrairement au contrôle isotypique. Plusieurs études récentes d’immunothérapie, suggèrent une implication de la macroautophagie dans la dégradation médiée par les anticorps. Nous montrons que les anticorps internalisés dans les neurones colocalisent avec différents marqueurs de la voie lysosomiale (NPC1, Lamp2) confirmant l’hypothèse d’une dégradation par le lysosome. Cependant, la barrière hémato encéphalique étant très sélective, il est fort probable que les anticorps générés restent en périphérie. [...] / Tau pathology is a common lesion observed in more than twenty neurological disorders, refered to as Tauopathies. It corresponds to the aggregation of the microtubule associated protein Tau hyper and abnormally phosphorylated into neurofibrillary tangles. In Alzheimer's disease (AD), the most common age-related dementia, the distribution and progression of Tau pathology have been reported to be well-correlated to the cognitive decline. Currently, treatments remain essentially symptomatic. However, several therapeutic approaches, including immunotherapy, are being developed to treat Tau pathology. The work presented in this thesis aim to investigate the effects of both active and passive Tau immunotherapy in a transgenic mouse model of Alzheimer’s disease-like Tau pathology. THY-Tau22 mice overexpress a double-mutated isoform of the human Tau protein, whose expression is under the control of a neuronal promoter. Several lines of evidence suggest that this transgenic mouse model is reproducing early stages of AD. Indeed, at three month old, the THY-Tau22 mouse model presents a progressive impairment of learning and memory without major neuronal loss, in parallel to Tau accumulation in the hippocampus. Moreover, Tau is hyper and abnormally phosphorylated at different sites including Ser422. Noteworthy, the pSer422 epitope is a single abnormal site of phosphorylation present only in pathological conditions, rendering it of particular interest for immunotherapy. In this present thesis work, we showed that early vaccination against the phosphorylated Ser422 Tau protein reduced spatial memory impairment as measured by the Y-maze test. Interestingly, this is associated with the decrease of the abnormal phosphorylation of Tau in the hippocampus and with a significant reduction of insoluble Tau species. Based on these results, we generated a new monoclonal antibody raised against pSer422 (2H9) to evaluate the effects of passive immunotherapy. The antibody was injected every week, intraperitoneally (5mg/kg and 10mg/kg of 2H9 or saline buffer) in the THY-Tau22 mice. We show that this approach can prevent the appearance of spatial memory deficits as measured by the Y maze and the Morris water maze tests. Immunohistochemical analysis also revealed a reduction of abnormally phosphorylated Tau proteins in the hippocampus. To investigate the mechanisms underlying Tau immunotherapy, we performed stereotaxic injections of anti-phosphoTau antibodies in the hippocampus of THY-Tau22 mice. We showed that, once in the brain, anti-phosphoTau antibodies were located inside the neurons in contrast to the isotype control. Several recent studies of immunotherapy, suggest an involvement of macroautophagy in the antibody-mediated degradation. We showed that internalized antibodies colocalized with different markers of the lysosomal pathway (NPC1, Lamp2) confirming this hypothesis of a lysosomal-mediated degradation. However, since the blood-brain barrier is highly selective, the antibodies may more likely act in the periphery. Indeed, we showed that peripheral administration of 250μg of the 2H9 generates a significant increase of Tau proteins levels in plasma, suggesting a peripheral sink mechanism as for Aβ immunotherapy. Vaccination, which generates a polyclonal response, leads to a greater increase of plasmatic Tau that confirms the peripheral degradation of Tau. Overall, the results of this PhD work confirmed the promising potential of Tau immunotherapy for Alzheimer\\\\\\\'s disease and other Tauopathies treatment. Moreover, it address a new hypothesis suggesting that the antibodies mediated degradation of Tau proteins might be through a peripheral sink such as for Aβ.

Hypothermia is potently neuroprotective, but the molecular basis of this effect remains obscure and the practical challenges of cooling have restricted its clinical use. This thesis was borne on the premise that considerable therapeutic potential may lie in a deeper understanding of the neuronal physiology of cooling. Rodent studies indicate that hypothermia can elicit preconditioning wherein a subtoxic stress confers resistance to an otherwise lethal injury. This cooling-induced tolerance requires de novo protein synthesis – a fundamental arm of the cold-shock response, for which data in human neurons is lacking. Since cooling protects the human neonatal brain, experiments herein address the molecular effects of clinicallyrelevant cooling using functional, maturationally-comparable cortical neurons differentiated from human pluripotent stem cells (hCNs). Several core hypothermic phenomena are explored, with particular scrutiny of neuronal tau, since this protein is modified extensively in brains that are resistant to injury. Mild-to-moderate hypothermia produces an archetypal cold-shock response in hCNs and protects them from oxidative and excitotoxic stress. Principal features of human cortical tau development are recapitulated during hCN differentiation, and subsequently reversed by cooling, returning tau transcriptionally and post-translationally to an earlier foetallike state. These findings provide the first evidence of cold-stress-mediated ontogenic reversal in human neurons. Furthermore, neuroprotective hypothermia induces mild endoplasmic reticulum (ER) stress in hCNs, with subsequent activation of the unfolded protein response (UPR). Reciprocal modulation of both tau phosphorylation and the ER-UPR cascade suggests that cold-induced hyperphosphorylation of tau and ER-hormesis (preconditioning) represent significant components of hypothermic neuroprotection. Cooling thus modifies proteostatic pathways in a manner that supports neuronal viability. Historically, hypothermic preconditioning has been limited to the acute injury setting, and tau hyperphosphorylation is an established hallmark of chronic neural demise. More recently however, preconditioning has been proposed as a target for neurodegenerative disease and neuroprotective roles of phospho-tau have emerged. To date, hypothermia has protected hCNs against oxidative, excitotoxic and ER stress, all of which have been implicated in traumatic as well as degenerative processes. This ‘cross-tolerance’ effect places exponential value on the molecular neurobiology of cooling, with the potential to extract multiple therapeutic targets for an unmet need.

Two searches for the production of Higgs bosons decaying into tau and mu leptons, using data collected with the DZero detector at the Fermilab Tevatron p-bar(p)collider, are presented. A search for the pair production of doubly charged Higgs bosons in the processq-bar(q) to H^{++}H^{--}, where H^{++} decays to tau-tau, mu-mu or tau-mu lepton pairs, with an integrated luminosity of up to L= 7.0 fb^{-1}, is presented. No significant excess of data over the expected SM background is observed and the results are used to set 95\% C.L. limits on the pair production cross section of doubly charged Higgs bosons in the range 90 < M_{H^{++}} < 200 GeV. A second search for the production of the Standard Model Higgs boson in the final state tau-tau-mu+X is presented, using an integrated luminosity of L=8.6 fb^{-1}. Again no significant excess of data is observed over the background expectation and 95% C.L. limits are set on the observed cross section relative to the Standard Model prediction, in the range 100 < M_{H} < 200 GeV.

Im Rahmen dieser Arbeit sollte versucht werden neue Proteine zu identifizieren, welche in einem zellulären Modellsystem der Alzheimer Krankheit durch Tau-vermittelte Zytotoxizität signifikant beeinflusst worden sind, um die Signalkette zwischen abnormaler Tau-Phosphorylierung und neuronalem Zelltod besser verstehen zu können. Hierzu wurden die Proteine hwt- und hPHP-Tau exprimierender PC12-Zellen nach 24 Stunden NGF-Behandlung mit Hilfe eines phosphoproteomischen Ansatzes analysiert. Nach Entwicklung, Etablierung und Optimierung des Ansatzes konnten die Tyrosinhydroxylase, Hsp84, Chromogranin B, ATP Citrat Lyase, Annexin A2 und TRAP1 spezifisch als Kandidaten identifiziert werden. Dabei wurde vor allem Hsp84 in darauffolgenden Experimenten eingehender untersucht. Über massenspektrometrische Analysen konnten zwei Phosphorylierungsstellen von Hsp84 nachgewiesen werden, wobei Serin 261 differentiell nur in hwt-Tau exprimierenden PC12-Zellen phosphoryliert und Serin 255 in beiden Zelllinien phosphoryliert vorlag. Serin 261 ist innerhalb der geladenen "Linker-Domäne" von Hsp84 lokalisiert und möglicherweise funktionell über die Interaktion mit Apaf-1 mit einem apoptotischen Verhalten der Zelle verbunden. Des Weiteren zeigte eine altersabhängige Untersuchung in Mäusehirnlysaten eine erhöhte Expression von Hsp84 in 20-22 Monate alten Kontrolltieren, welche verstärkt bei den männlichen aber auch weiblichen hwt- und hPHP-Tau transgenen Tieren signifikant ausblieb. Rekombinant hergestelltes Annexin A2 hatte in vitro mit steigender Konzentration inhibierende Wirkung auf die Nukleation Tau-vermittelter Mikrotubuli-Polymerisation. Bezüglich der anderen identifizierten Kandidaten wurden bislang keine weiteren Studien durchgeführt. Mit Hilfe des phosphoproteomischen Ansatzes ist es möglich signifikant beeinflusste Proteine qualitativ und quantitativ reproduzierbar nachzuweisen. Dabei lässt der Anwendungsbereich ein breites Spektrum an Ausgangsmaterial und äußeren Bedingungen zu.

Indiana University-Purdue University Indianapolis (IUPUI) / Alzheimer’s disease (AD) is a progressive neurodegenerative disorder characterized by the post-mortem deposition of amyloid-beta (Aβ) containing neuritic plaques and tau-loaded tangles. According to the amyloid hypothesis, the generation of Aβ via the cleavage of Aβ precursor protein (APP) by β-APP site-cleaving enzyme 1 (BACE1) is a causative step in the development of AD. Therefore, targeting the production and/or clearance of Aβ peptide (by Aβ-degrading enzymes such as Neprilysin) would help understand the disorder as well as serves as therapeutic potential to treat the disorder. MicroRNA are small, noncoding RNA capable of modulating protein expression by primarily targeting their 3’UTR. Therefore, identifying miRNA which target APP, BACE1 and Neprilysin (NEP) would elucidate the complicated regulatory mechanisms involved in protein turnover and provide novel drug targets. We identified miR-20b as a modulator of APP and soluble Aβ. We also identified the target site for miR-20b’s binding on the APP 3’UTR. Further, miR-20b exerts influence on neuronal morphology, likely due to its APP reduction. We also identified miR-298 as a dual regulator of APP and BACE1 and confirmed miR-298’s targeting of both 3’UTRs. We also showed that miR-298 overexpression reduced levels of both soluble Aβ40 and Aβ42 peptides. Additionally, we identified two SNPs in proximity to the MIR298 gene, which are associated with AD-related biomarkers. Based on these results, we showed miR-298 targets a specific isoform of tau by putatively binding a non-canonical target site on the MAPT 3’UTR. Finally, the insertion of the NEP 3’UTR into a reporter vector increases reporter expression; suggesting regulatory elements targeting the 3’UTR. We subsequently identified miR-216 as reducing NEP 3’UTR-mediated luciferase activity. We also measured levels of NEP protein in various mammalian tissue – such as rodent and human fetal tissue, and subsequently showed measurable Aβ levels in correlation with NEP expression. Therefore, herein, we have identified miRNA involved in the regulation of proteins implicated in the pathogenesis of AD.

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