Thèse numérisée par la Direction des bibliothèques de l'Université de Montréal.

Etwa 25 % der Nierenzellkarzinome (RCC) weisen bei Diagnosestellung bereits Metastasen auf. Aufgrund der schlechten Prognose des metastasierten RCC besteht ein dringender Bedarf an neuen Therapieformen sowie an prognostischen und diagnostischen Markern. microRNAs (miRNAs) bieten sich dabei als vielversprechende molekulare Biomarker an. Für den klarzelligen RCC-Subtypen (ccRCC) wurde bereits ein umfangreiches miRNA Expressionsprofil erstellt, mit dem ccRCC-relevante, vorwiegend herunterregulierte miRNAs identifiziert werden konnten. In der vorliegenden Arbeit wurde gezeigt, dass die Expression der miR-141 und miR-145 in RCC-Zelllinien durch epigenetische Mechanismen gehemmt ist und die Promotorbereiche dieser miRNAs stark methyliert vorliegen. In RCC-Zellen konnte eine tumorsuppressive Wirkung dieser miRNAs durch Hemmung der Migration (beide) und Invasion (miR-141) nachgewiesen werden. Durch die gleichzeitige Überexpression der beiden miRNAs kam es zu einer kooperativen Wirkung und so zu einer verstärkten Hemmung der Zellmigration. Weitere Untersuchungen konnten eine Reihe neuer onkogener Targets der miR 141 und miR 145 identifizieren. Dabei zeigte sich ein kooperativer Effekt durch Kombination beider miRNAs auf die Expression der Targets HS6ST2 und LOX. Die Targets LOX und MAP4K4 waren in ccRCC Gewebe auf mRNA-Ebene stark überexprimiert im Vergleich zum umliegenden Normalgewebe. Bei der anschließenden Tissue-Mikroarray-Analyse der Expression auf Proteinebene zeigte sich zudem ein prognostisches Potenzial der Targets LOX und MAP4K4 für das Gesamtüberleben von ccRCC Patienten. Diese Daten verdeutlichen den enormen Einfluss von epigenetisch dysregulierten miRNAs und deren spezifischen Targets auf tumorassoziierte Prozesse. Zudem bietet das Netzwerk aus Epigenetik, miRNAs und deren jeweiligen Targets nicht nur eine Reihe von diagnostischen und prognostischen Möglichkeiten, sondern liefert auch viele Ansatzpunkte für die Entwicklung von neuen therapeutischen Strategien. / Approximately 25 % of diagnosed renal cell carcinoma (RCC) have already metastasized. Due to poor prognosis of metastatic RCC, there is an urgent need for new therapies and prognostic and diagnostic markers to identify high-risk patients. Here microRNAs (miRNAs) might be promising new molecular biomarkers. For the clear cell RCC subtype (ccRCC) a comprehensive miRNA expression profile was already established. In this profiling several ccRCC-associated, predominantly down-regulated miRNAs were identified. In the present study, epigenetic mechanisms were identified to play a significant role in the down regulation of miR-141 and miR-145 in RCC cell lines. In addition, a strong methylation of the corresponding promoter regions was detected at molecular level. In RCC cells a tumor suppressive effect of these miRNAs was shown by decreasing migration (both) and invasion (miR-141) and furthermore, co overexpression of both miRNAs resulted in a cooperative effect with increased inhibition of cell migration. Several new oncogenic targets of miR-141 and miR-145 were identified by further investigations. Here the two miRNAs again showed a cooperative effect, as demonstrated by a significantly increased inhibition of HS6ST2 and LOX expression. In ccRCC tissue the expression of LOX and MAP4K4 was strongly enhanced on mRNA level compared to normal tissue. In the subsequent tissue microarray analysis of protein expression, LOX and MAP4K4 showed a prognostic impact for the overall survival of patients with ccRCC. These results illustrate a huge impact of epigenetically dysregulated miRNAs and of their specific targets on tumor-associated processes. Furthermore, the network of epigenetics, miRNAs and their respective targets will offer a number of diagnostic and prognostic capabilities, but will also provide many opportunities for the development of new therapeutic strategies.

O Glioblastoma (GBM) é um astrocitoma grau IV, representando o glioma de maior malignidade e o tumor cerebral primário mais frequente em humanos. A terapia indicada para o GBM é a ressecção cirúrgica, quimioterapia e radioterapia, todas com baixíssima eficiência devido a agressividade e as características do GBM. Consequentemente, a sobrevida dos pacientes indicados aos tratamentos convencionais é pouco mais de um ano. A inflamação é, sabidamente, uma das características que participa de modo decisivo do desenvolvimento tumoral, incluindo do GBM, e as relações entre mediadores inflamatórios e câncer são alvos de pesquisa nos últimos anos. Diversos estudos apontam um papel da via dos eicosanoides na modulação de processos patológicos envolvidos na inflamação e no câncer. Os eicosanoides são mediadores lipídicos bioativos, envolvidos em diversos processos fisiológicos e patológicos, em especial os associados à resposta inflamatória. As principais vias de produção de eicosanoides (ciclooxigenases, lipoxigenases, e citocromo P450), assim como seus produtos, são frequentemente alterados em diversos tipos de tumor, associados ao crescimento e a progressão tumoral. Contudo, o perfil dessas vias é consideravelmente pouco compreendido em GBM. O objetivo deste estudo foi analisar in vitro o perfil e o papel das três vias de eicosanoides e seus produtos (eicosanoides ou não) nas linhagens de GBM (U251-MG, U87-MG, A172, T98G e U138-MG), modulando a atividade de enzimas chaves com drogas especificas para, então, analisar parâmetros de proliferação, migração e morte celular. Nossos resultados mostram, em todas as linhagens analisadas, um perfil heterogêneo das enzimas e receptores chaves das três vias. O perfil lipídico evidencia a produção de 13-HODE, produto de 15-lipoxigenase-1 em todas as linhagens, bem como ausência de leucotrienos e 5-HETE do eixo de 5-lipoxigenase. Os inibidores farmacológicos para 15-LOX e 12-LOX/15-LOX foram capazes de reduzir o crescimento, modular o ciclo celular e a migração celular das linhagens U251-MG, U87-MG e A172. O mesmo é visto com a inibição de mPGES-1. A inibição de 5-LOX por outro lado não afetou nenhum parâmetro nas mesmas linhagens. Todos os resultados apontam, portanto, para um papel do eixo de 15-LOX e COX no crescimento e na migração das células de GBM humano. / Glioblastoma (GBM) is a grade IV astrocytoma, the most malignant and the most frequent primary brain tumour in humans. Standard therapies for treating GBM are surgical resection, chemotherapy and radiotherapy, all with low efficiency due to GBM aggressiveness and characteristics. Therefore, patient survival after conventional treatments is about one year. Inflammation is one of the main characteristics that plays a decisive role in tumour development, including in GBM. The relationships between inflammatory mediators and cancer have been the subject of research in recent years. Several studies point to the eicosanoid pathways as modulators of pathological processes between inflammation and cancer. Eicosanoids are bioactive lipid mediators, involved in various physiological and pathological processes, especially those associated with the inflammatory response. The major eicosanoid pathways (cyclooxygenases, lipoxygenases, and cytochrome P450) as well as their products, are frequently altered in several tumours, associated with tumour growth and progression. However, the profile of these pathways is poorly understood in GBM. The objective of this study was to analyse, in vitro, the profile and role of eicosanoid pathways and their products (eicosanoids or not) in GBM cell lines (U251-MG, U87-MG, A172, T98G and U138-MG), modulating the key enzymes with inhibitors, analysing proliferation, migration and cell death. Our results show, in all analysed cell lines, a heterogeneous profile for the key enzymes and receptors of the three pathways. The lipid profile shows the production of 13-HODE, a product of 15-lipoxygenase-1, as well the absence of leukotrienes and 5-HETE of the 5-lipoxygenase axis. The pharmacological inhibitors for 15-LOX and 12-LOX / 15-LOX led to changes in cell cycle, reduced growth, reduced migration of U251-MG, U87-MG and A172 cell lines. The same was seen with the inhibition of mPGES-1. Inhibition of 5-LOX, on the other hand, did not affect any of these parameters in the same cell lines. All results, therefore, point to an important role of 15-LOX and COX pathways in the growth and migration of human GBM cells.

Le nouveau moteur cryogénique de l'étage supérieur du nouveau lanceur Ariane présente la particularité d'être plusieurs fois réallumable, une fois la mise en orbite du lanceur. Le réallumage d'un moteur est particulièrement difficile durant les conditions de vol spatial. Ce moteur est composé d'un dôme LOX alimenté en oxygène liquide (LOX) qui est approvisionné par une vanne à boisseau positionnée en entrée d'une canne d'alimentation. Le mélange liquide / gaz formé dans le dôme LOX est injecté dans la chambre de combustion à travers des injecteurs reliant le dôme à la chambre. En conséquence, la distribution de l'écoulement diphasique en sortie des injecteurs revêt une importance particulière en terme d'allumage, de l'ouverture à la fermeture de cette vanne. La prise en compte de ces conditions de vol est primordiale pour qualifier le moteur. Cependant ces conditions ne peuvent pas être reproduites de façon représentative au cours d’essais au sol. Dans le cadre de ces études, un programme de recherche a été mis en place par le CNES (Centre National d'Études Spatiales) et SAFRAN Snecma pour étudier le remplissage du dôme LOX, via des études expérimentales et numériques. L'objectif est de connaître les conditions aux limites en sortie des injecteurs qui sont déterminantes pour appréhender la phase d'allumage dans la chambre de combustion. Des expériences ont été menées au LEGI (Laboratoire des Écoulements Géophysiques et Industriels) avec des fluides de substitution (de l'eau et de l'air), sans transfert de masse et de chaleur, sur la maquette du dôme d’alimentation d'un moteur de fusée. Les travaux présentés, menés à l'IMFT (Institut de Mécanique des Fluides de Toulouse), tentent de reproduire les expériences réalisées à l'aide de simulations numériques 3D incompressibles diphasiques. La géométrie du domaine de calcul est représentative de la maquette expérimentale, qui est composée d'une canne d'alimentation, d'un dôme, d'un allumeur et d'un grand nombre d'injecteurs. Le but de cette étude est de démontrer la faisabilité d'un calcul 3D instationnaire diphasique du remplissage du dôme oxygène du moteur-fusée avec le code industriel NEPTUNE_CFD, en prenant en compte la géométrie réelle et les phénomènes physiques prépondérants. La comparaison des prédictions avec les résultats expérimentaux est réalisée afin d'évaluer la capacité du code à prédire l'écoulement à l'aide des modèles de fermeture disponibles. Enfin, plusieurs études de sensibilité sur les modèles de fermeture sont menées pour estimer leur influence sur les résultats des simulations. Un travail important a été effectué pour imposer les mêmes conditions d'entrée que dans les expériences. Des études ont également été conduites sur un injecteur isolé. / New generation cryogenic upper-stage rocket engines are planned to be restartable during the orbit mission. The re-ignition of the engine is particularly difficult in space flight conditions. The engine contains a LOX dome fed with liquid oxygen (LOX) supplied by a bushel valve through a pipe. The gas / liquid mixture forming in the dome is injected into the combustion chamber through a number of injectors. Therefore the two-phase flow distribution at injectors outlet carries a real importance in terms of the ignition from the opening to the closing phases of the main valve. These flight conditions are of paramount importance, however, they are truly difficult to reproduce by experimental ground tests. In the framework of these studies, a research program set up by CNES (the French Space Agency) and SAFRAN Snecma, tries to study the LOX dome filling, through experiments and numerical studies. The aim is to identify the phenomena at sake to know the limit conditions at injectors, which will determine the ignition stage in the combustion chamber. Experiments are carried out at LEGI (Geophysical and Industrial Flows Laboratory in Grenoble) with substitution fluids (air and water), without heat and mass transfer on a rocket engine mockup. The work presented here, conducted at IMFT (Fluid Mechanics Institute in Toulouse), intends to reproduce the experimental results using incompressible two-phase flow simulations. The geometry used is representative of the experimental mockup composed of a feeding pipe, a dome, an igniter pipe and injectors. The aim of this study is to demonstrate the feasibility of a 3D unsteady two-phase flow calculation with the industrial code NEPTUNE_CFD, to simulate the LOX dome filling of the rocket engine, by taking into account the real geometry and the preponderant physical phenomena. The comparison of the predictions with the experimental results is carried out in order to estimate the code capability to predict the flow behavior, according to available closure laws. Finally, several sensitivity studies on the closure laws have been conducted to assess their influence on the numerical results. An important work has been carried out to obtain the proper inlet conditions to be imposed in the code in coherence with the experiments. Studies have equally been conducted on an isolated injector.

Computational Fluid Dynamics (CFD) simulations are presented within this study for super-cooled liquid oxygen atomisation and gasification in a subcritical chamber operating at 1MPa. Relatively low cost simulation techniques have been used and their accuracy evaluated. Gasification efficiency expected from theory is compared with simulation results and physical limitation in addition to modelling limitations are discussed. Impinging jets have been used within the simulations with the intent of atomising the incoming liquid oxygen, followed by injection of hot water vapour perpendicularly, to increase turbulent mixing, residence time and in turn expected gasification efficiency. A computational fluid dynamics heating analysis is also included in order to highlight constraints on the chamber geometry imposed by transient rapid oxidation material limits. 316 stainless steel and 3D printed Inconel 718 were investigated experimentally to identify their transient macroscopic rapid oxidation limits. This information supplements existing published literature for operation at high temperatures for a transient period of time in oxygen rich environments. ANSYS Fluent 2020R1, and its newly included Volume of Fluid to Discrete Particle (VOF-DPM) Model, is used for CFD simulation of LOx atomisation and vaporisation. The CFD simulation technique is discussed in detail in order to allow the reader to gain knowledge into areas where computational power can be saved while still allowing assessment of trends for conducting relatively quick feasibility reviews e.g. for different chamber configurations. The CFD simulation results are compared with published experimental data and its accuracy when extended to this application is discussed. Results indicate that gasification of LOx within a compact chamber may be feasible if sufficient turbulence, resulting in longer residence times is present providing sufficient time for heat and mass transfer from the continuous phase. Simulations indicate that due to the mixing and gasification process the LOx particles within the chamber that have not entered the gaseous phase are smaller than that from pure atomisation and therefore more susceptible to gasification if injected into the main motor combustion chamber. Results hint at the potential benefit of swirl injection of hot gases to increase residence time and in turn the gasification efficiency, therefore, this is recommended for the topic of future research. / Computational Fluid Dynamics (CFD) simuleringar presenteras i denna studie för superkyld flytande syreförstoftning och förgasning i en underkritisk kammare som arbetar vid SI 1 MPa. Relativt billiga simuleringstekniker har använts och deras noggrannhet utvärderats. Förgasningseffektivitet som förväntas från teorin jämförs med simuleringsresultat och fysisk begränsning utöver detta diskuteras modelleringsberäkningarna. Stötstrålar har använts inom simuleringarna med avsikt att finfördela det inkommande flytande syret, följt av injektion av varm vattenånga vinkelrätt, för att öka turbulent blandning, uppehållstid och i sin tur förväntad förgasningseffektivitet. En beräkningsenhetsanalys för uppvärmningsdynamik ingår också för att belysa begränsningar för kammargeometri som införs genom övergående gränser för snabb oxidation. 316 rostfritt stål och 3D-printad Inconel 718 undersöktes experimentellt för att identifiera deras övergående makroskopiska snabba oxidationsgränser. Denna information kompletterar befintlig publicerad litteratur för drift vid höga temperaturer under en kort tid i syrgasrika miljöer. ANSYS Fluent 2020R1, och dess nyligen inkluderade volym av vätska till diskret partikel (VOF-DPM) -modell, används för CFD-simulering av LOxatomisering och förångning. CFD-simuleringstekniken diskuteras i detalj för att göra det möjligt för läsaren att få kunskap om områden där beräkningskraft kan sparas medan man fortfarande tillåter bedömning av trender för att göra relativt snabba genomförbarhetsgranskningar, t.ex. för olika kammarkonfigurationer. CFD-simuleringsresultaten jämförs med publicerade experimentella data och dess noggrannhet när den utvidgas till denna applikation diskuteras. Resultaten indikerar att förgasning av LOx i en kompakt kammare kan vara möjlig vid tillräcklig turbulens, vilket resulterar i längre uppehållstider är närvarande som ger tillräcklig tid för värme och massöverföring från den kontinuerliga fasen. Simuleringar indikerar att på grund av blandnings- och förgasningsprocessen är LOx-partiklarna i kammaren som inte har gått in i gasfasen mindre än den från ren förgasning och därför mer mottagliga för förgasning om de injiceras i huvudmotorns förbränningskammare. Resultat antyder den potentiella fördelen med virvelinjektion av heta gaser för att öka uppehållstiden och i sin tur förgasningseffektivitet, därför rekommenderas detta för ämnet för framtida forskning.

A better understanding of these structural changes that occur before Hypertension (HTN) could ultimately result in a treatment that can prevent or reverse this disease state before its onset. T cells have been shown essential for the development of HTN. The aim of these murine studies was to investigate the role of the T-helper CD4⁺ lymphocytes in initiating vascular remodeling and HTN in the absence of an increased mechanical load and to investigate the role of T-helper 17 (Th17) CD4⁺ lymphocyte initiating vascular remodeling and HTN by stimulating Lysyl Oxidase (LOX). LOX is known to cross-link collagen and elastin and. Excess synthesis of collagen and elastin results in a stiffer artery and hypertension. We established L-NAME-induce HTN in wild type (WT) mice. CD4⁺ splenic lymphocytes were isolated from these mice and adoptively transferred into naïve syngeneic severe combined immunodeficient (SCID) mice. The SCID mice receiving these cells became hypertensive. Cytokine analysis demonstrated an increase in both Th1 and Th17 cytokine in HTN donor mice and of lymphocytes in the aortic infiltrates of the HTN recipient mice. The increased collagen and LOX expression in recipient mice suggest that the adoptively transfer CD4⁺ lymphocytes are associated with vascular extracellular matrix remodeling. Furthermore we examined the role of Th17 lymphocyte in aortic LOX regulation in Angiotensin II-induced hypertension. The Increase in blood pressure and Velocity Time Integral (VTI) was measured in WT Angiotensin II treated mice whereas no change was detected in the Th17 deficient (RORT KO) Angiotensin II treated group. When compared to the control group the WT group infused with Angiotensin II had higher LOX protein expression, LOX fluorescent Immunohistochemical stain and LOX activity. This group also had increased hydroxyproline levels, collagen stain, hyperplasia and aortic thickening. In contrast, the Th17 deficient mice Angiotensin II treated group had no changes in these parameters. The results provide evidence that IL-17 mediates Angiotensin II-induced hypertension and vascular dysfunction by the overstimulation of LOX. Potentially targeting T17 cells will allow for a drug-specific therapeutic approach and delay the progression of hypertension pathology.

Several members of the lysyl oxidase family of copper-dependent amine oxidases have been implicated in tumor development. The Iysyl oxidase (LOX) and LOX-like 2 (LOXL2) genes have been mapped to chromosomal regions affected by loss of heterozygosity (LOH) in several cancers, including those of the colon and esophagus. Indeed, there have been numerous reports of reduced LOX and a few reports of reduced LOXL2 expression in various cancers. Identification of microsatellite markers within the LOX locus and the LOXL2 gene allowed for evaluation ofthe status of these gene alleles in colon and esophageal tumors. There was significant LOH of the LOX locus in colon tumors that was accompanied by reduced mRNA expression and a spectrum of alterations and mutations affecting the LOX gene. This study demonstrated, for the first time, that genetic events, namely LOH, deletions and mutations ofthe LOX gene, were responsible, at least partly, for the reduction of LOX gene expression. There was also significant LOH of the LOXL2 gene in both colon and esophageal tumors. However, instead of a reduction of LOXL2 expression, there was increased expression that correlated with less differentiated tumors and absent elastosis, both indicators of poor prognosis. Further studies indicated that both LOX and LOXL2 are absent in non-invasive tumor cell lines but re-expressed in invasive cell lines, likely as part of the thelial-mesenchymal transition that occurs in the last steps of tumorigenesis to facilitate metastasis. The results presented and research strategy outlined in this dissertation will define the importance of LOXL2 amine oxidase activity and protein interactions in the critical but poorly understood process oftumor cell migration and invasion.

No description available.

京都大学 / 新制・課程博士 / 博士(農学) / 甲第24669号 / 農博第2552号 / 新制||農||1099(附属図書館) / 学位論文||R5||N5450(農学部図書室) / 京都大学大学院農学研究科応用生命科学専攻 / (主査)教授 菅瀬 謙治, 教授 小川 順, 教授 森 直樹 / 学位規則第4条第1項該当 / Doctor of Agricultural Science / Kyoto University / DFAM

L’ostéopétrose est une pathologie osseuse caractérisée par des os denses et fragiles principalement due à l’incapacité des ostéoclastes, cellules d’origine hématopoïétique, à résorber le tissu osseux. La forme la plus sévère de cette maladie génétique est l’ostéopétrose autosomale récessive infantile due à une mutation du gène Ostm1 (Protéine transmembranaire de type 1 associée à l'ostéopétrose). Le gène Ostm1 est exprimé principalement dans la lignée des cellules hématopoïétiques, mais aussi dans le système nerveux central et les mélanocytes. Cette mutation développe plusieurs symptômes comme l’apparition d’une couleur de pelage gris chez la souris, une anémie sévère, une sensibilité aux infections et des troubles neuronaux chez l’homme et la souris. Afin de mieux comprendre cette maladie, nous avons généré des souris transgéniques sur un fond génétique grey-lethal (gl) dans lesquelles l’expression d’Ostm1 est ciblée à un tissu spécifique. Nous avons caractérisé le gène Ostm1 responsable de la mutation ostéopétrotique spontanée gl chez la souris. La complémentation fonctionnelle des défauts hématopoïétiques a été obtenue dans les souris transgéniques PU.1-Ostm1-gl/gl mais ces souris meurent prématurément avec une neurodegénérescence sévère. Cette perte cellulaire affecte le système nerveux central dans son ensemble incluant la rétine. Ce mémoire porte sur le but d’établir le profil d’expression du gène Ostm1 dans la rétine puisque la perte du gène entraine une dégénérescence rétinienne. Pour définir le rôle d’Ostm1 dans la rétine, nous avons caractérisé son expression dans ce tissu (organe). Des analyses PCR, démontrent une expression d’Ostm1 dans l’œil total et enrichie dans la neurorétine et dans l’épithélium pigmentée (RPE). Après avoir caractérisé avec des marqueurs protéiques spécifiques les sous populations cellulaires de la rétine, in situ hybridation détecte l’expression préférentielle d’Ostm1 dans l’épithélium pigmentée (RPE) et la couche nucléaire interne (INL). Basé sur ce profil d’expression, nous avons induit dans un premier temps la perte de fonction d’Ostm1 spécifiquement dans le RPE. Dans un premier temps nous avons vérifié que l’expression de la recombinasse Cre seule n’est pas toxique. Nous avons ensuite induit la perte d’expression d’Ostm1 dans ces cellules et démontré que la perte d’Ostm1 dans le RPE se traduit par une perte graduelle des photorécepteurs avec l’âge. Ces résultats préliminaires suggèrent que l’expression post-natale d’Ostm1 dans le RPE est essentielle au maintien de l’homéostasie des photorécepteurs dans la rétine. / Osteopetrosis is a disease characterized by high bone density and fragility principally caused by impaired activity of osteoclasts, which are cells that reside in bone and dissolve bone tissue. The most severe form of osteopetrosis is infantile autosomal recessive osteopetrosis (ARO) which is caused by mutations in genes Ostm1. As Ostm1(osteopetrosis-associated transmembrane protein 1) is expressed in multiple hematopoietic stem cell lineages, melanocytes and the nervous system, mutations in Ostm1 can cause coat color change in mice as well as bone fragility, anemia, infections and neuronal disorders in humans and mice. To further the understanding of these conditions linked with Ostm1 loss, multiple tissue specific Ostm1 transgenic mice over an Ostm1 knockout (gl/gl) background were constructed. To better understand this disease, we characterized the Ostm1 gene responsible for the spontaneous osteopetrotic mutation grey- lethal (gl) in mice. Functional complementation of hematopoietic defects was obtained in PU.1-Ostm1-gl/gl transgenic mice, but these mice die prematurely with severe neurodegeneration. This indicates that Ostm1 has a crucial role in neuronal and retinal health. As a result, we wished to establish an expression profile of Ostm1 in all the layers of the retina to further decipher the role of Ostm1 in the retina. Polymerase chain reaction (PCR) of reverse-transcribed mRNA of separated sections of the eye demonstrate that Ostm1 is expressed in the whole eye, neuroretina and retinal pigmented epithelium (RPE). Further specific expression analyses were performed by in- situ hybridization which showed that Ostm1 is expressed specifically in the inner nuclear layer of the neuroretina as well as in the RPE. Based on this tissue expression pattern, we have constructed, for the first time, an RPE specific knockdown of Ostm1 expression and verified that the expression of Cre recombinase in this tissue is not toxic. The reduction of Ostm1 in the RPE of the eye resulted in gradual loss of photoreceptors of the retina. These preliminary results suggest that the post-natal expression of Ostm1 in the RPE is essential for maintaining the homeostasis of the photoreceptors of the retina.