Abstract The amount and distribution of genetic diversity are likely to affect the evolutionary potential of a species. When feasible and cost-effective policies for management and monitoring of endangered populations or species are planned, knowledge of the spatial genetic structure and the type of population dynamics is of great concern. In this thesis the genetic diversity and population structures of two endangered arctic plant species was examined on different geographical scales in Northern Europe. The species were Siberian primrose (Primula nutans) and pendant grass (Arctophila fulva), which both grow in a seashore habitat and have similar distribution patterns on the shores of the Arctic Ocean and the Bothnian Bay. The goal of the present study was to provide basic population genetic information for the study species using microsatellite and AFLP markers. Both markers were used for the first time in these species, and species-specific microsatellite markers were developed during the study. A further aim was to interlink the population genetic processes of the species into distribution patterns at the regional and population levels and to compile a synthesis of the impact of hierarchical spatiotemporal processes and autocorrelation in genetic variation at different levels. The studies of this thesis provided new information on the diversity and population structure of the endangered study species and new markers that are useful in future genetic studies of primrose species. The diversity of Siberian primrose was low, and there was no dispersal between the study regions. In pendant grass, a relatively high amount of variation was found considering the evident clonal reproduction and gene flow that was detected between populations connected by waterways. The results suggested that both clonal and sexual reproduction are important in this species. On a local scale, pendant grass populations had characteristics of “stepping stone” and classical metapopulation models. The results indicated that on a long time scale, both species will continue to decline without efficient management efforts. Most critical for the persistence of the species is the conservation of suitable habitats. Translocations could be considered in order to enhance the diversity of existing populations and establish new populations. By examining the present day structure of Siberian primrose, it was possible to make inferences regarding the colonisation history of the species in the North European area. The current distribution of Siberian primrose seemed to result from a vicariant process that took place after the last ice-age, when the species colonised the area. It spread first to the White Sea area, probably from the east, and subsequently colonised the Bothnian Bay and the Barents Sea in the west. The isostatic land uplift after the retreat of the Eurasian ice sheet uncovered large areas of land from the Baltic Sea basin that previously were under water. These geological changes resulted in the current disjunct distribution of Siberian primrose.

L'instabilité microsatellitaire (MSI) résulte d'une déficience du système de réparation des mésappariements de l'ADN. Cette instabilité est observée dans 10-15% des tumeurs chez l'Homme, incluant les cancers colorectaux (CCR) et de l'estomac (CG). En 2011, notre laboratoire a rapporté la mutation de la chaperonne HSP110 dans les CCR MSI. Cette mutation affecte un microsatellite intronique de 17 thymidines (T17), localisé au niveau de l'intron 8. Les grandes délétions somatiques du T17 (? 5 paires de bases), représentant 25% des CCR MSI, conduisent à l'inactivation complète de la chaperonne HSP110 dans les CCR MSI. De manière remarquable, ces grandes délétions sont prédictives chez les patients d'une excellente réponse à la chimiothérapie adjuvante. Au cours de ma thèse, mes travaux ont visé à étudier l'impact de la mutation d'HSP110 dans les tumeurs gastro-intestinales MSI. Mes résultats démontrent que la mutation du microsatellite T17 d'HSP110 a pour conséquence une diminution de la prolifération cellulaire en partie lié à la diminution de la phosphorylation du facteur de transcription STAT3. En outre, mes résultats suggèrent que cette mutation serait un facteur prédictif de survie chez les patients atteint de CG, indiquant le potentiel théranostique d'HSP110. Enfin, je propose une approche thérapeutique innovante pour les patients atteints de CCR MSI, basée sur la potentialisation de l'expression de transcrits mutants, codant pour des protéines délétères pour la cellule tumorale, à l'instar du dominant négatif HSP110DE9 résultant de la mutation d'HSP110 dont l'ARN semble être régulé par le système NMD (Nonsense-Mediated mRNA Decay). / Microsatellite instability (MSI) results from impaired DNA mismatch repair, being observed in 10-15% of frequent tumors in human, e.g. Colorectal (CRC), Gastric Cancers (GC) and others. In 2011, frequent somatic mutations of the HSP110 chaperone have been reported in MSI CRC by my lab, affecting a T17 intronic DNA repeat located in intron 8. Large (≥ 5 base pairs) bi-allelic somatic deletions of this DNA repeat in tumor DNAs, as observed in about 25% of MSI CRC, lead to complete inactivation of HSP110 by exon 9 skipping and sensitization of tumor cells to chemotherapy. These large deletions are predictive of improved response to adjuvant chemotherapy in CRC patients. During my PhD thesis, I further investigated the role of HSP110 in MSI tumors. My results demonstrate that HSP110 mutation leads to cell proliferation decrease through the reduction of STAT3 transcription factor phosphorylation in CRC tumors (Berthenet*, Bokhari*, et al., Oncogene 2016). Furthermore, I showed that HSP110 mutation is also frequently observed in MSI gastric cancer, leading to very similar pathophysiological consequences during tumor progression and improved patient’s survival independently from tumor stage (Cervera*, Lagrange*, Bokhari* et al., submitted). Finally, I worked on an innovative therapeutic approach that consisted in inhibiting the NMD (Nonsense-Mediated mRNA Decay) system, an ubiquitous process recognizing and degrading mRNAs containing premature termination codons (PTC). The inhibition of NMD leads to the expression of deleterious MSI-driven mutant transcripts such as the HSP110DE9, coding for a dominant negative mutant, derived from HSP110 mutation in MSI cancer cells.

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / Millions of years ago many species had their populations isolated by geographic barriers, during the formation of the drainage systems of South America Rivers. Modifications in relief lead to the actual formation of the hydrographic basins from Rio Grande do Sul state. Since these basins do not have connection among them, species widely distributed, as Aegla uruguayana Schmitt, 1942, may have their genetic variability being influenced by the watersheds which do not allow gene flow to occur. The aim of this study was to verify the influence of geographic barriers to the genetic structure of different populations of A. uruguayana. The migration patterns of heteroduplex DNA were used to analyze two populations of A. uruguayana from two hydrographic regions of the state (East and West). A significant number of haplotypes was observed in each population, reflecting a high proportion of intrapopulation diversity in AMOVA (49,38%). The geographic barriers seem to be influencing the genetic differentiation of A. uruguayana populations, at least among the populations from Rivers Santa Maria, Ibirapuitã and Camaquã, which had FST values calculated. For the other populations it is necessary to analyze a greater number of individuals, what will make easier the verification of the genetic structure. The utilization of an ultra sensitive molecular marker, such as the microsatellites, will allow a more refined analysis about this question. Thus, microsatellite loci were isolated and characterized. The efficiency of the microsatellite isolation was high and primers were designed for three loci, two of these being polymorphic. Two loci, Au05 and Au13, successfully amplified and were highly polymorphic, with seven and eight alleles, respectively, making them promising for the evaluation of differences among A. uruguayana populations. These loci also successfully cross-amplificated in Aegla longirostri, and have potential for the evaluation of other species from the same gender. / Muitas espécies tiveram populações separadas por barreiras geográficas milhões de anos atrás durante a formação dos sistemas de drenagem dos Rios da América do Sul. Modificações no relevo levaram a atual formação das bacias hidrográficas do estado do Rio Grande do Sul. Como estas bacias não possuem ligação entre si, em espécies com distribuição ampla, como Aegla uruguayana Schmitt, 1942, os divisores de água podem estar influenciando a variabilidade genética das populações que não podem manter um fluxo gênico. O objetivo desta dissertação é verificar a influência das barreiras geográficas na estruturação genética de diferentes populações de A. uruguayana. Foi utilizado padrão de bandas de DNA heteroduplex para analisar populações de A. uruguayana de duas regiões hidrográficas do estado (Leste e Oeste). Foi observado um número significativo de haplótipos em cada população, refletindo uma alta proporção de diversidade intra-populacional na AMOVA (49,38%). As barreiras geográficas parecem estar influenciando na diferenciação genética das populações de A. uruguayana, ao menos entre as populações dos Rios Santa Maria, Ibirapuitã e Camaquã, as quais tiveram os valores de FST calculados. Para as outras populações será necessária a análise de um número maior de indivíduos, possibilitando a verificação da estrutura genética. A utilização de um marcador molecular ultra-sensível, como os microssatélites, permitirá uma análise mais refinada sobre esta questão. Para tanto, foram isolados e caracterizados locos de microssatélites. A eficiência do isolamento de microssatélites foi bastante alta, pois designamos primers para três locos, desses, dois se mostraram polimórficos. Os dois locos, Au05 e Au13, amplificaram com sucesso e foram bastante polimórficos, com 07 e 08 alelos respectivamente, o que os torna promissores para avaliar diferenças entre populações de A. uruguayana. Esses locos amplificaram com sucesso em Aegla longirostri, tendo potencial para avaliar outras espécies do mesmo gênero.

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / The Aeglidae are Decapoda crustaceans endemic from the neotropical region of South America. They are the only anomuran family entirely restricted to freshwater, occurring mainly in streams. Requirement for well conserved habitat has restricted the populations to springs, as an effect of the constant degradation of continental aquatic ecosystems. In the central region of Rio Grande do Sul state, the species Aegla longirostri, besides suffering the effects of habitat fragmentation, is subjected to a mountainous geographic barrier, which has been separating the basins of River Guaíba and River Uruguay for about 11 my. Microsatellite loci are molecular markers with high levels of heterozigosity which have been widely used in population genetics studies. Microsatellite markers previously isolated from A. longirostri genome were characterized and the levels of heterozigosity and allelic diversity were calculated for each locus. A total of seven polymorphic microsatellite loci were used to verify genetic variability among four different populations from central region of Rio Grande do Sul state concerned the two basins described above. Results show a great genetic differentiation among all populations and not only between populations isolated by the geographic barrier. Factors as lack of larval stage and low dispersion capacity are possibly contributing to such differentiation level. Anthropic actions, resulting in alteration of aquatic environments, can also be a more recent factor contributing to the genetic diversity among the populations studied, since aeglids are very strict in relation to the water quality. / Os aeglídeos são crustáceos decápodos endêmicos da região Neotropical da América do Sul. São os únicos anomuros de água doce, habitantes principalmente de cursos d água. A exigência por um habitat conservado tem restringido as populações de Aegla às nascentes, como efeito da constante degradação ambiental dos ecossistemas aquáticos continentais. Na região central do estado do Rio Grande do Sul destacamos a ocorrência de populações de Aegla longirostri, que além dos efeitos de fragmentação de habitat, estão expostas a uma barreira geográfica de formação montanhosa, que há cerca de 11 milhões de anos separa as bacias dos Rios Uruguai e Guaíba. Locos de microssatélites são marcadores que apresentam altos níveis de heterozigosidade e têm sido amplamente utilizados em estudos de genética de populações. Marcadores microssatélites previamente isolados do genoma de A. longirostri foram caracterizados determinando-se os níveis de heterozigosidade e a diversidade alélica para cada loco. Dos oito locos de microssatélites analisados, sete se mostraram polimórficos e foram empregados em indivíduos de quatro diferentes populações da região central do estado do Rio Grande do Sul para analisar a variabilidade genética entre as populações de A. longirostri em ambas as bacias. Os resultados mostram uma grande diferenciação genética entre todas as populações e não somente entre as populações isoladas pela barreira geográfica. Possivelmente, fatores como ausência de fase larval durante o desenvolvimento destes crustáceos, aliada a uma baixa capacidade de dispersão, podem estar contribuindo para esta diferenciação. A ação antrópica, resultando em degradação do ambiente aquático, também pode ser um fator recente a contribuir para a diferenciação genética entre as populações estudadas, visto que os aeglídeos são bem exigentes em relação à qualidade da água onde vivem.

Parasitic copepods including Lepeophtheirus salmonis have been the focus of strong concern for the health of wild and farmed salmonids in the Pacific and Atlantic Oceans. Salmon are highly valuable species from both socioeconomic and ecological perspectives. The host-parasite dynamics of Lepeophtheirus salmonis and the Atlantic and Pacific salmonids have changed over evolutionary time to the point that both Atlantic and Pacific salmon and Atlantic and Pacific Lepeophtheirus salmonis are genetically distinct. Recent human interference with the natural population dynamics of this parasite and its hosts may have altered the population genetic structure of Lepeophtheirus salmonis, particularly because salmon farms may provide more stable conditions for parasite population growth. High abundance of Lepeophtheirus salmonis on salmon farms causes damage to the farmed salmon and leads to increased infection intensities in nearby wild hosts. Some Atlantic Lepeophtheirus salmonis have developed resistance to the anti-parasitic drugs they are repeatedly exposed to. No drug resistance has yet been detected within the Pacific Ocean, where only one drug is available, and heavily relied on, to treat Lepeophtheirus salmonis infections. Control of Lepeophtheirus salmonis abundance on Pacific salmon farms is important to maintain the health of farmed salmon and is also important to protect wild salmonids from increased infections originating from salmon farms. The goal of this thesis was to characterize and employ a large suite of molecular markers to assess the population structure of Lepeophtheirus salmonis in the Pacific Ocean. Until this point, the primary focus of Lepeophtheirus salmonis population genetics research has been limited to the Atlantic Ocean and has relied on a small number of available molecular markers. Available expressed sequence tag DNA libraries were screened to identify putative polymorphic loci, which were then experimentally evaluated. We characterized 22 novel microsatellite loci and 87 single nucleotide polymorphisms within 25 nuclear loci for Lepeophtheirus salmonis. We used these genetic markers, as well as 5 microsatellite loci previously developed for use in Atlantic Lepeophtheirus salmonis population studies, to genotype 562 Lepeophtheirus salmonis that were collected from12 Pacific Ocean sampling locations. We compared Lepeophtheirus salmonis genotypes among: (1) seven wild host populations and five farmed host populations within the Pacific Ocean; (2) geographically separated wild host populations, ranging from the Bering Sea to the southwest end of Vancouver Island, British Columbia; and (3) temporally separated cohorts of farmed Atlantic salmon from two geographically distant farm locations on the northwest coast of Vancouver Island and the Campbell River area east of central Vancouver Island. Our analyses failed to resolve significant population structure among sampled Pacific Lepeophtheirus salmonis and, therefore, supports a hypothesis of high gene flow throughout the Northeast Pacific Ocean. It is important to understand the biology and population dynamics of Lepeophtheirus salmonis because it is a consequential parasite of wild and farmed salmonids in the Pacific Ocean. Both the molecular tools developed for this study and the population genetics information generated from this study have contributed to our overall understanding of the evolutionary history and population dynamics of Lepeophtheirus salmonis. / Graduate

Broad whitefish, Coregonus nasus, is an important subsistence fish species in Arctic North America, yet virtually nothing is known regarding the genetic population structure of Nearctic populations of this species. In this thesis, microsatellite DNA variation was assayed among 1213 broad whitefish from 47 localities throughout North America, with emphasis on the Mackenzie River system, Northwest Territories. Specifically, I examined geographic variation in allele frequencies to assess how historical factors (Pleistocene glaciations) have shaped the current structuring of genetic variability and population differentiation. Microsatellite data was also used to resolve the relative contributions of broad whitefish populations to subsistence fisheries in the Mackenzie River system. Overall, broad whitefish exhibit relatively high intrapopulation microsatellite variation (average 12.29 alleles/locus, average HE = 0.58) and there were declines in these measures of genetic diversity with distance from putative refugia suggesting historical factors, namely post-glacial dispersal, have influenced current microsatellite variation. Interpopulation divergence was low (overall FST = 0.07), but the main regions assayed in this study (Russia, Alaska, Mackenzie River and Travaillant Lake systems) are genetically differentiated. Strong isolation-by-distance among samples was resolved when including only those populations occupying former Beringia, but not when assaying those at the periphery of the range in the Mackenzie River system, suggesting that broad whitefish in the Mackenzie system have not occupied the region long enough since their invasion post-glacially to have approached equilibrium between gene flow and drift. Mixture analysis indicated that most fish from the lower Mackenzie River subsistence fishery originated from the Peel River, highlighting the importance of this tributary. Additionally the mixture analysis provides evidence for a putative riverine life history form in the Mackenzie River. My results indicate that glaciation and post-glacial colonization have been important in shaping the current genetic population structure of North American broad whitefish. They also illustrate the utility of microsatellite DNA to delineate population structure and patterns of genetic diversity in recently founded populations in addition to resolving contributions to fisheries. My data also support the hypothesis that there are several designatable units of conservation among broad whitefish populations and that management strategies should be implemented accordingly. / Science, Faculty of / Zoology, Department of / Graduate

L'inactivation du système MMR (mismatch repair) favorise un processus oncogénique d'instabilité des microsatellites du génome (MSI). Au cours de ma thèse, j'ai étudié d'une part le rôle de l'azathioprine (Aza) dans l'induction de tumeurs MSI chez la souris. Des études épidémiologiques avaient rapporté une corrélation entre l'émergence de cancers MSI tardifs chez l'homme, et la prise au long cours de cet immunosuppresseur dont la cytotoxicité in vitro est médiée par l'activité MMR. Dans une étude dose-réponse, j'ai observé l'émergence de rares lymphomes MSI de survenue tardive chez la souris de génotype sauvage traitée par l'Aza, mais pas par la ciclosporine (autre immunosuppresseur utilisé en comparaison). Ces résultats permettent d'établir in vivo que l'Aza est un facteur de risque pour l'émergence de tumeurs MSI lors d'une exposition prolongée. D'autre part, je me suis intéressée au syndrome CMMRD (constitutional MMR deficiency), une prédisposition majeure et rare, aux cancers MSI. Les patients atteints étant porteurs de mutations germinales bialléliques d'un gène MMR, le diagnostic repose sur le génotypage constitutionnel, une méthode non-contributive quand un variant de signification inconnue est détecté (30% patients). Dans ce contexte, j'ai développé une méthode d'aide au dépistage de ce syndrome chez les sujets à risque, l'hypothèse étant que 2 caractéristiques fonctionnelles des cellules tumorales MMR-déficientes, le phénotype MSI et la tolérance aux agents génotoxiques tels que l'Aza, pouvaient être objectivées dans les tissus sains des patients CMMRD. Mes travaux proposent un test diagnostique sensible et spécifique qui répond aux limites de l'analyse génétique. / Inactivation of the MMR (mismatch repair) system promotes the oncogenic process of microsatellite instability (MSI). During my PhD, I firstly investigated the role of azathioprine (Aza) in the induction of MSI tumors in mice. Epidemiological studies reported a correlation between the occurrence of late MSI cancers in humans and long-term treatment with this immunosuppressant whose cytotoxicity was shown in vitro to be mediated by MMR activity. Using a dose-response study, I observed the occurrence of rare late-onset MSI lymphomas in wild-type mice treated with Aza, but not with ciclosporin (another immunosuppressant used for comparison). These results established in vivo that long-term Aza exposure is a risk factor for the emergence of MSI tumors. Secondly, I was interested in the CMMRD syndrome (constitutional MMR deficiency), a major and rare predisposition to MSI cancers. Since CMMRD patients are carriers of biallelic germline mutations of a MMR gene, diagnosis is based on constitutional genotyping, a method that was found non-contributory when a variant of unknown significance is detected (30% patients). In this context, I developed a complementary approach for the detection of this syndrome in at-risk patients, based on the hypothesis that two functional features of MMR-deficient tumor cells, i.e. the MSI phenotype and the tolerance to genotoxic agents such as Aza, can be demonstrated in non-neoplastic tissues of CMMRD patients. We provided a sensitive and specific method that may constitute a valuable tool when diagnosis of CMMRD could not be confirmed by genetic testing.

Orientadora: Profa. Dra. Cibele Biondo / Dissertação (mestrado) - Universidade Federal do ABC, Programa de Pós-Graduação em Evolução e Diversidade, São Bernardo do Campo, 2018. / A queixada (Tayassu pecari) é uma espécie considerada vulnerável globalmente e criticamente ameaçada em áreas de Mata Atlântica, devido à pressão de caça e fragmentação de habitat. A fragmentação de habitat devido a atividades antrópicas pode acarretar em estruturação genética das populações, além perda de diversidade genética e aumento de endogamia, o que pode contribuir para o processo de extinção local. O presente estudo teve como objetivo estimar o status genético da população de queixadas da região do Pontal do Paranapanema, na Mata Atlântica de SP. Essa região foi altamente fragmentada por atividades antrópicas e, no momento da realização da amostragem para este trabalho, a espécie se encontrava em apenas três dos fragmentos remanescentes (Parque Estadual do Morro do Diabo ¿ PEMD, Fazenda Ponte Branca ¿ FPB e Fazenda Santa Mônica ¿ FSM). Atualmente, é considerada extinta na região. Foram analisadas 47 amostras de sangue, coletadas entre os anos de 1999 e 2005, que foram genotipadas para 11 locos de microssatélites. Observou-se uma estruturação genética das três localidades amostradas em duas subpopulações. Duas das três localidades foram agrupadas em uma única subpopulação (PEMD e FPB) e houve diferenciação genética significativa apenas entre duas delas (PEMD e FSM). Os índices de diversidade genética foram similares para as três localidades e não foram encontradas evidências significativas de endogamia. Esses resultados podem ser explicados de duas formas: 1) existência de fluxo gênico, mesmo que baixo, entre as localidades amostradas (principalmente entre PEMD e FPB); 2) diferenciação recente das subpopulações. Conforme esperado para populações fragmentadas, houve evidências de gargalo populacional recente nas três localidades. Os resultados aqui obtidos podem ser indicativos do papel da paisagem antropizada na estruturação de populações de queixadas mesmo em pequena escala, já que o mesmo não tem sido registrado para regiões mais conservadas. Desta forma, estas evidências podem servir de direcionamento para planos de manejos e ações conservacionistas para outras populações de queixadas que se encontram em condições semelhantes a esta, visando diminuir as chances de extinção local em outras regiões. / The white-lipped peccary (Tayassu pecari) is considered vulnerable throughout its distribution and it is critically endangered in the Atlantic Forest, due to hunting pressure and habitat fragmentation. Habitat fragmentation caused by anthropic activities usually lead to population genetic structure, loss of genetic diversity and higher levels of inbreeding, which contributes to local extinctions. In this study, we addressed the genetic status of a white-lipped peccary population from Pontal do Paranapanema region, located in the Atlantic Forest of São Paulo State in Brazil. This area was highly fragmented by anthropic activities and white-lipped peccaries were found only in three remnant fragments (Parque Estadual do Morro do Diabo ¿ PEMD, Fazenda Ponte Branca ¿ FPB e Fazenda Santa Mônica ¿ FSM) when they were sampled for this work. Currently, they are considered extinct in the region. A total of 47 blood samples were collected between 1999 and 2005 and were genotyped for 11 microsatellite loci. We found that the three fragments sampled were structured in two subpopulations, with two of them being grouped into a single subpopulation (PEMD and FPB). We observed significant genetic differentiation only between two fragments (PEMD and FSM). The genetic diversity indices were similar for the three fragments and no indicators of inbreeding were found. These results can be explained in two ways: 1) existence of gene flow, even if low, between fragments (mainly between PEMD and FPB); 2) recent differentiation of the subpopulations. As expected for fragmentated populations, we found evidence of a population bottleneck for all the fragments sampled. These results can be indicative of the role of anthropized landscape in the structuration of white-lipped peccary populations even on a small scale, since it has not been recorded for well conserved regions. These finds can serve to delineate management plans and conservation actions for other white-lipped peccary populations that are in similar conditions, aiming to reduce the chances of local extinction in other regions.

Microsatellites, or simple tandem repeat sequences, occur naturally in the human genome and have important roles in genome evolution and function. However, the expansion of microsatellites is associated with over two dozen neurological diseases. A common denominator among the majority of these disorders is the expression of expanded tandem repeat-containing RNA, referred to as xtrRNA, which can mediate molecular disease pathology in multiple ways. Frontotemporal Dementia (FTD) and Amyotrophic Lateral Sclerosis (ALS) are two fatal neurodegenerative diseases with significant clinical, neurological and genetic overlap thus referred to as C9FTD/ALS. Currently, gaps in the study of the underlying disease mechanisms persist, which can aid in the identification of promising therapeutic approaches. Access to simple models of neurological repeat expansion disease is critical for investigating biochemical mechanisms and for early therapeutic discovery. To better understand the molecular pathology of C9FTD/ALS repeat expansion disorder, we cloned GGGGCC repeats, which are the leading genetic cause of C9FTD/ALS. We employed a recursive directional ligation (RDL) technique to build multiple GGGGCC repeat-containing vectors and validated the cloning to facilitate step-by-step characterization of disease mechanisms at the cellular and molecular level using these vectors. In this study, we also differentiated C9FTD/ALS patient-derived induced pluripotent stem cells (iPSCs) to neural stem cells (NSCs) to be used as model systems. The use of iPSCs and NSCs to reveal important insights into the pathogenic mechanisms and to generate multiple neural cell types presents an excellent opportunity for researchers to model neurodegenerative diseases for cell therapy and drug discovery. We further investigated potential nuclear export mechanisms for C9FTD/ALS xtrRNA. The nuclear export mechanisms of xtrRNA in C9FTD/ALS are not well studied. ASOs and siRNAs were employed to knockdown genes of interest to study their involvement in the nuclear export of xtrRNA. We saw promising results on knockdown of TorsinA involved in nuclear export of xtrRNAs, corroborated by a substantial increase in the average number of xtrRNA foci in the nucleus. Our initial study provides evidence that TOR1A may be involved in the nuclear export of aberrant C9FTD/ALS repeat-containing RNAs. Due to the lack of reliable and robust assays to detect RAN translation products, the effect of the knockdown of TorsinA in these cell lines still remains to be explored. But the current study lays the groundwork for a deeper understanding of the less-studied nuclear export mechanisms in C9FTD/ALS and could reveal new therapeutic approaches to selectively block the nuclear export of xtrRNA through the use of RNAi and ASOs. The insights gained from this study will help us understand future events in the xtrRNA life cycle such as repeat translation mechanisms.

京都大学 / 0048 / 新制・課程博士 / 博士(医学) / 甲第21990号 / 医博第4504号 / 新制||医||1037(附属図書館) / 京都大学大学院医学研究科医学専攻 / (主査)教授 武藤 学, 教授 松田 文彦, 教授 小川 誠司 / 学位規則第4条第1項該当 / Doctor of Medical Science / Kyoto University / DFAM