Thesis (Ph. D.)--University OF MICHIGAN.

Thesis--University of Oklahoma. / Bibliography: leaves 90-93.

A leucemia mielóide aguda (LMA) acomete pacientes de todas as faixas etárias com freqüência aumentada em idosos. Nosso estado não apresenta registros sistemáticos da patologia, justificando-se desta forma o levantamento através dos centros de diagnóstico e tratamento, os quais concentram a maioria dos casos. Foram incluídos todos os pacientes com diagnóstico de LMA “de novo” levantados a partir dos registros dos centros de diagnóstico e tratamento da patologia no estado no período entre 1996 e 2000. Foram computados no período 532 pacientes (taxa estimada de 1,04 casos / 100.000 habitantes / ano). A idade média ao diagnóstico foi 41,0 anos e 46,6 % dos casos eram do sexo masculino. Houve distribuição semelhante nas 7 mesorregiões geográficas do estado. Em relação ao número de casos por faixa etária, observou-se uma taxa estimada em 0,5 – 1 caso / 100.000 habitantes até a faixa etária dos 45 anos, atingindo 3,5 casos / 100.000 habitantes na faixa etária igual ou superior a 70 anos. A sobrevida média considerando todos os casos foi de 17%, aumentando para 25% considerando a população abaixo de 60 anos e alcançando 40% na população entre 10 e 20 anos. Tais dados evidenciam uma distribuição semelhante por região, estimativa de incidência na faixa infantil semelhante a países desenvolvidos e bastante menor na população idosa. A mortalidade geral foi semelhante àquela relatada na literatura. / Acute myeloid leukemia (AML) affects patients in all age groups, but it is more frequent among elderly people. The state of Rio Grande do Sul does not have systematic data on AML; thus these data have to be collected from hospitals or centers where the disease is diagnosed and treated. Patients with de novo acute myeloid leukemia were included based on the information retrieved from hospital registries between 1996 and 2000. At the time, 532 patients had been registered (estimated rate of 1.04 cases per 100,000 inhabitants / year). Mean age at diagnosis was 41.0 years, and 46.6% of the patients were male. Cases were similarly distributed across the seven meso-regions of Rio Grande do Sul. There was an estimated incidence of 0.5 to 1 case per 100,000 inhabitants up to the age of 45 years, and of 3.5 cases per 100,000 inhabitants for individuals aged 70 years and older. The mean five-year survival rate was 17% for all cases, 25% for patients under the age of 60 years, and 40% for those aged between 10 and 20 years. There was a similar distribution of AML cases per region, an incidence rate in the pediatric population similar to that of developed countries, and a much lower incidence among elderly patients. Overall mortality was comparable to that reported in the literature.

A Leucemia Mielóide Crônica (LMC) é uma doença rara. No entanto, os avanços nas pesquisas básica e clínica nos últimos anos, colocaram a LMC em evidência sendo hoje uma neoplasia maligna potencialmente curável. O diagnóstico e tratamento desta doença são, no entanto, extremamente caros. Não havendo dados sistemáticos nem registros de incidência da LMC no Rio Grande do Sul ou no Brasil, o levantamento de dados baseado em registros dos centros de referência se justifica também para planejar ações em saúde. Entre 1996 e 2000, 276 casos foram diagnosticados. A estimativa de casos novos anuais foi de aproximadamente 0,6:100.000 habitantes, e a idade média no momento do diagnóstico foi 42 anos e 4 meses (±16 anos e 2 meses). Quanto ao tratamento e evolução destes pacientes, dos 257 avaliados, 56 (21,8%) foram submetidos ao transplante alogênico de medula óssea, com taxa de sobrevida em 5 anos de 75% e 27% para as fases crônica e acelerada/blástica, respectivamente. O tempo médio de sobrevida para os 257 pacientes foi de 47,7 meses (IC 43,3 - 52,1). Comparando ao relatado na literatura, encontramos um menor número anual de novos casos e também uma média de idade no diagnóstico mais baixa. Isto poderia ser explicado pela menor referência de idosos a serviços terciários de saúde. Para os pacientes transplantados, os resultados foram semelhantes aos relatados na literatura. / Although rare, the advances made in basic and clinical research throughout the last years have thrown a spotlight on CML. Diagnosis and treatment of CML is of high cost. Since there is no systematic data or information about the incidence of CML in Rio Grande do Sul or Brazil, the data obtained from reference centers serve to estimate the number of CML cases in our state to better plan health actions. Between 1996 and 2000, 276 cases were diagnosed. The annual estimate of new cases was approximately of 0,6:100,000 inhabitants, and the median age at diagnosis was 42 years and 4 months (±16 years and 2 months). The mean overall survival time for the 257 patients was 47,7 months (CI 43,3-52,1). That could be explained by the lack of referral for older patients. Regarding treatment and evolution, of the 257 valuable patients, 56 (21,8%) were submitted to allogeneic BMT with a five-year survival of 75% and 27% for chronic and accelerated/blastic phases, respectively. In conclusion, we found a lower estimated incidence and a lower median age at diagnosis. For the transplanted patients the results were similar to those reported in the literature.

Le facteur de transcription Ikaros est un régulateur essentiel de la lymphopoïèse. Ikaros est nécessaire à la différenciation des lymphocytes B et joue aussi un rôle important dans la suppression des LAL-T. Contrairement aux souris mutantes nulles pour Ikaros, les souris mutantes hypomorphes IkL/L développent des lymphocytes B matures après la naissance. Avec l’âge, toutes les souris IkL/L développent des leucémies T Notch dépendantes avec des mutations similaires à celles trouvées chez les patients atteints de LAL-T. La souris IkL/L est donc un excellent modèle pour étudier l’activation des cellules B matures et la pathogenèsedes LAL-T. Nous avons montré que la délétion spécifique du promoteur et de l’exon 1 de Notch1 dans les cellules T conduit à l’activation de promoteurs cryptiques dans la région 3’ du gène, qui génèrent des transcrits codant pour des protéines Notch1 constitutivement actives qui accélèrent la leucémogenèse dans la souris IkL/L. De plus, nous mettrons en évidence l’existence de cellules initiatrices de leucémie dans les tumeurs IkL/L puisque nous avons trouvé que des cellules ayant la capacité de s’auto-renouveler représentent 1 sur 500. Enfin, nous avons montré que les cellules B IkL/L ont une activation excessive d’ERK et dep38 après la stimulation du BCR, ce qui résulte en une hyper-prolifération et une production d’autoanticorps liés au lupus systémique érythémateux. Nos résultats suggèrent qu’Ikaros est un régulateur négatif de l’activation des lymphocytes B. / The Ikaros transcription factor is a critical regulator of lymphopoeisis. Ikaros is needed for the differentiation of B cell and plays an important role in tumor suppression in T cells. Contrary to the other Ikaros targeted mutant mice, the IkL/L mouse develops mature B cells after birth. With age, it develops Notch dependent tumors with similar mutations as thosefound in human T cell acute lymphoblastic leukemia (T-ALL). Thus, the IkL/L mouse is an excellent model to study mature B cell activation and T-ALL tumor development. Here, we have shown that the deletion of the promoter and exon1 of the Notch1 gene in IkL/L T cells activates a cryptic promoter in the 3’ region which leads to the production of truncated, constitutively activated Notch1 proteins that accelerate tumorigenesis in the IkL/L mice. Moreover, we give evidence for the existence of LICs in the IkL/L tumors as we have found a specific subpopulation of cells with a frequency of 1 in 500 which show self-renewal capacity. In addition, we demonstrated that some tumor cells have the ability to efflux the Hoechst dye and that this side population is enriched in quiescent cells. Finally, we elucidate that IkL/L B cells display an enhanced activation of ERK and p38 after BCR stimulation that results in hyper-proliferation and the production of autoantibodies relatedto systemic lupus erythematosus (SLE). Our results suggest that Ikaros is a negative regulator of B cell activation.

A Leucemia Mielóide Crônica (LMC) é uma doença rara. No entanto, os avanços nas pesquisas básica e clínica nos últimos anos, colocaram a LMC em evidência sendo hoje uma neoplasia maligna potencialmente curável. O diagnóstico e tratamento desta doença são, no entanto, extremamente caros. Não havendo dados sistemáticos nem registros de incidência da LMC no Rio Grande do Sul ou no Brasil, o levantamento de dados baseado em registros dos centros de referência se justifica também para planejar ações em saúde. Entre 1996 e 2000, 276 casos foram diagnosticados. A estimativa de casos novos anuais foi de aproximadamente 0,6:100.000 habitantes, e a idade média no momento do diagnóstico foi 42 anos e 4 meses (±16 anos e 2 meses). Quanto ao tratamento e evolução destes pacientes, dos 257 avaliados, 56 (21,8%) foram submetidos ao transplante alogênico de medula óssea, com taxa de sobrevida em 5 anos de 75% e 27% para as fases crônica e acelerada/blástica, respectivamente. O tempo médio de sobrevida para os 257 pacientes foi de 47,7 meses (IC 43,3 - 52,1). Comparando ao relatado na literatura, encontramos um menor número anual de novos casos e também uma média de idade no diagnóstico mais baixa. Isto poderia ser explicado pela menor referência de idosos a serviços terciários de saúde. Para os pacientes transplantados, os resultados foram semelhantes aos relatados na literatura. / Although rare, the advances made in basic and clinical research throughout the last years have thrown a spotlight on CML. Diagnosis and treatment of CML is of high cost. Since there is no systematic data or information about the incidence of CML in Rio Grande do Sul or Brazil, the data obtained from reference centers serve to estimate the number of CML cases in our state to better plan health actions. Between 1996 and 2000, 276 cases were diagnosed. The annual estimate of new cases was approximately of 0,6:100,000 inhabitants, and the median age at diagnosis was 42 years and 4 months (±16 years and 2 months). The mean overall survival time for the 257 patients was 47,7 months (CI 43,3-52,1). That could be explained by the lack of referral for older patients. Regarding treatment and evolution, of the 257 valuable patients, 56 (21,8%) were submitted to allogeneic BMT with a five-year survival of 75% and 27% for chronic and accelerated/blastic phases, respectively. In conclusion, we found a lower estimated incidence and a lower median age at diagnosis. For the transplanted patients the results were similar to those reported in the literature.

A leucemia mielóide aguda (LMA) acomete pacientes de todas as faixas etárias com freqüência aumentada em idosos. Nosso estado não apresenta registros sistemáticos da patologia, justificando-se desta forma o levantamento através dos centros de diagnóstico e tratamento, os quais concentram a maioria dos casos. Foram incluídos todos os pacientes com diagnóstico de LMA “de novo” levantados a partir dos registros dos centros de diagnóstico e tratamento da patologia no estado no período entre 1996 e 2000. Foram computados no período 532 pacientes (taxa estimada de 1,04 casos / 100.000 habitantes / ano). A idade média ao diagnóstico foi 41,0 anos e 46,6 % dos casos eram do sexo masculino. Houve distribuição semelhante nas 7 mesorregiões geográficas do estado. Em relação ao número de casos por faixa etária, observou-se uma taxa estimada em 0,5 – 1 caso / 100.000 habitantes até a faixa etária dos 45 anos, atingindo 3,5 casos / 100.000 habitantes na faixa etária igual ou superior a 70 anos. A sobrevida média considerando todos os casos foi de 17%, aumentando para 25% considerando a população abaixo de 60 anos e alcançando 40% na população entre 10 e 20 anos. Tais dados evidenciam uma distribuição semelhante por região, estimativa de incidência na faixa infantil semelhante a países desenvolvidos e bastante menor na população idosa. A mortalidade geral foi semelhante àquela relatada na literatura. / Acute myeloid leukemia (AML) affects patients in all age groups, but it is more frequent among elderly people. The state of Rio Grande do Sul does not have systematic data on AML; thus these data have to be collected from hospitals or centers where the disease is diagnosed and treated. Patients with de novo acute myeloid leukemia were included based on the information retrieved from hospital registries between 1996 and 2000. At the time, 532 patients had been registered (estimated rate of 1.04 cases per 100,000 inhabitants / year). Mean age at diagnosis was 41.0 years, and 46.6% of the patients were male. Cases were similarly distributed across the seven meso-regions of Rio Grande do Sul. There was an estimated incidence of 0.5 to 1 case per 100,000 inhabitants up to the age of 45 years, and of 3.5 cases per 100,000 inhabitants for individuals aged 70 years and older. The mean five-year survival rate was 17% for all cases, 25% for patients under the age of 60 years, and 40% for those aged between 10 and 20 years. There was a similar distribution of AML cases per region, an incidence rate in the pediatric population similar to that of developed countries, and a much lower incidence among elderly patients. Overall mortality was comparable to that reported in the literature.

A leucemia mielóide aguda (LMA) acomete pacientes de todas as faixas etárias com freqüência aumentada em idosos. Nosso estado não apresenta registros sistemáticos da patologia, justificando-se desta forma o levantamento através dos centros de diagnóstico e tratamento, os quais concentram a maioria dos casos. Foram incluídos todos os pacientes com diagnóstico de LMA “de novo” levantados a partir dos registros dos centros de diagnóstico e tratamento da patologia no estado no período entre 1996 e 2000. Foram computados no período 532 pacientes (taxa estimada de 1,04 casos / 100.000 habitantes / ano). A idade média ao diagnóstico foi 41,0 anos e 46,6 % dos casos eram do sexo masculino. Houve distribuição semelhante nas 7 mesorregiões geográficas do estado. Em relação ao número de casos por faixa etária, observou-se uma taxa estimada em 0,5 – 1 caso / 100.000 habitantes até a faixa etária dos 45 anos, atingindo 3,5 casos / 100.000 habitantes na faixa etária igual ou superior a 70 anos. A sobrevida média considerando todos os casos foi de 17%, aumentando para 25% considerando a população abaixo de 60 anos e alcançando 40% na população entre 10 e 20 anos. Tais dados evidenciam uma distribuição semelhante por região, estimativa de incidência na faixa infantil semelhante a países desenvolvidos e bastante menor na população idosa. A mortalidade geral foi semelhante àquela relatada na literatura. / Acute myeloid leukemia (AML) affects patients in all age groups, but it is more frequent among elderly people. The state of Rio Grande do Sul does not have systematic data on AML; thus these data have to be collected from hospitals or centers where the disease is diagnosed and treated. Patients with de novo acute myeloid leukemia were included based on the information retrieved from hospital registries between 1996 and 2000. At the time, 532 patients had been registered (estimated rate of 1.04 cases per 100,000 inhabitants / year). Mean age at diagnosis was 41.0 years, and 46.6% of the patients were male. Cases were similarly distributed across the seven meso-regions of Rio Grande do Sul. There was an estimated incidence of 0.5 to 1 case per 100,000 inhabitants up to the age of 45 years, and of 3.5 cases per 100,000 inhabitants for individuals aged 70 years and older. The mean five-year survival rate was 17% for all cases, 25% for patients under the age of 60 years, and 40% for those aged between 10 and 20 years. There was a similar distribution of AML cases per region, an incidence rate in the pediatric population similar to that of developed countries, and a much lower incidence among elderly patients. Overall mortality was comparable to that reported in the literature.

Orientadores: Fabiola Traina, Sara Teresinha Olalla Saad / Dissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Ciências Médicas / Made available in DSpace on 2018-08-17T21:51:25Z (GMT). No. of bitstreams: 1 MachadoNeto_JoaoAgostinho_M.pdf: 23849071 bytes, checksum: df78354ffdd25c98c274422937e03f93 (MD5) Previous issue date: 2011 / Resumo: A ocorrência da leucemia aguda resulta de uma combinação de mutações e alterações em funções protéicas que conferem a capacidade de proliferação, defeito na diferenciação e apoptose celular. Síndromes mielodisplásicas (SMD) são desordens hematopoéticas resultantes de alterações na célula pluripotente, caracterizadas por hematopoese ineficaz e alta taxa de evolução para leucemia mieloide aguda (LMA). Células leucêmicas expressam uma variedade de receptores de fatores de crescimento e citocinas, como o receptor do Insulin-like growth factor 1 (IGF-1R). A via de sinalização do IGF-1 inicia-se através da ativação de seu receptor e subsequente ativação de seus substratos, como os substratos do receptor de insulina (IRS). Algumas evidências indicam a participação das proteínas IRS em doenças hematológicas: (1) IRS1 foi descrito como constitutivamente fosforilado e associado ao BCR-ABL em células K562; (2) a expressão de IRS1 foi relacionada com pior prognóstico em leucemia linfóide aguda (LLA) BCR-ABL positiva; (3) IRS2 associa-se ao receptor de eritropoetina; (4) a expressão de IRS2 foi modulada durante estímulos com eritropoetina e IGF-1 e em processos de diferenciação em células hematopoéticas normais e leucêmicas. Neste estudo, foi observada a presença da expressão gênica e protéica de IRS1 e IRS2 em células hematopoéticas normais, mielodisplásicas e leucêmicas, entretanto, o padrão de expressão das duas proteínas foi diferente. Em linhagens celulares de leucemia aguda, IRS1 foi expresso em linhagens de leucemia aguda mieloide (P39, K562, NB4, KG-1, e HL60) e linfoide (MOLT4, Jurkat, Raji e Daudi), enquanto que IRS2 foi expresso preferencialmente em linhagens mieloides. Em células hematopoéticas primárias, não houve diferença na expressão de IRS1 entre células hematopoéticas de pacientes com SMD e LMA e controles normais, e a expressão gênica de IRS1 apresentou-se aumentada em amostras de medula óssea de pacientes com LLA em relação aos controles normais. A expressão de IRS2 foi menor nas amostras de medula óssea de pacientes com SMD, LMA e LLA em relação aos controles normais, e a expressão de IRS2 foi menor em pacientes com SMD de alto risco se comparados com SMD baixo risco, de acordo com a classificação FAB, WHO e com número de citopenias. A participação de IRS2 na diferenciação eritróide de células hematopoéticas normais e mielodisplásicas foi evidenciada através da avaliação da expressão de IRS2 durante a diferenciação eritroide de células progenitoras de medula óssea de doadores normais e de pacientes com SMD. O estudo evidenciou o aumento da expressão de IRS2 durante a diferenciação eritroide, sendo que nas células mielodisplásicas, IRS2 apresentou um menor aumento se comparado às células hematopoéticas normais. Em células BCR-ABL positivas, a inibição da expressão de IRS1 (realizada através do uso de shRNA mediado por lentivírus específico para IRS1) resultou em inibição da proliferação celular e crescimento clonal, acúmulo de células na fase G0/G1 e redução de células na fase S do ciclo celular. A inibição de IRS1 resultou na inibição da fosforilação das proteínas Akt, P70S6K e ERK. Entretanto, a inibição de IRS1 não modulou a apoptose e as proteínas BCL2, BAX e BAD, assim como não modulou a fosforilação de BCR-ABL e CRKL e não apresentou sinergismo quando associado ao inibidor de tirosina quinase do BCR-ABL (imatinib). Estes dados indicam que IRS1 participa dos processos celulares de proliferação celular e clonogenicidade em células BCR-ABL positivas, através da modulação de Akt, P70S6K e ERK. Os achados aqui descritos sugerem que IRS1 é expresso em células hematopoéticas normais, mielodisplásicas e leucêmicas, destacando-se sua elevada expressão em células de LLA e sua participação na via de sinalização BCR-ABL. Estes dados indicam que IRS1 pode ser um alvo terapêutico em LLA e leucemia mieloide crônica (LMC), especialmente nas leucemias BCR-ABL positivas e resistentes a inibidores da atividade tirosina quinase do BCR-ABL. Adicionalmente, IRS2 é expresso em células hematopoéticas, destacando-se a sua expressão reduzida em células mielodisplásicas e leucêmicas quando comparadas às células hematopoéticas normais. A reduzida expressão de IRS2 em células hematopoéticas de pacientes com SMD de alto risco quando comparados aos de baixo risco e o reduzido aumento da expressão de IRS2 na diferenciação eritroide de progenitores de pacientes com SMD sugerem que a expressão de IRS2 participa da fisiopatologia das SMD e pode ser um marcador prognóstico nesta doença / Abstract: Acute leukemia results from a combination of mutations and changes in protein functions that confer the ability of proliferation, and defect in differentiation and apoptosis. Myelodysplastic syndromes (MDS) are hematopoietic disorders caused by alterations in pluripotent cells, characterized by ineffective hematopoiesis and a high rate of progression towards acute myeloid leukemia (AML). Leukemia cells express a variety of receptors for growth factors and cytokines, such as Insulin-like growth factor 1 (IGF-1R). The signaling pathway is initiated by activating its receptor and subsequent activation of its substrates such as insulin receptor substrate (IRS). There is evidence that suggests an involvement of IRS proteins in hematopoeitic disease: (1) IRS1 was described as constitutively phosphorylated and associated with BCR-ABL in K562 cells, (2) the IRS1 expression was associated with a poorer prognosis in BCR-ABL positive acute lymphoblastic leukemia (ALL), (3) IRS2 bind to erythropoietin receptors, (4) IRS2 expression was modulated during stimulation with erythropoietin and IGF-1 during cell differentiation in normal and leukemia hematopoietic cells. In this study, we observed the presence of the gene and protein of IRS1 and IRS2 in normal hematopoietic, leukemia, and myelodysplastic cells, however, the expression pattern of these proteins was different. In acute leukemia cell lines, IRS1 was expressed in myeloid leukemia cells (P39, K562, NB4, KG-1 e HL60) and lymphoid leukemia cells (MOLT4, Junkat, Raji e Daudi), whereas IRS2 expression was more evident in myeloid cell lines. In primary hematopoietic cells, no difference was observed in IRS1 expression between normal, MDS and AML cells, and the IRS1 expression was increased in bone marrow samples from ALL patients compared to normal controls. IRS2 expression was lower in bone marrow samples from patients with MDS, AML and ALL compared to normal controls, and the IRS2 expression was lower in high risk compared with low risk MDS patients, according to FAB and WHO classification, and number of cytopenias. The participation of IRS2 in erythroid differentiation of normal and myelodysplastic hematopoietic cells was evidenced by evaluating IRS2 expression during erythroid differentiation of progenitor cells from the bone marrow of normal donors and patients with MDS. The study demonstrated an increase in IRS2 expression during erythroid differentiation, whereas in myelodysplastic cells, IRS2 showed a smaller increase compared to normal hematopoietic cells. In BCR-ABL positive cells, IRS1 inhibition (by lentivirus-mediated shrunk specific for IRS1) resulted in inhibition of cell proliferation and clonal growth, accumulation of the cells in G0/G1 phase and reduction of cells in S phase of cell cycle. The IRS1 silencing resulted in inhibition of Akt, P70S6K and ERK phosphorylation. However, IRS1 inhibition did not modulate apoptosis; and the proteins BCL2, BAX and BAD, did not modulate the phosphorylation of BCR-ABL and CRKL, nor did they show synergism when combined with tyrosine kinase inhibitor of BCR-ABL (Imatinib). These data indicate that IRS1 participates in the proliferation and clonogenic of BCRABL positive cells by modulation of Akt, P70S6K and ERK. The findings reported herein suggest that IRS1 is expressed in normal hematopoietic, leukemia and myelodysplastic cells, highlighting its high expression in ALL cells and involvement in the BCR-ABL pathway. These data indicate that IRS1 may be a therapeutic target in ALL and chronic myeloid leukemia (CML), especially in BCR-ABL positive leukemias resistant to inhibitors of tyrosine kinase activity of BCRABL. In addition, IRS2 is expressed in hematopoietic cells, highlighting its reduced expression in myelodysplastic and leukemia cells compared to normal hematopoietic cells. The reduced IRS2 expression in high risk when compared to low risk MDS and the lower increase in IRS2 expression during the erythroid differentiation of progenitor cells from MDS patients suggest that the expression of IRS2 participates in the pathophysiology of MDS and may be a prognostic marker in this disease / Mestrado / Ciencias Basicas / Mestre em Clinica Medica

<p>It has been long known that many types of cancers have high metabolic requirements and use reprogrammed metabolism to support cellular activities. The first identified metabolic alteration in cancer cells was elevated glucose uptake, glycolysis activity and lactate production even in the presence of oxygen. This metabolic program, termed aerobic glycolysis or the Warburg effect, provides cells with energy as well as biosynthetic substrates to sustain cell survival and rapid cell proliferation. Cancer metabolism is closely linked to genetic mutations and oncogenic signaling pathways, such as PI3K/Akt, cMyc and HIF pathways. These oncogenic signals can direct metabolic reprogramming while changes in metabolic status can regulate activities of these signaling pathways in turn. In addition to glucose, later studies also found utilization of alternate nutrients in cancer cells, including glutamine and lipids. Glutamine is the second major metabolic fuel and can be converted to various substrates to support cell bioenergetics needs and biosynthetic reactions. Usage of metabolic fuels in cancer cells, however, is variable. While certain cancers display addiction to one type of nutrient, others are capable of using multiple nutrients. </p><p>The unique metabolic features of cancer cells raise the possibility of targeting metabolism as a novel therapeutic approach for cancer treatment. Using pharmacological inhibitors, previous research has provided corroborating evidence that metabolic stress can impact survival and growth of proliferative cancer cells by regulating cell apoptotic machinery and cell cycle checkpoints. Due to lack of genetic tools and side effects from these inhibitors, however, mechanistic understanding of cell response to metabolic inhibition was limited in these studies. More importantly, how metabolic stress affects cancer progression in a physiological condition has not yet been well investigated. Lastly, current research has not examined metabolic program in indolent cancers and the metabolic requirements and activities in less proliferative cells also remain to be understood.</p><p>This work examines nutrients utilization in B cell derived acute and chronic leukemia (B-ALL and B-CLL). B-ALL is an aggressive form of leukemia. Using cell lines and primary patient samples, we found B-ALL cells primarily used glucose through aerobic glycolysis, similar to other proliferative cancer cells. B-ALL cells were also more sensitive to inhibition of glycolysis than normal B cells. Employing an untargeted metabolomics profiling in combination with isotope labeled glucose tracing approach, we show in a B-ALL model that genetic ablation of glucose transporter Glut1 partially reduced glucose uptake, sufficiently hindered anabolic pathways and promoted catabolic metabolism. This metabolic shift led to sharply curtailed B-ALL proliferation in vitro and reduced leukemic burden in vivo. Furthermore, this partial inhibition of glucose metabolism sensitized B-ALL cells to apoptotic stimuli and non-cytotoxic metabolic inhibition significantly enhanced efficacy of a tyrosine kinase inhibitor to eliminate B-ALL cells in vitro and in vivo. Thus, partial inhibition of glucose metabolism can provide a plausible adjuvant therapy to treat cancers that depend on glycolysis for survival and proliferation. </p><p>In contrast to B-ALL, B-CLL is an indolent form of cancer. Most B-CLL cells exhibited low glucose metabolic activities that were comparable with normal B cells at resting stage. Similar to chronically stimulated and anergic B cells, these B-CLL cells also failed to upregulate glucose metabolism in response to IgM stimulation. We also observed an altered amino acid and acyl-carnitine profile and increased glutaminase mRNA in B-CLL relative to normal B cells, suggesting the capability of using alternate nutrients such as glutamine in these cells. Finally, we explored the possibility of suppressing mitochondria metabolism to induce B-CLL cell death through inhibition of the nuclear hormone receptor and metabolic regulator ERRalpha. ERRalpha is known to regulate mitochondrial metabolism and was expressed higher in B-CLL than normal B cells. ERRalpha inhibition decreased viability of oncogene transformed pro-B cells, suggesting ERRalpha as a potential target for B-CLL treatment.</p><p> Collectively, this work investigates metabolic phenotype in two forms of leukemia derived from B cells. It reveals different metabolic requirements and activities in aggressive and indolent leukemia and explores different approaches to suppress metabolism in these cancers. Findings of this work shed light on how to potentially design metabolic approach to improve cancer treatment.</p> / Dissertation