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http://hdl.handle.net/11375/25141
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DC Field | Value | Language |
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dc.contributor.advisor | Bhatia, Mick | - |
dc.contributor.author | Golubeva, Diana | - |
dc.date.accessioned | 2020-01-03T15:42:56Z | - |
dc.date.available | 2020-01-03T15:42:56Z | - |
dc.date.issued | 2019 | - |
dc.identifier.uri | http://hdl.handle.net/11375/25141 | - |
dc.description.abstract | Leukemia is a cancer of the blood and bone marrow where acute myeloid leukemia (AML) accounts for the most leukemic deaths annually. Only 5-10% of patients over 60 reach complete remission and the median survival for elderly patients is 8.5 months. Elderly patients succumb to the disease due to inability to tolerate conventional chemotherapy regimens. Despite advances in sequencing technologies supporting mutation identification in AML patients, it is still unclear which mutations are disease drivers and specific treatments for almost all patients are not available. This is in part because there are no known targets that distinguishing leukemic cells from their healthy counterparts. Using patient samples to derive new therapies has been challenging due to the technical inability to identify healthy from leukemic cells in a sample. Reprogramming of somatic cells into induced pluripotent stem cells (iPSCs) uniquely enables the separation of healthy and leukemic cells from AML patients through the derivation of AML and healthy isogenic iPSC cell lines. This holds potential for disease modelling and elucidation of functional driver mutation in AML. Leukemic iPSC lines have not yet been studied comprehensively due to the lack of a robust method to reprogram AML. Deriving iPSCs from leukemia patient samples has proven to be challenging, where all combined studies are limited by capturing a total of four patients spanning only two of fourteen genetic classes. Here, we for the first time interrogate strategies to selectively reprogram AML and healthy cells from patients. Using fluorescently activated cell sorting, we show that AML and healthy reprograming is restricted by the differentiation state. Further, we demonstrate that the use of myeloid markers in deriving AML-iPSCs is disadvantageous as AML clones reside in a variety of compartments. We present, the most successful to date, derivation of mutant and healthy variety of AML-iPSCs from three patients, capturing a wide variety of genetic aberrations. AML-iPSCs demonstrate pluripotency features with the ability to model AML through hematopoietic differentiation. The derived AML-iPSC model has further potential to use gene expression and chromatic availability signatures compared to isogenic controls to identify novel genetic and epigenetic targets in AML. | en_US |
dc.language.iso | en | en_US |
dc.title | Deriving an Induced Pluripotent Stem Cell Model of Acute Myeloid Leukemia | en_US |
dc.title.alternative | Deriving an Induced Pluripotent Stem Cell Model of Acute Myeloid Leukemia to Capture and Isolate Disease Heterogeneity | en_US |
dc.type | Thesis | en_US |
dc.contributor.department | Biochemistry and Biomedical Sciences | en_US |
dc.description.degreetype | Thesis | en_US |
dc.description.degree | Master of Science (MSc) | en_US |
dc.description.layabstract | Leukemia is the cancer of the blood system, in which leukemic cells divide too quickly and fail to mature into functional blood cells. These cells overcrowd healthy blood cells and prevent their normal function. Acute myeloid leukemia (AML) is a subtype of leukemia where only 5-15% of patients over 60 achieve remission and the median survival for elderly patients is less than a year. Elderly patients succumb to the disease due to their inability to tolerate aggressive and non-selective treatments. The lack of specific treatments is in part, due to a wide variety of genomic mutations in AML leading to variable responses to standardized therapy. In addition, there are no therapies to exclusively eliminate leukemic but not healthy blood cells. In the laboratory, many cancers are studied by using cell or animal models of the disease. Unfortunately, in the case of AML, these models are flawed as they capture less than half of genomic variations present in patients. When using patient samples directly it is often impossible to discern healthy from leukemic cells making it problematic to derive therapies specific to cancer cells. Reprogramming cancer cells into induced pluripotent stem cells (iPSCs) is a way to capture patient specific genetic mutations and physically separate healthy and mutated cells to be able to study them separately. This model has the potential to dissect variable patient mutations to identify which are root causes of the disease. It has been shown that AML is difficult to reprogram, and here we dissect reprogramming strategies used to the past to for the first time. We then demonstrate the most successful to date capture of three AML patients in an iPSC model and interrogate the model’s ability to capture and recapitulate AML disease. | en_US |
Appears in Collections: | Open Access Dissertations and Theses |
Files in This Item:
File | Description | Size | Format | |
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Golubeva_Diana_A_201912_MSc.pdf | 4.78 MB | Adobe PDF | View/Open |
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