Gfi1-controlled transcriptional circuits in normal and malignant hematopoiesis

Gfi1-controlled transcriptional circuits in normal and malignant hematopoiesis

Bibliographic Details
Main Author: Muench, David
Language:English
Published: University of Cincinnati / OhioLINK 2019
Subjects:
Molecular Biology
Hematopoiesis
MDS
SCN
Single Cell RNA Sequencing
Mouse Models
Online Access:http://rave.ohiolink.edu/etdc/view?acc_num=ucin1553250015825734
id ndltd-OhioLink-oai-etd.ohiolink.edu-ucin1553250015825734
record_format oai_dc
collection NDLTD
language English
sources NDLTD
topic Molecular Biology
Hematopoiesis
MDS
SCN
Single Cell RNA Sequencing
Mouse Models
spellingShingle Molecular Biology
Hematopoiesis
MDS
SCN
Single Cell RNA Sequencing
Mouse Models
Muench, David
Gfi1-controlled transcriptional circuits in normal and malignant hematopoiesis
author Muench, David
author_facet Muench, David
author_sort Muench, David
title Gfi1-controlled transcriptional circuits in normal and malignant hematopoiesis
title_short Gfi1-controlled transcriptional circuits in normal and malignant hematopoiesis
title_full Gfi1-controlled transcriptional circuits in normal and malignant hematopoiesis
title_fullStr Gfi1-controlled transcriptional circuits in normal and malignant hematopoiesis
title_full_unstemmed Gfi1-controlled transcriptional circuits in normal and malignant hematopoiesis
title_sort gfi1-controlled transcriptional circuits in normal and malignant hematopoiesis
publisher University of Cincinnati / OhioLINK
publishDate 2019
url http://rave.ohiolink.edu/etdc/view?acc_num=ucin1553250015825734
work_keys_str_mv AT muenchdavid gfi1controlledtranscriptionalcircuitsinnormalandmalignanthematopoiesis
_version_ 1719455069261266944
spelling ndltd-OhioLink-oai-etd.ohiolink.edu-ucin15532500158257342021-08-03T07:09:37Z Gfi1-controlled transcriptional circuits in normal and malignant hematopoiesis Muench, David Molecular Biology Hematopoiesis MDS SCN Single Cell RNA Sequencing Mouse Models Lifelong hematopoiesis requires balanced hematopoietic stem cell (HSC) self-renewal and differentiation into mature blood populations. Such cell fate decisions are often initiated in response to extracellular cues (cytokines) to induce distinct transcriptional programs through coordinated teams of transcription factors that vary between cell states. Inherited or acquired genetic mutations in transcription factors or signaling molecules disrupts this balance to initiate any number of hematologic diseases that each arise from a single or a concise set of cell states. More specifically, cytopenias are life-threatening conditions characterized by low levels of mature hematopoietic cells that are associated with a diverse number of hematopoietic disorders in turn caused by distinct genetic alterations. For example, severe congenital neutropenia (SCN) presents early in life and is characterized by the relative absence of neutrophils, which predisposes these patients to life threatening infections. In contrast to SCN, myelodysplastic syndrome (MDS) typically affects the elderly, is usually accompanied by anemia (can include neutropenia) and presents with abnormal/dysplastic cells. The proper understanding of these conditions is hampered by a lack of accurate models and techniques to enable proper analyses of heterogeneous populations. Furthermore, elucidating the sequence of normal cell state transitions that occur during steady-state hematopoietic development at the single cell level is prerequisite to understanding the corresponding disease states.In this dissertation, I explore the cell states and signaling pathways of murine hematopoietic stem cells (HSC) and granulocytic progenitors and precursors that are disrupted by genetic mutations found in humans with neutropenia or MDS. This was achieved by utilizing novel murine models of disease, many single cell analytical procedures, novel bioinformatics algorithms and techniques, and global proteomics. My findings from three separate studies highlight the utility of modeling patient-derived mutations in mice, the necessity of single cell analysis (transcriptional and functional), and the importance of protein-level investigation to appropriately determine the impact made by genetic mutations upon the normal cell states and signaling pathways in hematopoietic development. First, I conclude from analysis of primary MDS patient samples and mouse modeling that miR-21-mediated loss of SKI in early stage MDS activates TGF-beta signaling and alternative splicing to impair normal HSC while it may act to promote the expansion of MDS-genic clones. Next, global phosphoproteomics was used to determine the impact of neutropenia-patient-relevant CSF3R mutations and identified BTK as a potential therapeutic target (with an available FDA-approved drug) for this class of patients, which currently lack effective treatment options. Finally, the generation and extensive analysis of the first accurate mouse models of human SCN (by introducing SCN-patient mutations in transcription factor Gfi1) facilitated the identification of two independent neutrophil defects in the disease and led to the insight that most Gfi1-target genes are differentially impacted in different cell states despite the presence of only a single mutation. Overall, my work here provides useful reference datasets to the field, highlights the importance of making proper comparisons at the cell-state level, and exemplifies broadly-applicable workflows to model and understand the global impact of human disease-relevant mutations. 2019-06-11 English text University of Cincinnati / OhioLINK http://rave.ohiolink.edu/etdc/view?acc_num=ucin1553250015825734 http://rave.ohiolink.edu/etdc/view?acc_num=ucin1553250015825734 unrestricted This thesis or dissertation is protected by copyright: all rights reserved. It may not be copied or redistributed beyond the terms of applicable copyright laws.

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