Nprl3 is a highly conserved regulator of erythropoiesis
<p>The neighbouring genomic positions of Nprl3 and a-globin, which share a topologically associated domain, have been maintained for over 500 million years. Nprl3 contains 4 of the 5 a-globin enhancers in mice, and 3 of 4 in humans. However, the potential functional requirement of this genomic...
| मुख्य लेखक: | |
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| अन्य लेखक: | |
| स्वरूप: | थीसिस |
| भाषा: | English |
| प्रकाशित: |
2022
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| विषय: |
| _version_ | 1826312486478938112 |
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| author | Preston, AE |
| author2 | Drakesmith, A |
| author_facet | Drakesmith, A Preston, AE |
| author_sort | Preston, AE |
| collection | OXFORD |
| description | <p>The neighbouring genomic positions of Nprl3 and a-globin, which share a topologically associated domain, have been maintained for over 500 million years. Nprl3 contains 4 of the 5 a-globin enhancers in mice, and 3 of 4 in humans. However, the potential functional requirement of this genomic linkage has not been investigated until now.</p>
<p>As part of the Gator-1 complex, the canonical signalling of Nprl3 has been defined in cell lines. Gator-1 provides negative regulation of mTORC1, a critical controller of cellular metabolism. We aimed to confirm this signalling in erythroid cells, and to investigate whether Nprl3 is required during erythropoiesis at steady state. Analysis of Nprl3-/- fetal livers revealed severe impairment of erythroid development. Autophagic flux was significantly dampened in Nprl3-/- erythroblasts, which is a possible driver of the observed block in erythropoiesis.</p>
<p>To discover whether NPRL3 is also required for human erythropoiesis, NPRL3-knockout was induced in primary human CD34+ progenitors using an RNP-editing system. These progenitors demonstrated reduced ability to produce enucleated erythroid cells compared to their negative control counterparts. We also used this model to investigate whether NPRL3 is particularly important in the context of fluctuating nutrients or growth factors. NPRL3-knockout erythroblasts demonstrated defective mTORC1 signalling responses to iron deficiency, amino acid withdrawal and erythropoietin stimulation. This suggested an intriguing role for NPRL3 as an environmental interpreter, which could be particularly important in the highly metabolically active and nutrient-demanding process of erythropoiesis.</p>
<p>We further aimed to characterise the role of Nprl3 in broader haematopoiesis, and to observe whether Nprl3 influences erythroid lineage commitment. In competitive bone marrow - fetal liver chimera experiments, Nprl3-/- fetal liver cells contributed poorly to erythroid lineage reconstitution of irradiated adult bone marrow. An intriguing competitive disadvantage was also observed at the HSC level.</p>
<p>Importantly, Nprl3 expression is known to be elevated in erythroid cells, increasing ~30-fold during erythroid lineage commitment due to a-globin enhancer activity. We sought to discover whether this erythroid-specific increase in Nprl3 expression is required for the functionality outlined above. This was achieved using transgenic mouse models to eliminate all interactions between Nprl3 and the a-globin enhancers, which resulted in impaired erythropoiesis reminiscent of Nprl3-/-.</p>
<p>Overall, in this work we show that Nprl3 is functionally important in erythroid cells, and that this is supported by transcriptional influence of the a-globin enhancers. We hypothesize that the deep evolutionary coupling of a-globin and Nprl3 has enabled the a-globin enhancers to control metabolism, as well as a-globin production, in developing erythroid cells.</p> |
| first_indexed | 2024-03-07T07:39:55Z |
| format | Thesis |
| id | oxford-uuid:c5c6d676-5564-47c4-a8c2-f3fb93b8e9e5 |
| institution | University of Oxford |
| language | English |
| last_indexed | 2024-04-09T03:55:18Z |
| publishDate | 2022 |
| record_format | dspace |
| spelling | oxford-uuid:c5c6d676-5564-47c4-a8c2-f3fb93b8e9e52024-03-11T08:56:28ZNprl3 is a highly conserved regulator of erythropoiesisThesishttp://purl.org/coar/resource_type/c_db06uuid:c5c6d676-5564-47c4-a8c2-f3fb93b8e9e5HematologyMolecular biologyGenomicsEnglishHyrax Deposit2022Preston, AEDrakesmith, ARoy, NMilne, TWeiss, M<p>The neighbouring genomic positions of Nprl3 and a-globin, which share a topologically associated domain, have been maintained for over 500 million years. Nprl3 contains 4 of the 5 a-globin enhancers in mice, and 3 of 4 in humans. However, the potential functional requirement of this genomic linkage has not been investigated until now.</p> <p>As part of the Gator-1 complex, the canonical signalling of Nprl3 has been defined in cell lines. Gator-1 provides negative regulation of mTORC1, a critical controller of cellular metabolism. We aimed to confirm this signalling in erythroid cells, and to investigate whether Nprl3 is required during erythropoiesis at steady state. Analysis of Nprl3-/- fetal livers revealed severe impairment of erythroid development. Autophagic flux was significantly dampened in Nprl3-/- erythroblasts, which is a possible driver of the observed block in erythropoiesis.</p> <p>To discover whether NPRL3 is also required for human erythropoiesis, NPRL3-knockout was induced in primary human CD34+ progenitors using an RNP-editing system. These progenitors demonstrated reduced ability to produce enucleated erythroid cells compared to their negative control counterparts. We also used this model to investigate whether NPRL3 is particularly important in the context of fluctuating nutrients or growth factors. NPRL3-knockout erythroblasts demonstrated defective mTORC1 signalling responses to iron deficiency, amino acid withdrawal and erythropoietin stimulation. This suggested an intriguing role for NPRL3 as an environmental interpreter, which could be particularly important in the highly metabolically active and nutrient-demanding process of erythropoiesis.</p> <p>We further aimed to characterise the role of Nprl3 in broader haematopoiesis, and to observe whether Nprl3 influences erythroid lineage commitment. In competitive bone marrow - fetal liver chimera experiments, Nprl3-/- fetal liver cells contributed poorly to erythroid lineage reconstitution of irradiated adult bone marrow. An intriguing competitive disadvantage was also observed at the HSC level.</p> <p>Importantly, Nprl3 expression is known to be elevated in erythroid cells, increasing ~30-fold during erythroid lineage commitment due to a-globin enhancer activity. We sought to discover whether this erythroid-specific increase in Nprl3 expression is required for the functionality outlined above. This was achieved using transgenic mouse models to eliminate all interactions between Nprl3 and the a-globin enhancers, which resulted in impaired erythropoiesis reminiscent of Nprl3-/-.</p> <p>Overall, in this work we show that Nprl3 is functionally important in erythroid cells, and that this is supported by transcriptional influence of the a-globin enhancers. We hypothesize that the deep evolutionary coupling of a-globin and Nprl3 has enabled the a-globin enhancers to control metabolism, as well as a-globin production, in developing erythroid cells.</p> |
| spellingShingle | Hematology Molecular biology Genomics Preston, AE Nprl3 is a highly conserved regulator of erythropoiesis |
| title | Nprl3 is a highly conserved regulator of erythropoiesis |
| title_full | Nprl3 is a highly conserved regulator of erythropoiesis |
| title_fullStr | Nprl3 is a highly conserved regulator of erythropoiesis |
| title_full_unstemmed | Nprl3 is a highly conserved regulator of erythropoiesis |
| title_short | Nprl3 is a highly conserved regulator of erythropoiesis |
| title_sort | nprl3 is a highly conserved regulator of erythropoiesis |
| topic | Hematology Molecular biology Genomics |
| work_keys_str_mv | AT prestonae nprl3isahighlyconservedregulatoroferythropoiesis |