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Molecular atlas of fetal and adult human liver sinusoidal endothelial cells: a F8 secreting cell [expression array]

ABSTRACT: Molecular atlas of fetal and adult human liver sinusoidal endothelial cells: a F8 secreting cell [expression array]

PROVIDER: PRJNA587922 | ENA |

REPOSITORIES: ENA

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Molecular atlas of fetal and adult human liver sinusoidal endothelial cells: a F8 secreting cell [expression array]

Project description:In human F8 deficiency leads to hemophilia A and is largely synthesized and secreted by the sinusoidal endothelial cells of the liver (LSECs). However, the specificity and characteristics of these cells is not well known. In this study, we performed genome wide expression and CpG methylation profiling of fetal and adult human LSECs together with other fetal endothelial cells, from lung (micro-vascular and arterial), and heart (micro-vascular). Our results reveal plenty of expression and methylation markers distinguishing LSECs at both fetal and adult stages. Differential gene expression of fetal LSECs in comparison to other fetal endothelial cells pointed to several differential regulated pathways and functions in fetal LSECs. We used targeted bisulfite re-sequencing to confirm selected top differentially methylated regions. We further designed an assay where we used the selected methylation markers to test the degree of similarity of in house iPSs generated vascular endothelial cells to fetal and adult LSECs; a higher similarity was found to the fetal than to adult LSECs. Our molecular profiling study provides a guide to test the effectiveness of production of in vitro differentiated LSECs that could be used in cellular therapies. In-silico analysis to identify molecular signature of liver sinusoidal endothelial cells in comparison to other endohtelial cells

2020-10-25 | GSE139993 | GEO

Molecular atlas of fetal and adult human liver sinusoidal endothelial cells: a F8 secreting cell [methylation II]

Project description:Molecular atlas of fetal and adult human liver sinusoidal endothelial cells: a F8 secreting cell [methylation II]

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Molecular atlas of fetal and adult human liver sinusoidal endothelial cells: a F8 secreting cell [methylation I]

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Molecular atlas of fetal and adult human liver sinusoidal endothelial cells: a F8 secreting cell [methylation III]

Project description:Molecular atlas of fetal and adult human liver sinusoidal endothelial cells: a F8 secreting cell [methylation III]

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Molecular atlas of fetal and adult human liver sinusoidal endothelial cells: a F8 secreting cell [methylation II]

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Molecular atlas of fetal and adult human liver sinusoidal endothelial cells: a F8 secreting cell [methylation I]

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Molecular atlas of fetal and adult human liver sinusoidal endothelial cells: a F8 secreting cell [methylation III]

Project description:In-silico analysis to identify molecular signature of liver sinusoidal endothelial cells in comparison to other endohtelial cells

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F8 intein precise CodopN6 reconstitution

Project description:Liver gene therapy with adeno-associated viral (AAV) vectors is under clinical investigation for hemophilia A (HemA), the most common inherited X-linked bleeding disorder. Major limitations are the large size of the F8 transgene, which makes packaging in a single AAV vector a challenge, as well as the development of circulating anti-F8 antibodies which neutralize F8 activity. Taking advantage of split inteins-mediated protein trans-splicing, we divided the coding sequence of the large and highly secreted F8-N6 variant in two separate AAV split-inteins vectors whose co-administration to HemA mice results in F8 protein reconstitution and expression of therapeutic levels of F8 over time without anti-F8 antibodies development.

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The Role of microRNAs in Defining LSECs Cellular Identity and in Regulating F8 Gene Expression

Project description:Coagulation Factor VIII (FVIII) plays a pivotal role within the coagulation cascade, and deficiencies in its levels, as seen in Hemophilia A, can lead to significant health implications. Liver sinusoidal endothelial cells (LSECs) are the main producers and contributors of FVIII in blood, a fact we have previously elucidated through mRNA expression profiling when comparing these cells to other endothelial cell types. Our current investigation delves into small microRNAs, analyzing their distinct expression patterns across various endothelial cells and hepatocytes. The outcome of this exploration underscores the discernible microRNA expression differences that set LSECs apart from both hepatocytes (193 microRNAs at p < 0.05) and other endothelial cells (72 microRNAs at p < 0.05). Notably, the 134 and 35 overexpressed microRNAs in LSECs compared to hepatocytes and other endothelial cells, respectively, shed light on the unique functions of LSECs in the liver. Our investigation identified a panel of 10 microRNAs (miR-429, miR-200b-3p, miR-200a-3p, miR-216b-5p, miR-1185-5p, miR-19b-3p, miR-192-5p, miR-122-5p, miR-30c-2-3p, and miR-30a-5p) that distinctly define LSEC identity. Furthermore, our scrutiny extended to microRNAs implicated in F8 regulation, revealing a subset - miR-122-5p, miR-214-3p, miR-204-3p, and miR-2682-5p - whose expression intricately correlates with F8 expression within LSECs. This microRNA cohort emerges as a crucial modulator of F8, both directly through suppression and indirect effects on established F8-related transcription factors. The above miRNA emerged as potential targets for innovative therapies in Hemophilia A patients.

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