Identifying regulators of hepatic leukemia factor (HLF) expression in hematopoietic cells using a fluorescent reporter transgene

Le facteur de leucémie hépatique (HLF) est un facteur de transcription et l’un des gènes les plus sélectivement exprimés par les cellules souches hématopoïétiques (CSHs) humaines et de souris. L’expression de HLF identifie l’activité fonctionnelle des CSHs et est un gène clé du caractère souche maintenant la quiescence et l’auto-renouvellement. Une surexpression de HLF a été observée dans les sous-groupes de leucémie myéloïde aiguë (LMA) comportant des mutations des gènes NPM1 (NPM1c) et FLT3 (FLT3-ITD), et il a été suggéré que ceci protège contre la mort cellulaire et les traitements chimio-toxiques. Cependant, la voie de signalisation responsable de la surexpression de HLF dans les leucémies avec mutation NPM1c reste inconnue. Puisque l’expression de HLF est associée à l’activité CSH et diminue au moment de la différentiation, nous avons émis l’hypothèse que la fusion du gène HLF avec un gène rapporteur fluorescent (HLF-ZsG) dans une lignée de cellules leucémiques (IMS-M2) pourrait être utilisée pour identifier les gènes qui régulent l’expression de HLF dans les CSHs normales. La protéine NPM1 réside dans le noyau et régule la réparation de l’ADN, la croissance cellulaire et la prolifération. Des mutations qui entraînent sa localisation cytoplasmique (NPM1c) ont été observées dans la LMA, entraînant une prolifération et une différenciation aberrante des cellules. La relocalisation nucléaire de la protéine NPM1c entraîne la régulation à la baisse des gènes homéotiques dans les cellules IMS-M2. Nous avons validé l’expression de la fusion HLF-ZsG dans les cellules IMS-M2 et confirmé la dépendance de l’expression de HLF à NPM1c. De plus, nous avons identifié MEIS1, HOXA10 et NKX2-3 comme étant des régulateurs potentiels de HLF. Nous avons également identifié un réseau régulant l’expression de HLF en aval de NPM1c aidant à comprendre les mécanismes de régulation de HLF dans les CSHs normales.Hepatic leukemia factor (HLF) is a transcription factor and one of the most selectively expressed genes in human and mouse hematopoietic stem cells (HSCs). HLF identifies functional HSC activity and is a key “stemness” gene that maintains quiescence and self-renewal. Upregulation of HLF has been observed in NPM1c/FLT3-ITD-mutated acute myeloid leukemia (AML) and is thought to protect cells from death and chemotoxic insults. However, the pathways which cause HLF expression in NPM1c-mutated leukemia are unknown. Since HLF expression is associated with HSC activity and decreases upon differentiation, we hypothesized that an HLF-ZsG fluorescent reporter in a leukemia cell line (IMS-M2) could be used as a readout to identify genes that regulate HLF in normal HSCs. The NPM1 protein resides in the nucleus and regulates DNA repair, cell growth and proliferation. Mutations which result in its cytoplasmic localization (NPM1c) have been observed in AML and lead to aberrant cell proliferation and differentiation. Nuclear relocalization of NPM1c results in the downregulation of homeobox genes in IMS-M2 cells. We validated the HLF-ZsG reporter in IMS-M2 cells and confirmed the dependency of HLF expression on NPM1c. We then identified MEIS1, HOXA10 and NKX2-3 as potential regulators of HLF. We have identified a preliminary network of HLF expression downstream of NPM1c, which may also help us understand HLF regulation in normal HSCs

How to (re) value Indigenous performing arts

In their paper on &ldquo;Excellence and access: Indigenous performing arts&rdquo; the problem that Hilary Glow and Katya Johanson describe is discursive, one that also has a discursive solution. An alternative can come from thinking through the process by which &ldquo;value&rdquo; is created, supported and circulated in the art world. Strategies to re-value Indigenous performance can thereby be directed to the various sites of institutional value which have long characterized Australian cultural policy &ndash; its importance to national identity, in the connection between culture and economics as well as to the debate over access and excellence &ndash; and come from artists as well as the communities they serve.<br /

Maternity Dress

1966https://digitalcommons.risd.edu/risdmuseum_channel/1038/thumbnail.jp

Diversity for design: a framework for involving neurodiverse children in the technology design process

The neurodiversity movement seeks to positively reframe certain neurological conditions, such as autism spectrum disorders (ASD) and dyslexia, by concentrating on their strengths. In recent years, neurodiverse children have increasingly been involved in the technology design process, but the design approaches adopted have focused mostly on overcoming difficulties of working with these children, leaving their strengths untapped. We present a new participatory design (PD) framework, Diversity for Design (D4D), which provides guidance for technology designers working with neurodiverse children in establishing PD methods that capitalize on children’s strengths and also support potential difficulties. We present two case studies of use of the D4D framework, involving children with ASD and dyslexia, showing how it informed the development and refinement of PD methods tailored to these populations. In addition, we show how to apply the D4D framework to other neurodiverse populations

Having Our Say: Examining Career Trajectories of Black Male Educators in P-12 Education

Achieving a diverse and inclusive workforce within P-12 education is critical to ensure that students receive a robust, quality educational experience. However, overcoming the shortage of educators of color has been a major dilemma for our nation's schools for decades. And, despite students of color comprising over 50% of current classroom populations and the United States Census Bureau's prediction that people of color will become the "majority-minority" in the overall United States population by 2043, these trends fail to correlate with representations of educators of color in P-12 education, especially for new cohorts of Black male teachers. In fact, according to the National Center for Education Statistics (NCES), male educators comprise only 23% of the public school workforce and, more troubling, Black male teachers represent less than 2% of the total teacher population