scholarly journals NUPR1 is a critical repressor of ferroptosis

2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Jiao Liu ◽  
Xinxin Song ◽  
Feimei Kuang ◽  
Qiuhong Zhang ◽  
Yangchun Xie ◽  
...  
Keyword(s):  
Cell Death ◽  
Transcription Factor ◽  
Transcription Factors ◽  
Mouse Models ◽  
Iron Metabolism ◽  
Knockout Mice ◽  
Multiple Roles ◽  
Conditional Knockout ◽  
Regulated Cell Death ◽  
Nanostring Technology

AbstractFerroptosis is a type of iron-dependent regulated cell death, representing an emerging disease-modulatory mechanism. Transcription factors play multiple roles in ferroptosis, although the key regulator for ferroptosis in iron metabolism remains elusive. Using NanoString technology, we identify NUPR1, a stress-inducible transcription factor, as a driver of ferroptosis resistance. Mechanistically, NUPR1-mediated LCN2 expression blocks ferroptotic cell death through diminishing iron accumulation and subsequent oxidative damage. Consequently, LCN2 depletion mimics NUPR1 deficiency with respect to ferroptosis induction, whereas transfection-enforced re-expression of LCN2 restores resistance to ferroptosis in NUPR1-deficient cells. Pharmacological or genetic blockade of the NUPR1-LCN2 pathway (using NUPR1 shRNA, LCN2 shRNA, pancreas-specific Lcn2 conditional knockout mice, or the small molecule ZZW-115) increases the activity of the ferroptosis inducer erastin and worsens pancreatitis, in suitable mouse models. These findings suggest a link between NUPR1-regulated iron metabolism and ferroptosis susceptibility.

Abstract MP12: The Mef2c Transcription Factor Regulates The Renin Cell Identity

Hypertension ◽  
2021 ◽  
Vol 78 (Suppl_1) ◽  
Author(s):  
Omar E Guessoum ◽  
Kristyna Kupkova ◽  
Nathan Sheffield ◽  
Maria Luisa Sequeira Lopez ◽  
Roberto A Gomez
Keyword(s):  
Transcription Factor ◽  
Transcription Factors ◽  
Consensus Sequence ◽  
Renin Angiotensin System ◽  
Fold Increase ◽  
Conditional Knockout ◽  
Motif Analysis ◽  
Regulation Of Expression ◽  
Cell Identity

Introduction: The Renin-Angiotensin-System is essential to maintain blood pressure and fluid electrolyte homeostasis. Because precise regulation of expression and release of renin is critical for survival, understanding the molecular regulation of the renin cell identity is a vital area of study. Advances in epigenetics have enabled finer dissection of chromatin factors which maintain the identity of the renin cell. By studying genes with heightened accessibility profiles that are unique to the JG cell, we now have the capacity to unravel the determinants of the renin cell identity. Hypothesis: That transcription factors central to the governance of renin cell identity can be identified through the Assay for Transposase Accessible Chromatin (ATAC-seq) differential accessibility analysis. Methods: Native renin cell ATAC-seq was compared to existing ENCODE ATAC-seq datasets from 40 other cell types to define regions/peaks which characterize the JG program. Peaks with high intensity and ≥2-fold increase in signal were selected for Motif analysis to search for transcription factors (TFs) whose consensus sequence is enriched in those regions. Identified TFs were then selected for validation by in-situ hybridization and conditional deletion in renin cells. Results: 1) The Mef2c transcription factor was identified as having a consensus sequence in regulatory regions unique to the JG cell. It has clear expression in RNA-seq of renin cells (65 transcripts per million, n=3) and a predicted binding site in the renin gene. These results were validated by in-situ hybridization where signal localized at the JG area was detected in concordance with our in-silico results. 2) We generated Mef2c conditional knockout animals using our Ren1d-Cre mouse to study the effect in renin expression and identity. These mice displayed reduced renin immunostaining at the JG area and a 40% reduction in renin mRNA expression by qPCR from kidney cortices relative to wild-type (n=2, preliminary data). Conclusions: Our studies identified Mef2c as a TF target which likely has an essential role in maintaining and preserving renin cell identity. Experiments involving transcriptomics and epigenomics are ongoing to understand the changes wrought by Mef2c deletion in renin cells.

scholarly journals Conditional knockout mice demonstrate function of Klf5 as a myeloid transcription factor

Blood ◽  
2016 ◽  
Vol 128 (1) ◽  
pp. 55-59 ◽  
Author(s):  
Nur Hezrin Shahrin ◽  
Sonya Diakiw ◽  
Lindsay A. Dent ◽  
Anna L. Brown ◽  
Richard J. D’Andrea
Keyword(s):  
Bone Marrow ◽  
Transcription Factor ◽  
Knockout Mice ◽  
Conditional Knockout ◽  
Conditional Knockout Mice ◽  
Key Points

Key Points Klf5 functions in hematopoiesis to regulate HSC and progenitor proliferation and localization in the bone marrow. Klf5 is required in the granulocyte lineage and positively affects neutrophil output at the expense of eosinophil production.

scholarly journals The transcription factor Zeb2 regulates development of conventional and plasmacytoid DCs by repressing Id2

2016 ◽  
Vol 213 (6) ◽  
pp. 897-911 ◽  
Author(s):  
Charlotte L. Scott ◽  
Bieke Soen ◽  
Liesbet Martens ◽  
Nicolas Skrypek ◽  
Wouter Saelens ◽  
...  
Keyword(s):  
Dendritic Cells ◽  
Transcription Factor ◽  
Transcription Factors ◽  
Chromatin Immunoprecipitation ◽  
Conditional Knockout ◽  
Altered Expression ◽  
E Box ◽  
A Cell ◽  
Plasmacytoid Dcs ◽  
Direct Target

Plasmacytoid dendritic cells (DCs [pDCs]) develop from pre-pDCs, whereas two lineages of conventional DCs (cDCs; cDC1s and cDC2s) develop from lineage-committed pre-cDCs. Several transcription factors (TFs) have been implicated in regulating the development of pDCs (E2-2 and Id2) and cDC1s (Irf8, Id2, and Batf3); however, those required for the early commitment of pre-cDCs toward the cDC2 lineage are unknown. Here, we identify the TF zinc finger E box–binding homeobox 2 (Zeb2) to play a crucial role in regulating DC development. Zeb2 was expressed from the pre-pDC and pre-cDC stage onward and highly expressed in mature pDCs and cDC2s. Mice conditionally lacking Zeb2 in CD11c+ cells had a cell-intrinsic reduction in pDCs and cDC2s, coupled with an increase in cDC1s. Conversely, mice in which CD11c+ cells overexpressed Zeb2 displayed a reduction in cDC1s. This was accompanied by altered expression of Id2, which was up-regulated in cDC2s and pDCs from conditional knockout mice. Zeb2 chromatin immunoprecipitation analysis revealed Id2 to be a direct target of Zeb2. Thus, we conclude that Zeb2 regulates commitment to both the cDC2 and pDC lineages through repression of Id2.

scholarly journals The Snail transcription factor CES-1 regulates glutamatergic behavior in C. elegans

PLoS ONE ◽  
2021 ◽  
Vol 16 (2) ◽  
pp. e0245587
Author(s):  
Lidia Park ◽  
Eric S. Luth ◽  
Kelsey Jones ◽  
Julia Hofer ◽  
Irene Nguyen ◽  
...  
Keyword(s):  
Cell Death ◽  
Transcription Factor ◽  
Transcription Factors ◽  
Synaptic Plasticity ◽  
Ampa Receptor ◽  
Active Form ◽  
Cell Lineage ◽  
Receptor Expression ◽  
Loss Of Function ◽  
Major Mechanism

Regulation of AMPA-type glutamate receptor (AMPAR) expression and function alters synaptic strength and is a major mechanism underlying synaptic plasticity. Although transcription is required for some forms of synaptic plasticity, the transcription factors that regulate AMPA receptor expression and signaling are incompletely understood. Here, we identify the Snail family transcription factor ces-1 in an RNAi screen for conserved transcription factors that regulate glutamatergic behavior in C. elegans. ces-1 was originally discovered as a selective cell death regulator of neuro-secretory motor neuron (NSM) and I2 interneuron sister cells in C. elegans, and has almost exclusively been studied in the NSM cell lineage. We found that ces-1 loss-of-function mutants have defects in two glutamatergic behaviors dependent on the C. elegans AMPA receptor GLR-1, the mechanosensory nose-touch response and spontaneous locomotion reversals. In contrast, ces-1 gain-of-function mutants exhibit increased spontaneous reversals, and these are dependent on glr-1 consistent with these genes acting in the same pathway. ces-1 mutants have wild type cholinergic neuromuscular junction function, suggesting that they do not have a general defect in synaptic transmission or muscle function. The effect of ces-1 mutation on glutamatergic behaviors is not due to ectopic cell death of ASH sensory neurons or GLR-1-expressing neurons that mediate one or both of these behaviors, nor due to an indirect effect on NSM sister cell deaths. Rescue experiments suggest that ces-1 may act, in part, in GLR-1-expressing neurons to regulate glutamatergic behaviors. Interestingly, ces-1 mutants suppress the increased reversal frequencies stimulated by a constitutively-active form of GLR-1. However, expression of glr-1 mRNA or GFP-tagged GLR-1 was not decreased in ces-1 mutants suggesting that ces-1 likely promotes GLR-1 function. This study identifies a novel role for ces-1 in regulating glutamatergic behavior that appears to be independent of its canonical role in regulating cell death in the NSM cell lineage.

scholarly journals Transcription factor CHF1/Hey2 regulates EC coupling and heart failure in mice through regulation of FKBP12.6

2012 ◽  
Vol 302 (9) ◽  
pp. H1860-H1870 ◽  
Author(s):  
Yonggang Liu ◽  
F. Steven Korte ◽  
Farid Moussavi-Harami ◽  
Man Yu ◽  
Maria Razumova ◽  
...  
Keyword(s):  
Heart Failure ◽  
Transcription Factor ◽  
Ryanodine Receptor ◽  
Knockout Mice ◽  
Contractile Function ◽  
Conditional Knockout ◽  
Calcium Handling ◽  
Western Society ◽  
Calcium Stores ◽  
Calcium Cycling

Heart failure is a leading cause of morbidity and mortality in Western society. The cardiovascular transcription factor CHF1/Hey2 has been linked to experimental heart failure in mice, but the mechanisms by which it regulates myocardial function remain incompletely understood. The objective of this study was to determine how CHF1/Hey2 affects development of heart failure through examination of contractility in a myocardial knockout mouse model. We generated myocardial-specific knockout mice. At baseline, cardiac function was normal, but, after aortic banding, the conditional knockout mice demonstrated a greater increase in ventricular weight-to-body weight ratio compared with control mice (5.526 vs. 4.664 mg/g) and a significantly decreased ejection fraction (47.8 vs. 72.0% control). Isolated cardiac myocytes from these mice showed decreased calcium transients and fractional shortening after electrical stimulation. To determine the molecular basis for these alterations in excitation-contraction coupling, we first measured total sarcoplasmic reticulum calcium stores and calcium-dependent force generation in isolated muscle fibers, which were normal, suggesting a defect in calcium cycling. Analysis of gene expression demonstrated normal expression of most genes known to be involved in myocardial calcium cycling, with the exception of the ryanodine receptor binding protein FKBP12.6, which was expressed at increased levels in the conditional knockout hearts. Treatment of the isolated knockout myocytes with FK506, which inhibits the association of FKBP12.6 with the ryanodine receptor, restored contractile function. These findings demonstrate that conditional deletion of CHF1/Hey2 in the myocardium leads to abnormalities in calcium handling mediated by FKBP12.6 that predispose to pressure overload-induced heart failure.

scholarly journals The Molecular Mechanisms of Iron Metabolism and Its Role in Cardiac Dysfunction and Cardioprotection

2020 ◽  
Vol 21 (21) ◽  
pp. 7889 ◽  
Author(s):  
Tanya Ravingerová ◽  
Lucia Kindernay ◽  
Monika Barteková ◽  
Miroslav Ferko ◽  
Adriana Adameová ◽  
...  
Keyword(s):  
Cell Death ◽  
Iron Metabolism ◽  
Molecular Mechanisms ◽  
Ischemia Reperfusion ◽  
Physiological Role ◽  
Cellular Respiration ◽  
Protective Mechanisms ◽  
Alcoholic Fatty Liver ◽  
Regulated Cell Death ◽  
Wide Range

Iron is an essential mineral participating in different functions of the organism under physiological conditions. Numerous biological processes, such as oxygen and lipid metabolism, protein production, cellular respiration, and DNA synthesis, require the presence of iron, and mitochondria play an important role in the processes of iron metabolism. In addition to its physiological role, iron may be also involved in the adaptive processes of myocardial “conditioning”. On the other hand, disorders of iron metabolism are involved in the pathological mechanisms of the most common human diseases and include a wide range of them, such as type 2 diabetes, obesity, and non-alcoholic fatty liver disease, and accelerate the development of atherosclerosis. Furthermore, iron also exerts potentially deleterious effects that may be manifested under conditions of ischemia/reperfusion (I/R) injury, myocardial infarction, heart failure, coronary artery angioplasty, or heart transplantation, due to its involvement in reactive oxygen species (ROS) production. Moreover, iron has been recently described to participate in the mechanisms of iron-dependent cell death defined as “ferroptosis”. Ferroptosis is a form of regulated cell death that is distinct from apoptosis, necroptosis, and other types of cell death. Ferroptosis has been shown to be associated with I/R injury and several other cardiac diseases as a significant form of cell death in cardiomyocytes. In this review, we will discuss the role of iron in cardiovascular diseases, especially in myocardial I/R injury, and protective mechanisms stimulated by different forms of “conditioning” with a special emphasis on the novel targets for cardioprotection.

Circadian rhythms in the absence of the clock gene Bmal1

Science ◽  
2020 ◽  
Vol 367 (6479) ◽  
pp. 800-806 ◽  
Author(s):  
Sandipan Ray ◽  
Utham K. Valekunja ◽  
Alessandra Stangherlin ◽  
Steven A. Howell ◽  
Ambrosius P. Snijders ◽  
...  
Keyword(s):  
Transcription Factor ◽  
Transcriptional Regulation ◽  
Transcription Factors ◽  
Circadian Rhythms ◽  
Molecular Clock ◽  
Knockout Mice ◽  
Clock Gene ◽  
Skin Fibroblasts ◽  
Temperature Cycles ◽  
Ets Family

Circadian (~24 hour) clocks have a fundamental role in regulating daily physiology. The transcription factor BMAL1 is a principal driver of a molecular clock in mammals. Bmal1 deletion abolishes 24-hour activity patterning, one measure of clock output. We determined whether Bmal1 function is necessary for daily molecular oscillations in skin fibroblasts and liver slices. Unexpectedly, in Bmal1 knockout mice, both tissues exhibited 24-hour oscillations of the transcriptome, proteome, and phosphoproteome over 2 to 3 days in the absence of any exogenous drivers such as daily light or temperature cycles. This demonstrates a competent 24-hour molecular pacemaker in Bmal1 knockouts. We suggest that such oscillations might be underpinned by transcriptional regulation by the recruitment of ETS family transcription factors, and nontranscriptionally by co-opting redox oscillations.

scholarly journals Determination of the Potential Tumor-Suppressive Effects of Gsdme in a Chemically Induced and in a Genetically Modified Intestinal Cancer Mouse Model

Cancers ◽  
2019 ◽  
Vol 11 (8) ◽  
pp. 1214 ◽  
Author(s):  
Lieselot Croes ◽  
Erik Fransen ◽  
Marieke Hylebos ◽  
Kimberly Buys ◽  
Christophe Hermans ◽  
...  
Keyword(s):  
Cell Death ◽  
Mouse Model ◽  
Mouse Models ◽  
Tumor Biology ◽  
Suppressor Gene ◽  
Intestinal Cancer ◽  
Cancer Model ◽  
Inflammatory Microenvironment ◽  
Regulated Cell Death ◽  
Chemically Induced

Gasdermin E (GSDME), also known as deafness autosomal dominant 5 (DFNA5) and previously identified to be an inducer of regulated cell death, is frequently epigenetically inactivated in different cancer types, suggesting that GSDME is a tumor suppressor gene. In this study, we aimed to evaluate the tumor-suppressive effects of GSDME in two intestinal cancer mouse models. To mimic the silencing of GSDME by methylation as observed in human cancers, a Gsdme knockout (KO) mouse was developed. The effect of GSDME on tumorigenesis was studied both in a chemically induced and in a genetic intestinal cancer mouse model, as strong evidence shows that GSDME plays a role in human colorectal cancer and representative mouse models for intestinal cancer are available. Azoxymethane (AOM) was used to induce colorectal tumors in the chemically induced intestinal cancer model (n = 100). For the genetic intestinal cancer model, Apc1638N/+ mice were used (n = 37). In both experiments, the number of mice bearing microscopic proliferative lesions, the number and type of lesions per mouse and the histopathological features of the adenocarcinomas were compared between Gsdme KO and wild type (WT) mice. Unfortunately, we found no major differences between Gsdme KO and WT mice, neither for the number of affected mice nor for the multiplicity of proliferative lesions in the mice. However, recent breakthroughs on gasdermin function indicate that GSDME is an executioner of necrotic cell death. Therefore, it is possible that GSDME may be important for creating an inflammatory microenvironment around the tumor. This is in line with the trend towards more severe inflammation in WT compared to Gsdme KO mice, that we observed in our study. We conclude that the effect of GSDME in tumor biology is probably more subtle than previously thought.

scholarly journals Novel Insights into the Genetic Controls of Primitive and Definitive Hematopoiesis from Zebrafish Models

2012 ◽  
Vol 2012 ◽  
pp. 1-13 ◽  
Author(s):  
Raman Sood ◽  
Paul Liu
Keyword(s):  
Transcription Factors ◽  
Knockout Mice ◽  
Model Organism ◽  
Conditional Knockout ◽  
Current Status ◽  
Ideal Model ◽  
Zebrafish Model ◽  
Hematopoietic Stem ◽  
Adult Transition ◽  
Definitive Hematopoiesis

Hematopoiesis is a dynamic process where initiation and maintenance of hematopoietic stem cells, as well as their differentiation into erythroid, myeloid and lymphoid lineages, are tightly regulated by a network of transcription factors. Understanding the genetic controls of hematopoiesis is crucial as perturbations in hematopoiesis lead to diseases such as anemia, thrombocytopenia, or cancers, including leukemias and lymphomas. Animal models, particularly conventional and conditional knockout mice, have played major roles in our understanding of the genetic controls of hematopoiesis. However, knockout mice for most of the hematopoietic transcription factors are embryonic lethal, thus precluding the analysis of their roles during the transition from embryonic to adult hematopoiesis. Zebrafish are an ideal model organism to determine the function of a gene during embryonic-to-adult transition of hematopoiesis since bloodless zebrafish embryos can develop normally into early larval stage by obtaining oxygen through diffusion. In this review, we discuss the current status of the ontogeny and regulation of hematopoiesis in zebrafish. By providing specific examples of zebrafish morphants and mutants, we have highlighted the contributions of the zebrafish model to our overall understanding of the roles of transcription factors in regulation of primitive and definitive hematopoiesis.

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