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1.
Cell Metab ; 33(11): 2189-2200.e3, 2021 11 02.
Article in English | MEDLINE | ID: mdl-34508697

ABSTRACT

Aging leads to profound changes in glucose homeostasis, weight, and adiposity, which are considered good predictors of health and survival in humans. Direct evidence that these age-associated metabolic alterations are recapitulated in animal models is lacking, impeding progress to develop and test interventions that delay the onset of metabolic dysfunction and promote healthy aging and longevity. We compared longitudinal trajectories, rates of change, and mortality risks of fasting blood glucose, body weight, and fat mass in mice, nonhuman primates, and humans throughout their lifespans and found similar trajectories of body weight and fat in the three species. In contrast, fasting blood glucose decreased late in life in mice but increased over the lifespan of nonhuman primates and humans. Higher glucose was associated with lower mortality in mice but higher mortality in nonhuman primates and humans, providing a cautionary tale for translating age-associated metabolic changes from mice to humans.


Subject(s)
Blood Glucose , Fasting , Adiposity , Animals , Blood Glucose/metabolism , Longevity , Mice , Obesity/metabolism
2.
Geroscience ; 43(5): 2129-2137, 2021 10.
Article in English | MEDLINE | ID: mdl-34297313

ABSTRACT

Healthspan is a complex trait, influenced by many genes and environmental factors that accelerate or delay aging, reduce or increase disease risk, and extend or reduce lifespan. Thus, assessing the role of genetic variation in aging requires an experimental strategy capable of modeling the genetic and biological complexity of human populations. The goal of the The Jackson Laboratory Nathan Shock Center (JAX NSC) is to provide research resources and training for geroscience investigators that seek to understand the role of genetics and genetic diversity on the fundamental process of aging and diseases of human aging using the laboratory mouse as a model system. The JAX NSC has available novel, deeply characterized populations of aged mice, performs state-of-the-art phenotyping of age-relevant traits, provides systems genetics analysis of complex data sets, and provides all of these resources to the geroscience community. The aged animal resources, phenotyping capacity, and genetic expertise available through the JAX NSC benefit the geroscience community by fostering cutting-edge, novel lines of research that otherwise would not be possible. Over the past 15 years, the JAX NSC has transformed aging research across the geroscience community, providing aging mouse resources and tissues to researchers. All JAX NSC data and tools are publicly disseminated on the Mouse Phenome Database and the JAX NSC website, thus ensuring that the resources generated and expertise acquired through the Center are readily available to the aging research community. The JAX NSC will continue to enhance its ability to perform innovative research using a mammalian model to illuminate novel genotype-phenotype relationships and provide a rational basis for designing effective risk assessments and therapeutic interventions to boost longevity and disease-free healthspan.


Subject(s)
Geroscience , Laboratories , Aging/genetics , Animals , Genetic Variation , Longevity/genetics , Mice
3.
Blood Cells Mol Dis ; 87: 102524, 2021 03.
Article in English | MEDLINE | ID: mdl-33341069

ABSTRACT

Inherited bone marrow failure syndromes (IBMFS) are heterogeneous disorders characterized by dysregulated hematopoiesis in various lineages, developmental anomalies, and predisposition to malignancy. The scat (severe combined anemia and thrombocytopenia) mouse model is a model of IBMFS with a phenotype of pancytopenia cycling through crises and remission. Scat carries an autosomal recessive missense mutation in Rasa3 that results in RASA3 mislocalization and loss of function. RASA3 functions as a Ras-GTPase activating protein (GAP), and its loss of function in scat results in increased erythroid RAS activity and reactive oxygen species (ROS) and altered erythroid cell cycle progression, culminating in delayed terminal erythroid differentiation. Here we sought to further resolve the erythroid cell cycle defect in scat through ex vivo flow cytometric analyses. These studies revealed a specific G0/G1 accumulation in scat bone marrow (BM) polychromatophilic erythroblasts and scat BM Ter119-/c-KIT+/CD71lo/med progenitors, with no changes evident in equivalent scat spleen populations. Systematic analyses of RNAseq data from megakaryocyte-erythroid progenitors (MEPs) in scat crisis vs. scat partial remission reveal altered expression of genes involved in the G1-S checkpoint. Together, these data indicate a precise, biphasic role for RASA3 in regulating the cell cycle during erythropoiesis with relevance to hematopoietic disease progression.


Subject(s)
Erythroid Cells/cytology , Erythropoiesis , GTPase-Activating Proteins/metabolism , Animals , Cell Cycle , Cells, Cultured , Erythroid Cells/metabolism , GTPase-Activating Proteins/genetics , Mice, Inbred BALB C , Mutation, Missense , ras Proteins/metabolism
4.
PLoS Genet ; 16(12): e1008857, 2020 12.
Article in English | MEDLINE | ID: mdl-33370780

ABSTRACT

Studies of the severely pancytopenic scat mouse model first demonstrated the crucial role of RASA3, a dual RAS and RAP GTPase activating protein (GAP), in hematopoiesis. RASA3 is required for survival in utero; germline deletion is lethal at E12.5-13.5 due to severe hemorrhage. Here, conditional deletion in hematopoietic stem and progenitor cells (HSPCs) using Vav-iCre recapitulates the null phenotype demonstrating that RASA3 is required at the stem and progenitor level to maintain blood vessel development and integrity and effective blood production. In adults, bone marrow blood cell production and spleen stress erythropoiesis are suppressed significantly upon induction of RASA3 deficiency, leading to pancytopenia and death within two weeks. Notably, RASA3 missense mutations in two mouse models, scat (G125V) and hlb381 (H794L), show dramatically different hematopoietic consequences specific to both genetic background and molecular variant. The mutation effect is mediated at least in part by differential effects on RAS and RAP activation. In addition, we show that the role of RASA3 is conserved during human terminal erythropoiesis, highlighting a potential function for the RASA3-RAS axis in disordered erythropoiesis in humans. Finally, global transcriptomic studies in scat suggest potential targets to ameliorate disease progression.


Subject(s)
GTPase-Activating Proteins/genetics , Genetic Background , Hematopoiesis , Mutation , Pancytopenia/genetics , Animals , Cells, Cultured , Female , GTPase-Activating Proteins/metabolism , Humans , Male , Mice , Mice, Inbred BALB C
5.
Aging Cell ; 19(11): e13263, 2020 11.
Article in English | MEDLINE | ID: mdl-33105070

ABSTRACT

Epidemiological studies of human longevity found two interesting features, robust advantage of female lifespan and consistent reduction of lifespan variation. To help understand the genetic aspects of these phenomena, the current study examined sex differences and variation of longevity using previously published mouse data sets including data on lifespan, age of puberty, and circulating insulin-like growth factor 1 (IGF1) levels in 31 inbred strains, data from colonies of nuclear-receptor-interacting protein 1 (Nrip1) knockout mice, and a congenic strain, B6.C3H-Igf1. Looking at the overall data for all inbred strains, the results show no significant difference in lifespan and lifespan variation between sexes; however, considerable differences were found among and within strains. Across strains, lifespan variations of female and male mice are significantly correlated. Strikingly, between sexes, IGF1 levels correlate with the lifespan variation and maximum lifespan in different directions. Female mice with low IGF1 levels have higher variation and extended maximum lifespan. The opposite is detected in males. Compared to domesticated inbred strains, wild-derived inbred strains have elevated lifespan variation due to increased early deaths in both sexes and extended maximum lifespan in female mice. Intriguingly, the sex differences in survival curves of inbred strains negatively associated with age of female puberty, which is significantly accelerated in domesticated inbred strains compared to wild-derived strains. In conclusion, this study suggests that genetic factors are involved in the regulation of sexual disparities in lifespan and lifespan variation, and dissecting the mouse genome may provide novel insight into the underlying genetic mechanisms.


Subject(s)
Genetic Variation/genetics , Longevity/genetics , Animals , Female , Male , Mice , Phenotype
6.
Cell Rep ; 32(9): 108091, 2020 09 01.
Article in English | MEDLINE | ID: mdl-32877673

ABSTRACT

Genetic mechanisms underlying age-related cognitive decline and dementia remain poorly understood. Here, we take advantage of the Diversity Outbred mouse population to utilize quantitative trait loci mapping and identify Dlgap2 as a positional candidate responsible for modifying working memory decline. To evaluate the translational relevance of this finding, we utilize longitudinal cognitive measures from human patients, RNA expression from post-mortem brain tissue, data from a genome-wide association study (GWAS) of Alzheimer's dementia (AD), and GWAS results in African Americans. We find an association between Dlgap2 and AD phenotypes at the variant, gene and protein expression, and methylation levels. Lower cortical DLGAP2 expression is observed in AD and is associated with more plaques and tangles at autopsy and faster cognitive decline. Results will inform future studies aimed at investigating the cross-species role of Dlgap2 in regulating cognitive decline and highlight the benefit of using genetically diverse mice to prioritize novel candidates.


Subject(s)
Alzheimer Disease/genetics , Cognitive Dysfunction/genetics , Dementia/genetics , Nerve Tissue Proteins/metabolism , Black or African American/genetics , Age Factors , Female , Genome-Wide Association Study , Humans , Male , Middle Aged , Species Specificity
7.
Geroscience ; 42(1): 183-199, 2020 02.
Article in English | MEDLINE | ID: mdl-31811527

ABSTRACT

Circular RNAs (circRNAs) are an emerging class of non-coding RNA molecules that are thought to regulate gene expression and human disease. Despite the observation that circRNAs are known to accumulate in older organisms and have been reported in cellular senescence, their role in aging remains relatively unexplored. Here, we have assessed circRNA expression in aging human blood and followed up age-associated circRNA in relation to human aging phenotypes, mammalian longevity as measured by mouse median strain lifespan and cellular senescence in four different primary human cell types. We found that circRNAs circDEF6, circEP300, circFOXO3 and circFNDC3B demonstrate associations with parental longevity or hand grip strength in 306 subjects from the InCHIANTI study of aging, and furthermore, circFOXO3 and circEP300 also demonstrate differential expression in one or more human senescent cell types. Finally, four circRNAs tested showed evidence of conservation in mouse. Expression levels of one of these, circPlekhm1, was nominally associated with lifespan. These data suggest that circRNA may represent a novel class of regulatory RNA involved in the determination of aging phenotypes, which may show future promise as both biomarkers and future therapeutic targets for age-related disease.


Subject(s)
MicroRNAs , RNA, Circular , Aged , Aging/genetics , Animals , Cellular Senescence/genetics , Hand Strength , Humans , Longevity/genetics , Mice , Phenotype
8.
Sci Rep ; 8(1): 12793, 2018 08 24.
Article in English | MEDLINE | ID: mdl-30143664

ABSTRACT

Anemic Nan mice carry a mutation (E339D) in the second zinc finger of erythroid transcription factor KLF1. Nan-KLF1 fails to bind a subset of normal KLF1 targets and ectopically binds a large set of genes not normally engaged by KLF1, resulting in a corrupted fetal liver transcriptome. Here, we performed RNAseq using flow cytometric-sorted spleen erythroid precursors from adult Nan and WT littermates rendered anemic by phlebotomy to identify global transcriptome changes specific to the Nan Klf1 mutation as opposed to anemia generally. Mutant Nan-KLF1 leads to extensive and progressive transcriptome corruption in adult spleen erythroid precursors such that stress erythropoiesis is severely compromised. Terminal erythroid differentiation is defective in the bone marrow as well. Principle component analysis reveals two major patterns of differential gene expression predicting that defects in basic cellular processes including translation, cell cycle, and DNA repair could contribute to disordered erythropoiesis and anemia in Nan. Significant erythroid precursor stage specific changes were identified in some of these processes in Nan. Remarkably, however, despite expression changes in large numbers of associated genes, most basic cellular processes were intact in Nan indicating that developing red cells display significant physiological resiliency and establish new homeostatic set points in vivo.


Subject(s)
Aging/pathology , Anemia/genetics , Anemia/pathology , Cell Differentiation/genetics , Erythropoiesis/genetics , Kruppel-Like Transcription Factors/genetics , Mutation/genetics , Transcriptome/genetics , Animals , Base Sequence , Cell Cycle/genetics , DNA Damage , Erythroid Cells/metabolism , Female , Gene Expression Regulation, Developmental , Gene Ontology , Kruppel-Like Transcription Factors/metabolism , Liver/embryology , Liver/metabolism , Mice , Mice, Mutant Strains , Mitophagy/genetics , Molecular Sequence Annotation , Principal Component Analysis , Reactive Oxygen Species/metabolism , Spleen/embryology , Spleen/metabolism
9.
Front Physiol ; 9: 689, 2018.
Article in English | MEDLINE | ID: mdl-29922180

ABSTRACT

RASA3 is a Ras GTPase activating protein that plays a critical role in blood formation. The autosomal recessive mouse model scat (severe combined anemia and thrombocytopenia) carries a missense mutation in Rasa3. Homozygotes present with a phenotype characteristic of bone marrow failure that is accompanied by alternating episodes of crisis and remission. The mechanism leading to impaired erythropoiesis and peripheral cell destruction as evidenced by membrane fragmentation in scat is unclear, although we previously reported that the mislocalization of RASA3 to the cytosol of reticulocytes and mature red cells plays a role in the disease. In this study, we further characterized the bone marrow failure in scat and found that RASA3 plays a central role in cell cycle progression and maintenance of reactive oxygen species (ROS) levels during terminal erythroid differentiation, without inducing apoptosis of the precursors. In scat mice undergoing crises, there is a consistent pattern of an increased proportion of cells in the G0/G1 phase at the basophilic and polychromatophilic stages of erythroid differentiation, suggesting that RASA3 is involved in the G1 checkpoint. However, this increase in G1 is transient, and either resolves or becomes indiscernible by the orthochromatic stage. In addition, while ROS levels are normal early in erythropoiesis, there is accumulation of superoxide levels at the reticulocyte stage (DHE increased 40% in scat; p = 0.02) even though mitochondria, a potential source for ROS, are eliminated normally. Surprisingly, apoptosis is significantly decreased in the scat bone marrow at the proerythroblastic (15.3%; p = 0.004), polychromatophilic (8.5%; p = 0.01), and orthochromatic (4.2%; p = 0.02) stages. Together, these data indicate that ROS accumulation at the reticulocyte stage, without apoptosis, contributes to the membrane fragmentation observed in scat. Finally, the cell cycle defect and increased levels of ROS suggest that scat is a model of bone marrow failure with characteristics of aplastic anemia.

10.
Sci Rep ; 7: 44620, 2017 03 17.
Article in English | MEDLINE | ID: mdl-28304372

ABSTRACT

MicroRNAs (miRNAs) are small non-coding RNA species that have been shown to have roles in multiple processes that occur in higher eukaryotes. They act by binding to specific sequences in the 3' untranslated region of their target genes and causing the transcripts to be degraded by the RNA-induced silencing complex (RISC). MicroRNAs have previously been reported to demonstrate altered expression in several aging phenotypes such as cellular senescence and age itself. Here, we have measured the expression levels of 521 small regulatory microRNAs (miRNAs) in spleen tissue from young and old animals of 6 mouse strains with different median strain lifespans by quantitative real-time PCR. Expression levels of 3 microRNAs were robustly associated with strain lifespan, after correction for multiple statistical testing (miR-203-3p [ß-coefficient = -0.6447, p = 4.8 × 10-11], miR-664-3p [ß-coefficient = 0.5552, p = 5.1 × 10-8] and miR-708-5p [ß-coefficient = 0.4986, p = 1.6 × 10-6]). Pathway analysis of binding sites for these three microRNAs revealed enrichment of target genes involved in key aging and longevity pathways including mTOR, FOXO and MAPK, most of which also demonstrated associations with longevity. Our results suggests that miR-203-3p, miR-664-3p and miR-708-5p may be implicated in pathways determining lifespan in mammals.


Subject(s)
Longevity/genetics , MicroRNAs/genetics , Animals , Gene Expression Profiling , Gene Expression Regulation , Mice, Inbred Strains , MicroRNAs/metabolism , RNA, Messenger/genetics , RNA, Messenger/metabolism , Signal Transduction/genetics
11.
Development ; 144(3): 430-440, 2017 02 01.
Article in English | MEDLINE | ID: mdl-28143845

ABSTRACT

Transcription factor control of cell-specific downstream targets can be significantly altered when the controlling factor is mutated. We show that the semi-dominant neonatal anemia (Nan) mutation in the EKLF/KLF1 transcription factor leads to ectopic expression of proteins that are not normally expressed in the red blood cell, leading to systemic effects that exacerbate the intrinsic anemia in the adult and alter correct development in the early embryo. Even when expressed as a heterozygote, the Nan-EKLF protein accomplishes this by direct binding and aberrant activation of genes encoding secreted factors that exert a negative effect on erythropoiesis and iron use. Our data form the basis for a novel mechanism of physiological deficiency that is relevant to human dyserythropoietic anemia and likely other disease states.


Subject(s)
Anemia, Neonatal/genetics , Kruppel-Like Transcription Factors/genetics , Mutation , Amino Acid Substitution , Anemia, Neonatal/blood , Anemia, Neonatal/embryology , Animals , Animals, Newborn , Cytokines/blood , DNA/genetics , DNA/metabolism , Disease Models, Animal , Erythrocytes/metabolism , Erythropoiesis/genetics , Gene Expression Regulation, Developmental , Heterozygote , Humans , Kruppel-Like Transcription Factors/blood , Kruppel-Like Transcription Factors/deficiency , Mice , Mice, Knockout , Mice, Mutant Strains , Models, Biological , Muscle Proteins/blood , Mutant Proteins/blood , Mutant Proteins/genetics
12.
Nucleic Acids Res ; 45(3): 1130-1143, 2017 02 17.
Article in English | MEDLINE | ID: mdl-28180284

ABSTRACT

The rules of engagement between zinc finger transcription factors and DNA have been partly defined by in vitro DNA-binding and structural studies, but less is known about how these rules apply in vivo. Here, we demonstrate how a missense mutation in the second zinc finger of Krüppel-like factor-1 (KLF1) leads to degenerate DNA-binding specificity in vivo, resulting in ectopic transcription and anemia in the Nan mouse model. We employed ChIP-seq and 4sU-RNA-seq to identify aberrant DNA-binding events genome wide and ectopic transcriptional consequences of this binding. We confirmed novel sequence specificity of the mutant recombinant zinc finger domain by performing biophysical measurements of in vitro DNA-binding affinity. Together, these results shed new light on the mechanisms by which missense mutations in DNA-binding domains of transcription factors can lead to autosomal dominant diseases.


Subject(s)
DNA/metabolism , Kruppel-Like Transcription Factors/genetics , Kruppel-Like Transcription Factors/metabolism , Mutant Proteins/genetics , Mutant Proteins/metabolism , Transcriptome/genetics , Zinc Fingers/genetics , Animals , Cell Line , Cell Survival/genetics , Erythroid Cells/metabolism , Erythropoiesis/genetics , Humans , Kruppel-Like Transcription Factors/chemistry , Mice , Models, Genetic , Models, Molecular , Mutant Proteins/chemistry , Mutation, Missense , Protein Binding
13.
Aging Cell ; 15(5): 903-13, 2016 10.
Article in English | MEDLINE | ID: mdl-27363602

ABSTRACT

Dysregulation of splicing factor expression and altered alternative splicing are associated with aging in humans and other species, and also with replicative senescence in cultured cells. Here, we assess whether expression changes of key splicing regulator genes and consequent effects on alternative splicing are also associated with strain longevity in old and young mice, across 6 different mouse strains with varying lifespan (A/J, NOD.B10Sn-H2(b) /J, PWD.Phj, 129S1/SvlmJ, C57BL/6J and WSB/EiJ). Splicing factor expression and changes to alternative splicing were associated with strain lifespan in spleen and to a lesser extent in muscle. These changes mainly involved hnRNP splicing inhibitor transcripts with most changes more marked in spleens of young animals from long-lived strains. Changes in spleen isoform expression were suggestive of reduced cellular senescence and retained cellular proliferative capacity in long-lived strains. Changes in muscle isoform expression were consistent with reduced pro-inflammatory signalling in longer-lived strains. Two splicing regulators, HNRNPA1 and HNRNPA2B1, were also associated with parental longevity in humans, in the InCHIANTI aging study. Splicing factors may represent a driver, mediator or early marker of lifespan in mouse, as expression differences were present in the young animals of long-lived strains. Changes to alternative splicing patterns of key senescence genes in spleen and key remodelling genes in muscle suggest that correct regulation of alternative splicing may enhance lifespan in mice. Expression of some splicing factors in humans was also associated with parental longevity, suggesting that splicing regulation may also influence lifespan in humans.


Subject(s)
Alternative Splicing/genetics , Longevity/genetics , RNA Splicing Factors/genetics , Animals , Genetic Variation , Humans , Mice, Inbred C57BL , Mice, Inbred NOD , Muscles/metabolism , Protein Isoforms/genetics , Protein Isoforms/metabolism , RNA Splicing Factors/metabolism , RNA, Messenger/genetics , RNA, Messenger/metabolism , Spleen/metabolism
14.
Cell Rep ; 15(3): 490-498, 2016 Apr 19.
Article in English | MEDLINE | ID: mdl-27068466

ABSTRACT

The actin-binding protein adducin was recently identified as a component of the neuronal subcortical cytoskeleton. Here, we analyzed mice lacking adducin to uncover the function of this protein in actin rings. α-adducin knockout mice presented progressive axon enlargement in the spinal cord and optic and sciatic nerves, followed by axon degeneration and loss. Using stimulated emission depletion super-resolution microscopy, we show that a periodic subcortical actin cytoskeleton is assembled in every neuron type inspected including retinal ganglion cells and dorsal root ganglia neurons. In neurons devoid of adducin, the actin ring diameter increased, although the inter-ring periodicity was maintained. In vitro, the actin ring diameter adjusted as axons grew, suggesting the lattice is dynamic. Our data support a model in which adducin activity is not essential for actin ring assembly and periodicity but is necessary to control the diameter of both actin rings and axons and actin filament growth within rings.


Subject(s)
Axons/metabolism , Calmodulin-Binding Proteins/metabolism , Microfilament Proteins/metabolism , Animals , Axonal Transport , Cytoskeleton/metabolism , Growth Cones/metabolism , Hippocampus/pathology , Mice, Inbred C57BL , Mice, Knockout , Nerve Degeneration/metabolism , Nerve Degeneration/pathology
15.
J Gerontol A Biol Sci Med Sci ; 71(2): 170-7, 2016 Feb.
Article in English | MEDLINE | ID: mdl-25533306

ABSTRACT

Understanding the source of genetic variation in aging and using this variation to define the molecular mechanisms of healthy aging require deep and broad quantification of a host of physiological, morphological, and behavioral endpoints. The murine model is a powerful system in which to understand the relations across age-related phenotypes and to identify research models with variation in life span and health span. The Jackson Laboratory Nathan Shock Center of Excellence in the Basic Biology of Aging has performed broad characterization of aging in genetically diverse laboratory mice and has placed these data, along with data from several other major aging initiatives, into the interactive Mouse Phenome Database. The data may be accessed and analyzed by researchers interested in finding mouse models for specific aging processes, age-related health and disease states, and for genetic analysis of aging variation and trait covariation. We expect that by placing these data in the hands of the aging community that there will be (a) accelerated genetic analyses of aging processes, (b) discovery of genetic loci regulating life span, (c) identification of compelling correlations between life span and susceptibility for age-related disorders, and (d) discovery of concordant genomic loci influencing life span and aging phenotypes between mouse and humans.


Subject(s)
Aging/genetics , Databases, Genetic , Genetic Variation , Longevity/genetics , Mice, Inbred Strains/genetics , Animals , Genomics , Genotype , Mice , Mice, Inbred Strains/anatomy & histology , Mice, Inbred Strains/classification , Models, Animal , Phenotype , Polymorphism, Single Nucleotide , Quantitative Trait Loci
16.
Curr Protoc Mouse Biol ; 5(2): 95-133, 2015 Jun 01.
Article in English | MEDLINE | ID: mdl-26069080

ABSTRACT

Despite the dramatic increase in human lifespan over the past century, there remains pronounced variability in "health-span," or the period of time in which one is generally healthy and free of disease. Much of the variability in health-span and lifespan is thought to be genetic in origin. Understanding the genetic mechanisms of aging and identifying ways to boost longevity is a primary goal in aging research. Here, we describe a pipeline of phenotypic assays for assessing mouse models of aging. This pipeline includes behavior/cognition testing, body composition analysis, and tests of kidney function, hematopoiesis, and immune function, as well as physical parameters. We also describe study design methods for assessing lifespan and health-span, and other important considerations when conducting aging research in the laboratory mouse. The tools and assays provided can assist researchers with understanding the correlative relationships between age-associated phenotypes and, ultimately, the role of specific genes in the aging process.


Subject(s)
Aging , Disease Models, Animal , Laboratory Animal Science/methods , Mice , Aging/physiology , Aging/psychology , Animal Structures/physiology , Animals , Behavior, Animal , Humans , Longevity , Mice/genetics , Mice/physiology , Research Design
17.
Nature ; 522(7557): 474-7, 2015 Jun 25.
Article in English | MEDLINE | ID: mdl-25970251

ABSTRACT

Many acute and chronic anaemias, including haemolysis, sepsis and genetic bone marrow failure diseases such as Diamond-Blackfan anaemia, are not treatable with erythropoietin (Epo), because the colony-forming unit erythroid progenitors (CFU-Es) that respond to Epo are either too few in number or are not sensitive enough to Epo to maintain sufficient red blood cell production. Treatment of these anaemias requires a drug that acts at an earlier stage of red cell formation and enhances the formation of Epo-sensitive CFU-E progenitors. Recently, we showed that glucocorticoids specifically stimulate self-renewal of an early erythroid progenitor, burst-forming unit erythroid (BFU-E), and increase the production of terminally differentiated erythroid cells. Here we show that activation of the peroxisome proliferator-activated receptor α (PPAR-α) by the PPAR-α agonists GW7647 and fenofibrate synergizes with the glucocorticoid receptor (GR) to promote BFU-E self-renewal. Over time these agonists greatly increase production of mature red blood cells in cultures of both mouse fetal liver BFU-Es and mobilized human adult CD34(+) peripheral blood progenitors, with a new and effective culture system being used for the human cells that generates normal enucleated reticulocytes. Although Ppara(-/-) mice show no haematological difference from wild-type mice in both normal and phenylhydrazine (PHZ)-induced stress erythropoiesis, PPAR-α agonists facilitate recovery of wild-type but not Ppara(-/-) mice from PHZ-induced acute haemolytic anaemia. We also show that PPAR-α alleviates anaemia in a mouse model of chronic anaemia. Finally, both in control and corticosteroid-treated BFU-E cells, PPAR-α co-occupies many chromatin sites with GR; when activated by PPAR-α agonists, additional PPAR-α is recruited to GR-adjacent sites and presumably facilitates GR-dependent BFU-E self-renewal. Our discovery of the role of PPAR-α agonists in stimulating self-renewal of early erythroid progenitor cells suggests that the clinically tested PPAR-α agonists we used may improve the efficacy of corticosteroids in treating Epo-resistant anaemias.


Subject(s)
Erythroid Precursor Cells/cytology , Erythropoiesis , PPAR alpha/metabolism , Receptors, Glucocorticoid/metabolism , Acute Disease , Anemia/drug therapy , Anemia/metabolism , Anemia/pathology , Anemia, Hemolytic/metabolism , Animals , Butyrates/pharmacology , Butyrates/therapeutic use , Cell Culture Techniques , Cells, Cultured , Chromatin/genetics , Chromatin/metabolism , Chronic Disease , Disease Models, Animal , Erythroid Precursor Cells/drug effects , Erythroid Precursor Cells/metabolism , Erythropoiesis/drug effects , Erythropoietin/pharmacology , Female , Fenofibrate/pharmacology , Glucocorticoids/pharmacology , Humans , Liver/cytology , Liver/drug effects , Liver/embryology , Mice , PPAR alpha/agonists , PPAR alpha/deficiency , Phenylhydrazines/pharmacology , Phenylurea Compounds/pharmacology , Phenylurea Compounds/therapeutic use , Signal Transduction/drug effects
18.
J Clin Invest ; 125(4): 1419-32, 2015 Apr.
Article in English | MEDLINE | ID: mdl-25705885

ABSTRACT

The small GTPase RAP1 is critical for platelet activation and thrombus formation. RAP1 activity in platelets is controlled by the GEF CalDAG-GEFI and an unknown regulator that operates downstream of the adenosine diphosphate (ADP) receptor, P2Y12, a target of antithrombotic therapy. Here, we provide evidence that the GAP, RASA3, inhibits platelet activation and provides a link between P2Y12 and activation of the RAP1 signaling pathway. In mice, reduced expression of RASA3 led to premature platelet activation and markedly reduced the life span of circulating platelets. The increased platelet turnover and the resulting thrombocytopenia were reversed by concomitant deletion of the gene encoding CalDAG-GEFI. Rasa3 mutant platelets were hyperresponsive to agonist stimulation, both in vitro and in vivo. Moreover, activation of Rasa3 mutant platelets occurred independently of ADP feedback signaling and was insensitive to inhibitors of P2Y12 or PI3 kinase. Together, our results indicate that RASA3 ensures that circulating platelets remain quiescent by restraining CalDAG-GEFI/RAP1 signaling and suggest that P2Y12 signaling is required to inhibit RASA3 and enable sustained RAP1-dependent platelet activation and thrombus formation at sites of vascular injury. These findings provide insight into the antithrombotic effect of P2Y12 inhibitors and may lead to improved diagnosis and treatment of platelet-related disorders.


Subject(s)
GTPase-Activating Proteins/physiology , Platelet Activation/physiology , rap1 GTP-Binding Proteins/antagonists & inhibitors , Animals , Cellular Senescence , Clopidogrel , GTPase-Activating Proteins/genetics , Guanine Nucleotide Exchange Factors/deficiency , Hemostasis , Lymphopenia/genetics , Mice , Mice, Inbred BALB C , Mice, Inbred C57BL , Mice, Knockout , Platelet Activation/drug effects , Platelet Activation/genetics , Platelet Aggregation Inhibitors/pharmacology , Proto-Oncogene Proteins c-akt/physiology , Receptors, Purinergic P2Y12/physiology , Saphenous Vein/injuries , Splenectomy , Thrombocytopenia/genetics , Thrombocytopenia/surgery , Thrombopoiesis , Ticlopidine/analogs & derivatives , Ticlopidine/pharmacology , rap1 GTP-Binding Proteins/physiology
19.
J Gerontol A Biol Sci Med Sci ; 70(7): 817-26, 2015 Jul.
Article in English | MEDLINE | ID: mdl-25070661

ABSTRACT

We previously reported that insulin-like growth factor 1 (IGF1) was involved in coregulating female sexual maturation and longevity. To understand the underlying genetic mechanisms, based on the strain survey assays of development and aging traits, we crossed two mouse strains, KK/HIJ and PL/J, and produced 307 female F2 mice. We observed the age of vaginal patency (AVP) and the life span of these females. We also measured circulating IGF1 level at 7, 16, 24, 52, and 76 weeks. IGF1 level at 7 weeks significantly correlated with AVP. IGF1 levels at ages of 52 and 76 weeks negatively correlated with longevity (p ≤ .05). A gene mapping study found 22, 4 ,and 3 quantitative trait loci for IGF1, AVP, and life span, respectively. Importantly, the colocalization of IGF1, AVP, and life span quantitative trait loci in the distal region of chromosome 2 suggests this locus carries gene(s) that could regulate IGF1, AVP, and life span. In this region, proprotein convertase subtilisin/kexin type 2 has been found to be associated with female sexual maturation in a human genome-wide association study. We verified the roles of proprotein convertase subtilisin/kexin type 2 in regulating IGF1 and AVP by showing that depletion of proprotein convertase subtilisin/kexin type 2 significantly reduced IGF1 and delayed AVP in mice, suggesting that it also might be involved in the regulation of aging.


Subject(s)
Insulin-Like Growth Factor I/metabolism , Longevity/physiology , Proprotein Convertase 2/genetics , Quantitative Trait Loci/genetics , Sexual Maturation/physiology , Animals , Chromosome Mapping , Female , Lod Score , Mice , Mice, Inbred Strains , Sexual Maturation/genetics
20.
Front Physiol ; 5: 375, 2014.
Article in English | MEDLINE | ID: mdl-25324783

ABSTRACT

The lengths of the sarcomeric thin filaments vary in a skeletal muscle-specific manner and help specify the physiological properties of skeletal muscle. Since the extent of overlap between the thin and thick filaments determines the amount of contractile force that a sarcomere can actively produce, thin filament lengths are accurate predictors of muscle-specific sarcomere length-tension relationships and sarcomere operating length ranges. However, the striking uniformity of thin filament lengths within sarcomeres, specified during myofibril assembly, has led to the widely held assumption that thin filament lengths remain constant throughout an organism's lifespan. Here, we rigorously tested this assumption by using computational super-resolution image analysis of confocal fluorescence images to explore the effects of postnatal development and aging on thin filament length in mice. We found that thin filaments shorten in postnatal tibialis anterior (TA) and gastrocnemius muscles between postnatal days 7 and 21, consistent with the developmental program of myosin heavy chain (MHC) gene expression in this interval. By contrast, thin filament lengths in TA and extensor digitorum longus (EDL) muscles remained constant between 2 mo and 2 yr of age, while thin filament lengths in soleus muscle became shorter, suggestive of a slow-muscle-specific mechanism of thin filament destabilization associated with aging. Collectively, these data are the first to show that thin filament lengths change as part of normal skeletal muscle development and aging, motivating future investigations into the cellular and molecular mechanisms underlying thin filament adaptation across the lifespan.

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