ABSTRACT
Background: Long non-coding RNAs (lncRNAs) play crucial roles in gene regulation and are implicated in neurodegenerative diseases, including frontotemporal dementia (FTD). However, their expression patterns and potential as biomarkers in genetic FTD involving Chromosome 9 Open Reading Frame (C9ORF72), Microtubule Associated Protein Tau (MAPT), and Progranulin (GRN) genes are not well understood. Objective: This study aimed to profile the expression levels of lncRNAs in peripheral blood mononuclear cells collected within the GENetic Frontotemporal dementia Initiative (GENFI). Methods: Fifty-three lncRNAs were analyzed with the OpenArray Custom panel, in 131 patients with mutations in C9ORF72, MAPT, and GRN, including 68 symptomatic mutation carriers (SMC) and 63 presymptomatic mutation carriers (PMC), compared with 40 non-carrier controls (NC). Results: Thirty-eight lncRNAs were detectable; the relative expression of NEAT1 and NORAD was significantly higher in C9ORF72 SMC as compared with NC. GAS5 expression was instead significantly lower in the GRN group versus NC. MAPT carriers showed no significant deregulations. No significant differences were observed in PMC. Disease duration did not correlate with lncRNA expression. Conclusions: NEAT1 and NORAD are upregulated in C9ORF72 SMC and GAS5 levels are downregulated in GRN SMC, underlining lncRNAs' relevance in FTD and their potential for biomarker development. Further validation and mechanistic studies are crucial for clinical implications.
Subject(s)
C9orf72 Protein , Frontotemporal Dementia , Progranulins , RNA, Long Noncoding , tau Proteins , Humans , Frontotemporal Dementia/genetics , RNA, Long Noncoding/genetics , Female , Male , Middle Aged , C9orf72 Protein/genetics , Progranulins/genetics , tau Proteins/genetics , Aged , Mutation , Biomarkers/bloodABSTRACT
Helicobacter pylori (H. pylori) infection is the primary risk factor for the progress of gastric diseases. The persistent stomach colonization of H. pylori is closely associated with the development of gastritis and malignancies. Although the involvement of progranulin (PGRN) in various cancer types has been well-documented, its functional role and underlying mechanisms in gastric cancer (GC) associated with H. pylori infection remain largely unknown. This report demonstrated that PGRN was up-regulated in GC and associated with poor prognosis, as determined through local and public database analysis. Additionally, H. pylori induced the up-regulation of PGRN in gastric epithelial cells both in vitro and in vivo. Functional studies have shown that PGRN promoted the intracellular colonization of H. pylori. Mechanistically, H. pylori infection induced autophagy, while PGRN inhibited autophagy to promote the intracellular colonization of H. pylori. Furthermore, PGRN suppressed H. pylori-induced autophagy by down-regulating decorin (DCN) through the mTOR pathway. In general, PGRN inhibited autophagy to facilitate intracellular colonization of H. pylori via the PGRN/mTOR/DCN axis. This study provides new insights into the molecular mechanisms underlying the progression of gastric diseases, suggesting PGRN as a potential therapeutic target and prognostic predictor for these disorders.
Subject(s)
Autophagy , Epithelial Cells , Gastric Mucosa , Helicobacter Infections , Helicobacter pylori , Progranulins , Stomach Neoplasms , TOR Serine-Threonine Kinases , Progranulins/metabolism , TOR Serine-Threonine Kinases/metabolism , Humans , Epithelial Cells/microbiology , Epithelial Cells/metabolism , Helicobacter Infections/microbiology , Helicobacter Infections/metabolism , Animals , Stomach Neoplasms/microbiology , Stomach Neoplasms/metabolism , Stomach Neoplasms/pathology , Gastric Mucosa/microbiology , Gastric Mucosa/metabolism , Mice , Signal TransductionABSTRACT
Background and Objectives: Diabetes is a significant health problem, prompting the search for new therapeutic strategies. Recently, researchers have focused on identifying novel markers for the progression of this condition. It is well established that adipokines, such as progranulin and vaspin, play crucial roles in regulating lipid and carbohydrate metabolism. Materials and Methods: This single-center cross-sectional study aimed to assess serum progranulin and vaspin levels in 80 children diagnosed with type 1 diabetes (T1D) and to examine their correlation with body mass index (BMI), glycated hemoglobin, and lipid profile. The cohort included 40 children newly diagnosed with diabetes, 40 children with long-term diabetes (20 well-controlled and 20 poorly controlled), and 14 non-diabetic children as a control group. Progranulin and vaspin levels were determined using a sandwich enzyme-linked immunosorbent assay. Results: There were no significant differences in the progranulin and vaspin concentrations in the studied groups (p = 0.246 and p = 0.095, respectively). No statistically significant differences were noted in the levels of both adipokines among boys and girls within the T1D, well-controlled T1D, and poorly controlled T1D groups. We did not find any differences in the progranulin and vaspin levels among all children with T1D and healthy controls when divided based on BMI percentiles. A negative correlation was observed between progranulin concentration and the age of children in the T1D, well-controlled T1D, and healthy groups. Furthermore, progranulin correlated negatively with BMI among children with T1D. In contrast, vaspin concentration correlated positively with age among healthy children. Conclusions: Our study provides novel insights into the status of progranulin and vaspin among pediatric participants with varying levels of type 1 diabetes control. However, further research involving larger patient cohorts and different stages of sexual maturation is warranted.
Subject(s)
Biomarkers , Body Mass Index , Diabetes Mellitus, Type 1 , Progranulins , Serpins , Humans , Female , Male , Serpins/blood , Progranulins/blood , Diabetes Mellitus, Type 1/blood , Diabetes Mellitus, Type 1/complications , Child , Biomarkers/blood , Cross-Sectional Studies , Adolescent , Glycated Hemoglobin/analysis , Enzyme-Linked Immunosorbent AssayABSTRACT
In systemic sclerosis (SSc), fibrosis of the myocardium along with ongoing autoimmune inflammation can alter the electric function of the cardiac myocytes, which may increase the risk for ventricular arrhythmias and sudden cardiac death. We analyzed the electrocardiographic (ECG) variables describing ventricular repolarization such as QT interval, QT dispersion (QTd), T wave peak-to-end interval (Tpe), and arrhythmogeneity index (AIX) of 26 patients with SSc and 36 healthy controls. Furthermore, echocardiographic and laboratory parameters were examined, with a focus on inflammatory proteins like C-reactive ptotein (CRP), soluble intracellular adhesion molecule-1 (sICAM-1), soluble vascular adhesion molecule-1 (sVCAM-1), and progranulin (PGRN). The CRP, sICAM-1, and sVCAM-1 levels were positively correlated with the length of the QT interval. Although the serum PGRN levels were not increased in the SSc group compared to the controls, in SSc patients, the PGRN levels were positively correlated with the QT interval and the AIX. According to our results, we conclude that there may be a potential association between autoimmune inflammation and the risk for ventricular arrhythmias in patients with SSc. We emphasize that the measurement of laboratory parameters of inflammatory activity including CRP, PGRN, sVCAM-1, and sICAM-1 could be helpful in the prediction of sudden cardiac death in patients with SSc.
Subject(s)
Arrhythmias, Cardiac , Intercellular Adhesion Molecule-1 , Progranulins , Scleroderma, Systemic , Vascular Cell Adhesion Molecule-1 , Humans , Scleroderma, Systemic/blood , Scleroderma, Systemic/complications , Female , Male , Middle Aged , Vascular Cell Adhesion Molecule-1/blood , Intercellular Adhesion Molecule-1/blood , Arrhythmias, Cardiac/etiology , Arrhythmias, Cardiac/blood , Progranulins/blood , Electrocardiography , Adult , Biomarkers/blood , Case-Control Studies , Aged , Risk Factors , C-Reactive Protein/metabolism , C-Reactive Protein/analysisABSTRACT
Heterozygous loss-of-function mutations in the GRN gene are a major cause of hereditary frontotemporal dementia. The mechanisms linking frontotemporal dementia pathogenesis to progranulin deficiency are not well understood, and there is currently no treatment. Our strategy to prevent the onset and progression of frontotemporal dementia in patients with GRN mutations is to utilize small molecule positive regulators of GRN expression to boost progranulin levels from the remaining functional GRN allele, thus restoring progranulin levels back to normal within the brain. This work describes a series of blood-brain-barrier-penetrant small molecules which significantly increase progranulin protein levels in human cellular models, correct progranulin protein deficiency in Grn+/- mouse brains, and reverse lysosomal proteome aberrations, a phenotypic hallmark of frontotemporal dementia, more efficiently than the previously described small molecule suberoylanilide hydroxamic acid. These molecules will allow further elucidation of the cellular functions of progranulin and its role in frontotemporal dementia and will also serve as lead structures for further drug development.
Subject(s)
Frontotemporal Dementia , Haploinsufficiency , Lysosomes , Progranulins , Proteome , Progranulins/metabolism , Progranulins/genetics , Animals , Humans , Frontotemporal Dementia/genetics , Frontotemporal Dementia/metabolism , Frontotemporal Dementia/drug therapy , Proteome/metabolism , Mice , Lysosomes/metabolism , Lysosomes/drug effects , Brain/metabolism , Brain/drug effects , Vorinostat/pharmacologyABSTRACT
Alzheimer's Disease (AD) and Frontotemporal Dementia (FTD) are the two major neurodegenerative diseases with distinct clinical and neuropathological profiles. The aim of this report is to conduct a population-based investigation in well-characterized APP, PSEN1, PSEN2, MAPT, GRN, and C9orf72 mutation carriers/pedigrees from the north, the center, and the south of Italy. We retrospectively analyzed the data of 467 Italian individuals. We identified 21 different GRN mutations, 20 PSEN1, 11 MAPT, 9 PSEN2, and 4 APP. Moreover, we observed geographical variability in mutation frequencies by looking at each cohort of participants, and we observed a significant difference in age at onset among the genetic groups. Our study provides evidence that age at onset is influenced by the genetic group. Further work in identifying both genetic and environmental factors that modify the phenotypes in all groups is needed. Our study reveals Italian regional differences among the most relevant AD/FTD causative genes and emphasizes how the collaborative studies in rare diseases can provide new insights to expand knowledge on genetic/epigenetic modulators of age at onset.
Subject(s)
Alzheimer Disease , Frontotemporal Dementia , Mutation , tau Proteins , Humans , Alzheimer Disease/genetics , Alzheimer Disease/epidemiology , Italy/epidemiology , Frontotemporal Dementia/genetics , Frontotemporal Dementia/epidemiology , Frontotemporal Dementia/pathology , Female , Male , Middle Aged , Aged , tau Proteins/genetics , Age of Onset , C9orf72 Protein/genetics , Presenilin-2/genetics , Retrospective Studies , Amyloid beta-Protein Precursor/genetics , Presenilin-1/genetics , Progranulins/genetics , Adult , Aged, 80 and over , Genetic Predisposition to DiseaseABSTRACT
Purpose: This study aimed to investigate the expression levels of progranulin (PGRN) in the tears of patients with diabetic retinopathy (DR) versus healthy controls. Additionally, we sought to explore the correlation between PGRN levels and the severity of ocular surface complications in patients with diabetes. Methods: In this prospective, single-visit, cross-sectional study, patients with DR (n = 48) and age-matched healthy controls (n = 22) were included and underwent dry eye examinations. Tear fluid was collected, and its components were analyzed using the Luminex assay. The subbasal nerve plexus of all participants was evaluated by in vivo confocal microscopy. Results: Patients with DR exhibited more severe dry eye symptoms, along with a reduction in nerve fiber density, length, and branch density within the subbasal nerve plexus, accompanied by an increase in the number of dendritic cells. Tear PGRN levels were also significantly lower in patients with diabetes than in normal controls, and the levels of some inflammatory factors (TNF-α, IL-6, and MMP-9) were higher in patients with DR. Remarkably, the PGRN level significantly correlated with nerve fiber density (R = 0.48, P < 0.001), nerve fiber length (R = 0.65, P < 0.001), and nerve branch density (R = 0.69, P < 0.001). Conclusions: Tear PGRN levels might reflect morphological changes in the corneal nerve plexus under diabetic conditions, suggesting that PGRN itself is a reliable indicator for predicting the advancement of neurotrophic keratopathy in patients with diabetes. Translational Relevance: PGRN insufficiency on the ocular surface under diabetic conditions was found to be closely associated with nerve impairment, providing a novel perspective to discover the pathogenesis of diabetic complications, which could help in developing innovative therapeutic strategies.
Subject(s)
Biomarkers , Cornea , Diabetes Mellitus, Type 2 , Diabetic Retinopathy , Progranulins , Tears , Humans , Tears/metabolism , Tears/chemistry , Male , Female , Progranulins/metabolism , Middle Aged , Cross-Sectional Studies , Prospective Studies , Biomarkers/analysis , Biomarkers/metabolism , Diabetes Mellitus, Type 2/complications , Diabetes Mellitus, Type 2/metabolism , Cornea/innervation , Cornea/metabolism , Cornea/pathology , Diabetic Retinopathy/metabolism , Diabetic Retinopathy/diagnosis , Diabetic Retinopathy/pathology , Aged , Microscopy, Confocal , Dry Eye Syndromes/metabolism , Dry Eye Syndromes/diagnosis , Dry Eye Syndromes/etiology , Dry Eye Syndromes/pathology , Nerve Fibers/pathology , Nerve Fibers/metabolismABSTRACT
PROBLEM: To explore the clinical utility of nine inflammatory immune-, adhesion-, and extracellular matrix-related mediators in the plasma for predicting intraamniotic inflammation and/or microbial invasion of the amniotic cavity (IAI/MIAC) and composite neonatal morbidity and/or mortality (CNMM) in women with preterm premature rupture of membranes (PPROM) when used alone or in combination with conventional blood-, ultrasound-, and clinical-based factors. METHODS OF STUDY: This retrospective cohort comprised 173 singleton pregnant women with PPROM (24 + 0 - 33 + 6 weeks), who underwent amniocentesis. Amniotic fluid was cultured for microorganisms and assayed for IL-6 levels. Plasma levels of AFP, CXCL14, E-selectin, Gal-3BP, kallistatin, progranulin, P-selectin, TGFBI, and VDBP were determined by ELISA. Ultrasonographic cervical length (CL) and neutrophil-to-lymphocyte ratio (NLR) were measured. RESULTS: Multivariate logistic regression analyses revealed significant associations between (i) decreased plasma kallistatin levels and IAI/MIAC and (ii) decreased plasma progranulin levels and increased CNMM risk after adjusting for baseline variables (e.g., gestational age at sampling [or delivery] and parity). Using stepwise regression analysis, noninvasive prediction models for IAI/MIAC and CNMM risks were developed, which included plasma progranulin levels, NLR, CL, and gestational age at sampling, and provided a good prediction of the corresponding endpoints (area under the curve: 0.79 and 0.87, respectively). CONCLUSIONS: Kallistatin and progranulin are potentially valuable plasma biomarkers for predicting IAI/MIAC and CNMM in women with PPROM. Particularly, the combination of these plasma biomarkers with conventional blood-, ultrasound-, and clinical-based factors can significantly support the diagnosis of IAI/MIAC and CNMM.
Subject(s)
Biomarkers , Fetal Membranes, Premature Rupture , Progranulins , Serpins , Humans , Female , Pregnancy , Progranulins/blood , Biomarkers/blood , Adult , Serpins/blood , Retrospective Studies , Fetal Membranes, Premature Rupture/blood , Infant, Newborn , Amniotic Fluid/microbiology , Amniotic Fluid/metabolism , Chorioamnionitis/blood , Chorioamnionitis/immunology , Intercellular Signaling Peptides and Proteins/blood , Inflammation/bloodABSTRACT
Hematopoietic stem cell transplantation can deliver therapeutic proteins to the central nervous system (CNS) through transplant-derived microglia-like cells. However, current conditioning approaches result in low and slow engraftment of transplanted cells in the CNS. Here we optimized a brain conditioning regimen that leads to rapid, robust, and persistent microglia replacement without adverse effects on neurobehavior or hematopoiesis. This regimen combines busulfan myeloablation and six days of Colony-stimulating factor 1 receptor inhibitor PLX3397. Single-cell analyses revealed unappreciated heterogeneity of microglia-like cells with most cells expressing genes characteristic of homeostatic microglia, brain-border-associated macrophages, and unique markers. Cytokine analysis in the CNS showed transient inductions of myeloproliferative and chemoattractant cytokines that help repopulate the microglia niche. Bone marrow transplant of progranulin-deficient mice conditioned with busulfan and PLX3397 restored progranulin in the brain and eyes and normalized brain lipofuscin storage, proteostasis, and lipid metabolism. This study advances our understanding of CNS repopulation by hematopoietic-derived cells and demonstrates its therapeutic potential for treating progranulin-dependent neurodegeneration.
Subject(s)
Busulfan , Microglia , Progranulins , Animals , Microglia/metabolism , Microglia/drug effects , Progranulins/metabolism , Progranulins/genetics , Mice , Busulfan/pharmacology , Hematopoietic Stem Cell Transplantation , Aminopyridines/pharmacology , Brain/metabolism , Pyrroles/pharmacology , Mice, Inbred C57BL , Hematopoietic Stem Cells/metabolism , Hematopoietic Stem Cells/drug effects , Hematopoietic Stem Cells/cytology , Bone Marrow Transplantation , Male , Central Nervous System/metabolism , Mice, Knockout , Transplantation Conditioning/methods , Single-Cell Analysis , Cytokines/metabolism , Receptors, Granulocyte-Macrophage Colony-Stimulating Factor/metabolism , Receptors, Granulocyte-Macrophage Colony-Stimulating Factor/genetics , Receptors, Granulocyte-Macrophage Colony-Stimulating Factor/antagonists & inhibitorsABSTRACT
INTRODUCTION: Frontotemporal lobar degeneration (FTLD) encompasses behavioral variant frontotemporal dementia (bvFTD), progressive supranuclear palsy, corticobasal syndrome/degeneration, and primary progressive aphasias (PPAs). We cross-validated fluid biomarkers and neuroimaging. METHODS: Seven fluid biomarkers from cerebrospinal fluid and serum were related to atrophy in 428 participants including these FTLD subtypes, logopenic variant PPA (lvPPA), Alzheimer's disease (AD), and healthy subjects. Atrophy was assessed by structural magnetic resonance imaging and atlas-based volumetry. RESULTS: FTLD subtypes, lvPPA, and AD showed specific profiles for neurofilament light chain, phosphorylated heavy chain, tau, phospho-tau, amyloid beta1-42 from serum/cerebrospinal fluid, and brain atrophy. Neurofilaments related to regional atrophy in bvFTD, whereas progranulin was associated with atrophy in semantic variant PPA. Ubiquitin showed no effects. DISCUSSION: Results specify biomarker and atrophy patterns in FTLD and AD supporting differential diagnosis. They identify neurofilaments and progranulin in interaction with structural imaging as promising candidates for monitoring disease progression and therapy. HIGHLIGHTS: Study cross-validated neuroimaging and fluid biomarkers in dementia. Five kinds of frontotemporal lobar degeneration and two variants of Alzheimer's disease. Study identifies disease-specific fluid biomarker and atrophy profiles. Fluid biomarkers and atrophy interact in a disease-specific way. Neurofilaments and progranulin are proposed as biomarkers for diagnosis and therapy.
Subject(s)
Alzheimer Disease , Atrophy , Biomarkers , Brain , Frontotemporal Lobar Degeneration , Magnetic Resonance Imaging , Neurofilament Proteins , Progranulins , tau Proteins , Humans , Biomarkers/cerebrospinal fluid , Biomarkers/blood , Frontotemporal Lobar Degeneration/pathology , Male , Female , Atrophy/pathology , Aged , Middle Aged , Neurofilament Proteins/cerebrospinal fluid , Neurofilament Proteins/blood , tau Proteins/cerebrospinal fluid , Brain/pathology , Brain/diagnostic imaging , Alzheimer Disease/pathology , Alzheimer Disease/cerebrospinal fluid , Amyloid beta-Peptides/cerebrospinal fluidABSTRACT
Progranulin (PGRN) haploinsufficiency is a major risk factor for frontotemporal lobar degeneration with TAR DNA-binding protein 43 (TDP-43) pathology (FTLD-GRN). Multiple therapeutic strategies are in clinical development to restore PGRN in the CNS, including gene therapy. However, a limitation of current gene therapy approaches aimed to alleviate FTLD-associated pathologies may be their inefficient brain exposure and biodistribution. We therefore developed an adeno-associated virus (AAV) targeting the liver (L) to achieve sustained peripheral expression of a transferrin receptor (TfR) binding, brain-penetrant (b) PGRN variant [AAV(L):bPGRN] in two mouse models of FTLD-GRN, namely, Grn knockout and GrnxTmem106b double knockout mice. This therapeutic strategy avoids potential safety and biodistribution issues of CNS-administered AAVs and maintains sustained concentrations of PGRN in the brain after a single dose. AAV(L):bPGRN treatment reduced several FTLD-GRN-associated pathologies including severe motor function deficits, aberrant TDP-43 phosphorylation, dysfunctional protein degradation, lipid metabolism, gliosis, and neurodegeneration in the brain. The potential translatability of our findings was tested in an in vitro model using cocultured human induced pluripotent stem cell (hiPSC)-derived microglia lacking PGRN and TMEM106B and wild-type hiPSC-derived neurons. As in mice, aberrant TDP-43, lysosomal dysfunction, and neuronal loss were ameliorated after treatment with exogenous TfR-binding protein transport vehicle fused to PGRN (PTV:PGRN). Together, our studies suggest that peripherally administered brain-penetrant PGRN replacement strategies ameliorate FTLD-GRN relevant phenotypes including TDP-43 pathology, neurodegeneration, and behavioral deficits. Our data provide preclinical proof of concept for the use of this AAV platform for treatment of FTLD-GRN and potentially other CNS disorders.
Subject(s)
Brain , Dependovirus , Disease Models, Animal , Frontotemporal Lobar Degeneration , Mice, Knockout , Progranulins , Animals , Humans , Mice , Brain/metabolism , Brain/pathology , Dependovirus/metabolism , DNA-Binding Proteins/metabolism , DNA-Binding Proteins/genetics , Frontotemporal Lobar Degeneration/metabolism , Frontotemporal Lobar Degeneration/pathology , Genetic Therapy , Phosphorylation , Progranulins/metabolism , Progranulins/genetics , Receptors, Transferrin/metabolismABSTRACT
The aim of the present study was to determine the effects of the adipokines progranulin and omentin on the basic functions of feline ovarian cells. For this purpose, we investigated the effects of the addition of progranulin and omentin (0, 0.1, 1, or 10 ng/ml) on the proliferation (accumulation of PCNA and cyclin B1), apoptosis (accumulation of Bax and caspase 3) and progesterone release of cultured feline ovarian granulosa cells by quantitative immunocytochemistry and enzyme-linked immunosorbent assays (ELISAs). Both progranulin and omentin increased cell proliferation and decreased apoptosis. Both progranulin and omentin promoted progesterone release. The present findings demonstrate that the adipokines progranulin and omentin can directly regulate basic feline ovarian cell functions.
Subject(s)
Apoptosis , Cell Proliferation , Granulosa Cells , Animals , Female , Cats , Granulosa Cells/metabolism , Granulosa Cells/drug effects , Apoptosis/drug effects , Cell Proliferation/drug effects , Progesterone/metabolism , Progesterone/pharmacology , Progranulins/metabolism , Cytokines/metabolism , Cells, Cultured , Lectins/metabolism , Lectins/pharmacologyABSTRACT
Fungal keratitis (FK) is a severe corneal condition caused by pathogenic fungi and is associated with the virulence of fungi and an excessive tissue inflammatory response. Progranulin (PGRN), functioning as a multifunctional growth factor, exerts a pivotal influence on the regulation of inflammation and autophagy. The aim of our research was to analyze the role of PGRN in Aspergillus fumigatus (A. fumigatus) keratitis. We found that PGRN expression was increased in the mouse cornea with A. fumigatus keratitis. In our experiments, corneas of mice with FK were treated with 100 ng/mL of PGRN. In vitro, RAW 264.7 cells were treated with 10 ng/mL of PGRN before A. fumigatus stimulation. The findings suggested that PGRN effectively alleviated corneal edema and decreased the expression of pro-inflammatory cytokines in mice. In stimulated RAW 264.7 cells, PGRN treatment suppressed the expression of pro-inflammatory cytokines IL-6 and TNF-α but promoted the expression of the anti-inflammatory cytokines IL-10. PGRN treatment significantly upregulated the expression of autophagy-related proteins LC3, Beclin-1, and Atg-7. 3-Methyladenine (3-MA, autophagy inhibitor) reversed the regulation of inflammatory cytokines by PGRN. In addition, our study demonstrated that PGRN also enhanced phagocytosis in RAW 264.7 cells. In summary, PGRN attenuated the inflammatory response of A. fumigatus keratitis by increasing autophagy and enhanced the phagocytic activity of RAW 264.7 cells. This showed that PGRN had a protective effect on A. fumigatus keratitis.
Subject(s)
Aspergillosis , Aspergillus fumigatus , Autophagy , Cytokines , Keratitis , Progranulins , Animals , Mice , Autophagy/drug effects , Keratitis/microbiology , Keratitis/drug therapy , RAW 264.7 Cells , Aspergillosis/drug therapy , Aspergillosis/microbiology , Cytokines/metabolism , Cornea , Inflammation/drug therapy , Disease Models, AnimalABSTRACT
BACKGROUND: Prior research has shown that granulin precursor (GRN, also termed PGRN) is closely linked to aphasia. However, there has been little research on the mechanism of action of GRN in post-stroke aphasia (PSA). METHODS: In this study, RT-qPCR was used to identify variations in gene expression, while RNA sequencing (RNA-seq) was utilized to acquire transcriptional profiles. The Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) databases were employed for bioinformatics analysis. RESULTS: GRN was considerably more active in PSA subjects. After silencing the GRN, 197 transcripts had differential expression, and 237 alternative splicing events (ASEs) were substantially affected. The analysis of differentially expressed genes (DEGs) using GO and KEGG approaches showed that these genes have various molecular functions and are significantly enriched in metabolic signaling pathways. Regarding Alternative Splicing (AS), the GO and KEGG analyses revealed numerous functional genes involved in transcription and metabolism. CONCLUSIONS: The knockdown of GRN has been shown to be associated with alterations in transcription, metabolism, and ASEs, potentially impacting transcriptional and metabolic pathways through its involvement in AS. Furthermore, GRN knockdown is associated with nervous system disease-related gene transcription and AS processes, as well as its involvement in G protein-coupled receptor (GPCR) and wingless/integrated (Wnt) signaling pathways, which impact the initiation and resolution of PSA.
Subject(s)
Alternative Splicing , Aphasia , Progranulins , Progranulins/genetics , Alternative Splicing/genetics , Humans , Animals , Rats , Aphasia/genetics , Aphasia/metabolism , PC12 Cells , Gene Knockdown Techniques , Male , Stroke/geneticsABSTRACT
ADAR1-mediated RNA editing establishes immune tolerance to endogenous double-stranded RNA (dsRNA) by preventing its sensing, primarily by MDA5. Although deleting Ifih1 (encoding MDA5) rescues embryonic lethality in ADAR1-deficient mice, they still experience early postnatal death, and removing other MDA5 signaling proteins does not yield the same rescue. Here, we show that ablation of MDA5 in a liver-specific Adar knockout (KO) murine model fails to rescue hepatic abnormalities caused by ADAR1 loss. Ifih1;Adar double KO (dKO) hepatocytes accumulate endogenous dsRNAs, leading to aberrant transition to a highly inflammatory state and recruitment of macrophages into dKO livers. Mechanistically, progranulin (PGRN) appears to mediate ADAR1 deficiency-induced liver pathology, promoting interferon signaling and attracting epidermal growth factor receptor (EGFR)+ macrophages into dKO liver, exacerbating hepatic inflammation. Notably, the PGRN-EGFR crosstalk communication and consequent immune responses are significantly repressed in ADAR1high tumors, revealing that pre-neoplastic or neoplastic cells can exploit ADAR1-dependent immune tolerance to facilitate immune evasion.
Subject(s)
Adenosine Deaminase , ErbB Receptors , Hepatocytes , Interferon-Induced Helicase, IFIH1 , Liver , Macrophages , Mice, Knockout , Progranulins , Animals , Adenosine Deaminase/metabolism , Adenosine Deaminase/genetics , ErbB Receptors/metabolism , Macrophages/metabolism , Macrophages/immunology , Progranulins/metabolism , Progranulins/genetics , Liver/metabolism , Liver/immunology , Liver/pathology , Hepatocytes/metabolism , Mice , Interferon-Induced Helicase, IFIH1/metabolism , Interferon-Induced Helicase, IFIH1/genetics , Signal Transduction , RNA-Binding Proteins/metabolism , RNA-Binding Proteins/genetics , Humans , Intercellular Signaling Peptides and Proteins/metabolism , Intercellular Signaling Peptides and Proteins/genetics , Mice, Inbred C57BL , RNA, Double-Stranded/metabolism , RNA EditingABSTRACT
Most of the heritability in frontotemporal dementia (FTD) is accounted for by autosomal dominant hexanucleotide expansion in the chromosome 9 open reading frame 72 (C9orf72), pathogenic/likely pathogenic variants in progranulin (GRN), and microtubule-associated protein tau (MAPT) genes. Until now, there has been no systematic analysis of these genes in the Serbian population. Herein, we assessed the frequency of the C9orf72 expansion, pathogenic/likely pathogenic variants in GRN and MAPT in a well-characterized group of 472 subjects (FTD, Alzheimer's disease - AD, mild cognitive impairment - MCI, and unspecified dementia - UnD), recruited in the Memory Center, Neurology Clinic, University Clinical Center of Serbia. The C9orf72 repeat expansion was detected in 6.98% of FTD cases (13.46% familial; 2.6% sporadic). In the UnD subgroup, C9orf72 repeat expansions were detected in 4.08% (8% familial) individuals. Pathogenic variants in the GRN were found in 2.85% of familial FTD cases. Interestingly, no MAPT pathogenic/likely pathogenic variants were detected, suggesting possible geographical specificity. Our findings highlight the importance of wider implementation of genetic testing in neurological and psychiatric practice managing patients with cognitive-behavioral and motor symptoms.
Subject(s)
C9orf72 Protein , Frontotemporal Dementia , Progranulins , tau Proteins , Humans , tau Proteins/genetics , C9orf72 Protein/genetics , Progranulins/genetics , Female , Male , Aged , Frontotemporal Dementia/genetics , Middle Aged , Serbia/epidemiology , DNA Repeat Expansion/genetics , Cognitive Dysfunction/genetics , Cognitive Dysfunction/epidemiology , Alzheimer Disease/genetics , Intercellular Signaling Peptides and Proteins/genetics , Aged, 80 and overABSTRACT
Loss-of-function mutations in the progranulin (GRN) gene are an autosomal dominant cause of Frontotemporal Dementia (FTD). These mutations typically result in haploinsufficiency of the progranulin protein. Grn+/- mice provide a model for progranulin haploinsufficiency and develop FTD-like behavioral abnormalities by 9-10 months of age. In previous work, we demonstrated that Grn+/- mice develop a low dominance phenotype in the tube test that is associated with reduced dendritic arborization of layer II/III pyramidal neurons in the prelimbic region of the medial prefrontal cortex (mPFC), a region key for social dominance behavior in the tube test assay. In this study, we investigated whether progranulin haploinsufficiency induced changes in dendritic spine density and morphology. Individual layer II/III pyramidal neurons in the prelimbic mPFC of 9-10 month old wild-type or Grn+/- mice were targeted for iontophoretic microinjection of fluorescent dye, followed by high-resolution confocal microscopy and 3D reconstruction for morphometry analysis. Dendritic spine density in Grn+/- mice was comparable to wild-type littermates, but the apical dendrites in Grn+/- mice had a shift in the proportion of spine types, with fewer stubby spines and more thin spines. Additionally, apical dendrites of Grn+/- mice had longer spines and smaller thin spine head diameter in comparison to wild-type littermates. These changes in spine morphology may contribute to altered circuit-level activity and social dominance deficits in Grn+/- mice.
Subject(s)
Dendritic Spines , Haploinsufficiency , Prefrontal Cortex , Progranulins , Animals , Dendritic Spines/metabolism , Prefrontal Cortex/pathology , Prefrontal Cortex/metabolism , Progranulins/deficiency , Progranulins/genetics , Mice , Pyramidal Cells/metabolism , Pyramidal Cells/pathology , Male , Mice, Inbred C57BLABSTRACT
GRN mutations cause progranulin haploinsufficiency, which eventually leads to frontotemporal dementia (FTD-GRN). PR006 is an investigational gene therapy delivering the granulin gene (GRN) using an adeno-associated virus serotype 9 (AAV9) vector. In non-clinical studies, PR006 transduced neurons derived from induced pluripotent stem cells of patients with FTD-GRN, resulted in progranulin expression and improvement of lipofuscin, lysosomal and neuroinflammation pathologies in Grn-knockout mice, and was well tolerated except for minimal, asymptomatic dorsal root ganglionopathy in non-human primates. We initiated a first-in-human phase 1/2 open-label trial. Here we report results of a pre-specified interim analysis triggered with the last treated patient of the low-dose cohort (n = 6) reaching the 12-month follow-up timepoint. We also include preliminary data from the mid-dose cohort (n = 7). Primary endpoints were safety, immunogenicity and change in progranulin levels in cerebrospinal fluid (CSF) and blood. Secondary endpoints were Clinical Dementia Rating (CDR) plus National Alzheimer's Disease Coordinating Center (NACC) Frontotemporal Lobar Degeneration (FTLD) rating scale and levels of neurofilament light chain (NfL). One-time administration of PR006 into the cisterna magna was generally safe and well tolerated. All patients developed treatment-emergent anti-AAV9 antibodies in the CSF, but none developed anti-progranulin antibodies. CSF pleocytosis was the most common PR006-related adverse event. Twelve serious adverse events occurred, mostly unrelated to PR006. Deep vein thrombosis developed in three patients. There was one death (unrelated) occurring 18 months after treatment. CSF progranulin increased after PR006 treatment in all patients; blood progranulin increased in most patients but only transiently. NfL levels transiently increased after PR006 treatment, likely reflecting dorsal root ganglia toxicity. Progression rates, based on the CDR scale, were within the broad ranges reported for patients with FTD. These data provide preliminary insights into the safety and bioactivity of PR006. Longer follow-up and additional studies are needed to confirm the safety and potential efficacy of PR006. ClinicalTrials.gov identifier: NCT04408625 .
Subject(s)
Dependovirus , Frontotemporal Dementia , Genetic Therapy , Progranulins , Humans , Frontotemporal Dementia/genetics , Frontotemporal Dementia/therapy , Frontotemporal Dementia/cerebrospinal fluid , Progranulins/genetics , Genetic Therapy/adverse effects , Genetic Therapy/methods , Dependovirus/genetics , Middle Aged , Female , Male , Aged , Intercellular Signaling Peptides and Proteins/genetics , Intercellular Signaling Peptides and Proteins/cerebrospinal fluid , Genetic Vectors , Animals , Treatment Outcome , Translational Research, Biomedical , Mice , Neurofilament Proteins/genetics , Neurofilament Proteins/cerebrospinal fluid , Neurofilament Proteins/bloodABSTRACT
Critical reprogramming factors resided predominantly in the oocyte or male pronucleus can enhance the efficiency or the quality of induced pluripotent stem cells (iPSCs) induction. However, few reprogramming factors exist in the male pronucleus had been verified. Here, we demonstrated that granulin (Grn), a factor enriched specifically in male pronucleus, can significantly improve the generation of iPSCs from mouse fibroblasts. Grn is highly expressed on Day 1, Day 3, Day 14 of reprogramming induced by four Yamanaka factors and functions at the initial stage of reprogramming. Transcriptome analysis indicates that Grn can promote the expression of lysosome-related genes, while inhibit the expression of genes involved in DNA replication and cell cycle at the early reprogramming stage. Further verification determined that Grn suppressed cell proliferation due to the arrest of cell cycle at G2/M phase. Moreover, ectopic Grn can enhance the lysosomes abundance and rescue the efficiency reduction of reprogramming resulted from lysosomal protease inhibition. Taken together, we conclude that Grn serves as an activator for somatic cell reprogramming through mitigating cell hyperproliferation and promoting the function of lysosomes.
Subject(s)
Cell Proliferation , Cellular Reprogramming , Fibroblasts , Induced Pluripotent Stem Cells , Lysosomes , Animals , Lysosomes/metabolism , Cellular Reprogramming/genetics , Male , Mice , Induced Pluripotent Stem Cells/metabolism , Fibroblasts/metabolism , Granulins , Progranulins/metabolism , Progranulins/genetics , Cell Nucleus/metabolismABSTRACT
Frontotemporal dementia (FTD) is a debilitating neurodegenerative disorder with currently no disease-modifying treatment options available. Mutations in GRN are one of the most common genetic causes of FTD, near ubiquitously resulting in progranulin (PGRN) haploinsufficiency. Small molecules that can restore PGRN protein to healthy levels in individuals bearing a heterozygous GRN mutation may thus have therapeutic value. Here, we show that epigenetic modulation through bromodomain and extra-terminal domain (BET) inhibitors (BETi) potently enhance PGRN protein levels, both intracellularly and secreted forms, in human central nervous system (CNS)-relevant cell types, including in microglia-like cells. In terms of potential for disease modification, we show BETi treatment effectively restores PGRN levels in neural cells with a GRN mutation known to cause PGRN haploinsufficiency and FTD. We demonstrate that BETi can rapidly and durably enhance PGRN in neural progenitor cells (NPCs) in a manner dependent upon BET protein expression, suggesting a gain-of-function mechanism. We further describe a CNS-optimized BETi chemotype that potently engages endogenous BRD4 and enhances PGRN expression in neuronal cells. Our results reveal a new epigenetic target for treating PGRN-deficient forms of FTD and provide mechanistic insight to aid in translating this discovery into therapeutics.