mRNA analysis of genes responsible for idiopathic hypogonadotrophic hypogonadism

Background. Hypogonadotropic hypogonadism (HH) is a disorder characterized by delayed or absent pubertal development due to pathology of the hypothalamic-pituitary-gonadal axis. HH may be both congenital (Kallmann’s syndrome) and sporadic. Congenital or isolated HH is divided into with anosmia/hyposmia (KS) and with normal olfaction (nIHH). Nowadays several tens of genes involved in the functioning of the reproductive axis are known. However DNA lesions can be found just in 5-15% of such cases of HH.Aim. So we decided to measure mRNA expression of several genes which can be found in leukocytes of peripheral blood - namely GNRHR and GNRH1 (are necessary for adequate biological effect of GnRH); PROK2 and CHD7 (are responsible for the migration of GnRH neurons), WDR11 and DUSP6 (are involved in normal sexual development).Methods. A quantitative determination of mRNA expression of these genes were comlpeted in the fresh peripheral blood sample by PCR in real time.Results. Examined patients: 9 women with hypogonadotropic hypogonadism (age from 18 to 28 y.o.); duration of the disease from 2 to 15 years; 3 of them – amenorrhea I and 6 – amenorrhea II. Reasons of amenorrhea II were: stress, excessive exercises, rapid body weight loss, past use of oral contraceptives. The control group: 19 healthy women; age from 19 to 37 y.o.; with regular ovalutory menstrual cycle, some of them have children. mRNA expression of examined genes differed from normal patterns in each case of hypogonadotropic hypogonadism. Changes in GNRHR, GNRH1 and DUSP6 mRNA expression were found in most of cases. However variations of mRNA expression were multidirectional in each case and there was no similarity among expression profiles of patients according to amenorrhea type or anamnestic factors.Conclusions. According to our preliminary results, in women with hypogonadotropic hypogonadism the functional activity damage of “reproductive-responsible” genes could be found in each case. Probably mRNA expression measuring could be a perspective method for proving hypothalamo-pituitary level of reproductive disorders and may help to determine which genes should be tested for DNA impairment.

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Deregulated microRNA and mRNA expression profiles in the peripheral blood of patients with Marfan syndrome

Journal of Translational Medicine ◽

10.1186/s12967-018-1429-3 ◽

2018 ◽

Vol 16 (1) ◽

Cited By ~ 9

Author(s):

Masood Abu-Halima ◽

Mustafa Kahraman ◽

Dominic Henn ◽

Tanja Rädle-Hurst ◽

Andreas Keller ◽

...

Keyword(s):

Mrna Expression ◽

Marfan Syndrome ◽

Peripheral Blood ◽

Expression Profiles

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Exploring the association of long noncoding RNA expression profiles with intracranial aneurysms, based on sequencing and related bioinformatics analysis

BMC Medical Genomics ◽

10.1186/s12920-020-00805-x ◽

2020 ◽

Vol 13 (1) ◽

Author(s):

Yi Sun ◽

Yeying Wen ◽

Qishuang Ruan ◽

Le Yang ◽

Shuna Huang ◽

...

Keyword(s):

Risk Factors ◽

Logistic Regression ◽

Inflammatory Response ◽

Mrna Expression ◽

Peripheral Blood ◽

Expression Profiles ◽

Peripheral Blood Leukocytes ◽

Multivariate Logistic Regression ◽

Blood Leukocytes ◽

Lncrna Malat1

Abstract Background The present study aims to investigate the complete long non-coding RNA (lncRNA) and messenger RNA (mRNA) expression profiles in Intracranial aneurysm (IA) patients and controls by RNA sequencing, which reveals the lncRNA with predictive value for IA risk. Methods The comprehensive lncRNA and mRNA expression profiles were detected by RNA-Seq in human IA walls and superficial temporal arteries (STAs), followed by bioinformatics analyses, such as GO analysis, KEGG pathway analysis, and CNC network construction. Subsequently, qRT-PCR was used to profile the expression levels of selected lncRNA (lncRNA ENST000000576153, lncRNA ENST00000607042, lncRNA ENST00000471220, lncRNA ENST00000478738, lncRNA MALAT1, lncRNA ENST00000508090 and lncRNA ENST00000579688) in 30 (small) or 130 (large) peripheral blood leukocytes, respectively. Multivariate logistic regression was utilized to analyze the effects of lncRNA on IA. Receiver operating characteristic (ROC) curve was further drawn to explore the value of lncRNA in predicting IA. Results Totally 900 up-regulated and 293 down-regulated lncRNAs, as well as 1297 up-regulated and 831 down-regulated mRNAs were discovered in sequencing. Enrichment analyses revealed that they were actively involved in immune/inflammatory response and cell adhesion/extracellular matrix. Co-expression analysis and further enrichment analyses showed that five candidate lncRNAs might participate in IA’s inflammatory response. Besides, after controlling other conventional risk factors, multivariate logistic regression analysis disclosed that low expression of lncRNA ENST00000607042, lncRNA ENST00000471220, lncRNA ENST00000478738, lncRNA MALAT1 in peripheral blood leukocytes were independent risk factors for IA. LncRNA ENST00000607042 has superior diagnostic value for IA. Conclusions This study reveals the complete lncRNAs expression profiles in IA. The inflammatory response was closely related to IA. Besides, lncRNA ENST00000607042 might be a novel biomarker for IA risk.

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Microarray study of gene expression profiles in peripheral blood samples from lung cancer patients before and after erlotinib treatment

Journal of Clinical Oncology ◽

10.1200/jco.2009.27.15_suppl.e19072 ◽

2009 ◽

Vol 27 (15_suppl) ◽

pp. e19072-e19072

Author(s):

A. Irigoyen ◽

C. Olmedo ◽

J. Valdivia ◽

A. Comino ◽

C. Cano ◽

...

Keyword(s):

Gene Expression ◽

Lung Cancer ◽

Growth Factor ◽

Healthy Volunteers ◽

Cancer Patients ◽

Peripheral Blood ◽

Expression Profiles ◽

Control Group ◽

Blood Samples ◽

Lung Cancer Patients

e19072 Background: The gene expression profile in peripheral blood samples from lung cancer patients is a potential predictor to treatment response. Methods: The study has been developed using 10 healthy volunteers as the control group and 10 lung cancer patients (stage IV). Written informed consent was obtained being the protocol approved by the local Clinical Research and Ethics Committee. Peripheral blood samples were obtained from lung cancer patients before (T0) and after treatment (T15d). RNA from peripheral blood samples was extracted and purified selecting 28S/18S ratios>1.5 to obtain cDNA and cRNA for hybridization of the 20,000 genes included in Human 20K CodeLink. An array from each participant was obtained in duplicate. For each array, 2 μg of cRNA was compared to 2 μg of healthy cRNA.. Significant genes were found using Significance Analysis of Microarrays which uses repeated permutations of the data. Results: The selected genes were expressed >3-fold with a false discovery rate =0.05. Before treatment (T0) when patients were compared to healthy volunteers there was an increase in the expression of: histone 1 H4c, transforming growth factor beta 2, endothelial cell growth factor 1 (platelet-derived), glucose-6-phosphatase catalytic 2, Relaxin 3 receptor 1, Insulin-like growth factor binding protein 2, RAS-like family 11 member B, and ELK4. After treatment (T15d), when each lung cancer patient's results were compared to their own before treatment results (T0), there was an increase in the expression of: Bcl2, myosin light polypeptide 4; interferon alpha-inducible protein 27; interferon gamma receptor 1; RASSF5, ARHGEF6, IGFBP5, tumor protein p53 inducible nuclear protein 1, peroxisome proliferative activated receptor gamma. Conclusions: The data presented identifies biologically relevant over-expressed genes in lung cancer. A validation of these results and the analysis of the genes that identify patients who will respond positively to erlotinib treatment is being carried out. No significant financial relationships to disclose.

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Analysis of miRNA-mRNA expression profiles in maternal peripheral blood natural killer cells during pregnancy

Journal of Reproductive Immunology ◽

10.1016/j.jri.2015.09.006 ◽

2015 ◽

Vol 112 ◽

pp. 121

Author(s):

Yoichi Ishida ◽

Dongwei Zhao ◽

Akihide Ohkuchi ◽

Tomoyuki Kuwata ◽

Shigeki Matsubara ◽

...

Keyword(s):

Natural Killer Cells ◽

Mrna Expression ◽

Natural Killer ◽

Peripheral Blood ◽

Expression Profiles ◽

Killer Cells ◽

Maternal Peripheral Blood

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Comparative expression analysis of human endogenous retrovirus elements in peripheral blood of children with specific language impairment

Balkan Journal of Medical Genetics ◽

10.2478/bjmg-2019-0014 ◽

2019 ◽

Vol 22 (1) ◽

pp. 49-56

Author(s):

DS Minchev ◽

NT Popov ◽

SI Naimov ◽

IN Minkov ◽

TI Vachev

Keyword(s):

Dna Sequences ◽

Language Impairment ◽

Specific Language Impairment ◽

Peripheral Blood ◽

Expression Profiles ◽

Endogenous Retrovirus ◽

Human Endogenous Retrovirus ◽

Control Group ◽

Healthy Children ◽

Specific Language

AbstractSpecific language impairment (SLI) is a psychiatric condition with a complex etiology and a substantial genetic basis that affects children's verbal communication abilities. In this study, we examined the expression of five different human endogenous retrovirus elements (HERVs) in a cohort of 25 children with SLI and 25 healthy children in the control group. Human endogenous retrovirus elements, a diverse group of repetitive DNA sequences, can potentially cause considerable genetic heterogeneity. They had been integrated in the genome of our ancestors throughout evolution and now consist of about 8.0% of the human genome. Several HERV loci are transcribed in various cell types. Their expression in peripheral blood and in the brain is altered in many neurological and psychiatric diseases. To date, HERV expression profiles have never been studied in patients with SLI. This study aimed to elucidate differentially regulated human endogenous retroelements in peripheral blood of children with SLI, in comparison with healthy controls, through quantitative reverse tran-scription-polymerase chain reaction (qRT-PCR) methodology. Our results show that two genes: HERV-K (HLM-2) gag and HERV-P env were expressed at lower levels in the blood samples from SLI children in comparison with those in the control group.

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Integrating the lncRNA and mRNA expression profiles of TLR4-primed mesenchymal stem cells in ankylosing spondylitis

10.21203/rs.3.rs-45893/v1 ◽

2020 ◽

Author(s):

Yuxi Li ◽

Ming Li ◽

Jiajun Huang ◽

Yuwei Liang ◽

Junshen Huang ◽

...

Keyword(s):

Ankylosing Spondylitis ◽

Mrna Expression ◽

High Throughput Sequencing ◽

Mononuclear Cells ◽

Expression Profiles ◽

Functional Networks ◽

Control Group ◽

Mrna Levels ◽

Peripheral Blood Mononuclear ◽

Qrt Pcr

Abstract Background Our previous study found that the toll-like receptor 4 (TLR4) expression of ankylosing spondylitis (AS) patients was significantly different from that of healthy donors. The goals of this study were to explore the expression profiles and functional networks of lncRNAs and mRNAs in TLR4-primed mesenchymal stromal cells from AS patients (AS-MSCs) and to clarify the mechanisms by which TLR4-primed MSCs exert immunoregulatory effects in AS. Methods Firstly, the immunoregulatory effects of MSCs were determined after TLR4 activation. Then, the differentially expressed (DE) lncRNAs and mRNAs between the control group (AS-MSCs without stimulation) and experimental group (AS-MSCs stimulated with lipopolysaccharide) were identified through high-throughput sequencing followed by qRT-PCR confirmation. Finally, bioinformatic analyses were performed to identify the critical biological functions, signalling pathways and associated functional networks involved in the TLR4-primed immunoregulatory function of AS-MSCs. Results TLR4-primed AS-MSCs showed a strong ability to inhibit the proliferation of peripheral blood mononuclear cells (PBMCs) with 1 µg/ml LPS stimulation for 4 hours. A total of 147 DE lncRNAs and 698 DE mRNAs were identified between TLR4-primed AS-MSCs and unstimulated AS-MSCs. Significant fold changes in lncRNA and mRNA levels were confirmed by qRT-PCR. GO and KEGG analysis demonstrated that the DE mRNAs and lncRNAs were highly associated with the inflammatory response. Cis-regulation prediction revealed 9 novel lncRNAs while trans-regulation prediction revealed 15 lncRNAs, respectively. Conclusions Our research describes the lncRNA and mRNA expression profiles and functional networks in TLR4-primed AS-MSCs, which is supposed to enhance the understanding of the pathogenesis of AS-MSC immunoregulatory dysfunction.

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Clarification of the Role of miR-9 in the Angiogenesis, Migration, and Autophagy of Endothelial Progenitor Cells Through RNA Sequence Analysis

Cell Transplantation ◽

10.1177/0963689720963936 ◽

2020 ◽

Vol 29 ◽

pp. 096368972096393

Author(s):

Jian Zhu ◽

Li-Li Sun ◽

Wen-Dong Li ◽

Xiao-Qiang Li

Keyword(s):

Sequence Analysis ◽

Mrna Expression ◽

Progenitor Cells ◽

Endothelial Progenitor Cells ◽

Expression Profiles ◽

Treatment Strategies ◽

Control Group ◽

Endothelial Progenitor ◽

Rna Sequence ◽

Autophagy Pathway

We have previously reported that miR-9 promotes the homing, proliferation, and angiogenesis of endothelial progenitor cells (EPCs) by targeting transient receptor potential melastatin 7 via the AKT autophagy pathway. In this way, miR-9 promotes thrombolysis and recanalization following deep vein thrombosis (DVT). However, the influence of miR-9 on messenger RNA (mRNA) expression profiles of EPCs remains unclear. The current study comprises a comprehensive exploration of the mechanisms underlying the miR-9-regulated angiogenesis of EPCs and highlights potential treatment strategies for DVT. We performed RNA sequence analysis, which revealed that 4068 mRNAs were differentially expressed between EPCs overexpressing miR-9 and the negative control group, of which 1894 were upregulated and 2174 were downregulated. Gene ontology and Kyoto Encyclopedia of Genes and Genomes pathway analyses indicated that these mRNAs were mainly involved in regulating cell proliferation/migration processes/pathways and the autophagy pathway, both of which represent potential EPC-based treatment strategies for DVT. Reverse transcriptase quantitative polymerase chain reaction confirmed the changes in mRNA expression related to EPC angiogenesis, migration, and autophagy. We also demonstrate that miR-9 promotes EPC migration and angiogenesis by regulating FGF5 directly or indirectly. In summary, miR-9 enhances the expression of VEGFA, FGF5, FGF12, MMP2, MMP7, MMP10, MMP11, MMP24, and ATG7, which influences EPC migration, angiogenesis, and autophagy. We provide a comprehensive evaluation of the miR-9-regulated mRNA expression in EPCs and highlight potential targets for the development of new therapeutic interventions for DVT.

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Integration Analysis of MicroRNA and mRNA Expression Profiles in Human Peripheral Blood Lymphocytes Cultured in Modeled Microgravity

BioMed Research International ◽

10.1155/2014/296747 ◽

2014 ◽

Vol 2014 ◽

pp. 1-16 ◽

Cited By ~ 23

Author(s):

C. Girardi ◽

C. De Pittà ◽

S. Casara ◽

E. Calura ◽

C. Romualdi ◽

...

Keyword(s):

Cell Proliferation ◽

Inflammatory Response ◽

Mrna Expression ◽

Peripheral Blood ◽

Human Peripheral Blood ◽

Expression Profiles ◽

Microgravity Condition ◽

Peripheral Blood Lymphocytes ◽

Blood Lymphocytes ◽

Human Peripheral Blood Lymphocytes

We analyzed miRNA and mRNA expression profiles in human peripheral blood lymphocytes (PBLs) incubated in microgravity condition, simulated by a ground-based rotating wall vessel (RWV) bioreactor. Our results show that 42 miRNAs were differentially expressed in MMG-incubated PBLs compared with 1 g incubated ones. Among these, miR-9-5p, miR-9-3p, miR-155-5p, miR-150-3p, and miR-378-3p were the most dysregulated. To improve the detection of functional miRNA-mRNA pairs, we performed gene expression profiles on the same samples assayed for miRNA profiling and we integrated miRNA and mRNA expression data. The functional classification of miRNA-correlated genes evidenced significant enrichment in the biological processes of immune/inflammatory response, signal transduction, regulation of response to stress, regulation of programmed cell death, and regulation of cell proliferation. We identified the correlation of miR-9-3p, miR-155-5p, miR-150-3p, and miR-378-3p expression with that of genes involved in immune/inflammatory response (e.g., IFNG and IL17F), apoptosis (e.g., PDCD4 and PTEN), and cell proliferation (e.g., NKX3-1 and GADD45A). Experimental assays of cell viability and apoptosis induction validated the results obtained by bioinformatics analyses demonstrating that in human PBLs the exposure to reduced gravitational force increases the frequency of apoptosis and decreases cell proliferation.

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Mutational analysis of TAC3 and TACR3 genes in patients with idiopathic central pubertal disorders

Arquivos Brasileiros de Endocrinologia & Metabologia ◽

10.1590/s0004-27302012000900008 ◽

2012 ◽

Vol 56 (9) ◽

pp. 646-652 ◽

Cited By ~ 26

Author(s):

Cintia Tusset ◽

Sekoni D. Noel ◽

Ericka B. Trarbach ◽

Letícia F. G. Silveira ◽

Alexander A. L. Jorge ◽

...

Keyword(s):

Genomic Dna ◽

Rare Variants ◽

Mutational Analysis ◽

Pubertal Development ◽

Hypogonadotropic Hypogonadism ◽

Central Precocious Puberty ◽

Control Group ◽

Loss Of Function ◽

Coding Region ◽

Constitutional Delay

OBJECTIVE: To investigate the presence of variants in the TAC3 and TACR3 genes, which encode NKB and its receptor (NK3R), respectively, in a large cohort of patients with idiopathic central pubertal disorders. SUBJECTS AND METHODS: Two hundred and thirty seven patients were studied: 114 with central precocious puberty (CPP), 73 with normosmic isolated hypogonadotropic hypogonadism (IHH), and 50 with constitutional delay of growth and puberty (CDGP). The control group consisted of 150 Brazilian individuals with normal pubertal development. Genomic DNA was extracted from peripheral blood and the entire coding region of both TAC3 and TACR3 genes were amplified and automatically sequenced. RESULTS: We identified one variant (p.A63P) in NKB and four variants, p.G18D, p.L58L (c.172C>T), p.W275* and p.A449S in NK3R, which were absent in the control group. The p.A63P variant was identified in a girl with CPP, and p.A449S in a girl with CDGP. The known p.G18D, p.L58L, and p.W275* variants were identified in three unrelated males with normosmic IHH. CONCLUSION: Rare variants in the TAC3 and TACR3 genes were identified in patients with central pubertal disorders. Loss-of-function variants of TACR3 were associated with the normosmic IHH phenotype. Arq Bras Endocrinol Metab. 2012;56(9):646-52

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