scholarly journals Focal Adhesion Kinase As a Potential Target in AML and MDS

Blood ◽  
2015 ◽  
Vol 126 (23) ◽  
pp. 3680-3680
Author(s):  
Bing Z Carter ◽  
Steven M. Kornblau ◽  
Po Yee Mak ◽  
Hui Yang ◽  
Yihua Qiu ◽  
...  
Keyword(s):  
Gene Expression ◽  
Growth Factor ◽  
Focal Adhesion Kinase ◽  
Focal Adhesion ◽  
Leukemia Cells ◽  
Mrna Levels ◽  
Newly Diagnosed ◽  
Prior Chemotherapy ◽  
Expression Levels ◽  
Cd34 Cells

Abstract Focal adhesion kinase (FAK), a non-receptor tyrosine kinase is overexpressed/activated in several solid cancers. It controls cell growth, survival, invasion, metastasis, cell movement, gene expression, as well as stem cell self-renewal. Integrin ligation and growth factor/receptor interaction activate FAK leading to SRC phosphorylation and subsequent FAK/SRC phosphorylation at multiple sites which relays the external signal into cells by activating multiple cell proliferating/survival pathways. Although it is extensively studies in solid tumors, the expression and function of FAK in myeloid leukemia are not well investigated. We determined FAK expression by reversed-phase protein array in samples from a large cohort of newly diagnosed AML (n = 511) and MDS (n = 279). We found that in AML, FAK expression is associated with unfavorable cytogenetic group (P = 2x10-4). The lowest expression was seen in patients with inv16 (n = 19), t8;21 (n = 15) and t15;17 (n = 20) chromosome translocations. Patients with -5, -7, +8 (n = 100) expressed relatively higher FAK. FAK expression is higher in relapsed compared to paired newly diagnosed samples (n = 47, P = 0.02). FAK expression was significantly lower in patients with FLT-ITD (n=83, P = 0.0024) or RAS mutation (n = 64, P = 0.05) suggesting functional redundancy of these signaling pathways and therefore mutually exclusive expression patterns. FAK expression is highly correlated with SRC/p-SRC and ITGb3/ITGa2 levels among over 200 proteins analyzed suggesting a role of intergrin/FAK/SRC signal in AML cells. We also observed overexpression of FAK in MDS. FAK expression levels were significantly higher in both CD34+ (P = 5.42e-20) and CD34+ CD38- MDS cells (P = 7.62e-9) than in normal CD34+ cells (n = 16). Patients with higher expression of FAK in CD34+ cells had a trend towards better overall survival (P = 0.05) in newly diagnosed MDS. Furthermore, we determined the FAK mRNA expression levels in bone marrow CD34+ cells from 63 patients (57 MDS and 6 AML). In this cohort of patients, fifty (79%) were previously untreated with chemotherapy (46 MDS and 4 AML). No significant difference in FAK gene expression was observed when comparing patients with or without prior chemotherapy in either MDS or AML patients. Aberrant up-regulation (≥ 2 fold) was observed in 23% and 50% of the patients respectively compared to normal CD34+ cells (n = 5). In patients who received no prior chemotherapy (n = 50), aberrant up-regulation (≥2 fold) was observed in 22% and 50% of the patients respectively. FAK protein levels by immunohistochemical analysis were correlated with mRNA levels. Growth factor GM-CSF and co-culture with mesenchymal stem cells (MSCs) increased the expression of FAK in leukemia cells suggesting that the microenvironment modulates leukemia cell function in part via activating FAK signaling. Inhibition of FAK with VS-4718, a potent selective FAK inhibitor decreased viable cells and induced apoptosis in various AML cell lines and in co-cultures with MSCs. Inhibition of FAK decreased adhesion and migration of OCI-AML3 to MSCs suggesting FAK signaling promotes leukemia/stromal interactions. Furthermore, VS-4718 induced cell death in bulk, CD34+, and CD34+ CD38- cells from patients with AML even in co-cultures with MSCs. In conclusion, our results demonstrate that FAK is expressed in AML patient samples and that high expressions associate with unfavorable patient characteristics. Overexpression of FAK in MDS suggests that FAK signaling may be involved in pathogenesis of the disease. FAK, activated by tumor microenvironment, supports the survival of leukemia cells and in turn promotes leukemia/stromal interaction. FAK inhibition can induce apoptosis in leukemia cells. FAK may hence be a potential therapeutic target in AML and MDS. Disclosures Carter: PrismBiolab: Research Funding. Andreeff:Oncoceutics, Inc.: Membership on an entity's Board of Directors or advisory committees.

Caveolin-1 promotes trophoblast cell invasion through the focal adhesion kinase (FAK) signalling pathway during early human placental development

2019 ◽  
Vol 31 (6) ◽  
pp. 1057 ◽  
Author(s):  
Zhihui Dai ◽  
Fei Sheng ◽  
Ningxia Sun ◽  
Yixuan Ji ◽  
Qiuying Liao ◽  
...  
Keyword(s):  
Gene Expression ◽  
Focal Adhesion Kinase ◽  
Focal Adhesion ◽  
Cell Invasion ◽  
Trophoblast Cell ◽  
Signalling Pathway ◽  
Placental Development ◽  
Human Trophoblast ◽  
Expression Levels ◽  
Caveolin 1

Normal implantation and placental development depend on the appropriate differentiation and invasion of trophoblast cells. Inadequate trophoblast cell invasion results in pregnancy-related disorders, which endanger both mother and fetus; however, the mechanism of early placental development has not been fully explained. In this study we conducted gene expression profile analysis using mouse placental tissues at different developmental stages (embryonic day (E)7.5, E14.5 and E19.5) using series tests of cluster (STC) and Kyoto Encyclopaedia of Genes and Genomes (KEGG) pathway analyses. Focal adhesion kinase (FAK) signalling pathway-related gene expression levels were verified using quantitative reverse transcription polymerase chain reaction and western blot. The results showed that caveolin-1 (Cav1) was downregulated in the placenta of unexplained spontaneous abortion subjects compared with that of induced abortion. Furthermore, by modulating CAV1 expression levels, CAV1 was shown to promote human trophoblast cell proliferation, migration and invasion by activating the FAK signalling pathway. These results indicate that CAV1 and the FAK signalling pathway are crucial for early placental development, which sheds new light on our understanding of the mechanisms of human trophoblast cell invasion and early development of the placenta.

RASSF2 Functions As a Tumor Suppressor in t(8;21) AML Via Promotion of Apoptosis

Blood ◽  
2014 ◽  
Vol 124 (21) ◽  
pp. 3590-3590
Author(s):  
Samuel A Stoner ◽  
Russell Dekelver ◽  
Miao-Chia Lo ◽  
Dong-Er Zhang
Keyword(s):  
Gene Expression ◽  
Tumor Suppressor ◽  
Cell Lines ◽  
Hematopoietic Cells ◽  
Leukemia Cell ◽  
Leukemia Cells ◽  
Mrna Levels ◽  
Human Cd34 ◽  
Leukemia Cell Lines ◽  
Cd34 Cells

Abstract The t(8;21) chromosomal translocation is one of the most common chromosomal translocations associated with acute myeloid leukemia (AML), found in approximately 12% of de novo AML cases. The majority of these cases are classified as FAB-subtype M2 AML. The t(8;21) results in the stable fusion of the AML1 (RUNX1) and ETO (RUNX1T1) genes. The AML1-ETO fusion protein is composed of the N-terminal portion of AML1, which includes the DNA-binding Runt-homology domain, and nearly the full-length ETO protein. The primary accepted mechanism by which AML1-ETO promotes leukemia development is through the aberrant recruitment of transcriptional repression/activation complexes to normal AML1 target genes. Therefore, the identification of individual genes or biological pathways that are specifically disrupted in the presence of AML1-ETO will provide further molecular insight into the pathogenesis of t(8;21) AML and lead to the possibility for improved treatment for these patients. We identified RASSF2 as a gene that is specifically downregulated in (2-4 fold) in total bone marrow of t(8;21) patients compared to non-t(8;21) FAB-subtype M2 AML patients by analyzing publicly available gene expression datasets. Similarly, using a mouse model of t(8;21) AML we found Rassf2 mRNA levels to be nearly 30-fold lower in t(8;21) leukemia cells compared to wild-type Lin-Sca-cKit+ (LK) myeloid progenitors. Gene expression analysis by RT-qPCR in leukemia cell lines confirmed that RASSF2 mRNA levels are significantly downregulated (8-10-fold) in both Kasumi-1 and SKNO-1 t(8;21) cell lines as compared to a similar non-t(8;21) HL-60 cell line and to primary human CD34+ control cells. In addition, expression of AML1-ETO in HL-60 or CD34+ cells results in a decrease in RASSF2 mRNA expression, which further suggests that RASSF2 is a target for regulation by AML1-ETO. Assessment of published ChIP-seq data shows that AML1-ETO binds the RASSF2 gene locus at two distinct regions in both primary t(8;21) AML patient samples and in the Kasumi-1 and SKNO-1 cell lines. These regions are similarly bound by several important hematopoietic transcription factors in primary human CD34+ cells, including AML1, ERG, FLI1, and TCF7L2, implicating these two regions as important for the regulation of RASSF2 expression during blood cell differentiation. Overexpression of RASSF2 in human leukemia cell lines using an MSCV-IRES-GFP (MIG) construct revealed that RASSF2 has a strong negative effect on leukemia cell proliferation and viability. The overall percentage of GFP-positive cells in MIG-RASSF2 transduced cells markedly decreased compared to MIG-control transduced cells over a period of 14 days. This effect was primarily due to significantly increased apoptosis in the RASSF2 expressing cell populations. Similarly, we found that expression of RASSF2 significantly inhibits the long-term self-renewal capability of hematopoietic cells transduced with AML1-ETO in a serial replating/colony formation assay. AML1-ETO transduced hematopoietic cells were normally capable of serial replating for more than 6 weeks. However, AML1-ETO transduced cells co-expressing RASSF2 consistently had reduced colony number and lost their ability to replate after 3-4 weeks. This was due to a dramatically increased rate of apoptosis in RASSF2 expressing cells. RASSF2 is reported to be a tumor suppressor that is frequently downregulated at the transcriptional level by hypermethylation in primary tumor samples, but not healthy controls. Here we have identified RASSF2 as a target for repression, and demonstrated its tumor suppressive function in t(8;21) leukemia cells. Further insights into the molecular mechanisms of RASSF2 function in AML will continue to be explored. Disclosures No relevant conflicts of interest to declare.

MO453SERUM AND VASCULAR FIBROBLAST GROWTH FACTOR 23 (FGF23) ARE ASSOCIATED WITH VASCULAR CALCIFICATION

2021 ◽  
Vol 36 (Supplement_1) ◽  
Author(s):  
Javier Donate-Correa ◽  
Ernesto Martín-Núñez ◽  
Carolina Hernández-Carballo ◽  
Carla Ferri ◽  
Miguel Arévalo-Gómez ◽  
...  
Keyword(s):  
Gene Expression ◽  
Growth Factor ◽  
Fibroblast Growth Factor ◽  
Vascular Calcification ◽  
Fibroblast Growth Factor 23 ◽  
Case Group ◽  
Mrna Levels ◽  
Fibroblast Growth ◽  
Expression Levels ◽  
Vascular Expression

Abstract Background and Aims Mineral metabolism imbalances and inflammation are related to the development of vascular calcification (VC). Fibroblast growth factor-23 (FGF23) is the main regulator of phosphate homeostasis and various studies have shown the existence of an association between elevated levels of FGF23 and the appearance of cardiovascular disease (CVD). We conducted a case-control study to test the hypothesis that serum and vascular levels of FGF23 are associated with the presence of VC. In addition, we determined the influence of inflammation in these levels. Method One hundred and thirty-three patients diagnosed with clinical atherosclerotic disease undergoing elective vascular surgery, and 20 cadaveric organ donors with no medical history of CVD, were included in this study. Serum levels of intact FGF23 (iFGF23), together with tumor necrosis factor-alpha (TNFα), interleukin (IL)10, were determined by ELISA. Vascular fragments of aorta, carotid and femoral arteries were obtained for assaying the gene expression of FGF23, TNF, IL10 and RUNX2 by qPCR. Immunohistochemical procedures were employed to determine vascular protein levels of FGF23, TNFα and IL10. VC was diagnosed by imaging techniques and confirmed by histological procedures including von Kossa staining. Results Case group presented a higher prevalence of hypertension, hypercholesterolemia and reduced estimated glomerular filtration rate, with no differences regarding other parameters. Serum iFGF23 and TNFα/IL10 ratio were higher in the case group (P<0.01 and P<0.001, respectively). Vascular expression of FGF23 was detected in 58.6% of CVD patients vs 35% of donors, with mean expression levels significantly higher in the first group (P<0.01). Vascular expression of TNF/IL10 was also increased (P<0.001) in CVD patients. FGF23 immunoreactivity was detected in 84% of CVD patients and only in 35% of controls. Immunoreactivity for FGF23 and TNFα/IL10 ratio were significantly higher in CVD patients (P<0.001 and P<0.0001, respectively). Stratified analysis according to serum iFGF23 levels showed a higher prevalence of VC in the upper tertiles. Patients with VC presented increased levels of all the FGF23 variables including serum [1.5 (1.2-1.6) vs. 1.4 (0.9-1.5) pg/mL, P<0.01], vascular mRNA [26.1 (14.3-67.4) vs. 18.8 (8.8-312.9) log AU, P<0.01] and vascular immunoreactivity [4.6 (3.8-4.9) vs. 3.7 (3.1-4.1) log µm2, P<0.05]. Moreover, FGF23 immunoreactivity was detected in 92.3% of fragments with VC and only in 53.6% of those without VC. Serum TNFα/IL10 and RUNX2 mRNA levels were also higher in this group (P<0.01 for both). Correlation analysis showed associations of serum iFGF23 with serum TNFα (r=0.375, P<0.001), neutrophil/lymphocyte (r=0.142, P<0.05), vascular RUNX2 mRNA (r=0.55, P<0.05), and vascular FGF23 immunoreactivity (r=0.281, P<0.05) in the CVD group. Vascular FGF23 expression correlated with RUNX2 mRNA (r=0.315, P<0.05) and FGF23 immunoreactivity (r=0.254, P<0.05). Multiple regression analysis showed that iFGF23 levels were determined by UAE, HDL, FGe, calcium and TNFα levels (adjusted R2= 0.473, P<0.0001) and that vascular FGF23 mRNA expression was determined by TNFα, PCR, glucose and age (adjusted R2= 0.795, P<0.0001). Multivariate logistic regression, with VC as dependent variable, showed that both iFGF23 and vascular mRNA constitute independent risk factors for the existence of VC [OR (95% CI): 1.05 and 1.12, P<0.05 for both]. Conclusion Patients with atherosclerosis and VC present significantly higher serum concentrations of FGF23, as well as higher immunoreactivity and gene expression levels in the vascular wall compared to patients without VC. Moreover, both serum and vascular mRNA levels of FGF23 are associated with the inflammatory status. Whether the increase in systemic and vascular FGF23 can directly promote or favor the calcifying process in the vascular bed is currently an issue under discussion.

scholarly journals Methylobacterium oryzae Influences Isoepoxydon Dehydrogenase Gene Expression and Patulin Production by Penicillium expansum

Water ◽  
2021 ◽  
Vol 13 (10) ◽  
pp. 1427
Author(s):  
Tiago Barros Afonso ◽  
Lúcia Chaves Simões ◽  
Nelson Lima
Keyword(s):  
Gene Expression ◽  
Distribution Systems ◽  
Water Distribution ◽  
Penicillium Expansum ◽  
Transcriptional Level ◽  
Positive Trend ◽  
Mrna Levels ◽  
Expression Levels ◽  
Production Values ◽  
Methylobacterium Oryzae

Biofilms can be considered the main source of microorganisms in drinking water distribution systems (DWDS). The ecology of a biofilm is dependent on a variety of factors, including the presence of microbial metabolites excreted by its inhabitants. This study reports the effect of the Gram-negative bacteria Methylobacterium oryzae on the idh gene expression levels and patulin production of Penicillium expansum mature biofilms. For this purpose, a RT-qPCR method to quantify idh mRNA levels was applied. In addition, the idh expression levels were compared with the patulin production. The results obtained revealed that the effect of the bacterium on pre-established P. expansum biofilms is dependent on the time of interaction. More mature P. expansum biofilms appear to be more resistant to the inhibitory effect that M. oryzae causes towards idh gene expression and patulin production. A positive trend was observed between the idh expression and patulin production values. The results indicate that M. oryzae affects patulin production by acting at the transcriptional level of the idh gene.

scholarly journals Gene Expression of Nucleic Acid-Sensing Pattern Recognition Receptors in Children Hospitalized for Respiratory Syncytial Virus-Associated Acute Bronchiolitis

2009 ◽  
Vol 16 (6) ◽  
pp. 816-823 ◽  
Author(s):  
Carolina Scagnolari ◽  
Fabio Midulla ◽  
Alessandra Pierangeli ◽  
Corrado Moretti ◽  
Enea Bonci ◽  
...  
Keyword(s):  
Gene Expression ◽  
Pattern Recognition ◽  
Viral Load ◽  
Respiratory Syncytial Virus ◽  
Pattern Recognition Receptors ◽  
Mrna Levels ◽  
Acute Bronchiolitis ◽  
Expression Levels ◽  
Rsv Infection ◽  
Syncytial Virus

ABSTRACT Given the critical role of pattern recognition receptors (PRRs) in acid nucleic recognition in the initiation of innate immunity and the orchestration of adaptive immunity, the aim of this study was to determine whether any heterogeneity of PRR expression in the airway tracts of infants with respiratory syncytial virus (RSV) infection might explain the broad clinical spectrum of RSV-associated bronchiolitis in infants. For this purpose, the levels of melanoma differentiation-associated protein-5 (MDA-5), retinoic acid inducible gene-1 (RIG-1), and Toll-like receptor 3 (TLR-3), TLR-7, TLR-8, and TLR-9 mRNAs were evaluated, using TaqMan quantitative reverse transcription-PCR, in cells from nasopharyngeal washes collected from 157 infants suffering from acute bronchiolitis whether or not they were associated with respiratory viruses. High interindividual variability was observed in both virus-positive and -negative infants; however, the relative gene expression levels of MDA-5, RIG-1, TLR-7, and TLR-8 were significantly higher in the virus-infected group, whereas the expression levels of TLR-3 and TLR-9 were not significantly different. The differences in the gene expression of MDA-5, RIG-1, TLR-7, and TLR-8 were more evident in infants with RSV infection than in those with bocavirus or rhinovirus infection. In RSV-infected infants, PRR-mRNA levels also were analyzed in relation to interferon protein levels, viral load, clinical severity, days of hospitalization, age, and body weight. A significant positive correlation was observed only between RSV viral load and RIG-1 mRNA levels. These findings provide the first direct evidence that, in infants with respiratory virus-associated bronchiolitis, especially RSV, there are substantial changes in PRR gene expression; this likely is an important determinant of the clinical outcome of bronchiolitis.

scholarly journals Heterogeneous recruitment abilities to RNA polymerases generate nonlinear scaling of gene expression with cell volume

2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Qirun Wang ◽  
Jie Lin
Keyword(s):  
Gene Expression ◽  
Cell Volume ◽  
Direct Consequence ◽  
Rna Polymerases ◽  
Mrna Levels ◽  
Expression Levels ◽  
Promoter Sequences ◽  
Biophysical Mechanism ◽  
Expression Model ◽  
Cycle Regulation

AbstractWhile most genes’ expression levels are proportional to cell volumes, some genes exhibit nonlinear scaling between their expression levels and cell volume. Therefore, their mRNA and protein concentrations change as the cell volume increases, which often have crucial biological functions such as cell-cycle regulation. However, the biophysical mechanism underlying the nonlinear scaling between gene expression and cell volume is still unclear. In this work, we show that the nonlinear scaling is a direct consequence of the heterogeneous recruitment abilities of promoters to RNA polymerases based on a gene expression model at the whole-cell level. Those genes with weaker (stronger) recruitment abilities than the average ability spontaneously exhibit superlinear (sublinear) scaling with cell volume. Analysis of the promoter sequences and the nonlinear scaling of Saccharomyces cerevisiae’s mRNA levels shows that motifs associated with transcription regulation are indeed enriched in genes exhibiting nonlinear scaling, in concert with our model.

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