SRC Kinase Signaling Inhibitor Research Articles

MicroRNA-150 Modulates Ischemia-Induced Neovascularization in Atherosclerotic Conditions.

Hypercholesterolemia is an atherosclerotic condition that is associated with impaired neovascularization in response to ischemia. This study sought to define the role of microRNAs in that pathophysiology. Next-generation sequencing and quantitative reverse transcription polymerase chain reaction analyses identified miR-150 as a proangiogenic microRNA, which expression is significantly reduced in the ischemic muscles of hypercholesterolemic apolipoprotein E-deficient (ApoE-/-) mice, and in human umbilical vein endothelial cells exposed to oxidized low-density lipoprotein. Forced expression of miR-150 using a miR mimic could rescue oxidized low-density lipoprotein-mediated impairment of endothelial cell migration and tubule formation in vitro. In a mouse model of hindlimb ischemia, intramuscular injection of miR-150 mimic restored blood flow recuperation, vascular densities in ischemic muscles, and functional mobility in ApoE-/- mice. Treatment of ApoE-/- mice with miR-150 also increased the number and the activities of proangiogenic cells. miR-150 targets SRC kinase signaling inhibitor 1, an important regulator of Src (proto-oncogene tyrosine-protein kinase Src) activity. Here we found that hypercholesterolemia and oxidized low-density lipoprotein exposure are associated with increased SRC kinase signaling inhibitor 1 expression and decreased Src activity. However, treatment with miR-150 mimic reduces SRC kinase signaling inhibitor 1 expression and restores Src and downstream endothelial nitric oxide synthase and Akt (protein kinase B) activities both in vitro and in vivo. We also demonstrate the interrelation between miR-150 and SRC kinase signaling inhibitor 1 and their importance for endothelial cell angiogenic activities. Hypercholesterolemia is associated with reduced expression of miR-150, impaired Src signaling, and inefficient neovascularization in response to ischemia. Forced expression of miR-150 using a miR mimic could constitute a novel therapeutic strategy to improve ischemia-induced neovascularization in atherosclerotic conditions.

Abstract 13102: Microrna-150 Modulates Ischemia-induced Neovascularization in Atherosclerotic Conditions

Background: Hypercholesterolemia is an atherosclerotic condition that is associated with impaired neovascularization in response to ischemia. This study sought to define the role of microRNAs (miRNAs) in that pathophysiology. Methods and results: Next generation sequencing and qRT-PCR analyses identified miR-150 as a pro-angiogenic miRNA which expression is significantly reduced in the ischemic muscles of hypercholesterolemic ApoE-deficient (ApoE -/- ) mice, and in HUVECs exposed to oxLDL. Forced expression of miR-150 using a miR mimic could rescue oxLDL-mediated impairment of endothelial cell migration and tubule formation in vitro. In a mouse model of hindlimb ischemia, intramuscular injection of miR-150 mimic restored blood flow recuperation, vascular densities in ischemic muscles, and functional mobility in ApoE -/- mice. Treatment of ApoE -/- mice with miR-150 also increased the number and the activities of pro-angiogenic cells (PACs). miR-150 targets SRC kinase signaling inhibitor 1 (SRCIN1), an important regulator of Src activity. Here we found that hypercholesterolemia and oxLDL exposure are associated with increased SRCIN1 expression and decreased Src activity. However, treatment with miR-150 mimic reduces SRCIN1 expression and restores Src and downstream eNOS and Akt activities both in vitro and in vivo. We also demonstrate that the angiogenic effects of miR-150 are abolished when endothelial cells are treated with an inhibitor of Src. Conclusions: Hypercholesterolemia is associated with reduced expression of miR-150, impaired Src signaling and inefficient neovascularisation in response to ischemia. Forced expression of miR-150 using a miR mimic could constitute a novel therapeutic strategy to improve ischemia-induced neovascularization in atherosclerotic conditions.

Histone arginine methylation in cocaine action in the nucleus accumbens

Repeated cocaine exposure regulates transcriptional regulation within the nucleus accumbens (NAc), and epigenetic mechanisms-such as histone acetylation and methylation on Lys residues-have been linked to these lasting actions of cocaine. In contrast to Lys methylation, the role of histone Arg (R) methylation remains underexplored in addiction models. Here we show that protein-R-methyltransferase-6 (PRMT6) and its associated histone mark, asymmetric dimethylation of R2 on histone H3 (H3R2me2a), are decreased in the NAc of mice and rats after repeated cocaine exposure, including self-administration, and in the NAc of cocaine-addicted humans. Such PRMT6 down-regulation occurs selectively in NAc medium spiny neurons (MSNs) expressing dopamine D2 receptors (D2-MSNs), with opposite regulation occurring in D1-MSNs, and serves to protect against cocaine-induced addictive-like behavioral abnormalities. Using ChIP-seq, we identified Src kinase signaling inhibitor 1 (Srcin1; also referred to as p140Cap) as a key gene target for reduced H3R2me2a binding, and found that consequent Srcin1 induction in the NAc decreases Src signaling, cocaine reward, and the motivation to self-administer cocaine. Taken together, these findings suggest that suppression of Src signaling in NAc D2-MSNs, via PRMT6 and H3R2me2a down-regulation, functions as a homeostatic brake to restrain cocaine action, and provide novel candidates for the development of treatments for cocaine addiction.

Abstract 964: Circulating biomarker miRNA-150 promotes tumor cell proliferation and progression by targeting SRC kinase signaling inhibitor 1 (SRCIN1) in non-small cell lung cancer

Abstract Circulating microRNAs (c-miRNAs) expression ratio (ER) signatures were evaluated as prognostic biomarkers and therapeutic targets in early stage non-small cell lung cancer (NSCLC) patients. In a population study with 171 stage I and II NSCLC patients, the miR-150 and associated ER signature miR27A/miR-150 (ER ≥0.86, HR = 3.92, p = 1.46E-4) were significantly associated with 5-year survival. The upregulated expression of miR-150 was detected in primary tissue samples (n = 245) by miRNA microarray profiling analysis and significantly associated with cancer recurrence (p = 0.027) and overall survival (HR = 0.88, p = 0.04). Ectopic expression of miR-150 in NSCLC H1299, A549, and H2023 cells transfected by miR-150 expression plasmids promoted tumor cell proliferation, tumor cell-induced colony formation and migration/invasion, while blocking the action of miR-150 with a synthetic miR-150 inhibitor or transduction with a miRZip-150 inhibitor expressed by a lentiviral vector significantly inhibited these biological activities. The SRC kinase signaling inhibitor 1 (SRCIN1) was predicted to be a potential target of miR-150 by bioinformatics tools.miR-150-targeted binding and cleavage in the 3’-UTR of SRCIN1 mRNA were confirmed by a novel stem-loop array-reverse transcription-PCR assay. Both SRCIN1 gene and protein expression were significantly downregulated by ectopic expression of miR-150 and upregulated by miR-150 inhibitors in H1299 and H2023 NSCLC cells, as demonstrated by qRT-PCR and Western-blotting assays, respectively. Modulation of key elements in the SRCIN1 signaling pathway by ectopic expression or inhibition of miR-150 were further determined by qRT-PCR and predicted by ingenuity pathway analysis. Our findings suggest that circulating miR-150 and its associated ER signatures could serve as novel prognostic biomarkers for early stage NSCLC and function as an oncogenic miRNA that promotes lung cancer cell proliferation, invasion, and metastasis by direct targeting SRCIN1signaling pathway thus identifying SRCIN1 as a potential therapeutic target. Citation Format: Jing Lin, Liren Zhang, Yuanqing Ye, Taro Oba, Emanuela Gentile, Emanuela Gentile, Jing Wang, Yang Zhao, Jian Gu, Ignacio Wistuba, Jack A. Roth, Xifeng Wu, Lin Ji. Circulating biomarker miRNA-150 promotes tumor cell proliferation and progression by targeting SRC kinase signaling inhibitor 1 (SRCIN1) in non-small cell lung cancer. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 964.

REDUCED EXPRESSION OF MIR-150 LEADS TO IMPAIRMENT OF ISCHEMIA-INDUCED NEOVASCULARISATION IN HYPERCHOLESTEROLEMIC CONDITIONS

s S277 treatment of HC mice with miR-150 mimic can rescue the number of bone marrow and peripheral EPCs. Moreover, the angiogenic properties of EPCs (adhesion, migration and integration into tubules) are restored in HC mice treated with miR-150 mimic. In vitro, miR-150 rescues angiogenesis (migration, tube formation) in HUVECs exposed to oxLDL. miR-150 has previously been shown to target SRC kinase signaling inhibitor 1 (SRCIN1), an important regulator of SRC activity. We found that SRC activity is significantly reduced in HUVECs exposed to oxLDL. Moreover, the angiogenic effects of miR-150 mimic in HUVECs exposed to oxLDL are completely abolished when the cells are also treated with an inhibitor of SRC. CONCLUSION: Hypercholesterolemia is associated with reduced expression of miR-150, which leads to impaired neovascularization following ischemia. The mechanism involves beneficial effects on both angiogenesis and EPC activities. Our results suggest that SRC signaling is at least in part responsible for the pro-angiogenic effects of miR-150 in these conditions. IRSC 525 NEONATAL EXPOSURE TO HIGH OXYGEN LEVELS IS ASSOCIATED WITH IMPAIRED ISCHEMIA-INDUCED NEOVASCULARIZATION DURING ADULTHOOD R Mathieu, M Desjarlais, W Dhahri, A Cloutier, A Nuyt, A Rivard

MiR-211 promotes non-small cell lung cancer proliferation by targeting SRCIN1.

MicroRNAs (miRNAs) are a class of small non-coding RNAs that, when dysregulated, are involved in the initiation and progression of various cancers, including lung cancer, in humans. In the current study, qRT-PCR was performed to measure miR-211 expression in human non-small cell lung cancer (NSCLC) cell lines and tissues. Cell proliferation, cell cycle, colony formation, and invasion were performed to detect the functional role of miR-211 in human NSCLC cell line. We used luciferase reporter assay to find the potential target of miR-211. We found that miR-211 expression was upregulated in human non-small cell lung cancer (NSCLC) cell lines and tissues. The overexpression of miR-211 enhanced NSCLC cell proliferation, colony formation, and invasion. SRC kinase signaling inhibitor 1 (SRCIN1) was identified as a direct target of miR-211. SRCIN1 silencing promoted cell proliferation, and SRCIN1 expression was downregulated in human NSCLC cell lines. Thus, miR-211 may function as an oncogenic miRNA in NSCLC, partly by regulating SRCIN1, and the modulation of miR-211 expression represents a potential strategy for the treatment of NSCLC patients.

MiR-374a promotes cell proliferation, migration and invasion by targeting SRCIN1 in gastric cancer

MicroRNAs (miRNAs) play a prominent role in gastric cancer (GC) initiation and progression. In this study, we found that miR-374a expression was up-regulated in human GC cell lines and tissues. Inhibition of miR-374a suppressed GC cell proliferation, migration and invasion in vitro and slowed tumor growth in vivo. SRC kinase signaling inhibitor 1 (SRCIN1) was identified as a direct target of miR-374a. Silencing of SRCIN1 significantly enhanced cell proliferation, migration and invasion, whereas SRCIN1 reintroduction partially abrogated the oncogenic effects of miR-374a. Taken together, these findings suggest that miR-374a functions as a candidate oncogene in GC by directly targeting SRCIN1. miR-374a may therefore be useful as a promising therapeutic target for malignant GC.