aav9 negative control virus  (Genechem)

Journal: Frontiers in Cardiovascular Medicine

Article Title: microRNA-377 Signaling Modulates Anticancer Drug-Induced Cardiotoxicity in Mice

doi: 10.3389/fcvm.2021.737826

Figure Lengend Snippet: Doxorubicin upregulates miR-377 expression in mouse myocardium and human ventricular cardiomyocyte cell (AC16). (A) Quantification of miR-377 expression in mouse myocardium following 20 mg/kg DOX treatment for 3 days. miR-377 expression was normalized to U6 and values are shown as fold change. Data are represented as mean ± SEM ( n = 3, * p < 0.05 vs. vehicle control). (B) Quantification of miR-377 expression in human ventricular cardiomyocytes (AC16) following 1 μm DOX treatment for 24 h. miR-377 expression was normalized to U6 and values are shown as fold change. Data are represented as mean ± SEM ( n = 3, ** p < 0.01 vs. untreated control cells).

Article Snippet: Adeno-associated virus, serotype 9 (AAV9)-based murine shRNA-miR-377 and scrambled-negative control under a cytomegalovirus (CMV) promoter was procured from Vector Biolabs (Malvern, Pennsylvania).

Techniques: Expressing

Journal: Frontiers in Cardiovascular Medicine

Article Title: microRNA-377 Signaling Modulates Anticancer Drug-Induced Cardiotoxicity in Mice

doi: 10.3389/fcvm.2021.737826

Figure Lengend Snippet: Inhibition of miR-377 attenuates DOX-induced cardiomyocyte cell death. (A) Evaluation of transfection efficiency of miR-377 inhibitor in AC16 cell was performed by qPCR. Quantification of miR-377 expression in AC16 cells transfected with either anti-miR negative control (Scramble/Scr) or miR-377 inhibitor (anti-miR-377). miR-377 expression was normalized to U6 and values are shown as fold change. Data are represented as mean ± SEM ( n = 3, *** p < 0.001 vs. scrambled control cells), (B) Quantification of miR-377 expression in AC16 cells transfected with either anti-miR-377 or Scramble, following 1 μm DOX treatment for 24 h. miR-377 expression was normalized to U6 and values are shown as fold change. Data are represented as mean ± SEM ( n = 3, * p < 0.05, ** p < 0.01 vs. scrambled control cells). (C) Representative image of TUNEL assay in AC16 cells transfected with scramble control without DOX, scramble control with DOX, and anti-miR-377 with DOX; TUNEL-positive nuclei (red) and nuclear label Hoechst (blue).

Article Snippet: Adeno-associated virus, serotype 9 (AAV9)-based murine shRNA-miR-377 and scrambled-negative control under a cytomegalovirus (CMV) promoter was procured from Vector Biolabs (Malvern, Pennsylvania).

Techniques: Inhibition, Transfection, Expressing, Negative Control, TUNEL Assay

Journal: Frontiers in Cardiovascular Medicine

Article Title: microRNA-377 Signaling Modulates Anticancer Drug-Induced Cardiotoxicity in Mice

doi: 10.3389/fcvm.2021.737826

Figure Lengend Snippet: RNA sequencing and gene ontology analysis to show the effect of miR-377 inhibition on the transcriptome of DOX-stimulated AC16 cardiomyocytes. (A) The global transcriptional change across the compared groups was visualized by a volcano plot, with log2 fold change of each gene is represented on the -axis and the log10 of its adjusted p- value is on the y-axis. Genes with an adjusted p- value < 0.05 and a log2 fold change ≥1 are considered upregulated and indicated by red dots, whereas genes with an adjusted p- value < 0.05 and a log2 fold change ≤ -1 are considered downregulated and indicated by blue dots. (B) A bi-clustering heatmap representing the expression profile of the top 30 differentially expressed genes sorted by their adjusted p- value by plotting their log2 transformed expression values in between anti-miR-377 + DOX ( n = 2) and anti-miR negative control+DOX (scrambled control+DOX, n = 2) groups. (C) GO analysis: Gene ontology terms that are significantly enriched with an adjusted p- value < 0.05 in the differentially expressed genes. (D) List of cell survival genes that were differentially expressed in DOX-stimulated anti-miR377-treated cells. The list also shows if the differentially expressed genes are predicted targets of miR-377.

Article Snippet: Adeno-associated virus, serotype 9 (AAV9)-based murine shRNA-miR-377 and scrambled-negative control under a cytomegalovirus (CMV) promoter was procured from Vector Biolabs (Malvern, Pennsylvania).

Techniques: RNA Sequencing Assay, Inhibition, Expressing, Transformation Assay, Negative Control

Journal: Frontiers in Cardiovascular Medicine

Article Title: microRNA-377 Signaling Modulates Anticancer Drug-Induced Cardiotoxicity in Mice

doi: 10.3389/fcvm.2021.737826

Figure Lengend Snippet: AAV9-based inhibition of miR-377 in mouse myocardium and its effect on LV function (A) in vivo study design. (B) Quantification of miR-377 expression in the mouse myocardium at 28 days after AAV9 treatment. miR-377 expression was normalized to U6 and values are shown as fold change. Data are represented as mean ± SEM ( n = 2/group). (C) Representative echocardiography m-mode tracings 28 days post AAV9 administration. (D) Percent ejection fraction, (E) percent fractional shortening, (F) Left ventricular diameter at diastole, and (G) cardiac output of AAV9-Scr (anti-miR) and AAV9-anti-miR377 mice at the baseline and 28 days post-AAV9 administration. Data are represented as mean ± SEM ( n = 10/group).

Article Snippet: Adeno-associated virus, serotype 9 (AAV9)-based murine shRNA-miR-377 and scrambled-negative control under a cytomegalovirus (CMV) promoter was procured from Vector Biolabs (Malvern, Pennsylvania).

Techniques: Inhibition, In Vivo, Expressing

Journal: Frontiers in Cardiovascular Medicine

Article Title: microRNA-377 Signaling Modulates Anticancer Drug-Induced Cardiotoxicity in Mice

doi: 10.3389/fcvm.2021.737826

Figure Lengend Snippet: AAV9-anti-miR-377 injection in mice accelerates mortality following DOX administration and is associated with changes in gene expression. (A) Kaplan-Meier survival curve to show mortality in the mice pretreated with AAV9-scramble (anti-miR) control or AAV9-anti-miR-377 followed by DOX (20 mg/kg) administration ( n = 10/group). (B) Quantification of miR-377 expression in the mouse myocardium of AAV9-scramble (anti-miR) control or AAV9-anti-miR-377 at 28 days after AAV9 treatment. miR-377 expression was normalized to U6 and values are shown as fold change. Data are represented as mean ± SEM ( n = 3/group, **** p < 0.0001). (C) Representative Hematoxylin & Eosin-stained sections of left ventricle of AAV9-scramble-control or AAV9-anti-miR-377 at day 6. (D) Mean cross-sectional area of cardiomyocytes in the left ventricles of AAV9-scramble-control or AAV9-anti-miR-377 mice.

Article Snippet: Adeno-associated virus, serotype 9 (AAV9)-based murine shRNA-miR-377 and scrambled-negative control under a cytomegalovirus (CMV) promoter was procured from Vector Biolabs (Malvern, Pennsylvania).

Techniques: Injection, Expressing, Staining

Journal: Frontiers in Cardiovascular Medicine

Article Title: microRNA-377 Signaling Modulates Anticancer Drug-Induced Cardiotoxicity in Mice

doi: 10.3389/fcvm.2021.737826

Figure Lengend Snippet: Left ventricular function assessment in mice with AAV9-anti-miR-377 injection, 6 days after DOX administration. (A) Percent ejection fraction, (B) percent fractional shortening, (C) Left ventricular diameter at diastole, and (D) Left ventricular diameter at systole of AAV9-Scr (anti-miR) and AAV9-anti-miR377 mice after 6 days of DOX stimulation. Data are represented as mean ± SEM ( n = 3 in AAV9-Scr and n = 2 in AAV9-anti-miR-377).

Article Snippet: Adeno-associated virus, serotype 9 (AAV9)-based murine shRNA-miR-377 and scrambled-negative control under a cytomegalovirus (CMV) promoter was procured from Vector Biolabs (Malvern, Pennsylvania).

Techniques: Injection

Journal: Frontiers in Cardiovascular Medicine

Article Title: microRNA-377 Signaling Modulates Anticancer Drug-Induced Cardiotoxicity in Mice

doi: 10.3389/fcvm.2021.737826

Figure Lengend Snippet: Graphical representation showing the effect of miR-377 inhibition in-vitro and in-vivo on DOX-induced cardiotoxicity. In-vitro , miR-377 inhibition leads to upregulation of anti-oxidative stress response genes and therefore, inhibition of DOX-induced cardiomyocyte apoptosis. On the contrary, in-vivo inhibition of miR-377 (using AAV9-anti-miR-377) leads to increased mortality, potentially due to accelerated cell growth and upregulation of genes such as MET, E2F2, and GDF15 (that are involved in adverse cardiac remodeling).

Article Snippet: Adeno-associated virus, serotype 9 (AAV9)-based murine shRNA-miR-377 and scrambled-negative control under a cytomegalovirus (CMV) promoter was procured from Vector Biolabs (Malvern, Pennsylvania).

Techniques: Inhibition, In Vitro, In Vivo