miR-504 is downregulated in GBM and GSCs and inhibits the stemness and mesenchymal transformation of GSCs

via the miR-145/CTGF pathway

Simona Cazacu,1 Ariel Bier,2 Daniel Rand2, Hae Kyung Lee,1 Susan Finniss,1 Cunli Xiang,1 Wei Jiang1, Hiba Waldman Ben-Asher,2 Laila Poisson3 and Chaya Brodie1,2
1
Davidson Laboratory of Cell Signaling and Tumorigenesis, Hermelin Brain Tumor Center, Department of Neurosurgery, 2Everard and Mina Goodman Faculty of Life Sciences, Bar-Ilan
University, Ramat-Gan, Israel 3Department of Public Health Sciences
Henry Ford Hospital, Detroit, MI, USA

Abstract Expression of miR-504 in GBM GSCs and MiR-504 downregulate CTGF indirectly by
TCGA database upregulating miR-145 expression
Glioblastoma (GBM) is characterized by increased infiltration into the surrounding normal
tissue, therapeutic resistance, and poor prognosis. A major pathway contributing to these Figure 8
characteristics is GBM mesenchymal phenotype. A small subpopulation of glioma stem cells We reported miR-145 directly target CTGF in GSCs.

Relative miR-504
(GSCs) have been implicated in infiltration, radio-resistance and tumor recurrence. GSCs

Expression
share some similarities with neural stem cells (NSCs) but exhibit deregulated differentiation
ability and oncogenic potential. Recent studies documented miRNAs as important regulators
of GSC functions, malignancy and stemness. In this study we used miRNA microarray to
perform miRNA analysis of GSCs and compared to human NSCs and mesenchymal stromal 1.4
cells (MSCs) to identify significant miRNA pathways associated with the mesenchymal Healthy GBM
1.2
signature of GSCs. We identified 79 miRNAs (p<0.05) that were upregulated in GSCs and

CTGF Relative Expression

Con pre-
1
21 miRNAs (p<0.05) that were increased in MSCs. 26 miRNAs were downregulated in 4
miR
0.8
GSCs compared to NSCs and 21 miRNAs (p<0.05) from this group were further

Relative miR-504 Expression

3.5

downregulated in MSCs. The comparison of miRNA expression of GSCs and NSCs 3

0.6

identified gene clusters associated with GBM invasiveness, axonal guidance signaling and 2.5 0.4
2
TGF- signaling. miR-504 is one of the miRNAs that was significantly downregulated in

Relative miR-504
Fig 2,3,4 0.2

Expression
1.5

GSCs compared to NSCs (-7.8 is -5.9 ). The expression of miR-504 was also decreased in Legend and data comments 1 0
Con-premir premir-504 Con- Mir-145 Mir-145
mesenchymal and highly increased in the G-CIMP subset of GBM. miR-504 inhibited the 0.5 antagomir antagomir antagomir
premir-504
0
self-renewal, migration and the expression of the mesenchymal proteins:fibronectin, U87 HF HF Figure 9
MiR-145 inhibition abrogated miR-504 mediated CTGF silencing
vimentin, and CTGF in GSCs. The inhibition of miR-504 on the expression of CTGF was
Figure8 Is this correct or some labeling
indirectly by upregulating miR-145 expression that targeted CTGF expression. Moreover, N/A Classical G-CIMP MES Neural PN misyake? The abstract doesn’t mention
silencing of CTGF resulted in upregulation of both miR-145 and miR-504.(no data ?) In mir145 up-stream of 504 but opposite
conclusion, these results reveal novel miRNAs and potential target networks that play a role need mir 145 expression in pmir -504
in the oncogenic potential, stemness and mesenchymal transformation of GSCs and identify samples
miR-504 as a potential therapeutic target for the inhibition the migration and mesenchymal
transformation of GBM and GSCs.
 
MiR-504 inhibits the mesenchymal phenotype Summary
Differential expression of miRNAs in human NSCs
selfrenewal and migration in glioma cell lines.
GSCs, and mesenchymal stem cells using miR
microarray analysis
Con pre-miR

Pre-miR-504

Con pre-miR Pre-miR-504

FN 0 Hrs
miR-504

CTGF

GFAP 17Hrs

GAPDH

MiR-504 downregulated CTGF expression indirectly by Mesenchymal transition

Figure5 MiR-504 dicreses Mesechymal markers CTGF and upregulating miR-145 expression
Fybronectin and elevates GFAP, (astrocyte marker)

120
120

miR-504 was the most significantly downregulated in GSCs 100

Self-Renewal - % of Control

100
compared with NSCs 80
80
% Migration

Objectives: Analyze the role and function of miR-504 in the 60

60

mesenchymal transformation and stemness of GSCs. 40 40

Figure1
Sample Bibliography? Conclusions
20 20

A 3D PCA scatter plot of the miRNA data that

characterize the trends exhibited by the 0
Con pre- Pre-miR- Con pre- Pre-miR-
0
Con pre-miR Pre-miR-504
miR 504 miR 504
expression profiles
HF2414 HF2355

Figure 6 miR-504 inhibits GSC selfrenwal Figure 7 MiR-504 inhibits migration of GSCs
These results reveal novel miRNAs and potential target networks that play a role in the oncogenic potential,
stemness and mesenchymal transformation of GSCs and identify miR-504 as a potential therapeutic target for the
inhibition the migration and mesenchymal transformation of GBM and GSCs