epigenetics & tb: can mirnas be harnessed as an hdt?...advancing host directed therapy for...
TRANSCRIPT
Advancing Host Directed Therapyfor Tuberculosis
April 15/16, 2014Rockville
Anca Dorhoi/Stefan H.E. Kaufmann
Epigenetics & TB: Can miRNAs be Harnessed as an HDT?
Exuberant inflammation
Tuberculosis: HDT directions
HDTin TB
Protective immunity Functions
Defined parameters
Systems Pathology
Resolution Processes ScarringOrgano-genesis
Exuberant inflammation
Tuberculosis: HDT directions
HDTin TB
Protective immunity Functions
Defined parameters
Systems Pathology
Resolution Processes ScarringOrgano-genesis
Balance/Fine-tune
Tuberculosis: miRNA directed HDT
miRNAlncRNA
Biological process
MicroRNA (miRNA): biogenesis and function
22 nt long
Encoded as single entities or in clusters
Transcribed by RNA Pol II
30% of all genes might be miRNA targets
Pulmonary TB: Blood miR Signature
Dorhoi , Iannaccone et al., JCI , 2013
DAPI MPOGiemsa acid-fast
WT
miR
-223
-/-
miR-223: Controls Susceptibility to TB
0 20 40 600
20
40
60
80
100WT
miR-223-/-
Days p.i.
Sur
viva
l (%
)p=0.0008
Dorhoi , Iannaccone et al., JCI , 2013
miR-223: Inflammatory Hub
Fold
cha
nge
com
pare
d to
wild
type
Cxcl2 Ccl3 il-6 Ccl2 Cxcl10 0
5
10
15 naiveMtb
Dorhoi , Iannaccone et al., JCI , 2013
miR-223: Targets CXCL2, CCL3, IL-6Lu
cife
rase
act
ivity
0.0
0.5
1.0
1.5 ***
Luci
fera
se a
ctiv
ity
0.0
0.5
1.0
1.5 **
CXCL2 + + + -CXCL2 mut - - - +
miR-223 - + - +scramble - - + -
IL-6 + + + -IL-6 mut - - - +miR-223 - + - +scramble - - + -
Luci
fera
se a
ctiv
ity
0.0
0.5
1.0
1.5 *
CCL3 + + + -CCL3 mut - - - +miR-223 - + - +scramble - - + -
3‘ accccauaaACUGU UU GACUGu 5‘ miR-223: :
328 5‘ augacaaagUCAUGAA C UGACa 3‘ Ccl3
3‘ accccAUAAA CUGUUUGACUGu 5‘ miR-223:
86 5‘ uguucUCUACGAAGAACUGACa 3‘ Il6
3' accccauaaaCUG UUUGACUGu 5‘ miR-223
769 5' ggggguggg GAC AA AUAGAUG 3' Cxcl2
Dorhoi , Iannaccone et al., JCI , 2013
miR-223: Cell Intrinsic Effects in PMNm
iR-2
23 f
old
chan
ge
AM PMN iM0
1
2
*** ***
miR
-223
fol
d ch
ange
0
2
4
6*
IgG control Ly6G
Ccl
3 fo
ld c
hang
e
0
5
10
15
AM PMN iM
*
WT
miR-223-/-
Cxc
l2 fo
ld c
hang
e
0
5
10
15
AM PMN iM
***
WTmiR-223-/-
Dorhoi , Iannaccone et al., JCI , 2013
miR-223: Autocrine Regulation of PMN Recruitment
0 200
20
40
60
80
100
30 40 60
miR-223-/- IgG controlWT
miR-223-/- CXCL2/CCL3/IL-6
Days p.i.
Sur
viva
l (%
)
p=0.0186
0 200
20
40
60
80
100
30 40 80
miR-223-/- Ly6G
WT
miR-223-/- IgG control
Days p.i.
Sur
viva
l (%
)
p=0.0359
Days p.i.
Sur
viva
l (%
)
0 10 20 30 40 500
20
40
60
80
100
miR-223-/-
miR-223-/-Cxcr2-/-
p=0.0009
Dorhoi , Iannaccone et al., JCI , 2013
miRNAs: Additional Examples
miR-125 TNF-α: Rajaram et al., 2011miR-29 IFN-γ: Ma et al., 2011
miR Clusters: Maertzdorf et al., 2012
Harapan et al., 2013
miRNAs: Applications for other Diseases
Rooiji et al., Circ Res , 2013
miRNAs: From Bench to Bedside
Hydbring and Badalian-Very, F100Res , 2013
miRNAs: Risks and Challenges
Risks
• immune effects (siRNA)• specificity
Challenges
• delivery• local effects desired
miRNA HDT
miRNAs: Delivery anti-miRNA (antagomir)
Broderck and Zamore, Gene Ther , 2011
miRNAs: Risks and Challenges
Anti-miR: LNAmiR-mimics
lipo/exosomes, Abslinked, lenti/adenovirus
Risks
• immune effects (siRNA)• specificity
Challenges
• delivery• local effects desired
Lessons:HCV (miR-122)
Myocarditis (miR-530-3b; miR-199a-3p)
Lung cancer (let-7)
Lessons:HCV (miR-122)
Myocarditis (miR-530-3b; miR-199a-3p)
Lung cancer (let-7)
Biotechnology:siRNA TB (TGFβ)Biotechnology:
siRNA TB (TGFβ)