O-081 - M. LEPRAE MODULATES GLUCOSE UPTAKE AND

O-081 - M. LEPRAE MODULATES GLUCOSE UPTAKE AND

M. leprae Modulates Glucose Uptake and
Metabolism in the Host Cell
OSWALDO CRUZ INSTITUTE
OSWALDO CRUZ FOUNDATION
MINISTRY OF HEALTH
Brazil
Rychelle C. A. Medeiros1; Karina Girardi1; Julio Jablonski1,
Mauro Sola-Penna2; Patricia S. Rosa3; Marcus O.
Fernandes4; Euzenir N. Sarno5; Milton O. Moraes5;
Maria Cristina Pessolani1 and Flavio A. Lara1.
1) Lab. de Microbiologia Celular, IOC, FioCruz;
2) Lab. de Enzimologia e Controle do Metabolismo, Fac. de Farmácia, UFRJ;
3) Coordenação de Controle de Doenças, Inst. Lauro de Souza Lima;
4) Lab. de Bioquímica de Resposta ao Estresse, Inst. de Bioquímica Medica, UFRJ;
5) Lab. de Hanseníase, IOC, FioCruz;
Objective
Describe human Schwann cell lineage ST8814 energy metabolism
modulation during M. leprae infection.
M. leprae is able to modulate glucose uptake in the host cell
NBDG
MOI 1:20
Microscopy fluorescence
***
Mean gray value (A.U.)
125
***
100
75
50
25
0
Control
MOI 1:20
MOI 1:50
MOI 1:50
Mean fluorescence intensity (A.U.)
Control
Flow cytometry
Control
ML 1:10
ML 1:100 Smeg 1:10 Smeg 1:100
Why M. leprae infected cells capture more glucose?
M. Leprae (50:1)
TMRE = mitochondria
Control
Controle
CCCP
M.Leprae
Leprae (50:1)
M.
(50:1)
CCCP
Mean gray value (A. U.)
Control
*
Control Control
CCCP
TMRE + CCCP
ML
ML
CCCP
Mitochondrial activity measured by TMRM fluorescence after 48h of infection
Flow cytometry
400000
*
**
200000
C
ol
P
ig
om
yc
in
C
C
M
yc
in
L
M
L
M
L
O
lig
om
on
tr
ol
C
C
C
P
0
C
TMRM MFI (A.U.)
600000
Schwann cells infected with M. leprae display low mitochondrial activity
Why cells infected with M. leprae present lower mitochondrial activity?
Mitochondrium complex IV immunolocalization
Control
Infected
Although apparently the host cell starts to ferment glucose, lactate
production drops after infection!
Schwann cells supernatant
Leprosy patients sera
nt
s(
LL
)
tie
Pa
nt
s(
B
L)
tie
Pa
nt
s(
B
T)
tie
Pa
ve
C
on
t
ro
l
Irr
M
L
ad
(5
ia
0:
te
1)
d
M
L
Li
(5
ve
0:
1)
B
C
G
Li
(5
ve
0:
Irr
Sm
1)
ad
e
ia
g
te
(5
d
0:
Sm
1)
eg
(5
0:
1)
0
ls
10
***
on
tr
o
*
20
90
80
70
60
50
40
30
20
10
0
C
30
(Lactate) mg/dl
40
Li
Lactate (mg/dL)
50
H
M
Hypothesys: drop of lactate synthesis in infected Schwann cell as a new
axon damage mechanism in leprosy.
Adapted from: Youngjin Lee, et al.
Nature 487, 443–448 (26 July 2012)
Glucose
Glucose 6P
6P gluconate
X
Fructose 1,6PP
Pentose Cycle
Glyceraldehyde 3P
X
Piruvate
The main cellular reducing power generation pathways were turned on
after M. leprae infection
Piruvate Carboxilase
Fold Increase
20
15
10
5
0
BT
LL
Why M. leprae host cell produces so much reducing power?
How these cells recycle NADPH to NADP+ with an inhibited Krebs cycle?
Infected cells increase free radicals control
Stabilized
molecule
Free
radical
0.045
0.040
0.035
0.030
0.025
0.020
0.015
0.010
0.005
0.000
**
Sm
eg
L
L
ve
Li
Irr
ad
ia
te
d
M
M
e
Li
v
on
tr
ol
*
C
citrate lyase activity
(mmol/min/mg ptn)
Infected cells increases lipids synthesis
From pentose Cycle
Free radicals
Energetic Metabolism
Model on Leprosy
malate
lactate
Free radicals
X
X
My Team
Doctoral student:
Márcia Moreira
Masters students:
Débora Silva
Lívia Lobato
Jéssica Ferreira
Rychelle Medeiros
Undergraduate
Students:
Rodrigo Dutra
Arthur Newman
Sabrina Alves
Karina Girardi
High school student:
Breno Veloso
Acknowledgements
Collaborators
Lab. de Microbiologia Celular, IOC / Fiocruz
Dra. Maria Cristina Pessolani
Lab. de Bioquímica de Artrópodes Hematófagos
Dr. Pedro Lagerblad / Dr. Marcus Fernandes / Dr. Marcos Sorgine
Lab. de Enz. e Cont. do Metabolismo, Fac. de Farmácia / UFRJ
Prof. Mauro Sola-Penna
Inst. Lauro de Sousa Lima, Depto de Biologia, Bauru
Dra. Patricia Sammarco Rosa
Lab. de Hanseníase, IOC / Fiocruz
Dra. Euzenir Nunes Sarno, Dr. Milton Moraes and Dr. José Nery
Thank you for your attention