Transcript IL-1β - Canvas

Mechanisms of Cancer Treatment
Related Fatigue
Lisa J. Wood PhD RN FAAN
Amelia Peabody Professor for Nursing Research
School of Nursing, MGH Institute of Health
Professions
Fatigue: Definition
“Cancer related fatigue is a distressing persistent
subjective sense of physical, emotional and/or
cognitive tiredness or exhaustion related to
cancer or cancer treatment that is not
proportional to recent activity and interferes with
usual functioning”
National Comprehensive Cancer Network, 2011
http://www.nccn.org/professionals/physician_gls/pdf/fatigue.pdf
Cancer Treatment Related Fatigue
•
Occurs in greater than 60% of cancer patients undergoing
treatment with cytotoxic chemotherapy, and/or radiation therapy,
or cytokine therapies.
•
Responsible for reduced QoL, reduced physical functioning, and
is perceived as more distressing than pain, nausea and vomiting.
•
Patterns of fatigue during treatment, worse in the days following
chemotherapy returning to baseline before the next infusion.
•
In approximately 20-30% of cancer survivors, fatigue can persist
for months or even years after treatment has ended.
•
•
Fatigue is an obstacle for physical activity.
Importance of understanding mechanisms.
Chemotherapy induced IL-1β as the trigger of
cancer chemotherapy related fatigue
Sickness Behavior
Anemia
Decreased
Appetite
Muscle Loss
Chemotherapy
IL-1β
Systemic
Inflammation
Fatigue
Difficulty
Thinking
Depression
Sleep
Problems
Decreased
Physical
Activity
Interleukin-1β triggers sickness behavior
Cytokines interact with
specific populations of
neurons in the brain to
cause the signs and
symptoms of sickness
behavior.
Harmful stimulus
i.e. bacteria, tissue injury,
Intracellular contents and
other “Danger Signals”
Release of mature IL-1β into
the periphery stimulates the
production of IL-1β and
other inflammatory cytokines
i.e. TNF-α.
Production of acute
phase reactants by
the liver i.e. CRP
and coagulation
factors.
Macrophage
Gene expression
IL-1 Receptor
Increased
production &
maturation of
immune cells
Wood LJ, and Weymann K. Inflammation and neural signaling: Etiologic mechanisms of the
cancer treatment related symptom cluster. Curr Opin Support Palliat Care, Curr Opin
Support Palliat Care. 2013 Mar;7(1):54-9
Identification of the “Fatigue” Neuron?
Grossberg AJ, Zhu X, Leinninger GM, Levasseur PR, Braun TP, Myers MG,Jr, et al. Inflammation-induced lethargy is
mediated by suppression of orexin neuron activity. J Neurosci 2011 Aug 3;31(31):11376-11386.
Orexin neurons regulate
arousal and wakefulness.
They are active during
wakefulness and silent
when asleep
When orexin Neurons are active
they make and secrete a
neurotransmitter called orexin.
Hypothalamus
Inflammation causes
orexin neurons to
produce less orexin.
Less orexin means more
fatigue. Injecting orexin into the
rodent brain decreases fatigue
in rodents with sickness
behavior1.
Hypothesis
Cancer chemotherapy causes fatigue by
inducing inflammation and decreasing the
activity of hypothalamic orexin neurons.
Does Chemotherapy trigger IL-1β production by
immune cells and if so how?
Incubate cells with different
Chemotherapy drugs and
inflammatory inhibitors.
Collect macrophages
From mice bone marrow
Collect the culture fluid
and cells and measure IL1β levels.
Macrophages from mice genetically engineered to lack specific proteins
can be used to determine the specific role of these proteins in chemotherapy
Induced IL-1β production.
Chemotherapy triggers IL-1β by macrophages
Widespread cell death
releases cell components
which act as danger signals
warning macrophages to
take action.
Toll-Like
Receptors
Cytotoxic
Chemotherapy
Chemotherapy also activates
the inflammasome which
cleaves pro-IL-1β to its mature
form.
Chemotherapy activates an
enzyme called ZAK which
increases production of proIL-1β
ZAK
ZAK inhibitors nilotinib
and sorafenib decrease IL-1β
production.
IL-1β
Inflammasome
macrophage

Gene expression
Pro
Pro
IL-1β
Pro
Pro
IL-1β
IL-1β
IL-1β
Sauter AD, Wood LJ, Wong J, Iordanov M, and Magun BE. Doxorubicin and daunorubicin induce processing and release of interleukin-1β through activation of the NLRP3 inflammasome. Cancer Biol Ther. 2011 Jun 15;11(12):1008-16.
Wong J, Smith LB, Magun EA, Engstrom T, Kelley-Howard K, Jandhyala DM, Thorpe CM, Magun BE, Wood LJ. Small molecule kinase inhibitors block the ZAK-dependent inflammatory effects of doxorubicin.Cancer Biol Ther. 2013 Jan;14(1):56-63
Does Chemotherapy Increase Circulating IL-1β?
CAF or NS by
Intraperitoneal
injection
Collect peripheral blood at 1, 3, 6, 14,
and 24 hrs after injection.
Measure serum levels of inflammatory
cytokines and chemokines using bead
based immunoassays (Milliplex,
Millipore Inc.)
CAF: Cytoxan (167mg/kg) + Adriamycin 4mg/kg) + 5-fluorouracil (167mg/kg)
NS: Normal Saline
CAF increases serum levels of IL-1β, TNF-α,
and IL-6
Figure 1. CAF-induced IL-1β, TNF-α,
and IL-6 production in mice. A) Fold
increase in serum levels of IL-1β, TNF-α,
and IL-6 in CAF-treated relative to NStreated mice sacrificed at 1, 3, 6, 14 and
24 hours post-injection.
Smith LB, Leo MC, Anderson C, Wright TJ, Weymann KB, & Wood LJ. The role of IL-1β and TNF-α signaling in the genesis of cancer treatment related
symptoms (CTRS); a study using cytokine receptor-deficient mice. Brain, Behavior & Immunity, 2014 Jan 7. [Epub ahead of print].
Elsea CR, Kneiss JK, & Wood LJ. Induction of IL-6 by cytotoxic chemotherapy is associated with cachexia in tumor-free female mice. (Acepted with
revisions)
Is CAF induced inflammation associated with
fatigue?
Assess voluntary wheel running activity (VWRA), food intake and
body weight in female mice administered 4 doses of CAF at 21-day
intervals.
Minimitter, Bend Oregon
Patterns of change in fatigue, body weight, & food
intake in mice administered CAF or NS
CAF-induced changes in CTRS
in mice. A.) Daily VWRA, body
weight, and food intake during
baseline (B), and 4 cycles (C1C4) of CAF (n= 17, filled square)
or NS (n=17, open diamond).
Each data point represents the
mean of each value. *p<0.05
Acute
No
Persistent
MCP-1
KC
IL-7
IL-9
GCSF
IP10
-
-
Blocking the CAF-induced inflammatory
response
• IL-1 type 1 receptor (IL1R1: B6.129S7-Il1r1tm1Imx)
• IL1R1 and TNF-α p55 (type 1) receptor (TNFR1:
B6;129S-Tnfrsf1atm1Imx Il1r1tm1Imx/J)
• Jackson Laboratories, Bar Harbor Maine.
www.Jax.org
Blocking IL-1β and TNF-α activity dampens
CAF-induced inflammation
12
Fold increase (Relative to Controls)
10
Normal (7,7)
IL-1 receptor deficient (5,5)
IL-1 and TNF-deficient (7,7)
***
***
8
6
*
***
***
4
*
***
**
**
*
2
0
GCSF
IL-6
KC
MCP-1
Fold increase in serum levels of GCSF, IL6, KC and MCP-1 in CAF-treated relative to
NS-treated WT, IL-1R1- and IL1R1/TNFR1-deficient mice sacrificed 16
hours post-injection. The number of mice in
the NS and CAF treated groups for each
genotype are indicated in parentheses
respectively. (*p < 0.05, **p<0.001,
***p<0.0001).
Blockade of IL-1β and TNF-α signaling
reduces survival in CAF treated mice
80
80
60
p=0.197
40
20
0
Normal Saline (n=9)
Normal Chemo (n=9)
IL-1R1null Saline (n=9)
IL-1R1null Chemo (n=9)
1
2
3
Treatment Cycle
% Survival
100
% Survival
100
60
40
20
4
p=0.003
0
Normal Saline (n=9)
Normal Chemo (n=9)
TNF/IL-1Rnull Saline (n=9)
TNF/IL-1Rnull Chemo (n=9)
1
2
3
Treatment Cycle
4
Targeting the “Fatigue” Neuron?
Inflammation
Hypothalamus
Awake
Fatigued
Grossberg AJ, Zhu X, Leinninger GM, Levasseur PR, Braun TP, Myers MG,Jr, et al. Inflammation-induced
lethargy is mediated by suppression of orexin neuron activity. J Neurosci 2011 Aug 3;31(31):11376-11386.
CAF reduces CSF orexin-A
IHC
ip CAF/
sham
(OxA/cFos)
CSF-AM
lights off
CAF
1630
[OxA]-RIA
0600
1930
1800
Diurnal pattern: CSF Orexin-A highest
at end of active period
Rat CSF OxA (pg/mL)
CSF Ox-A
Orexin-A CSF
1000
800
600
***
400
200
0
saline CAF
Weyman KB, Wood LJ, Zhu X, & Marks DL. A role for orexin in cytotoxic chemotherapy-induced
fatigue. Brain, Behavior & Immunity (In Press).
CAF reduces c-Fos Expression in Orexin Neurons
PFA
LHA
Sham
DMH
CAF
3V
cFos/OxA co-IR
(% Ox neurons)
80
*
*
Sham
CAF
60
Orexin
cFos
40
20
0
All Nuclei
DMH
PFA
Orexin Ab stains
orexin neurons green
cFos Ab stains active
cells
LHA
Weyman KB, Wood LJ, Zhu X, & Marks DL. A role for orexin in cytotoxic
chemotherapy-induced fatigue. Brain, Behavior & Immunity (In Press).
Ox-A replacement restores activity
in CAF-treated rats
icv OxA
1730
ambulatory act.
(OxA/cFos)
lights off
1800
ipCAF 24 h
IHC
1930
CSF-AM
[OxA]-RIA
lights on
0600
OxA replacement restores
activity in CAF-treated rats
% of baseline
activity 18-1930
150
**
**
100
50
-O
xA
AF
C
Sh
am
-O
xA
C
AF
-a
C
SF
Sh
am
-a
C
SF
0
Weyman KB, Wood LJ, Zhu X, & Marks DL. A role for orexin in cytotoxic chemotherapy-induced
fatigue. Brain, Behavior & Immunity (In Press).
Suppressed orexin neuron activity in
persistently fatigued mice
% cFos+ Ox IR neurons
**
***
***
DMH
PFA
*
LHA
60
40
CAF
cFos/OxA co-IR
(% OxA neurons)
80
PFA
Sham
DMH
20
0
All Nuclei
LHA
Orexin
cFos
Orexin Ab stains
orexin neurons green
cFos Ab stains active
cells
Weyman KB, Wood LJ, Zhu X, & Marks DL. A role for orexin in cytotoxic
chemotherapy-induced fatigue. Brain, Behavior & Immunity (In Press).
Conclusions
• Chemotherapy triggers IL-1β production by immune cells
which leads to a systemic inflammatory response.
• Acute fatigue is associated with increased circulating levels of
inflammatory markers but not persistent fatigue.
• Blocking chemo-induced inflammation decreases survival.
• Targeting neural effectors of inflammatory cytokines may be a
more beneficial approach to decreasing chemo-related
fatigue.
• Acute chemo-related fatigue is associated with decreased
orexin in the CSF and orexin replacement decreases acute
chemo-related fatigue.
The Team
Lillian M. Nail PhD RN FAAN
Professor
OHSU SON
Logan Smith PhD
Kris B. Weymann PhD RN MS
Instructor
OHSU SON
John Wong PhD
Assistant Professor
IHP SON
Daniel Marks
Professor
OHSU SOM
Bruce Magun PhD
Professor, IHP SON
Acknowledgements
• Xinxia Zhu
• Caroline Anderson BSN
• Teresa Wright BSN
Funding
• NINR NR012479 (LJW)
• NRSA NINR F31NR013299 (KW, Nail/Wood Mentors)
Questions?