A little exercise goes a long way to deliver chemotherapy Keri Schadler May 21, 2015.
-
Upload
marianna-martin -
Category
Documents
-
view
214 -
download
1
Transcript of A little exercise goes a long way to deliver chemotherapy Keri Schadler May 21, 2015.
A little exercise goes a long way to deliver chemotherapy
Keri SchadlerMay 21, 2015
McDonald D and Choyke P. Nature Medicine 2003
Normal Vasculature Tumor Vasculature
Tumor vasculature is inefficient: Drug Delivery is Ineficient
Tumor vasculature can be ‘normalized’
Jain R. Science 2005
Goel S. et al., Physiol. Rev. 2011
Tumor vascular normalization enhances drug delivery
Increased perfusion after anti-angiogenic therapy correlates with survival in glioblastoma patients
AG Sorenson, et al.. Cancer Research 2012 Jan 15;72(2):402-7.
Limitations in the use of anti-angiogenic therapy for vascular normalization
Increased shear stress activates the endothelium and remodels the vasculature
network
• Shear stress is regulated by the rate of blood flow and the vessel diameter
Shear Stress Regulates Vascular Remodeling
Disturbed Shear (Tumor)- High EC Turnover:
proliferation & apoptosis
- High Permeability- Immature Vessels- Sporadic Blood Flow
Moderate Steady Shear– Quiescent ECs– Low permeability– Mature Vessels– Constant blood flow
Nat Rev Mol Cell Biol. 2009 January; 10(1): 53–62
Exercise increases shear stress
Suh et al., Ann Biomed Eng. 2011
Aerobic exercise
Heart rate
blood flow
shear stress
Mea
n w
all s
hear
stre
ss
(dyn
es/c
m2 )
Exercise is safe and feasible for cancer patients
Can Exercise Normalize Tumor Blood Vessels?
Jain R. Science 2005
The models: Mice, Cancer, and Exercise
HighModerateLow
50% 70%
12m/min ~ Brisk Walk45 min/day; 5 days/week
Begin when tumor is palpable
VO2 :
B16F10 Melanoma or PDAC-4662 Pancreatic Adenocarcinoma4T1 or E0771 Mammary Carcinoma
Voluntary wheel runningBegan day of tumor inoculation
Exercise Normalizes Tumor VasculaturePDAC
No Ex Ex
CD31; DAPI
4T1 Mammary
Vessel Length
*Lumens
*
B16F10 Melanoma
Pericyte Coverage Increases with Exercise
ExerciseNo Exercise4T1 Mammary
Exercise Increases Tumor Vessel Function
No Ex Ex
4T1 Mammary Tumors
Exercise Increases Tumor Vessel Function
CD31 Lectin Merge; DAPI
No Ex
Ex
*
PDAC tumors, n=5-6 per group
Exercise Induces Functional Endothelial Tubes in Matrigel
Lectin; DAPI Lectin;CD31;DAPI
No Ex
Ex
*
Shear Stress
Endothelial Cells in
Matrigel
Inject subcu; wait one day
Exercise (or not) 7 days Harvest
MatrigelShear Stress
Endothelial Cells in
Matrigel
Inject subcu; wait one day
Exercise (or not) 7 days Harvest
Matrigel
Dongha Bhang, UPenn
= Tumor Growth
Chemotherapy Delivery
Hypothesis: Exercise increases chemotherapy delivery and efficacy
The models: Mice, Cancer, and Exercise
12m/min ~ Brisk Walk45 min/day; 5 days/week
Begin when tumor is palpableDoxorubicin or Gemcitabine 2 or 3x weekly
B16F10 Melanoma or PDAC-4662 Pancreatic Adenocarcinoma4T1 or E0771 Mammary Carcinoma
Voluntary wheel runningBegan day of tumor inoculationCyclophosphamide days 7,9,11
Exercise Increases Chemo Efficacy
PDAC B16F10
4T1 Mammary
Chemotherapy delivery is increased in tumors from exercised mice
gH2AX DAPI
Gem Gem + Ex
*
Melanoma Lysates*
Functional vessels are made by balance of pro- and anti-angiogenic factors
4T1 whole tumor RNA: VEGF
Pro: VEGF Anti: TSP-1
TSP-1 is necessary for increased chemotherapy efficacy by exercise
PDAC tumors; Tsp1f/f ;VE-cad Cre mice
Gem + ExerTSP-1
b-actin
Whole Tumors
TSP-1
b-actin
Endothelial Cells
Hrs Shear: 0 24
Is normalization necessary, or is acute exercise enough?
= Tumor Growth
Chemotherapy Delivery
Melanoma: One exercise session does not increase doxorubicin delivery
B16F10 tumors; Dox assessed by spectrophotometry
Acute ExChronic Ex
*
How do tumor vessels respond to acute exercise?
5 min- Run 60 min - Rest
RatsMatLyLu prostate tumors
Treadmill: 15m/min, 10* incline (60% max capacity)
Danielle J. McCullough et al. JNCI J Natl Cancer Inst 2014;106:dju036
Acute exercise increases tumor blood flow and decreases vascular resistance
Blood Flow Vascular Resistance
Danielle J. McCullough et al. JNCI J Natl Cancer Inst 2014;106:dju036
Rest
Exer.
Acute exercise increases tumor blood volume
Danielle J. McCullough et al. JNCI J Natl Cancer Inst 2014;106:dju036
Tumor Arterioles Constrict Differently than Normal Vessels
Norepinephrine Constriction Intraluminal Pressure Response
Exercise Enhances Chemotherapeutic Efficacy
• Exercise Normalizes Tumor Vasculature• Exercise Causes Acute Increases in Tumor
Blood Flow• Blood Flow Promotes Tumor Growth…
=Tumor Growth
Blood Delivery
Is exercise a good thing when there’s no chemotherapy?
?
Exercise has cancer type-specific effects on tumor growth
*
B16F10 MelanomaPDAC-4662 Pancreatic Adeno.
4T1 Mammary Tumors
Allison S. Betof et al. JNCI J Natl Cancer Inst 2015;107:djv040
Allison S. Betof et al. JNCI J Natl Cancer Inst 2015;107:djv040
Exercise increases apoptosis in 4T1 mammary tumors
Exercise is Multi-faceted
Immune Response
Oxidative Stress/ROS
Metabolism
Cytokines / Growth Factors
IL-6IGF1
Work to be done…
• Acute vs Chronic?• What kind of exercise? What intensity?• Molecular Regulators?• Which patients will get the most benefit?• Combination with other therapies?• Effect on Metastasis? Overall survival?
Thank you…Collaborators Wanted!
In vitro studies of shear stress using 3D-lumenized blood vessel models
Chris Chen & Pete Galie,U Penn Bioengineering
Endothelial Tube
collagen
Pump Media Into Device Collect Media
Serum from Exercised Mice Inhibits EC Sprouting
No Exercise Serum Exercise Serum
*
Chris Chen & Pete Galie,U Penn Bioengineering
Shear Stress Conditioned Media Inhibits EC proliferation and tube formation
No shear C.M. Shear C.M.
WT mouse lung Endothelial Cells (ECs)*p<0.05, n =3
In this complex, PECAM-1 is hypothesized to be the primary mechanotransducer, where it is directly stimulated by force to initiate signaling. VE-cadherin functions as an adapter that brings VEGFR2 into proximity with PECAM-1, where VEGFR2 is activated by a src family kinase. Activated VEGFR2 then recruits PI3-kinase and mediates activation of downstream pathways. Recent work has revealed additional functions of these proteins in vascular remodeling and function, and shed new light on their involvement in mechanotransduction.
PLC
nucleus
PIP2
DAG
IP3
Ca2+
Endoplasimic Reticulum
NFATCalcineurin
P
P
Angiogenesis genes
NFAT
NFAT
TSP-1
Shear stress induces NFAT nuclear translocation
No
Shea
rSh
ear
NFATc1 NFATc1; DAPI
8 minutes shear stress, avg 10 dyne/cm2
*
Shear Stress induces NFAT gene targets
Rnd1Cox2
**
Hours Shear Stress
Cyclosporin (NFAT inhbition) abrogates exercise effect in B16F10 tumors
NFAT inhibition in DSCR1 Tg mice abrogates the exercise effect on PDAC tumors
Calcineurin
NFATp p
NFAT
DSCR1
Inhibition of NFAT prevents vascular normalization
Thrombospondin-1 (TSP-1): an NFAT regulated anti-angiogenic factor
TSP-1• Soluble, secreted factor• Produced by ECs,
stroma, immune• Receptor on ECs• Inhibits EC proliferation• Vessel Maturation
NFAT
IL-6Cox2Rnd1Ang2TSP-1
CD31; DAPI
CD31; DAPI
Tumor Growth
Chemotherapy Delivery
Inefficient Vasculature
“Normalized” Vasculature
Moderate Exercise
Shear Stressnucleus
Ca2
+
P
P
NFAT
TSP-1
NFAT
NFAT
Effects of aerobic exercise on angiogenesis and vascular maturity in primary 4T1 breast tumors.
Allison S. Betof et al. JNCI J Natl Cancer Inst 2015;107:djv040
© The Author 2015. Published by Oxford University Press.
Pharmacologic Increase in Shear Stress Also Enhances Gemcitabine Efficacy
*
Praz = Prazosin; PDAC tumors; 4-5 per group
p<0.05
Exercise inhibits Ewing’s sarcoma growth in mice
TC71 cells in nude mice, n = 3 -5 per group, Doxo = 4mg/kg/week
TSP-1 is needed for growth inhibition by shear stress conditioned media
Naïve WT cells
WT or Tsp1-/- ECconditioned media
Measure proliferation
Fluorescence photomicrograph of a prostate tumor cross-section, obtained from a MatLyLu Copenhagen rat prostate carcinoma after intraperitoneal injection of EF5 (30mg/kg) at rest
(A) and during acute exercise (B).
Danielle J. McCullough et al. JNCI J Natl Cancer Inst 2014;106:dju036
© The Author 2014. Published by Oxford University Press. All rights reserved. For Permissions, please e-mail: [email protected].