ROS production and angiogenic regulation by macrophages in response to heat therapy.
OBJECTIVE
It has been well established that inadequate blood supply combined with high metabolic rates of oxygen consumption results in areas of low oxygen tension (<1%) within malignant tumours and that elevating tumour temperatures above 39 degrees Celsius results in significant improvement in tumour oxygenation. Macrophages play a dual role in tumour initiation and progression having both pro-tumour and anti-tumour effects. However, the response of macrophages to heat within a hypoxic environment has not yet been clearly defined.


METHODS
Raw 264.7 murine macrophages were incubated under normoxia and chronic hypoxia at temperatures ranging from 37-43 degrees Celsius. Under normoxia at 41 degrees Celsius, macrophages start to release significant levels of superoxide. The combination of heat with hypoxia constitutes an additional stimulus leading to increased respiratory burst of macrophages.


RESULTS
The high levels of superoxide were found to be associated with changes in macrophage production of pro-angiogenic cytokines. While hypoxia alone (37 degrees Celsius) increased levels of hypoxia inducible factor-1alpha (HIF-1alpha) in macrophages, the combination of hypoxia and mild hyperthermia (39-41 degrees Celsius) induced a strong reduction in HIF-1alpha expression. The HIF-regulated vascular endothelial growth factor (VEGF) decreased simultaneously, revealing that heat inhibits both HIF-1alpha stabilization and transcriptional activity.


CONCLUSIONS
The data suggest that temperatures which are readily achievable in the clinic (39-41 degrees Celsius) might be optimal for maximizing hyperthermic response. At higher temperatures, these effects are reversed, thereby limiting the therapeutic benefits of more severe hyperthermic exposure.
Jackson
I L
IL
Department of Radiation Oncology, Duke University Medical Center, Durham, NC 27710, USA.
Batinic-Haberle
I
I
Sonveaux
P
P
Dewhirst
M W
MW
Vujaskovic
Z
Z
D000818:Animals; D002460:Cell Line; D016207:Cytokines / Q000378:metabolism; D006358:Hot Temperature; D006979:Hyperthermia, Induced; D000860:Hypoxia / Q000378:metabolism*; D051795:Hypoxia-Inducible Factor 1, alpha Subunit / Q000032:analysis / Q000378:metabolism / Q000502:physiology; D008264:Macrophages / Q000378:metabolism*; D051379:Mice; D009369:Neoplasms / Q000378:metabolism / Q000628:therapy; D018919:Neovascularization, Physiologic / Q000502:physiology*; D010101:Oxygen Consumption / Q000502:physiology; D017382:Reactive Oxygen Species / Q000378:metabolism*; D016897:Respiratory Burst / Q000502:physiology; D013481:Superoxides / Q000378:metabolism; D013696:Temperature; D042461:Vascular Endothelial Growth Factor A / Q000378:metabolism
D016207:Cytokines; C497443:Hif1a protein, mouse; D051795:Hypoxia-Inducible Factor 1, alpha Subunit; D017382:Reactive Oxygen Species; D042461:Vascular Endothelial Growth Factor A; D013481:Superoxides