Heat-inducible amplifier vector for high-level expression of granulocyte-macrophage colony-stimulating factor.
OBJECTIVE
In cytokine immunotherapy of cancer it is critical to deliver sufficiently high local cytokine concentrations in order to reach the therapeutic threshold needed for clinical efficacy. Simultaneously, for optimal clinical safety adverse effects caused by high systemic cytokine levels must be minimized. One of the most promising anti-cancer therapeutic cytokines, granulocyte-macrophage colony-stimulating factor (GM-CSF), has elicited anti-tumour immune responses in animal studies and clinical trials. However, the clinical efficacy has been limited, with local GM-CSF levels being therapeutically insufficient and systemic toxicity being a limiting factor.


METHODS
To address these problems we have developed a novel GM-CSF expression vector, pAD-HotAmp-GM-CSF, which can provide high levels of GM-CSF expression, and induction of cytokine expression to limited tissue areas. This expression system combines inducible and amplifying elements in a single multi-genic construct. The first transcriptional unit contains the inducible element, the heat shock protein 70B (HSP70B) promoter that regulates expression of the transcription-activating factor tat.


RESULTS
Upon the binding of tat to the second promoter, the HIV2 long terminal repeat amplifies downstream gene expression of the therapeutic cytokine GM-CSF. Moderate hyperthermia at 42 degrees C for 30 min induced GM-CSF expression in pAD-HotAmp-GM-CSF that was over 2.5- and 2.8-fold higher than levels reached with HSP70B promoter alone and the prototypical human cytomegalovirus promoter.


CONCLUSIONS
Thus, the inducible amplifier vector, pAD-HotAmp-GM-CSF, represents a novel system for regulated and enhanced GM-CSF expression, which enables both greater efficacy and safety in cytokine immunotherapy of cancer.
Dammeyer
Pascal
P
Department of Microbiology and Immunology, University of Arizona, Tucson, AZ, USA. pascal.dammeyer@ki.se
Jaramillo
Melba C
MC
Pipes
Brian L
BL
Badowski
Michael S
MS
Tsang
Tom C
TC
Harris
David T
DT
D000818:Animals; D045744:Cell Line, Tumor; D015696:Gene Products, tat / Q000378:metabolism*; D005822:Genetic Vectors / Q000096:biosynthesis*; D016178:Granulocyte-Macrophage Colony-Stimulating Factor / Q000008:administration & dosage / Q000096:biosynthesis* / Q000235:genetics; D018840:HSP70 Heat-Shock Proteins / Q000235:genetics / Q000378:metabolism*; D006979:Hyperthermia, Induced / Q000379:methods*; D008546:Melanoma, Experimental / Q000235:genetics / Q000378:metabolism / Q000628:therapy
D015696:Gene Products, tat; D018840:HSP70 Heat-Shock Proteins; D016178:Granulocyte-Macrophage Colony-Stimulating Factor