James P. Quigley
Lab Overview
Our laboratory uses model systems to study complex multi-step in vivo processes
such as tumor invasion, metastasis and angiogenesis. We are interested in detecting,
identifying, and characterizing cell surface and extracellular molecules that
functionally contribute to these processes. Subtractive immunization is often
used to generate unique function-blocking monoclonal antibodies that identify
the contributory molecules. Functional proteomics is also used to identify
contributory molecules. Specific enzyme activity chemical probes are also used
to identify contributory proteases. Then siRNA or shRNA approaches are used
to assess the specific functional contributions of the identified molecules.
The molecules we are presently examining include cell surface transmembrane
proteins, various secreted matrix metalloproteinases (MMPs) and some new membrane-anchored
serine proteases.
Highlight
The Role of Inflammatory Neutrophils and Human Neutrophil-Derived
MMP-9 in Physiologic and Tumor Angiogenesis.
Veronica Ardi, Elena Deryugina and James Quigley
The matrix metalloproteinases (MMPs) have been strongly linked to the angiogenic
switch, regarded as a critical event during tumor progression. MMP-9 (Gelatinase
B) represents one of the MMPs with pro-angiogenic characteristics. A major cellular
source of MMP-9 at the site of tumor development has been traced to inflammatory
leukocytes such as monocytes/macrophages and neutrophils. Although neutrophil-derived
MMP-9 is well characterized biochemically, the molecular mechanisms by which
MMP-9 from neutrophils facilitates angiogenesis are poorly understood. To analyze
for these mechanisms we employed a quantitative chick embryo angiogenesis model.
In this model system, collagen onplants supplemented with purified inflammatory
leukocytes and/or tumor cells are grafted on the chorioallantoic membrane of
the chick embryo. The levels of angiogenesis are determined within 3 days thus
allowing for efficient and quantitative evaluation of angiogenic potential of
various exogenous additives. We demonstrated that purified human neutrophils,
incorporated into standard collagen onplants, effectively induced physiologic
angiogenesis in a dose-dependent manner. A similar response was achieved when
the neutrophils were replaced with their contents, i.e. an induced releasate,
shown to contain MMP-9. However, when neutrophil releasate was specifically depleted
of MMP-9, it completely lost its pro-angiogenic property, implicating neutrophil
MMP-9 as the critical component responsible for of the angiogenic response. This
suggestion was confirmed when high levels of angiogenesis were induced by sub-nanogram
levels of affinity purified neutrophil MMP-9. Our findings also indicated that
the pro-MMP-9 zymogen, released by infiltrating neutrophils, underwent activation
which was abrogated by a specific activation-blocking antibody, and that the
generated proteolytic activity of the MMP-9 enzyme is responsible for the angiogenic
response. Importantly, purified neutrophil MMP-9 also efficiently facilitates
angiogenesis in onplants containing human tumor cells. When tumor-induced angiogenesis
was diminished by a potent anti-inflammatory drug Ibuprofen, which prevents infiltration
of endogenous leukocytes, purified neutrophil MMP-9 completely restored the diminished
angiogenesis levels. In addition, a non-disseminating human tumor cell variant,
HT-lo/diss, which exhibits only baseline levels of tumor angiogenesis, can be
induced to undergo high levels of angiogenesis by the addition of purified human
neutrophil MMP-9. Altogether, the results of this study indicate that inflammatory
neutrophils facilitate tumor angiogenesis by infiltrating sites of primary tumor
development where they are capable of rapid release of their pre-stored MMP-9,
now herein shown to be a potent pro-angiogenic protease. Further studies will
allow us to analyze mechanistically which neutrophil MMP-9 complex possesses
angiogenic potency and to identify in vivo target molecules/substrates of neutrophil
MMP-9 enzyme.
2006 Publications
Zijlstra A, Seandel M, Kupriyanova TA, Partridge JJ, Madsen
M, Hahn-Dantona EA, Quigley JP, Deryugina EI. Pro-angiogenic
role of neutrophil-like inflammatory heterophils during neovascularization
induced by growth factors and human tumor cells Blood 107:317-27
(2006).
Deryugina, E.I. and Quigley, J.P. Matrix metalloproteinases and
tumor metastasis. Cancer Metastasis Rev 25: 9–34 (2006).
Lewis JD, Destito G, Zijlstra A, Quigley JP, Manchester M, Stuhlmann
H. Viral nanoparticles (VNPs) as tools for in vivo vascular imaging.
Nat Med. 12(3):354-360 (2006).
Madsen MA, Deryugina EI, Niessen S, Cravatt B, Quigley JP. Activity
based protein profiling implicates urokinase (uPA) activation
as a key step in human fibrosarcoma intravasation. J. Biol. Chem.
281(23):15997-6005 (2006)
Zijlstra A, and Quigley JP. The DARC side of metastasis: Shining
a light on KAI1-mediated metastasis suppression in the vascular
tunnel. (Preview) Cancer Cell 10:177-178 (2006).
