inflammatory disease
2008/8/4
Cardiovasc
Res. 2008 Aug 5. [Epub ahead of print] Xanthoulea
S, Gijbels
MJ, van
der Made I, Mujcic
H, Thelen
M, Vergouwe
MN, Ambagts
MH, Hofker
MH, de
Winther MP. Department of Molecular
Genetics, Cardiovascular Research Institute Maastricht, Maastricht
University, Universiteitssingel 50, UNS 50/11, 6229ER Maastricht, The
Netherlands. AIMS: Tumour necrosis factor
(TNF) is a pivotal pro-inflammatory cytokine with a clear pathogenic role
in many chronic inflammatory diseases, and p55 TNF receptor (TNFR) mediates
the majority of TNF responses. The aim of the current study was to
investigate the role of p55 TNFR expression in bone marrow-derived cells,
in atherosclerotic lesion development. METHODS AND RESULTS: Irradiated
low-density lipoprotein receptor knock-out mice were reconstituted with
either p55 TNFR knock-out or control haematopoietic stem cells to generate
chimeras deficient or wild-type for p55 TNFR specifically in bone
marrow-derived cells, including macrophages. Upon high fat feeding, p55
TNFR knock-out transplanted mice developed smaller atherosclerotic lesions.
These lesions were characterized by the presence of smaller foam cells and
a reduced macrophage foam cell area. They did not differ in other
compositional characteristics as determined by quantification of
inflammatory T-cell and neutrophil influx, apoptotic and necrotic cell
death, and collagen content. In vitro studies confirmed a significant
defect in modified lipoprotein endocytosis by p55 TNFR knock-out
macrophages due to reduced scavenger receptor class A expression.
Interestingly, plasma cytokine/chemokine profile analysis indicated that
monocyte chemoattractant protein-1 (MCP-1) levels, a major chemokine
involved in atherogenesis, were consistently and significantly lower in p55
TNFR knock-out transplanted mice compared with controls, before and after
high fat feeding. CONCLUSION: p55 TNFR expression in bone marrow-derived
cells contributes to the development of atherosclerosis by enhancing
lesional foam cell formation and by promoting the expression of
pro-atherosclerotic chemokines such as MCP-1. J Immunol.
2008 Jul 15;181(2):1288-98. Pincheira
R, Castro
AF, Ozes
ON, Idumalla
PS, Donner
DB. Department of Surgery and Comprehensive
Cancer Center, University of California, San Francisco, CA 94115, USA.
pincheirar@surgery.ucsf.edu The type 1 TNFR (TNFR1)
contains a death domain through which it interacts with other death-domain
proteins to promote cellular responses. However, signaling through
death-domain proteins does not explain how TNFR1 induces the tyrosine
phosphorylation of intracellular proteins, which are important to cellular
responses induced by TNFR1. In this study, we show that TNFR1 associates
with Jak2, c-Src, and PI3K in various cell types. Jak2 and c-Src
constitutively associate with and are constitutively active in the TNFR1
complex. Stimulation with TNF induces a time-dependent change in the level
of Jak2, c-Src, and PI3K associated with TNFR1. The tyrosine kinase
activity of the complex varies with the level of tyrosine kinase associated
with TNFR1. TNFR1/c-Src plays a role in activating Akt, but not JNK or p38
MAPK, whereas TNFR1/Jak2 plays a role in activating p38 MAPK, JNK, and Akt.
TNFR1/c-Src, but not TNFR1/Jak2, plays an obligate role in the activation
of NF-kappaB by TNF, whereas TNFR1/Jak2, but not TNFR1/c-Src, plays an
obligate role in the activation of STAT3. Activation of TNFR1 increased the
expression of vascular endothelial growth factor, p21(WAF1/CIP1), and
manganese superoxide dismutase in MCF7 breast cancer cells, and increased
the expression of CCl2/MCP-1 and IL-1beta in THP-1 macrophages. Inhibitors
of Jak2 and c-Src impaired the induction of each of these target proteins.
These observations show that TNFR1 associates with and uses nonreceptor
tyrosine kinases to engage signaling pathways, activate transcription
factors, and modulate gene expression in cells. PMID: 18606683 [PubMed - in
process] Xi
Bao Yu Fen Zi Mian Yi Xue Za Zhi. 2007 Feb;23(2):172-5. [Article in Chinese] Yin
BJ, Yu
MX, Wang
J, Jiang
XD, Li
ZY. Department of Immunology,
Tongji Medical College of Huazhong University of Science and Technology,
Wuhan 430030, China. AIM: To investigate the biological
characteristics and effect of TNF-alpha binding peptide (TBP) and TNFR
blocking peptide (TRBP) in vitro. METHODS: The binding specificity of TBP
or TRBP was tested by competition experiment using GFP-TNF fusion protein
and detected by FCM and fluorescent microscope. The interaction between TBP
and TRBP was determined by non-denatured PAGE and the inhibitory effect of
TBP and TRBP on TNF-alpha cytotoxicity against U937 was carried out by MTT
colorimetry. The effects of TBP and TRBP on the functions of human
monocytes activated by TNF-alpha and IFN-gamma in vitro were detected by
nitrotetrazolium blue chloride (NBT) reduction assay for evaluating
respiratory burst and by RT-PCR for evaluating IL-1beta and IL-8 mRNA
transcription. RESULTS: It was showed that TBP and TRBP was able to block
the GFP-TNF binding to the TNFR on the surface of cells and no binding
interaction took place between TBP and TRBP. Both TBP and TRBP were able to
inhibit the TNF-alpha cytotoxicity against U937 and this inhibitory effect
was dose-dependent and the combination of TBP and TRBP (pep.38+X4) was able
to inhibit the TNF-alpha cytotoxicity more efficiently than the individual
peptide at all tested concentrations. The combination of TBP and TRBP was
able to obviously inhibit both respiratory burst of monocytes induced by
TNF-alpha and IFN-gamma and transcription level of IL-1beta and IL-8 mRNA
induced by TNF-alpha. CONCLUSION: TBP or TRBP can bind with their ligands
specifically and don't interact with each other. They can also block the
biological activity of the TNF-alpha in vitro, and the combination of TBP
and TRBP is able to inhibit the biological activity of the TNF-alpha more
efficiently. This research will provide an experimental basis for the
clinical treatment of the inflammatory disease. PMID: 17286914 [PubMed - in
process] Biochemistry.
2005 Nov 8;44(44):14509-18. Mol
Psychiatry. 2006 Jan;11(1):99-113. Pain.
2005 Oct;117(3):292-303. Neuroscience.
2006;137(1):287-99. Epub 2005 Nov 10. Gynecol
Oncol. 2006 Apr 20; [Epub ahead of print] : Neurochem
Int. 2006 Apr 16; [Epub ahead of print]
p55 Tumour necrosis factor receptor in bone
marrow-derived cells promotes atherosclerosis development in low-density
lipoprotein receptor knock-out mice.
Type 1 TNF receptor forms a complex with and
uses Jak2 and c-Src to selectively engage signaling pathways that regulate
transcription factor activity.
[Biological characteristics and effect of
TNF-alpha binding peptide and TNFR blocking peptide in vitro]
Expression of functional G
protein-coupled receptors in photoreceptors of transgenic Xenopus laevis.
Zhang
L, Salom
D, He
J, Okun
A, Ballesteros
J, Palczewski
K, Li
N.
Novasite Pharmaceuticals, Inc., San Diego, California 92121, USA.
G protein-coupled receptors (GPCRs) constitute the largest superfamily of
transmembrane signaling proteins; however, the only known GPCR crystal structure
is that of rhodopsin. This disparity reflects the difficulty in generating
purified GPCR samples of sufficient quantity and quality. Rhodopsin, the
light receptor of retinal rod neurons, is produced in large amounts of
homogeneous quality in the vertebrate retina. We used transgenic Xenopus
laevis to convert these retina rod cells into bioreactors to successfully
produce 20 model GPCRs. The receptors accumulated in rod outer segments and
were homogeneously glycosylated. Ligand and [(35)S]GTPgammaS binding assays
of the 5HT(1A) and EDG(1) GPCRs confirmed that they were properly folded
and functional. 5HT(1A)R was highly purified by taking advantage of the
rhodopsin C-terminal immunoaffinity tag common to all GPCR constructs. We
have also developed an automated system that can generate hundreds of
transgenic tadpoles per day. This expression approach could be extended to
other animal model systems and become a general method for the production
of large numbers of GPCRs and other membrane proteins for pharmacological
and structural studies.
PMID: 16262251 [PubMed - indexed for MEDLINE]
Overexpression of the Drosophila
vesicular monoamine transporter increases motor activity and courtship but
decreases the behavioral response to cocaine.
Chang
HY, Grygoruk
A, Brooks
ES, Ackerson
LC, Maidment
NT, Bainton
RJ, Krantz
DE.
Department of Psychiatry and Biobehavioral Sciences, Gonda (Goldschmied)
Center for Genetic and Neuroscience Research, Geffen School of
Medicine-UCLA, University of California at Los Angeles, 695 Charles Young
Drive, Los Angeles, CA 90095-1761, USA.
Aminergic signaling pathways have been implicated in a variety of
neuropsychiatric illnesses, but the mechanisms by which these pathways
influence complex behavior remain obscure. Vesicular monoamine transporters
(VMATs) have been shown to regulate the amount of monoamine
neurotransmitter that is stored and released from synaptic vesicles in
mammalian systems, and an increase in their expression has been observed in
bipolar patients. The model organism Drosophila melanogaster provides a
powerful, but underutilized genetic system for studying how dopamine (DA)
and serotonin (5HT) may influence behavior. We show that a Drosophila
isoform of VMAT (DVMAT-A) is expressed in both dopaminergic and
serotonergic neurons in the adult Drosophila brain. Overexpression of
DVMAT-A in these cells potentiates stereotypic grooming behaviors and
locomotion and can be reversed by reserpine, which blocks DVMAT activity,
and haloperidol, a DA receptor antagonist. We also observe a prolongation
of courtship behavior, a decrease in successful mating and a decrease in
fertility, suggesting a role for aminergic circuits in the modulation of
sexual behaviors. Finally, we find that DMVAT-A overexpression decreases
the fly's sensitivity to cocaine, suggesting that the synaptic machinery
responsible for this behavior may be downregulated. DVMAT transgenes may be
targeted to additional neuronal pathways using standard Drosophila
techniques, and our results provide a novel paradigm to study the
mechanisms by which monoamines regulate complex behaviors relevant to
neuropsychiatric illness.
PMID: 16189511 [PubMed - indexed for MEDLINE]
Spinal-supraspinal serotonergic circuits
regulating neuropathic pain and its treatment with gabapentin.
Suzuki
R, Rahman
W, Rygh
LJ, Webber
M, Hunt
SP, Dickenson
AH.
Department of Pharmacology Medical Sciences Building, University College
London, Gower Street, London WC1E 6BT, UK. ucklrsu@ucl.ac.uk
Not all neuropathic pain patients gain relief from current therapies that
include the anticonvulsant, gabapentin, thought to modulate calcium channel
function. We report a neural circuit that is permissive for the
effectiveness of gabapentin. Substance P-saporin (SP-SAP) was used to selectively
ablate superficial dorsal horn neurons expressing the neurokinin-1 receptor
for substance P. These neurons project to the brain as shown by retrograde
labelling and engage descending brainstem serotonergic influences that
enhance spinal excitability via a facilitatory action on 5HT(3) receptors.
We show the integrity of this pathway following nerve injury contributes to
the behavioural allodynia, neuronal plasticity of deep dorsal horn neurons
and the injury-specific actions of gabapentin. Thus SP-SAP attenuated the
tactile and cold hypersensitivity and abnormal neuronal coding (including
spontaneous activity, expansion of receptive field size) seen after spinal
nerve ligation. Furthermore the powerful actions of gabapentin after
neuropathy were blocked by either ablation of NK-1 expressing neurones or
5HT(3) receptor antagonism using ondansetron. Remarkably, 5HT(3) receptor
activation provided a state-dependency (independent of that produced by
neuropathy) allowing GBP to powerfully inhibit in normal uninjured animals.
This circuit is therefore a crucial determinant of the abnormal neuronal
and behavioural manifestations of neuropathy and importantly, the efficacy
of gabapentin. As this spino-bulbo-spinal circuit contacts areas of the
brain implicated in the affective components of pain, this loop may
represent a route by which emotions can influence the degree of pain in a
patient, as well as the effectiveness of the drug treatment. These
hypotheses are testable in patients.
PMID: 16150546 [PubMed - indexed for MEDLINE]
Serotonin1A autoreceptor activation by S
15535 enhances circadian activity rhythms in hamsters: evaluation of
potential interactions with serotonin2A and serotonin2C receptors.
Gannon
RL, Millan
MJ.
Department of Biology, Idle Hour Boulevard, Dowling College, Oakdale, NY
11769, USA. gannonr@dowling.edu
Mammalian circadian activity rhythms are generated by pacemaker cells in
the suprachiasmatic nucleus (SCN). As revealed by the actions of diverse
agonists, serotonergic input from raphe nuclei generally inhibits photic
signaling in the suprachiasmatic nucleus. In contrast, the serotonin
(5HT)1A partial agonist, 4-(benzodioxan-5-yl)1-(indan2-yl)piperazine (S
15535), was found to enhance the phase-shifting influence of light on
hamster circadian rhythms [Gannon, Neuroscience 119 (2003) 567]. Herein, we
extend this observation in showing that S 15535 (5.0 mg/kg, i.p.) markedly
(275%) enhanced the light-induced phase shift in circadian activity
rhythms: further, this action was dose-dependently abolished by the
highly-selective 5HT1A receptor antagonist, WAY 100,635
(N-[2-[4-(2-methoxyphenyl)-1-piperazinyl]ethyl]N-2-pyridinyl-cyclohexane-carboxamide
maleate) (0.1-0.5 mg/kg, i.p.). WAY 100,635, which was inactive alone,
shares the antagonist actions of S 15535 at postsynaptic 5HT1A sites, yet
blocks its effects at their presynaptic counterparts. Thus, 5HT1A
autoreceptor activation must be involved in this effect of S 15535 which
contrasts with the opposite, inhibitory influence upon phase shifts of the
"full" agonist, 8-OH-DPAT, which acts by stimulation of
postsynaptic 5HT1A receptors [Rea et al., J Neurosci 14 (1994) 3635].
Despite the occurrence of 5HT2A and 5HT2C receptors in the (rat)
suprachiasmatic nucleus, their influence on circadian rhythms is unknown
since actions of selective ligands have never been evaluated. This issue
was investigated with the most selective agents currently available.
However, the 5HT2A agonist, 1-(2,5-dimethoxy-4-iodophenyl)-2-aminopropane
(DOI) (0.25 and 0.5 mg/kg), and the 5HT2C agonist, alphaS-6-chloro-5-fluoro-a-methyl-1H-indole-1-ethanamine
fumarate (Ro-60-0175) (1.0 and 5.0 mg/kg), failed to affect light-induced
phase shifts in hamsters. Moreover, even over broad dose-ranges, the 5HT2A
antagonist, (+)-(2,3-dimethoxy-phenyl)-[1-[2-(4-fluoro-phenyl)-ethyl]-piperidin-4-yl]methanol
(MDL 100,907) (0.1-1.0 mg/kg), and the 5HT2C antagonist,
6-chloro-5-methyl-1-[6-(2-methylpyridin-3-yloxy)pyridin-3-yl
carbamoyl]indoline (SB 242,084) (1.0-10.0 mg/kg), were likewise inactive.
In view of evidence that 5HT2A and 5HT2C sites functionally interact with
5HT1A receptors, we also examined the influence of these agents upon the
actions of S 15535, but no significant alteration was seen in its
enhancement of rhythms. In conclusion, S 15535 elicits a striking enhancement
of light-induced phase shifts in circadian rhythms by specifically
recruiting 5HT1A autoreceptors, which leads to suppression of serotonergic
input to the suprachiasmatic nucleus. Surprisingly, no evidence for a role
of 5HT2A or 5HT2C sites was found, though comparable functional studies
remain to be undertaken in rats. Indeed, the present work underlines the
importance of comparative studies of circadian rhythms in various species,
as well as the need for further study of potential interactions among 5HT
receptor subtypes in their control.
Expression of chemokine CXCL12 and its receptor
CXCR4 in human epithelial ovarian cancer: An independent prognostic factor
for tumor progression.
Jiang
YP, Wu
XH, Shi
B, Wu
WX, Yin
GR.
Department of Obstetrics and Gynecology, the Second Hospital of Hebei
Medical University, Shijiazhuang, China.
OBJECTIVES.: Chemokine CXCL12 and its unique receptor CXCR4 have been
recently implicated in cancer metastasis. Our goal was to explore
expression of CXCL12 and CXCR4 protein in normal ovarian surface
epithelium, primary tumors and paired metastases of epithelial ovarian
cancer as well as its association with clinicopathological features. We also
wanted to test if expression of CXCR4 has prognostic value in epithelial
ovarian cancer patients. METHOD.: Sections from 6 normal ovarian surface
epithelium, 44 primary epithelial ovarian tumors and 30 paired metastatic
tumors in omentum were evaluated for CXCL12 and CXCR4 expression using
immunohistochemistry (IHC). RESULTS.: All samples of normal ovarian surface
epithelium were negative for CXCL12 and CXCR4 protein. Ovarian cancer cells
mainly showed cytoplasmic staining of CXCL12 and CXCR4. CXCL12 and CXCR4
staining were detected in 40/44 (91%) and 26/44 (59%) patients with primary
epithelial ovarian tumors respectively. CXCR4 expression in primary tumors
had no significant correlation with lymph nodes metastasis. However, if we
combined CXCR4 expression in primary tumors with metastatic tumors, a
significant correlation with lymph nodes metastasis was found (P = 0.018).
The intensity of CXCL12 staining correlated with ascites (P = 0.014). The
rate of CXCR4 expression in refractory and recurrent group (81% versus 28%,
P = 0.0008) was significantly higher than that in no-recurrent group. After
a median follow-up of 37 months, CXCR4 expression was found associated with
an unfavorable prognosis with significantly reduced median disease
progression-free survival and overall survival of 15 and 27 months (P =
0.0004, P = 0.017) respectively. Median time-to-event was not reached in
patients with negative CXCR4 staining. In multivariate analysis, CXCR4
expression and residual tumor size emerged as independent prognostic
factors in epithelial ovarian cancer patients. CONCLUSIONS.: This article
provides the first evidence that CXCR4 expression could be an independent
prognostic factor for epithelial ovarian cancer patients.
PMID: 16631235 [PubMed - as supplied by publisher]
Expression of CXC chemokine receptors 1-5
and their ligands in human glioma tissues: Role of CXCR4 and SDF1 in glioma
cell proliferation and migration.
Bajetto
A, Barbieri
F, Dorcaratto
A, Barbero
S, Daga
A, Porcile
C, Ravetti
JL, Zona
G, Spaziante
R, Corte
G, Schettini
G, Florio
T.
Department of Oncology, Biology and Genetics, University of Genova, Viale
Benedetto XV, 2, 16132 Genova, Italy.
Chemokines have been involved in cellular processes associated to malignant
transformation such as proliferation, migration and angiogenesis. The
expression of five CXC chemokine receptors and their main ligands was
analysed by RT-PCR in 31 human astrocytic neoplasms. The mRNAs for all the
receptors analysed were identified in a high percentage of tumours, while
their ligands showed lower expression. CXCR4 and SDF1 were the most
frequently mRNA identified (29/31 and 13/31 of the gliomas studied,
respectively). Thus, we further analysed the cell localization of CXCR4 and
SDF1 in immunohistochemistry experiments. We show a marked co-localization
of CXCR4 and SDF1 in tumour cells, mainly evident in psudolpalisade and
microcystic degeneration areas and in the vascular endothelium. In
addition, hSDF1alpha induced a significant increase of DNA synthesis in
primary human glioblastoma cell cultures and chemotaxis in a glioblastoma
cell line. These results provide evidence of the expression of multiple CXC
chemokines and their receptors in brain tumours and that in particular
CXCR4 and SDF1 sustain proliferation and migration of glioma cells to
promote malignant progression.
PMID: 16621164 [PubMed - as supplied by publisher]
2008/8/25