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REGULAR ARTICLES |
B Decoy Suppresses Experimental Crescentic Glomerulonephritis
*
Department of Geriatric Medicine, Osaka University Medical School, Suita,
Japan
Gene Therapy Science, Osaka University Medical School, Suita,
Japan
Department of Nephrology, Monash Medical Centre, Clayton, Victoria,
Australia
§
Dainippon Pharmaceutical Co., Ltd., Suita, Japan.
Correspondence to Dr. Toshio Ogihara, Department of Geriatric Medicine, Osaka University Medical School, 2-2 Yamada-oka, Suita 565, Japan. Phone : +81 6 6879 3852 ; Fax : +81 6 6879 3859 ; E-mail : ogihara{at}geriat.med.osakau.ac.jp
 |
Abstract |
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 Top Abstract IntroductionMaterials and MethodsResultsDiscussionReferences |
|---|
(TNF-), together with
leukocytic infiltration, are prominent features in crescentic
glomerulonephritis. Because these cytokines are targets for nuclear
transcription factor-
B (NF-B), the use of NF-B decoy
oligodeoxynucleotide (ODN) treatment was evaluated in an experimental disease
model. Crescentic glomerulonephritis was induced in primed Wistar rats by
injection of sheep antiglomerular basement membrane serum. Thirty minutes
after injection, rats were anesthetized and the left kidney was perfused with
NF-B decoy ODN or scrambled ODN control mixed with a virus-liposome
complex, and then killed 7 d later. Animals given the scrambled control ODN
developed severe glomerulonephritis by day 7 with heavy proteinuria,
glomerular crescents and interstitial lesions, marked leukocytic infiltration,
and upregulated renal expression of cytokines (IL-1 and TNF-) and
adhesion molecules (intercellular adhesion molecule-1). In contrast,
NF-B decoy ODN treatment substantially inhibited the disease with a 50%
reduction in proteinuria, a threefold reduction in histologic damage, a 50%
reduction in leukocytic infiltration, and a 50 to 80% reduction in the renal
expression of cytokines and leukocyte adhesion molecules. In conclusion, this
study has demonstrated that NF-B plays a key role in cytokine-mediated
renal injury and that NF-B decoy ODN treatment has clear therapeutic
potential in rapidly progressive glomerulonephritis. |
Introduction |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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[TNF-]), chemokines (monocyte chemoattractant protein-1), and leukocyte
adhesion molecules (intercellular adhesion molecule-1 [ICAM-1] and vascular
adhesion molecule-1)
(4,5,6,7,8).
Experimental models of crescentic glomerulonephritis, such as anti-glomerular
basement membrane (GBM) disease, have been shown to be leukocyte-dependent
(9). Furthermore, blocking the
action of IL-1 or TNF- in animal models of crescentic
glomerulonephritis has been shown to inhibit renal T cell and macrophage
infiltration and suppress renal injury, including crescent formation
(10,11,12,13,14).
The transcription factor nuclear factor-B (NF-B) is important
in the coordinated expression of various proinflammatory molecules, including
IL-1, TNF-, and ICAM-1
(15,
16). Therefore, we postulate
that inhibiting the action of NF-B using a synthetic decoy for the
cis-acting element in the promoter region of these genes can block
the underlying inflammatory response in crescentic glomerulonephritis. The
decoy approach to blocking transcription factor activity has been shown to be
an effective strategy for inhibiting specific gene expression in
vitro and in vivo
(17,18,19).
We have developed a gene therapy approach using oligodeoxynucleotides (ODN),
or plasmid DNA, complexed with hemagglutinating virus of Japan (HVJ) liposomes
that enables efficient transfection of various organs, such as the liver and
the vessel wall
(20,21,22,23).
This method has been refined to allow efficient transfection of glomerular
cells via perfusion of the renal artery
(24,
25). The aim of the current
study was to transfect glomerular cells with an NF-B decoy ODN to
determine whether NF-B is a key regulator of the inflammatory response
causing renal injury in an experimental model of crescentic
glomerulonephritis.
|
Materials and Methods |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
|---|
NF-B decoy ODN containing the NF-B consensus sequence
(underlined) :
Scrambled ODN :
The NF-B decoy ODN have been shown to bind to the NF-B
transcription factor (Figure
1b) (19).
Synthetic ODN were washed with 70% ethanol, dried, and dissolved in sterile
Tris-ethylenediaminetetra-acetic acid (EDTA) buffer (10 mM Tris, 1 mM EDTA).
The supernatant was purified over a Nick column (Pharmacia Biotech, Uppsala,
Sweden) and quantified by spectrophotometry. The single-stranded ODN were
annealed for 2 h while the temperature decreased from 80 to 25°C
(17,18,19).
FITC-labeled ODN were also prepared for the initial studies.
|
Gel Mobility Shift Assay
The cortex was isolated from the whole kidney, and a nuclear extract was
prepared as described previously
(17). Tissues were homogenized
using a Potte-Elvehjem homogenizer in 4 vol of ice-cold homogenization buffer
(10 mM Hepes, pH 7.5, 0.5 M sucrose, 0.5 mM spermidine, 0.15 mM spermin, 5 mM
EDTA, 0. 25 M ethyleneglycol-bis(ß-aminoethyl
ether)-N,N'-tetra-acetic acid, 7 mM ß-mercaptoethanol, and 1 mM
phenylmethylsulfonyl fluoride). After centrifugation at 12,000 x
g for 30 min at 4°C, the pellet was lysed in 1 vol of ice-cold
homogenization buffer containing 0.1% Nonidet P-40 using a Dounce homogenizer.
After centrifugation at 12, 000 x g for 30 min at 4°C,
nuclei were washed twice with ice-cold buffer containing 0.35 M sucrose. After
washing, nuclei were preextracted with 1 vol of ice-cold homogenization buffer
containing 0.05 M NaCl and 10% glycerol for 15 min at 4°C. The nuclei were
then extracted with homogenization buffer containing 0.3 M NaCl and 10%
glycerol for 1 h at 4°C, and the concentration of DNA was adjusted to 1
mg/ml. After pelleting the extracted nuclei at 12,000 x g for
30 min at 4°C, the supernatant was added to 45%
(NH4)6SO4 and stirred for 30 min at 4°C.
The precipitated protein was pelleted at 17,000 x g for 30 min,
resuspended in homogenization buffer containing 0.35 M sucrose, and stored in
aliquots at -70°C. NF-B and scrambled ODN probes were labeled with
32P using a 3' end-labeling kit (Clontech, Palo Alto, CA).
Labeled ODN probes were purified using a Nick column (Pharmacia Biotech).
Binding reactions (10 µl), including 32P-labeled probe (0.5 to 1
ng ; 10,000 to 15,000 cpm), and 1 µg of poly(deoxyinosinic-deoxycytidylic)
acid (Sigma Chemical Co., St. Louis, MO) were incubated with 10 µg of
nuclear extract for 30 min at room temperature, and then loaded onto a 5%
polyacrylamide gel. The gels were subjected to electrophoresis, drying, and
autoradiography.
Preparation of HVJ Liposomes
HVJ liposomes were prepared in a manner identical to that described
previously
(21,22,23,24).
Phosphatidylserine, phosphatidylcholine, and cholesterol were mixed in a
weight ratio of 1:4.8:2
(21,22,23,24).
The lipid mixture (10 mg) was deposited on the sides of a flask by removal of
tetrahydrofuran in a rotary evaporator. Dried lipid was hydrated in 200 µl
of balanced salt solution (137 mM NaCl, 5.4 mM KCl, 10 mM Tris-HCl, pH 7.6)
containing ODN. Liposomes were prepared by shaking and sonication. Purified
HVJ (Z strain) was inactivated by ultraviolet irradiation (110
erg/nm2 per s) for 3 min just before use. The liposome suspension
(0.5 ml, containing 10 mg of lipids) was mixed with HVJ (30,000
hemagglutinating units) in a total volume of 4 ml of balanced salt solution.
The mixture was incubated at 4°C for 5 min and then for 30 min with gentle
shaking at 37°C. Free HVJ was removed from the HVJ-liposomes by sucrose
density gradient centrifugation. The top layer of the sucrose gradient was
collected for use
(21,22,23,24).
Transfection of ODN-HVJ Liposomes into the Kidney
Male 8-wk-old Wistar rats weighing 150 g were purchased from Charles River
Japan (Osaka, Japan). Rats were anesthetized with pentobarbital, and the left
renal artery was surgically exposed. A cannula was introduced into the left
renal artery via the aorta. The artery was transiently isolated by temporary
ligatures, and the ODN-HVJ liposome complex was infused into the kidney over a
5-min period at room temperature. The infusion cannula was then removed, blood
flow to the renal artery was restored by release of the ligatures, and the
wound was closed. A series of preliminary studies was performed in normal rats
in which the left kidney was perfused with FITC-labeled ODN alone or
FITC-labeled ODN complexed with HVJ liposomes, and rats were killed after 1 h,
1 d, or 7 d ; the tissue was examined by immunofluorescence to determine
transfection of glomerular cells.
Rat Model of Crescentic Glomerulonephritis
A well characterized rat model of crescentic glomerulonephritis was used
(26). Disease was induced in
groups of six inbred male Wistar rats (150 g) by immunization with 5 mg of
normal rabbit IgG in Freund's complete adjuvant followed 5 d later (termed day
0) by intravenous injection of rabbit anti-GBM serum (50 µg IgG/100 g body
wt). Thirty minutes after the serum injection, the left kidney was perfused
with either NF-B decoy ODN or scrambled ODN complexed with HVJ
liposomes, as described above. Blood and 24-h urine collections were taken on
days 1, 3, 5, and 7. Animals were killed on day 7 and tissue was collected for
examination. A group of rats excluded from experimentation was used as the
normal control.
Twenty-four-hour urine collections were obtained from rats that were individually housed in metabolic cages. Rats were fasted during the collection period, but were allowed free access to water. Urine protein excretion was measured by a sulfosalicylic acid method.
Histologic Analysis and Immunohistochemistry Staining for
Leukocytes
Kidney tissue was fixed in 4% paraformaldehyde in phosphatebuffered saline
and embedded in paraffin. Hematoxylin and eosinstained 4-µm sections were
used to evaluate glomerular histology, as described previously
(26). Fifty glomeruli per
animal were assessed for glomerular cell number and crescent formation. The
degree of interstitial damage (fibrosis, tubular atrophy, and leukocytic
infiltration) was graded on a scale of 0 to 4 as described previously
(26). For detection of
leukocytes, snap-frozen tissue sections were fixed in 2% paraformaldehyde and
then stained with the OX-1 monoclonal antibody (anti-rat CD45)
(27), using a three-layer
peroxidase anti-peroxidase method with diaminobenzidine for color development
(26). The number of OX-1 +
leukocytes in 50 glomeruli per animal was counted under high power. In
addition, the number of interstitial OX-1 + cells in 20 high-power cortical
fields was counted (26). All
scoring was done on coded slides.
RNA Extraction and Northern Blot Analysis
Total cellular RNA was extracted from a half kidney using RNAzol (Tel-Test
Inc., Friendswood, TX). Samples (30 µg) were electrophoresed on a 1.5%
agarose-formaldehyde denaturing gel and transferred to a nitrocellulose
membrane (Amersham International, Buckinghamshire, United Kingdom). The filter
was baked, prehybridized, and hybridized with 32P-labeled
IL-1ß and TNF- oligonucleotide probes (Clontech), GAPDH
oligonucleotide probe (Clontech), and ICAM-1 cDNA probe (kindly donated by Dr.
Tetsuya Tomita, Osaka University, Osaka, Japan) as described previously
(28). Filters were washed
stringently and then exposed to x-ray film.
In Situ Hybridization
In situ hybridization was performed on 4-µm paraffin sections
using a microwave-based technology
(29). After dewaxing, sections
were placed in 0.01 M sodium citrate buffer, pH 6.0, and heated for 2 x
5 min in a microwave oven at 2450 MHz and a power output of 800 W. Sections
were then treated sequentially with 0.2 M HCl for 15 min, 1% Triton X-100 for
15 min, and digested for 20 min with 10 µg/ml proteinase K at 37°C
(Boehringer Mannheim, Mannheim, Germany). Sections were then washed in
2x SCC, prehybridized, and then hybridized with 0.3 ng/ml
digoxigenin-labeled sense or antisense IL-1ß and TNF- cRNA probe
overnight at 37°C in a hybridization buffer containing 50% deionized
formamide, 4x SSC, 2x Denhardt's solution, 1 mg/ml salmon sperm
DNA, and 1 mg/ml yeast tRNA. Sections were then washed in 0.1x SSC at
37°C, and the hybridized probe was detected using sheep anti-digoxigenin
antibody conjugated with alkaline phosphatase and color development with
nitroblue tetrazolium/X-phosphate (Boehringer Mannheim).
Statistical Analyses
All values are expressed as the mean ± SEM. ANOVA with subsequent
Dunnett test was used to determine significant differences in multiple
comparisons. P < 0.05 was considered significant.
|
Results |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
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B Decoy ODNB protein was easily detected in nuclear
extracts prepared from anti-GBM disease kidney, but was barely detectable in
normal rat kidney (Figure 1a).
The addition of excess unlabeled scrambled ODN had no effect on the binding of
labeled NF-B ODN to the extract from anti-GBM disease kidney, but this
binding was completely blocked by the addition of excess unlabeled NF-B
ODN (Figure 1b).
Transfection of Glomerular Cells using the HVJ Liposome
Technique
In preliminary experiments, the left kidney of normal rats was infused with
FITC-conjugated NF-B ODN complexed with HVJ liposomes. Animals were
killed at different times and the presence of ODN within the kidney was
assessed by immunofluorescence microscopy. As early as 1 h after infusion, 40
to 50% of glomeruli in the left kidney contained FITC-ODN. The ODN were still
present at 24 h (Figure 2), and
remained for 7 d (not shown). No glomerular staining was seen in the right
kidney, although some tubular uptake of FITC-ODN was evident
(Figure 2). Infusion of
FITC-ODN without HVJ liposomes was unable to transfect glomerular cells (not
shown).
|
NF-B Decoy ODN Inhibits Renal Injury in Crescentic
Glomerulonephritis
Rats infused with the scrambled ODN control developed crescentic
glomerulonephritis in terms of severe renal injury, as demonstrated by heavy
urinary protein excretion (Figure
3), and marked histologic damage, as shown by the presence of
glomerular hypercellularity, focal glomerular sclerosis, glomerular crescent
formation, and interstitial lesions (Figure
4 and Table 1). In
contrast, infusion with the NF-B decoy ODN caused an approximately 50%
reduction in urinary protein excretion over the 7-d period. There was also a
substantial reduction in histologic damage in the NF-B decoy
ODN-treated kidney compared with the scrambled ODN-treated kidney, including a
fivefold reduction in glomerular crescent formation
(Figure 4 and
Table 1).
|
|
|
NF-B Decoy ODN Inhibits Cytokine Production and Leukocytic
Infiltration in Crescentic Glomerulonephritis
Northern blot analysis showed that expression of IL-1ß, TNF-,
and ICAM-1 is increased in scrambled ODN-treated anti-GBM disease compared
with normal kidney, and reduced by treatment with the NF-B decoy ODN
(Figure 5). A more detailed
analysis of IL-1ß and TNF- expression was performed by in
situ hybridization. Constitutive expression of IL-1ß mRNA in normal
kidney was restricted to a small number of glomerular cells (mostly podocytes)
and a minority of tubules (Figures
6 and 7). There was
a dramatic increase in the number of glomerular cells expressing IL-1ß in
the scrambled ODN-treated anti-GBM disease (podocytes, infiltrating
leukocytes, and some mesangial cells), and most tubules showed IL-1ß mRNA
expression (Figures 6 and
7). Treatment with the
NF-B decoy ODN caused a substantial reduction in both glomerular and
tubular IL-1ß mRNA expression. The upregulation of TNF- mRNA
expression in anti-GBM disease was similar to that seen for IL-1ß, and
TNF- expression was also significantly reduced by NF-B decoy ODN
treatment (Figures 6 and
7).
|
|
|
Leukocyte infiltration in the kidney was assessed by immunoperoxidase
staining of tissue sections. The marked glomerular and interstitial leukocytic
infiltration seen in the scrambled ODN-treated animals was significantly
reduced, by approximately 50%, with NF-B decoy ODN treatment
(Figure 8).
|
|
Discussion |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
|---|
B plays a key role in regulating the inflammatory processes
underlying renal damage in experimental crescentic glomerulonephritis.
Treatment with the NF-B decoy ODN caused a 50% reduction in
proteinuria. Because the NF-B decoy ODN was infused into only one
kidney, this result documents a strong protective effect of decoy treatment on
urinary protein excretion. This is consistent with the marked reduction in
histologic damage, cytokine expression, and leukocyte infiltration in the
decoy-treated kidney versus the untreated kidney.
Previous studies in experimental anti-GBM crescentic glomerulonephritis
have shown that blocking the action of either IL-1 or TNF- inhibits
renal damage
(10,11,12,13,14).
These studies demonstrate that rather than being redundant, the cytokines IL-1
and TNF- act in an interdependent manner, presumably sharing a common
mechanism of action. This common mechanism may be the activation of the
nuclear transcription factor NF-B.
In vitro studies have clearly identified that many of the
proinflammatory actions of IL-1 and TNF- are mediated via activation of
NF-B
(16,30).
Furthermore, transcription of the genes for IL-1 and TNF- are also
regulated by NF-B, thereby providing an amplification mechanism during
the induction of an inflammatory response. A number of descriptive studies
have shown that NF-B is activated in experimental renal disease, and
that inhibition of renal injury in these models is associated with a reduction
in NF-B activation
(31,32).
The present study provides direct in vivo evidence that NF-B
is a key regulator of the underlying immune response in experimental
glomerulonephritis. Furthermore, this study provides direct evidence to
support the postulate that NF-B is a common pathway by which the
cytokines IL-1 and TNF- cause renal injury. Indeed, treatment of
anti-GBM disease with the NF-B decoy ODN produced a result similar to
that previously seen with IL-1 or TNF- blockade, i.e.,
suppression of leukocyte adhesion molecule expression, leukocytic
infiltration, and the associated renal damage
(10,11,12,13,14,
33).
Infusion of the renal artery with the NF-B decoy ODN
complex produced efficient transfection of glomerular cells, but relatively
little transfection of tubular epithelial cells. However, NF-B
decoy ODN treatment not only suppressed cytokine production and inflammation
within the glomerulus, but it also markedly inhibited tubular IL-1 ß and
TNF- expression, and the associated interstitial leukocytic infiltrate
and interstitial damage. This finding lends support to the hypothesis that
interstitial inflammation and damage in this disease model are secondary to
the release of glomerular cytokines
(34).
An interesting observation is that intrinsic renal cells, rather than just
leukocytes, are the major source of IL-1 and TNF- mRNA expression in
the kidney. This is consistent with studies of human crescentic
glomerulonephritis, in which IL-1 and TNF- have been shown to be
expressed by glomerular and tubular epithelial cells
(4,5,6).
This emphasizes the importance of direct transfection of resident glomerular
cells.
This study has shown that the NF-B decoy ODN is a powerful
inhibitor of the proinflammatory response in experimental crescentic
glomerulonephritis. This is a major step forward in the potential development
of molecular therapies for renal diseases. The kidney has proven to be a
difficult organ in which to achieve efficient cell transfection. Previous
attempts to transfect glomerular cells using adenoviral vectors have been
relatively unsuccessful (35).
Similarly, administration of naked ODN is unable to transfect glomerular
cells, resulting only in nonspecific tubular uptake. The use of HVJ liposomes
together with renal artery infusion has proven to be an effective means to
transfect glomerular cells with ODN, or plasmids, in this and previous studies
(24,25).
The next issues to address in developing molecular therapy for renal diseases
will be to prolong the stability of ODN within the transfected cells and to
enable repeated NF-B decoy ODN treatments.
The results of this study have implications for immune-mediated diseases
generally. Given that blockade of IL-1 or TNF- is an effective
treatment for a variety of different models of immune-mediated disease
(36), it is likely that
NF-B decoy ODN treatment will also be effective in such
diseases. This postulate is supported by the ability of antisense
oligonucleotides to the p65 subunit of NF-B to suppress
experimental inflammatory bowel disease
(37).
In summary, this study has demonstrated that in vivo treatment
with NF-B decoy ODN is both a feasible and effective treatment
to suppress renal damage in a model of rapidly progressive crescentic
glomerulonephritis. Furthermore, the study provides direct in vivo
evidence that activation of NF-B is a common mechanism
underlying inflammatory renal damage.
|
Acknowledgments |
|---|
|
References |
|---|
|
TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
|---|
, IL-1 ß and IL-2R in ANCA-positive
glomerulonephritis. Kidney Int43
: 682-692,1993[Medline]
expression in human glomerulonephritis. Nephrology3
: 329-337,1997
and IL-1 ß in human glomerulonephritides.
Kidney Int 52 :393
-403, 1997[Medline]
and a protease inhibitor reduce immune glomerular
injury. Kidney Int 40 :43
-51, 1991[Medline]
upregulates renal MIF expression in rat
crescentic glomerulonephritis. Mol Med3
: 136-144,1997[Medline]
B : A pleiotropic
mediator of inducible and tissue specific gene control.
Cell 58 :227
-229, 1989[Medline]
B : A pivotal
transcription factor in chronic inflammatory diseases. N Engl J
Med 336 :1066
-1071, 1997B binding site
prevents myocardial infarction. Nat Med3
: 894-899,1997[Medline]
B and AP-1 activation by glucocorticoids in
experimental glomerulonephritis in rats : Molecular mechanisms of
anti-nephritic action. Biochim Biophys Acta1362
: 252-262,1997[Medline]
B
activation in the kidney with ureteral obstruction. Kidney
Int 52 : 926-933,1997[Medline]
B abrogates established experimental colitis in mice.
Nat Med 2 :998
-1004, 1996[Medline]
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, scrambled ODN-treated rats ; •,
NF-
, normal rat kidney ; 