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Anti-monocyte chemoattractant protein-1 gene therapy inhibits vascular remodeling in rats: blockade of MCP-1 activity after intramuscular transfer of a mutant gene inhibits vascular remodeling induced by chronic blockade of NO synthesis

KENSUKE EGASHIRA¹, MASAMICHI KOYANAGI, SHIRO KITAMOTO, WEIHUA NI, CHU KATAOKA, RYUICHI MORISHITA, YASUFUMI KANEDA, CHIYUKI AKIYAMA, KEN-ICHI NISHIDA, KATSUO SUEISHI^* and AKIRA TAKESHITA

Department of Cardiovascular Medicine and
^* Pathology, Graduate School of Medical Sciences, Kyushu University, Fukuoka, Japan;
Division of Gene Therapy Sciences, Osaka University Medical School, Osaka, Japan; and
New Product Research Laboratories, Daiichi Pharmaceutical Co., Tokyo, Japan

¹Correspondence: Department of Cardiovascular Medicine, Graduate School of Medical Science, Kyushu University, 3-1-1, Maidashi, Higashi-ku, Fukuoka 812-8582, Japan. E-mail: egashira{at}cardiol.med.kyushu-u.ac.jp

	ABSTRACT

TOP ABSTRACT INTRODUCTION MATERIALS AND METHODS RESULTS Effects of the i.m.... DISCUSSION REFERENCES

 
Monocyte chemoattractant protein-1 (MCP-1) may play an essential part in the formation of arteriosclerosis by recruiting monocytes into the arterial wall. Thus, we devised a new strategy for anti-MCP-1 gene therapy against arteriosclerosis by transfecting an amino-terminal deletion mutant (missing the amino-terminal amino acids 2 to 8) of the human MCP-1 gene into a remote organ (skeletal muscles). Intramuscular transduction with the mutant MCP-1 gene blocked monocyte recruitment induced by a subcutaneous injection of recombinant MCP-1. In a rat model in which the chronic inhibition of endothelial nitric oxide synthesis induces early vascular inflammation as well as subsequent coronary vascular remodeling, this strategy suppressed monocyte recruitment into the coronary vessels and the development of vascular medial thickening, but did not reduce perivascular fibrosis. Thus, MCP-1 is necessary for the development of medial thickening but not for fibrosis in this model. This new strategy may be a useful and feasible gene therapy against arteriosclerosis.—Egashira, K., Koyanagi, M., Kitamoto, S., Ni, W., Kataoka, C., Morishita, R., Kaneda, Y., Akiyama, C., Nishida, K.-i., Sueishi, K., Takeshita, A. Anti-monocyte chemoattractant protein-1 gene therapy inhibits vascular remodeling in rats: blockade of MCP-1 activity after intramuscular transfer of a mutant gene inhibits vascular remodeling induced by chronic blockade of NO synthesis.

Key Words: endothelium-derived relaxing factors • remodeling • growth substances • inflammation • adhesion molecule • monocyte chemoattractant protein-1 • gene transfer

	INTRODUCTION

TOP ABSTRACT INTRODUCTION MATERIALS AND METHODS RESULTS Effects of the i.m.... DISCUSSION REFERENCES

 
ARTERIOSCLEROSIS IS RECOGNIZED to be a chronic inflammatory disease, because the infiltration and activation of mononuclear cells in blood vessels are critical steps in an early stage of atherosclerosis (1) . Unstable atheromas also represent such inflammatory lesions (2) . Monocyte chemoattractant protein-1 (MCP-1) is a member of the C-C chemokine family and a potent chemotactic factor for monocytes (3) . Recent studies have demonstrated that MCP-1 expression is increased in arteriosclerotic and atherosclerotic lesions (4 , 5) and that eliminating MCP-1 expression decreases atheroma formation in hypercholesterolemic mice (6 , 7) . These studies clearly indicate that MCP-1 plays a central part in atherogenesis and suggest that anti-MCP-1 therapy may be a potential form of gene therapy against arteriosclerosis or atherosclerosis.

We performed the present study to evaluate the use of gene therapy to block MCP-1 activity in vivo using an amino-terminal deletion mutant of MCP-1 called 7ND, which lacks the amino-terminal amino acids 2 to 8. This mutant MCP-1 has been shown to bind to the receptor for MCP-1 (CCR2) and subsequently block MCP-1-mediated monocyte chemotaxis (3 , 8) . We hypothesized that for this approach to work, the cells infected with 7ND must secrete 7ND protein into the circulating blood and the 7ND protein must bind to the MCP-1 receptor on monocytes or target cells in remote organs, and thus block the signal of MCP-1. Such blockade of MCP-1 activity would suppress MCP-1-mediated inflammation and thereby improve the function of the target organs. If this approach proves successful, direct gene transfer into the target organ will not be necessary. The use of skeletal muscle as a biofactory to produce a secreted protein has been reported previously (9) . Therefore, we tested the effectiveness of this new strategy in a rat model in which the chronic inhibition of nitric oxide synthesis (NO) by the administration of N-nitro-L-arginine methyl ester (L-NAME) induces early vascular inflammation (monocyte infiltration into the blood vessel wall and MCP-1 expression) and subsequently causes arteriosclerosis (medial thickening and fibrosis) (10 11 12 13) . Inhibition of NO synthesis has been shown to up-regulate cell adhesion molecules and/or MCP-1 in cultured endothelial cells through the increases in oxidative stress and/or the activity of nuclear factor B (14 15 16 17) .

	MATERIALS AND METHODS

TOP ABSTRACT INTRODUCTION MATERIALS AND METHODS RESULTS Effects of the i.m.... DISCUSSION REFERENCES

 
The expression vector and the preparation of HVJ–liposome complexes
Human 7ND cDNA with or without an epitope tag FLAG in the carboxyl-terminal was constructed by recombinant polymerase chain reaction using a wild type MCP-1 cDNA (a gift of Dr. Teizo Yoshimura, NCI, Frederick, Md.) as template and cloned into the BamHI(5') and NotI(3') sites of the pCDNA3 expression (Invitrogen) vector plasmid. Twenty four nucleotides encoding FLAG epitope(DYKDDDDK) were added directly at the 3' terminus of MCP-1 sequence. All sequences were confirmed by double-stranded DNA sequencing. HVJ-liposomes containing the plasmid DNA were prepared as described (9) .

An animal model of chronic inhibition of NO synthesis
Three groups of WKY rats were studied: The control rats received untreated chow and drinking water. The second group received L-NAME after an intramuscular (i.m.) injection of phosphate-buffered saline (PBS). The third group received L-NAME after an i.m. injection of HVJ–liposome complex containing the 7ND gene. On the 3rd day or 28th day of treatment, the systolic blood pressure was measured by the tail-cuff method. Venous blood was then collected and the rats were killed for morphometric, immunohistochemical, and biochemical analysis. The hearts were isolated and either fixed in a methacarn solution for histological analysis or snap-frozen in liquid nitrogen and stored at -80°C.

Histopathology and immunohistochemistry
Tissue sections were either stained with Masson-trichrome or subjected to immunostaining using antibodies against macrophage/monocyte (ED1, Serotec, Berlin, Germany), proliferating cell nuclear antigen (PCNA) (Dako, Carpinteria, Calif.), or non-immune mouse IgG (Zymed, San Francisco, Calif.). The slides were washed and incubated with biotinylated, affinity-purified goat anti-rabbit IgG. After avidin-biotin amplification, the slides were incubated with 3',3'-diaminobenzidine and counterstained with hematoxylin.

In situ hybridization
Four rats in the control and L-NAME+PBS groups were killed on the third day of treatment. Digoxigenin-labeled single-strand RNA probes (sense and antisense) were generated using a DIG RNA labeling kit (Boehringer Mannheim, Mannheim, Germany) according to the manufacturer’s protocol. Rat MCP-1 probes, a 665 or 930 base pair fragment of the rat MCP-1 cDNA, were used. In situ hybridization was performed on 2% paraformaldehyde-embedded sections as described (18) .

Morphometry and cell counting
Morphometry and cell counting were performed by a single observer who was blind to the treatment protocols. Each section (five per heart) stained with an antibody against ED1 or PCNA was scanned. The number of positive cells in each section was determined. The average number of positive cells per section was determined for each animal.

To evaluate the thickening of the coronary arterial wall and the extent of perivascular fibrosis, short-axis images of the coronary arteries were analyzed. (10 , 11 , 13) The inner border of the lumen and the outer border of the tunica media were traced from the Masson-trichrome stained sections. The wall-to-lumen ratio (the ratio of the medial thickness to the internal diameter) and the area of fibrosis (collagen deposition stained with aniline blue) immediately surrounding the blood vessels were then calculated. Perivascular fibrosis was estimated as the ratio of the area of fibrosis surrounding the vessel wall to the total vessel area.

Northern blot analysis
Total RNA was extracted from each sample, poly(A)+ RNA was purified, and then Northern blot hybridization was performed as we have described previously (11) . The cDNA probes used were human 7ND (8) and mouse GAPDH (American Type Culture Collection, Rockville, Md.).

Western blot analysis
After immunoprecipitation, the FLAG Western Detection kit (Stratagene, #200470) was used to detect FLAG protein in the serum.

Statistical analysis
Data are expressed as the mean ± SE. Statistical analysis of differences was compared by ANOVA and Bonferroni’s multiple comparison tests. A level of P < 0.05 was considered statistically significant.

	RESULTS

TOP ABSTRACT INTRODUCTION MATERIALS AND METHODS RESULTS Effects of the i.m.... DISCUSSION REFERENCES

 
Effects of the i.m. expression of 7ND on monocyte recruitment into the dermis induced by recombinant MCP-1
Male Wistar-Kyoto rats were anesthetized and their femoral muscle was exposed. An HVJ-liposome solution (5 µg of encapsulated human 7ND DNA) or PBS was injected into the muscles. We examined transgene expression after injection of 7ND or PBS into the femoral muscles (Fig. 1A ). Marked transgene expression with 7ND was observed in the femoral muscle (the site of injection) from the 7ND-transfected group. No such expression of the 7ND gene was detected in the muscles from the PBS control group. We further examined the 7ND protein production after i.m. injection of a carboxyl-terminal FLAG epitope-tagged 7ND gene. Western blot analysis of the serum for FLAG protein showed that FLAG/7ND protein was secreted into the serum (Fig. 1B ).

View larger version (49K): [in this window] [in a new window]   Figure 1. A) Northern blot analysis of the mutant MCP-1 (7ND) and GAPDH mRNA expression in femoral muscles after an i.m. injection of the 7ND gene or PBS. This is a representative assay from five separate experiments (n=5). The mRNA levels are assessed at the indicated times. B) Western blot analysis of the 7ND protein in the serum after an i.m. injection of a carboxyl-terminal epitope-tagged 7ND gene or PBS. This is a representative assay from five separate experiments (n=5). The FLAG/7ND protein levels are assessed at the indicated times. C) The number of ED1-positive monocytes infiltrating into the dermis induced by an intradermal injection of recombinant human MCP-1 (1 µg per 100 µl) or vehicle. *P < 0.01 vs. vehicle. P < 0.01 vs. the PBS-injected group. Mean ± SE (n=6 each) are shown.

Three days after rats were injected with the 7ND gene or PBS, recombinant human MCP-1 (1 µg/20 µl) or vehicle (PBS 20 µl) was injected into the dermis. Twenty-four hours after the intradermal injection, histopathologic sections of the injected sites were prepared and the ED1-positive monocytes that were recruited into the injected site were counted (19) . In the rats receiving PBS, the number of monocytes recruited into the dermis was significantly greater in the areas of MCP-1 injection than in the areas of vehicle injection (Fig. 1C ). This increase in ED1-positive monocytes was blocked by the i.m. injection of the 7ND gene.

	Effects of the i.m. expression of 7ND on monocyte recruitment and coronary vascular remodeling

TOP ABSTRACT INTRODUCTION MATERIALS AND METHODS RESULTS Effects of the i.m.... DISCUSSION REFERENCES

 
Rats were injected with either the 7ND gene or PBS, then treated with L-NAME in their drinking water (1 mg/ml). The control group received untreated chow and drinking water. Three days after the L-NAME administration was begun, the rats treated with L-NAME+PBS had a marked infiltration of ED1-positive monocytes into the intima and adventitia of their coronary arteries (Fig. 2 ). To assess proliferation, PCNA staining was performed in these cells. Nuclear staining for PCNA antibody was observed in some endothelial cells, vascular smooth muscle cells in the media, and monocytes or myofibroblast-like cells (Fig. 2) . No such inflammatory and proliferative changes were observed in the control group. In situ hybridization indicated that the MCP-1 induction was confined to the endothelium and some infiltrating monocytes in the L-NAME+PBS group (Fig. 2) . In rats treated with L-NAME+7ND, such inflammatory and proliferative changes were markedly suppressed (Fig. 2 and Fig. 3 ). This inhibitory effect was observed in all areas of the heart and vessels.

View larger version (53K): [in this window] [in a new window]   Figure 2. Histopathology, immunohistochemistry, and in situ hybridization of the coronary arteries. A) In situ hybridization of coronary artery sections with riboprobes for MCP-1 mRNA. The endothelium of the coronary arteries and the cells (possibly monocytes) infiltrating into the intima and adventitia from a rat receiving L-NAME stained strongly for MCP-1 mRNA with an antisense riboprobe. There was no staining with a sense riboprobe. Bar, 20 µm. B) Coronary artery sections from a control rat, a rat receiving L-NAME+PBS, and a rat receiving L-NAME+7ND after 3 days are stained with Hematoxylin-eosin (HE) or immunohistochemically for monocyte/macrophage (ED1), proliferating cells (PCNA), and nonimmune IgG (negative control). The bar indicates 100 µm. C) Coronary artery sections stained with Masson-trichrome stain from a control rat, a rat receiving L-NAME+PBS, and a rat receiving L-NAME+7ND at 28 days. The bar indicates 100 µm.

View larger version (18K): [in this window] [in a new window]   Figure 3. Effect of the i.m. transfer of the 7ND gene on the inflammatory and proliferative changes on day 3. A) The number of ED1-positive monocytes infiltrating into the coronary vessels and myocardium. B) The number of PCNA-positive cells appearing in the coronary vessels and myocardium. *P < 0.01 vs. the control group. P < 0.01 vs. the L-NAME+PBS group.

On day 28, significant structural changes in the coronary arteries were evident in rats treated with L-NAME+PBS. Treatment with L-NAME+7ND prevented any increase in medial thickening (the wall-to-lumen ratio) but progression of perivascular and cardiac fibrosis was unaffected (Fig. 2 and Fig. 4 ).

View larger version (19K): [in this window] [in a new window]   Figure 4. Effect of the i.m. transfer of the 7ND gene on vascular remodeling (medial thickening and perivascular fibrosis) on day 28. A) Medial thickening (the wall-to-lumen ratio) of the coronary arteries. B) Perivascular fibrosis of the coronary arteries. *P < 0.01 vs. the control group. P < 0.01 vs. the L-NAME+PBS group.

Compared with the control group, the L-NAME+ 7ND-transfected and L-NAME+PBS groups had a higher systolic arterial pressure on days 3 and 28 of treatment (Table 1 ). There was no significant difference in the white blood cell count in peripheral blood among the three groups (data not shown).

	DISCUSSION

TOP ABSTRACT INTRODUCTION MATERIALS AND METHODS RESULTS Effects of the i.m.... DISCUSSION REFERENCES

 
In the present study, we devised a new strategy to block the action of MCP-1 in vivo. The injection of the mutant 7ND into skeletal muscle suppressed monocyte recruitment to the dermis induced by recombinant MCP-1. Furthermore, this strategy blocked the early inflammatory and proliferative changes as well as prevented subsequent medial thickening of the coronary arteries in a rat model of chronic inhibition of NO synthesis. Rollins et al. (3 , 8) clearly demonstrated under in vitro conditions that the mutant MCP-1 forms inactive heterodimers with wild-type MCP-1 and exerts its inhibitory activity as a dominant-negative inhibitor. Thus, our new strategy can achieve an effective and sufficient blockade of MCP-1 activity in remote organs.

In contrast, the 7ND gene transfer could not reduce the development of perivascular fibrosis or arterial hypertension induced by the chronic inhibition of NO synthesis. These findings suggest that MCP-1 is essential for the formation of early inflammatory changes as well as the subsequent medial thickening but not for fibrogenesis. Furthermore, our data suggest that the observed effects of 7ND gene transfer are independent of the arterial hypertension induced by the blockade of NO synthesis in this experimental model. Since the recruitment and activation of monocytes as well as the proliferation of vascular smooth muscle cells are essential steps in the pathogenesis of many vascular diseases (1 , 2) , 7ND gene transfer would reduce the neointimal formation after vascular injury or inhibit the transformation from a stable to an unstable plaque prone to rupture. Therefore, our strategy could be a promising form of gene therapy against vascular diseases in humans such as restenosis after angioplasty or an unstable atheromatous plaque prone to rupture. Although no apparent side effects were observed during the period of this study, careful observation over a longer period of time needs to be done in future studies.

In conclusion, this study has shown that the i.m. expression of 7ND can effectively block MCP-1 activity in remote organs. This new strategy may be useful for clarifying the role of MCP-1 under pathophysiologic conditions in vivo, especially in organs into which direct gene transfer is difficult.

	ACKNOWLEDGMENTS

 
This study was supported by Grants-in-Aid for Scientific Research (11470164, 11158216, 11557056, 10307019, and 10177226) from the Ministry of Education, Science and Culture, Tokyo, Japan

Received for publication March 14, 2000. Revision received March 15, 2000.

	REFERENCES

TOP ABSTRACT INTRODUCTION MATERIALS AND METHODS RESULTS Effects of the i.m.... DISCUSSION REFERENCES

 

1. Ross, R. (1999) Atherosclerosis—an inflammatory disease. N. Engl. J. Med.. 340,115-126[Free Full Text]
2. Libby, P. (1995) Molecular bases of the acute coronary syndromes. Circulation 91,2844-2850[Medline]
3. Rollins, B. J. (1997) Chemokines. Blood 90,909-928[Free Full Text]
4. Takeya, M., Yoshimura, T., Leonard, E. J., Takahashi, K. (1993) Detection of monocyte chemoattractant protein-1 in human atherosclerotic lesions by an anti-monocyte chemoattractant protein-1 monoclonal antibody. Hum. Pathol. 24,534-539[Medline]
5. Yla-Herttuala, S., Lipton, B. A., Rosenfeld, M. E., Sarkioja, T., Yoshimura, T., Leonard, E. J., Witztum, J. L., Steinberg, D. (1991) Expression of monocyte chemoattractant protein 1 in macrophage-rich areas of human and rabbit atherosclerotic lesions. Proc. Natl. Acad. Sci. USA 88,5252-5256[Abstract/Free Full Text]
6. Boring, L., Gosling, J., Cleary, M., Charo, I. F. (1998) Decreased lesion formation in CCR2-/- mice reveals a role for chemokines in the initiation of atherosclerosis. Nature (London) 394,894-897[Medline]
7. Gu, L., Okada, Y., Clinton, S. K., Gerard, C., Sukhova, G. K., Libby, P., Rollins, B. J. (1998) Absence of monocyte chemoattractant protein-1 reduces atherosclerosis in low density lipoprotein receptor-deficient mice. Mol Cell. 2,275-281[Medline]
8. Zhang, Y., Rollins, B. J. (1995) A dominant negative inhibitor indicates that monocyte chemoattractant protein 1 functions as a dimer. Mol. Cell. Biol. 15,4851-4855[Abstract]
9. Ueki, T., Kaneda, Y., Tsutsui, H., Nakanishi, K., Sawa, Y., Morishita, R., Matsumoto, K., Nakamura, T., Takahashi, H., Okamoto, E., Fujimoto, J. (1999) Hepatocyte growth factor gene therapy of liver cirrhosis in rats. Nat Med. 5,226-230[Medline]
10. Takemoto, M., Egashira, K., Usui, M., Numaguchi, K., Tomita, H., Tsutsui, H., Shimokawa, H., Sueishi, K., Takeshita, A. (1997) Important role of tissue angiotensin-converting enzyme activity in the pathogenesis of coronary vascular and myocardial structural changes induced by long-term blockade of nitric oxide synthesis in rats. J. Clin. Invest. 99,278-287[Medline]
11. Tomita, H., Egashira, K., Kubo-Inoue, M., Usui, M., Koyanagi, M., Shimokawa, H., Takeya, M., Yoshimura, T., Takeshita, A. (1998) Inhibition of nitric oxide synthesis induces inflammatory changes and monocyte chemoattractant protein-1 expression in rat hearts and vessels. Arterioscler. Thromb. Vasc. Biol. 18,1456-1464[Abstract/Free Full Text]
12. Katoh, M., Egashira, K., Usui, M., Ichiki, T., Tomita, H., Shimokawa, H., Rakugi, H., Takeshita, A. (1998) Cardiac angiotensin II receptor is upregulated by long-term inhibition of nitric oxide synthesis in rats. Circ. Res. 83,743-751[Abstract/Free Full Text]
13. Usui, M., Egashira, K., Tomita, H., Koyanagi, M., Katoh, M., Shimokawa, H., Takeya, M., Yoshimura, T., Matsushima, K., Takeshita, A. (2000) Important role of local angiotensin II activity mediated via type 1 receptor in the pathogenesis of cardiovascular inflammatory changes induced by chronic blockade of nitric oxide synthesis in rats. Circulation 101,305-310[Medline]
14. Zeiher, A. M., Fisslthaler, B., Schray-Utz, B., Busse, R. (1995) Nitric oxide modulates the expression of monocyte chemoattractant protein 1 in cultural human endothelial cells. Circ. Res. 76,980-986[Abstract/Free Full Text]
15. Tsao, P. S., Wang, B., Buitrago, R., Shyy, J. Y., Cooke, J. P. (1997) Nitric oxide regulates monocyte chemotactic protein-1. Circulation 96,934-940[Medline]
16. De Caterina, R., Libby, P., Peng, H. B., Thannickal, V. J., Rajavashisth, T. B., Gimbrone, M. A., Jr, Shin, W. S., Liao, J. K. (1995) Nitric oxide decreases cytokine-induced endothelial activation. Nitric oxide selectively reduces endothelial expression of adhesion molecules and proinflammatory cytokines. J. Clin. Invest. 96,60-68
17. Peng, H. B., Libby, P., Liao, J. K. (1995) Induction and stabilization of I kappa B alpha by nitric oxide mediates inhibition of NF-kappa B. J. Biol. Chem. 270,14214-14219[Abstract/Free Full Text]
18. Kumar, A. G., Ballantyne, C. M., Michael, L. H., Kukielka, G. L., Youker, K. A., Lindsey, M. L., Hawkins, H. K., Birdsall, H. H., MacKay, C. R., LaRosa, G. J., Rossen, R. D., Smith, C. W., Entman, M. L. (1997) Induction of monocyte chemoattractant protein-1 in the small veins of the ischemic and reperfused canine myocardium. Circulation 95,693-700[Medline]
19. Yamashiro, S., Takeya, M., Kuratsu, J., Ushio, Y., Takahashi, K., Yoshimura, T. (1998) Intradermal injection of monocyte chemoattractant protein-1 induces emigration and differentiation of blood monocytes in rat skin. Int. Arch. Allergy Immunol. 115,15-23[Medline]

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				H. Shimizu, S. Maruyama, Y. Yuzawa, T. Kato, Y. Miki, S. Suzuki, W. Sato, Y. Morita, H. Maruyama, K. Egashira, et al. Anti-Monocyte Chemoattractant Protein-1 Gene Therapy Attenuates Renal Injury Induced by Protein-Overload Proteinuria J. Am. Soc. Nephrol., June 1, 2003; 14(6): 1496 - 1505. [Abstract] [Full Text] [PDF]

				K. Furuichi, T. Wada, Y. Iwata, K. Kitagawa, K.-i. Kobayashi, H. Hashimoto, Y. Ishiwata, N. Tomosugi, N. Mukaida, K. Matsushima, et al. Gene Therapy Expressing Amino-Terminal Truncated Monocyte Chemoattractant Protein-1 Prevents Renal Ischemia-Reperfusion Injury J. Am. Soc. Nephrol., April 1, 2003; 14(4): 1066 - 1071. [Abstract] [Full Text] [PDF]

				K. Egashira Molecular Mechanisms Mediating Inflammation in Vascular Disease: Special Reference to Monocyte Chemoattractant Protein-1 Hypertension, March 1, 2003; 41(3): 834 - 841. [Abstract] [Full Text] [PDF]

				S. Inoue, K. Egashira, W. Ni, S. Kitamoto, M. Usui, K. Otani, M. Ishibashi, K.-i. Hiasa, K.-i. Nishida, and A. Takeshita Anti-Monocyte Chemoattractant Protein-1 Gene Therapy Limits Progression and Destabilization of Established Atherosclerosis in Apolipoprotein E-Knockout Mice Circulation, November 19, 2002; 106(21): 2700 - 2706. [Abstract] [Full Text] [PDF]

				Y. Ikeda, Y. Yonemitsu, C. Kataoka, S. Kitamoto, T. Yamaoka, K.-I. Nishida, A. Takeshita, K. Egashira, and K. Sueishi Anti-monocyte chemoattractant protein-1 gene therapy attenuates pulmonary hypertension in rats Am J Physiol Heart Circ Physiol, November 1, 2002; 283(5): H2021 - H2028. [Abstract] [Full Text] [PDF]

				B. Kruger, B. Schroppel, R. Ashkan, B. Marder, C. Zulke, B. Murphy, B. K. Kramer, and M. Fischereder A Monocyte Chemoattractant Protein-1 (MCP-1) Polymorphism And Outcome After Renal Transplantation J. Am. Soc. Nephrol., October 1, 2002; 13(10): 2585 - 2589. [Abstract] [Full Text] [PDF]

				E. Mori, K. Komori, T. Yamaoka, M. Tanii, C. Kataoka, A. Takeshita, M. Usui, K. Egashira, and K. Sugimachi Essential Role of Monocyte Chemoattractant Protein-1 in Development of Restenotic Changes (Neointimal Hyperplasia and Constrictive Remodeling) After Balloon Angioplasty in Hypercholesterolemic Rabbits Circulation, June 18, 2002; 105(24): 2905 - 2910. [Abstract] [Full Text] [PDF]

				K. Egashira, Q. Zhao, C. Kataoka, K. Ohtani, M. Usui, I. F. Charo, K.-i. Nishida, S. Inoue, M. Katoh, T. Ichiki, et al. Importance of Monocyte Chemoattractant Protein-1 Pathway in Neointimal Hyperplasia After Periarterial Injury in Mice and Monkeys Circ. Res., June 14, 2002; 90(11): 1167 - 1172. [Abstract] [Full Text] [PDF]

				M. Kubo-Inoue, K. Egashira, M. Usui, M. Takemoto, K. Ohtani, M. Katoh, H. Shimokawa, and A. Takeshita Long-term inhibition of nitric oxide synthesis increases arterial thrombogenecity in rat carotid artery Am J Physiol Heart Circ Physiol, April 1, 2002; 282(4): H1478 - H1484. [Abstract] [Full Text] [PDF]

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