EBM Email Content Delivery
HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS
QUICK SEARCH:   [advanced]


     

This Article
Right arrow Abstract Freely available
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrow reprints & permissions
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Askari, F. K.
Right arrow Articles by Brewer, G. J.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Askari, F. K.
Right arrow Articles by Brewer, G. J.
Experimental Biology and Medicine 229:857-863 (2004)
© 2004 Society for Experimental Biology and Medicine

ORIGINAL RESEARCH ARTICLE

Tetrathiomolybdate Therapy Protects Against Concanavalin A and Carbon Tetrachloride Hepatic Damage in Mice

Fred K. Askari*, Robert Dick{dagger}, Maria Mao* and George J. Brewer*,{dagger},1

* Department of Internal Medicine and {dagger} Department of Human Genetics, University of Michigan Medical School, Ann Arbor, Michigan 48109

1To whom requests for reprints should be addressed at Department of Human Genetics, University of Michigan Medical School, 5024 Kresge Building II, Ann Arbor, MI 48109–0534. E-mail: brewergj{at}umich.edu


   Abstract
Top
 Abstract
Introduction
 Materials and Methods
 Results
 Discussion
 References
 
Tetrathiomolybdate, an anticopper drug, has been shown to protect mice against pulmonary fibrosis from bleomycin. Our hypothesis is that it does so by inhibiting fibrosis-inducing cytokines. Indeed, we have good evidence, not yet published, that tetrathiomolybdate inhibits pulmonary levels of transforming growth factor–ß and tumor necrosis factor-{alpha} expression in these bleomycin experiments. Herein, we evaluate tetrathiomolybdate’s effectiveness in mitigating hepatitis and fibrosis in mice from the hepatotoxins, concanavalin A and carbon tetrachloride, and its inhibition of cytokines as a possible mechanism. In short-term experiments, concanavalin A elevated serum amino leucine transferase levels several fold, and tetrathiomolybdate completely prevented this increase. In additional experiments, tetrathiomolybdate therapy reversed the elevated serum transaminase levels despite continued concanavalin A injections, with nearly significant serum interleukin-1ß inhibition. Concanavalin A given for 12 weeks produced mild fibrosis, whereas concomitant tetrathiomolybdate treatment resulted in normal histology. Carbon tetrachloride given for 12 weeks resulted in very high serum amino leucine transferase levels, high serum transforming growth factor–ß levels, cirrhosis as seen histologically, and increase in liver hydroxyproline, a measure of fibrosis. Concomitant tetrathiomolybdate partially and significantly protected against increases in amino leucine transferase and transforming growth factor–ß, fully protected against the increase in hydroxyproline, and resulted in normal histology. In conclusion, tetrathiomolybdate protects against the hepatitis and fibrosis produced by these hepatotoxins, probably by inhibiting the excessive increase in inflammatory and fibrotic cytokines.

Key Words: copper • hepatitis • cirrhosis • transforming growth factorß • interleukin-1ß


   Introduction
Top
 Abstract
 Introduction
Materials and Methods
 Results
 Discussion
 References
 
Tetrathiomolybdate (TM) was first developed as an anticopper drug for the initial treatment of patients with Wilson disease (1, 2). Tetrathiomolybdate is fast acting and relatively nontoxic. Subsequently, TM was shown to have anticancer activity in five rodent tumor models (3–6), in a study of spontaneous cancer in pet dogs (7), and in a phase 1/2 clinical study of 42 patients with metastatic and advanced cancer (8 and unpublished). The mechanism of the anticancer effect is through inhibition of angiogenesis (3, 9–11). Many cytokines that promote angiogenesis are copper dependent, such that if body copper levels are lowered with TM, angiogenic cytokine signaling is inhibited (3, 9–11). As long as copper levels are not lowered excessively, cellular copper requirements are met, avoiding toxic effects from copper deficiency (3–8). The level of ceruloplasmin (Cp), a serum protein that contains copper, is used as a surrogate marker of body copper status (3–8). The liver secretes Cp into the blood in an amount that depends on copper availability (12). The Cp level is maintained at midrange in both preclinical and clinical studies with TM therapy to provide an "antiangiogenic window."

Examination of the fibrotic pathway involving transforming growth factor–ß (TGFß) and connective tissue growth factor suggested to us that these cytokines might also be copper dependent. Overactivity and dysregulation of this pathway lead to fibrotic diseases in many organs (13, 14). Examples are pulmonary fibrosis, cirrhosis, renal fibrosis, and scleroderma (13, 15, 16). As a first test of the hypothesis that the fibrotic pathway was copper dependent, we used the bleomycin mouse model of pulmonary fibrosis (15). Therapy with TM completely prevented the inflammatory and fibrotic sequelae of intratracheal bleomycin instillation at the time of sacrifice 21 days after bleomycin treatment (17, 18). In work being prepared for publication, we have subsequently shown that TGFß levels, which are very high in the lungs of bleomycin controls at 21 days, are maintained at near-normal levels in the lungs of bleomycin animals treated with TM. In this work, we also found that the RNA expression levels of tumor necrosis factor-{alpha} (TNF-{alpha}), the major inflammatory cytokine involved in the inflammatory response to bleomycin, which is very elevated in the lungs of bleomycin controls at 7 days, was kept at near-normal levels at 7 days by TM treatment of bleomycin animals. By starting TM treatment after the inflammatory peak and then finding fibrosis inhibition by TM at the 21-day point, we were able to show that the fibrosis inhibition is an independent effect of TM not simply due to prior suppression of inflammation (18).

In this current work, we are extending our observations to the liver, using both the concanavalin A (Con A) and carbon tetrachloride (CT) models of liver injury in the mouse. Concanavalin A injected intravenously in the mouse produces acute hepatitis, marked by the release of the transaminase enzyme, amino leucine transferase (ALT), into the serum (19). Continued injection of Con A for weeks can produce some fibrosis, but a better model for this is CT treatment. Carbon tetrachloride injected intraperitoneally or given by oral gavage also produces acute hepatitis marked by an elevation of serum ALT level. However, continued injection for 12 weeks produces a well-established cirrhosis (20). Our objectives in the present work were to determine if TM therapy protects against these types of liver injury and, if so, whether inhibition of inflammatory and/or fibrotic cytokines in plasma could be detected.


   Materials and Methods
Top
 Abstract
 Introduction
 Materials and Methods
Results
 Discussion
 References
 
Mice.
The BALB/c (Experiments 1, 2, and 3) and CBA/J (Experiment 4) mice were 20-g females purchased from The Jackson Laboratory, Bar Harbor, ME. Experimental animals were housed in the University of Michigan Unit for Laboratory Animal Medicine facility and treated in accordance with a protocol approved by the University of Michigan Institutional Animal Care and Use Committee in accordance with the criteria outlined in the Guide for the Care and Use of Laboratory Animals prepared by the National Academy of Sciences and published by the National Institutes of Health (21). Animals were assigned to groups randomly, were provided food and water ad libitum, and kept on a 12:12-hr light:dark cycle.

Con A and CT Treatment.
Concanavalin A and CT were purchased from Sigma Chemical Co., St. Louis MO. In Experiments 1, 2, and 3, Con A–treated mice received a weekly intravenous (tail vein) injection of 15 mg/kg of body wt of Con A dissolved in 0.3 ml of 0.9% sodium chloride. Control mice received an intravenous injection of 0.3 ml of 0.9% sodium chloride alone. In Experiment 4, CT-treated mice received intraperitoneal injections of 1 ml/kg of body wt of CT in 200 µl of olive oil twice per week. Control animals received 200 µl of olive oil alone by intraperitoneal injection twice per week.

TM Treatment.
Tetrathiomolybdate was given in 0.25 ml of water by intragastric gavage once daily in the doses and times indicated in the various studies.

Con A Experiments.
Three types of experiments were performed. In Experiment 1, mice were divided into four groups: six to receive Con A only, four to receive Con A plus TM, six to receive saline instead of Con A, and three to receive TM only. In the TM-treated animals, TM was given in a daily dose of 0.9 mg, beginning 5 days before the first Con A injection. Concanavalin A was administered for 4 weeks. Twenty-four hours after the first three injections, blood was taken for ALT assay. Twenty-four hours after the fourth injection, all the animals were sacrificed and ALT and Cp measured on all.

In Experiment 2, mice were divided into three groups: six to receive Con A only, six to receive Con A plus TM, and four to receive saline instead of Con A. In the TM-treated animals, TM was withheld until after the fourth Con A injection and then initiated at a dose of 0.9 mg/day. Twenty-four hours after the fourth through 12th Con A injections, blood was taken for ALT assay from individual animals of the Con A and Con A plus TM groups and occasionally from a control animal. Different animals were used for these blood draws to avoid overbleeding. Concanavalin A injections were continued for 12 weeks and the animals sacrificed for histologic analysis of the livers.

In Experiment 3, mice were divided into two groups: 9 to receive Con A only and 10 to receive Con A plus TM. In TM-treated animals, TM treatment was begun 5 days before the first Con A injection at a daily dose of 0.9 mg. Concanavalin A was injected twice at weekly intervals. Two hours after the second injection (Week 2), the animals were sacrificed and cytokine measurements made in the blood.

CT Experiments.
In Experiment 4, mice were divided into four groups: five to receive CT only, five to receive CT plus TM, three to receive the vehicle for CT (olive oil), and three to receive TM only. Carbon tetrachloride was given in twice-weekly doses for 12 weeks. Tetrathiomolybdate treatment was begun in TM-treated animals after 4 weeks of CT injections at a dose of 0.9 mg once daily. After 12 weeks, the animals were sacrificed for histopathology studies of the liver, hydroxyproline assays of the liver, and ALT and TGFß assays in the serum.

Copper Status.
In the presence of TM therapy, serum copper cannot be used to assess copper status because of the accumulation in the blood of a tripartite complex of TM, copper, and albumin. We use serum Cp as a surrogate measure. Ceruloplasmin was assayed by its oxidase activity as previously described (5).

Hydroxyproline Assay in the Liver.
This assay was performed as previously described (18).

Serum ALT Assay.
This assay was performed using a quantitative colorimetric method commercially available from Sigma Diagnostics (Procedure 505), St. Louis, MO.

Cytokine Assays.
Interleukin-1ß (IL-1ß), TNF-{alpha}, and TGFß serum levels were determined by a quantitative sandwich enzyme immunoassay method using commercially available kits purchased from R & D Systems, Minneapolis, MN.

Statistics.
For comparisons of means, analysis of variance was used followed by the Scheffé test for multiple comparisons when appropriate.


   Results
Top
 Abstract
 Introduction
 Materials and Methods
 Results
Discussion
 References
 
Con A Studies.
In Experiment 1, where Con A was given for 4 weeks, serum ALT levels in single animals from each group (to avoid overbleeding) were measured 24 hrs after the first three injections (Table 1Go). Then all of the animals in each group were sacrificed and ALT assays performed on all the animals (Table 1Go; under fourth week of injection). The data are consistent in that Con A elevated ALT levels 4- to 6-fold, and TM strongly, and significantly at Week 4, protected against those elevations. The Cp levels at the time of sacrifice in the Con A plus TM groups averaged 47% in the Con A group, and the means were highly statistically significantly different (P = 0.0008, with the Scheffé correction) (data not shown).


View this table:
[in this window]
[in a new window]
  		Table 1. Serum ALT Results in Mice 24 hrs After Each of Four Weekly Serial Injections of Con A in Experiment 1a
 
In Experiment 2, Con A was given for 4 weeks before TM treatment was started in TM-treated animals. At Week 4, before any TM treatment, an individual animal from each group was bled 24 hrs after Con A injection for ALT assay. At 4 weeks, Con A–treated animals had an approximately 4-fold elevation of ALT levels compared with the saline control (Fig. 1Go). After that, at weekly intervals, different individual animals from the Con A and Con A plus TM groups were bled 24 hrs after Con A injection. Data for Weeks 5–7 are shown in Figure 1Go. Subsequent to TM therapy, ALT levels decreased to nearly normal during a 3-week period despite continued Con A injections. The Con A injections in these animals were continued for 12 weeks. The ALT levels in the six Con A animals bled during this period were statistically significantly higher than the six Con A plus TM animals bled during this same period (P = 0.03) and also significantly higher than the four control animals bled during this period (P = 0.04). The Con A plus TM animals were not different than controls. At 12 weeks, the animals were sacrificed and their livers examined histologically. The Con A animals had some inflammatory changes and minimal fibrosis, whereas the Con A plus TM animals had histologically normal livers (data not shown). The Cp levels at the time of sacrifice in the TM group averaged 18% of the Con A group, and the means were highly statistically significantly different (P = 0.0006) (data not shown).



View larger version (9K):
[in this window]
[in a new window]
  		Figure 1. Concanavalin A (Con A) was given for 4 weeks before initiation of tetrathiomolybdate (TM) in TM-treated animals in Experiment 2. An individual animal from each of the three groups was bled 24 hrs after Con A injection at Week 4 for amino leucine transferase (ALT) assay. Different individual animals were bled at Weeks 5, 6, and 7 from the Con A and Con A plus TM groups for ALT assay.

 
In Experiment 3, in blood taken 2 hrs after the second Con A injection, there were trends toward serum TNF-{alpha} being lower in the Con A plus TM samples than in the Con A samples, but the means were not statistically significantly different. Mean serum IL-1ß levels 2 hrs after the second Con A injection were 37% of the Con A mean in the Con A plus TM samples, and this difference was close to being statistically significant (P = 0.08) (data not shown).

CT Studies.
In Experiment 4, CT was given for 12 weeks in CT-treated animals, and TM treatment was started at the beginning of the fifth week in TM-treated animals. All animals were sacrificed at 12 weeks. Figure 2Go shows the serum ALT data at the 12-week point. (The two control groups were pooled, since their means were similar.) The CT treatment strongly elevates ALT levels over control levels, and TM partially and significantly (P = 0.05) protects against the CT-induced increase. At lower doses of CT and shorter time frames, this effect was even more pronounced (data not shown).



View larger version (10K):
[in this window]
[in a new window]
  		Figure 2. Mean and SD of serum amino leucine transferase data at 12 weeks in the carbon tetrachloride study (Experiment 4). Asterisk indicates means are statistically significantly different at P < 0.05.

 
Hydroxyproline levels at the 12-week point in Experiment 4 were elevated by CT treatment, and TM therapy almost completely and significantly (P = 0.03 with the Scheffé correction) prevented this CT-induced increase (Fig. 3Go). (The two control groups were pooled, since their means were similar.) The CT plus TM and control means were not significantly different.



View larger version (10K):
[in this window]
[in a new window]
  		Figure 3. Mean and SD of hydroxyproline content of the liver at 12 weeks in the carbon tetrachloride study (Experiment 4). Asterisk indicates means are statistically significantly different at P < 0.03.

 
In keeping with this biochemical evidence of TM protection against fibrosis, histopathology studies clearly documented TM’s protection against CT-induced cirrhosis (Fig. 4Go). The CT control animals had a well-developed cirrhosis, whereas the CT plus TM therapy animals were histologically normal (Fig. 4Go).



View larger version (65K):
[in this window]
[in a new window]
  		Figure 4. Masson trichrome stains of the liver of two mice treated with carbon tetrachloride (CT) for 12 weeks from Experiment 4. The control CT-treated animal is on the left and shows extensive fibrosis (blue staining). The CT-treated animal that received tetrathiomolybdate is on the right and shows normal histology.

 
Also in Experiment 4, TGFß levels were measured in the plasma at the 12-week point (Fig. 5Go). (The two control groups were pooled, since their means were similar.) The data show that CT treatment significantly elevates plasma TGFß levels (P < 0.02 with the Scheffé correction) and that TM therapy almost completely and statistically significantly (P = 0.02 with the Scheffé correction) prevents TGFß elevation in response to CT.



View larger version (10K):
[in this window]
[in a new window]
  		Figure 5. Mean and SD of plasma transforming growth factor–ß levels at 12 weeks in the carbon tetrachloride study (Experiment 4). Asterisk indicates means are statistically significantly different at P < 0.02.

 
The plasma Cp data from Experiment 4 are shown in Figure 6Go. The mean Cp level in CT-treated animals was markedly and statistically significantly lower in the TM-treated group. The Cp levels are statistically significantly elevated in CT animals vs controls (P = 0.0003), because Cp is an acute-phase reactant.



View larger version (8K):
[in this window]
[in a new window]
  		Figure 6. Mean and SD of plasma ceruloplasmin levels at 12 weeks in the carbon tetrachloride study (Experiment 4). Asterisk indicates means are statistically significantly different at P < 0.0002.

 

   Discussion
Top
 Abstract
 Introduction
 Materials and Methods
 Results
 Discussion
References
 
As was the case in the bleomycin lung injury study reported earlier (17, 18), TM strongly protects against liver injury from the two hepatotoxins studied herein. Tetrathiomolybdate protected against serum ALT level elevations from Con A (Table 1Go), even when TM treatment was started after several injections of Con A (Fig. 1Go). In the 12-week study with CT, TM treatment started 4 weeks after CT partially protected against CT-induced ALT level elevations (Fig. 2Go) and completely protected against fibrosis induced by CT as measured by hydroxyproline levels (Fig. 3Go) and as seen histologically (Fig. 4Go).

In our prior bleomycin study, the TM protection against lung injury appeared to be mediated by cytokine inhibition. In work being prepared for publication, we have shown TM inhibition of TGFß and TNF-{alpha} in the lung. Therefore, we postulate that the mechanism of TM protection against liver injury is also mediated by inhibition of these inflammatory and fibrotic cytokines. In the current study, our cytokine studies were limited to plasma and protein assays. (In the bleomycin study, we included assays in lung and also performed RNA assays.) In the Con A study, we saw trends toward a lower TNF-{alpha} protein level in the plasma of TM-treated Con A animals than in Con A animals, but the results were not statistically significant. Concanavalin A increased serum levels of IL-1ß protein, TM appeared to decrease these levels, and this effect approached statistical significance (P = 0.08). In the CT studies, CT elevated serum levels of the profibrotic cytokine, TGFß, and TM therapy clearly caused a significant decrease in these levels at the 12-week point (Fig. 5Go), when well-developed cirrhosis was occurring in CT controls.

It appears that TM is a somewhat general protectant against much of the organ injury produced by toxic agents. So far we have shown protection against bleomycin injury in the lung and, in this article, Con A and CT in the liver. Our working hypothesis is that these protections by TM do not involve inhibiting the initial toxic effect of the agent but rather inhibiting the subsequent excessive inflammatory response and the subsequent excessive harmful fibrosis. The best evidence for the lack of a TM effect on initial toxicity of these agents is our bleomycin work (18). In this situation, bleomycin is given only once and sets off the inflammatory and fibrotic cascade. Treatment with TM started on Day 7 after bleomycin, which brings the copper level down by approximately Day 10 or 11, still has a strong antifibrotic effect by Day 21 (18). Presumably, the bleomycin has been disposed of and is no longer producing new damage during the last 10 to 11 days in this type of study. In the present article, TM treatment started after damage from Con A or CT is also effective in inhibiting subsequent damage. However, these studies are less compelling as evidence against protection from initial damage, since the damaging agents continue to be administered in these liver models. The likely mechanism of the protective effect against subsequent excessive inflammation and fibrosis has already been discussed, namely, inhibition of excessive activity of inflammatory and fibrotic cytokines.

If this hypothesis is correct, it suggests that TM has the potential to be useful in preventing further damage from dysregulated inflammatory and fibrotic responses irrespective of the injury. This would mean that autoimmune diseases, which often have high levels of inflammatory and fibrotic cytokines, might be effectively treated with TM. These liver models, in which the injurious agent is repeatedly administered and TM protects against much of the injury, are likely very relevant to TM protection against the responses to autoimmune injury, which is also continuous.

The protection by TM against cirrhosis induced by CT and against pulmonary fibrosis induced by bleomycin suggests that TM deserves to be tested for efficacy in human disease of cirrhosis, pulmonary fibrosis, renal fibrosis, scleroderma, and other fibrotic diseases associated with excessive TGFß production (13, 14, 16). A clinical trial in idiopathic pulmonary fibrosis is under way, and trials are planned in scleroderma and primary biliary cirrhosis. Since these types of diseases are generally not treated effectively with current approaches, efficacy of TM therapy would be welcome.

Similarly, the protection by TM against excessive inflammation, as illustrated by the inhibition of hepatitis from Con A evidenced by the reduced ALT levels, suggests evaluating TM in diseases of excessive inflammation. Such trials are further suggested by TM inhibition of TNF-{alpha} and IL-1ß. Diseases of this type would include autoimmune diseases such as rheumatoid arthritis, systemic lupus erythematosus, vasculitis of various types, and Crohn disease.

Along these same lines, TNF-{alpha}–based antibodies have been shown to be effective in a series of diseases with TNF- {alpha} elevations, including rheumatoid arthritis (22, 23), Crohn disease (24, 25), psoriasis (26–28), and many others. Tetrathiomolybdate therapy, which is given orally, should be tried in diseases where injection of such antibodies has proven effective. Besides oral administration, a potential advantage of TM is that it does not inhibit cytokines below their constitutive levels (Fig. 5Go and unpublished data), while inhibiting much of the increase induced by injury. This may avoid some of the problems caused by antibody inhibition, such as tuberculosis reactivation by TNF-{alpha} antibodies (23), or activation of inflammation, shown by gene knockouts of TGFß in animals (29, 30).

The relative safety of TM therapy becomes important when discussing clinical applications. Obviously, copper levels cannot be pushed too low or copper deficiency problems will emerge. Our strategy has been to lower Cp levels to a midrange, which means that the liver still has enough copper to synthesize approximately half the normal amount of holoceruloplasmin. At these levels, cells in the body have enough copper to synthesize normal amounts of required cuproenzymes, such as lysyl oxidase, cytochrome oxidase, and superoxide dismutase, but cytokine signaling is inhibited in many cases. When Cp levels are pushed very low with TM therapy, the first adverse effect seen is bone marrow depression, because the bone marrow has a relatively high requirement for copper to make cells. The resulting anemia and/or leukopenia is relatively mild and quickly responsive to a dose reduction or drug holiday. If the dose is reduced when bone marrow effects are seen, other adverse effects from deepening copper deficiency are not seen. Tetrathiomolybdate has been safely used in this manner in a number of published (8, 31, 32) and ongoing clinical trials.


   Footnotes
 
The University of Michigan has recently licensed the antiangiogenic uses of tetrathiomolybdate to Attenuon LLC, and Dr. Brewer and Mr. Dick have equity in Attenuon LLC.

Received for publication January 20, 2004. Accepted for publication April 13, 2004.


   References
Top
 Abstract
 Introduction
 Materials and Methods
 Results
 Discussion
 References
 

  1. Brewer GJ, Dick RD, Yuzbasiyan-Gurkan V, Tankanow R, Young AB, Kluin KJ. Initial therapy of Wilson’s disease patients with tetrathiomolybdate. Arch Neurol 48:42–47, 1991.[Abstract]
  2. Brewer GJ, Hedera P, Kluin KJ, Carlson MD, Askari F, Dick RB, Sitterly JA, Fink JK. Treatment of Wilson’s disease with tetrathiomolybdate III: initial therapy in a total of 55 neurologically affected patients and follow-up with zinc therapy. Arch Neurol 60:378–385, 2003.
  3. Pan Q, Kleer CG, van Golen KL, Irani J, Bottema KM, Bias C, De Carvalho M, Mesri EA, Robins DM, Dick RD, Brewer GJ, Merajver SD. Copper deficiency induced by tetrathiomolybdate suppresses tumor growth and angiogenesis. Cancer Res 62:4854–4859, 2002.[Abstract/Free Full Text]
  4. Cox CD, Teknos TN, Barrios M, Brewer GJ, Dick RD, Merajver SD. The role of copper suppression as an antiangiogenic strategy in head and neck squamous cell carcinoma. Laryngoscope 111:696–701, 2001.[Medline]
  5. Khan MK, Miller MW, Taylor J, Gill NK, Dick RD, van Golen K, Brewer GJ, Merajver SD. Radiotherapy and antiangiogenic TM in lung cancer. Neoplasia 4:1–7, 2002.[Medline]
  6. van Golen K, Bao L, Brewer G, Pienta K, Karadt J, Livant D, Merajver S. Suppression of tumor recurrence and metastasis by a combination of the PHSCN sequence and the antiangiogenic compound tetrathiomolybdate in prostate carcinoma. Neoplasia 4:373–379, 2002.[Medline]
  7. Kent MS, Madewell BR, Dank G, Dick R, Merajver SD, Brewer GJ. An anticopper antiangiogenic approach for advanced cancer in spontaneously occurring tumors, using tetrathiomolybdate: a pilot study in a canine animal model. J Trace Elem Exp Med 17:9–20, 2004.
  8. Brewer GJ, Dick RD, Grover DK, LeClaire V, Tseng M, Wicha M, Pienta K, Redman BG, Thierry J, Sondak VK, Strawderman M, LeCarpentier G, Merajver SD. Treatment of metastatic cancer with tetrathiomolybdate, an anticopper, antiangiogenic agent, I: phase I study. Clin Cancer Res 6:1–10, 2000.[Abstract/Free Full Text]
  9. Brewer GJ. Copper control as an antiangiogenic anticancer therapy: lessons from treating Wilson’s disease. Exp Biol Med 226:665–673, 2001.[Abstract/Free Full Text]
  10. Brewer GJ. Tetrathiomolybdate anticopper therapy for Wilson’s disease inhibits angiogenesis, fibrosis, and inflammation. J Cell Mol Med 7:11–20, 2003.[Medline]
  11. Brewer GJ. Copper in medicine. Curr Opin Chem Biol 7:207–212, 2003.[Medline]
  12. Linder MC, Houle PA, Isaacs E, Moor JR, Scott LE. Copper regulation of ceruloplasmin in copper-deficient rats. Enzyme 24:23–35, 1979.[Medline]
  13. Border WA, Noble NA. Transforming growth factor beta in tissue fibrosis. N Engl J Med 331:1286–1292, 1994.[Free Full Text]
  14. Duncan MR, Frazier KS, Abramson S, Williams S, Klapper H, Huang X, Grotendorst GR. Connective tissue growth factor mediates transforming growth factor ß-induced collagen synthesis: down-regulation by cAMP. FASEB J 13:1774–1786, 1999.[Abstract/Free Full Text]
  15. Phan SH, Kunkel SL. Lung cytokine production in bleomycin-induced pulmonary fibrosis. Exp Lung Res 18:29–43, 1992.[Medline]
  16. Brigstock DR. The connective tissue growth factor/cysteinerich 61/nephroblastoma overexpressed (CCN) family. Endocr Rev 20:189–206, 1999.[Abstract/Free Full Text]
  17. Brewer GJ, Phan S. Tetrathiomolybdate anticopper therapy protects against bleomycin-pulmonary fibrosis in mice. J Invest Med 50:227A, 2002.
  18. Brewer GJ, Ullenbruch MR, Dick R, Olivarez L, Phan SH. Tetrathiomolybdate therapy protects against bleomycin-pulmonary fibrosis in mice. J Lab Clin Med 141:210–216, 2003.[Medline]
  19. Nakamura K, Okada M, Yoneda M, Takamoto S, Nakade Y, Tamori K, Aso K, Makino I. Macrophage inflammatory protein-2 induced by TNF-{alpha} plays a pivotal role in concanavalin A-induced liver injury in mice. J Hepatol 35:217–224, 2001.[Medline]
  20. Simeonova PP, Gallucci RM, Hulderman T, Wilson R, Kommineni C, Rao M, Luster MI. The role of tumor necrosis factor-{alpha} in liver toxicity, inflammation, and fibrosis induced by carbon tetrachloride. Toxicol Appl Pharmacol 177:112–120, 2001.[Medline]
  21. Public Health Service Policy on Humane Care and Use of Animals, the regulations of the Federal Animal Welfare Act, and University of Michigan Policy. Available at: http://grants.nih.gove/grants/olaw/olaw.htm and http://www.ucuca.umich.edu. Accessed July 1, 2004.
  22. Elliott MJ, Maini RN, Feldmann M, Long-Fox A, Charles P, Katsikis P, Brennan FM, Walker J, Bijl H, Ghrayeb J, Woody JN. Treatment of rheumatoid arthritis with chimeric monoclonal antibodies to TNF-{alpha}. Arthritis Rheum 36:1681–1690, 1993.[Medline]
  23. Shanahan CS, St. Clair EW. Tumor necrosis factor-{alpha} blockade: a novel therapy for rheumatic disease. Clin Immunol 103:231–242, 2002.[Medline]
  24. Present DH, Rutgeerts P, Targan S, Hanauer SB, Mayer L, van Hogezand RA, Podolsky DK, Sands BE, Braakman T, DeWoody KL, Schaible TF, van Deventer SJ. Infliximab for the treatment of fistulas in patients with Crohn’s disease. N Engl J Med 340:1398–1405, 1999.[Abstract/Free Full Text]
  25. D’haens G, Van Deventer S, van Hogezand R, Chalmers D, Kothe C, Baert F, Braakman T, Schaible T, Geboes K, Rutgeerts P. Endoscopic and histological healing with infliximab anti-tumor necrosis factor antibodies in Crohn’s disease: a European multi-center trial. Gastroenterology 116:1029–1034, 1999.[Medline]
  26. Ogilvie AL, Atoni C, Dechant C, Manger B, Kalden JR, Schuler G, Luftl M. Treatment of psoriatic arthritis with anti-tumor necrosis factor-alpha antibody clears skin lesions of psoriasis resistant to treatment with methotrexate. Br J Dermatol 144:587–589, 2001.[Medline]
  27. Chaudhari U, Roman P, Mulcahy LD, Dooley LT, Baker DG, Gottlieb AB. Efficacy and safety of infliximab monotherapy for plaque-type psoriasis: a randomized trial. Lancet 357:1842–1847, 2001.[Medline]
  28. Mease PJ, Goffe BS, Metz J, VanderStoep A, Finck B, Burge DJ. Etanercept in the treatment of psoriatic arthritis and psoriasis: a randomized trial. Lancet 35:385–390, 2000.
  29. Wahl SM, Orenstein JM, Chen W. TGF-ß influences the life and death decisions of T lymphocytes. Cytokine Growth Factor Rev 11:71–79, 2000.[Medline]
  30. Hahm KB, Im YH, Lee C, Parks WT, Bang YJ, Green JE, Kim SJ. Loss of TGF-ß signaling contributes to autoimmune pancreatitis. J Clin Invest 105:1057–1065, 2000.[Medline]
  31. Vine AK, Brewer GJ. Tetrathiomolybdate as an antiangiogenesis therapy for subfoveal choroidal neovascularization secondary to age-related macular degeneration. Trans Am Ophthalmol Soc 100:73–76, 2002.[Medline]
  32. Redman BG, Esper P, Pan Q, Dunn RL, Hussain HK, Chenevert T, Brewer GJ, Merajver SD. Phase II trial of tetrathiomolybdate in patients with advanced kidney cancer. Clin Cancer Res 9:1666–1672, 2003.[Abstract/Free Full Text]



This article has been cited by other articles:


Home page
 Exp. Biol. Med.Home page
C. Zeng, G. Hou, R. Dick, and G. J. Brewer
Tetrathiomolybdate Is Partially Protective Against Hyperglycemia in Rodent Models of Diabetes
Experimental Biology and Medicine, August 1, 2008; 233(8): 1021 - 1025.
[Abstract] [Full Text] [PDF]

Home page
 J. Pharmacol. Exp. Ther.Home page
M. Song, Z. Song, S. Barve, J. Zhang, T. Chen, M. Liu, G. E. Arteel, G. J. Brewer, and C. J. McClain
Tetrathiomolybdate Protects against Bile Duct Ligation-Induced Cholestatic Liver Injury and Fibrosis
J. Pharmacol. Exp. Ther., May 1, 2008; 325(2): 409 - 416.
[Abstract] [Full Text] [PDF]

Home page
 Exp. Biol. Med.Home page
G. J. Brewer
Iron and Copper Toxicity in Diseases of Aging, Particularly Atherosclerosis and Alzheimer's Disease
Experimental Biology and Medicine, February 1, 2007; 232(2): 323 - 335.
[Abstract] [Full Text] [PDF]

Home page
 Mol. Pharmacol.Home page
D. Gong, J. Lu, X. Chen, S. Y. Choong, S. Zhang, Y.-K. Chan, S. Glyn-Jones, G. D. Gamble, A. R. J. Phillips, and G. J. S. Cooper
Molecular Changes Evoked by Triethylenetetramine Treatment in the Extracellular Matrix of the Heart and Aorta in Diabetic Rats
Mol. Pharmacol., December 1, 2006; 70(6): 2045 - 2051.
[Abstract] [Full Text] [PDF]

This Article
Right arrow Abstract Freely available
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrow reprints & permissions
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Askari, F. K.
Right arrow Articles by Brewer, G. J.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Askari, F. K.
Right arrow Articles by Brewer, G. J.

HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS