C. G. J. (Fred) Sweep

Expression of the transcription factor Ets-1 is an independent prognostic marker for relapse-free survival in breast cancer.

The transcription factor Ets-1 regulates the expression of several angiogenic and extracellular matrix remodeling factors, and might be implicated in disease progression of breast cancer. In the present study, the prognostic value of Ets-1 expression was assessed by quantitative real-time fluorescence RT–PCR in 123 sporadic primary breast cancer samples of patients with a median follow-up time of 62 months. Ets-1 expression levels correlated significantly with VEGF and PAI-1 in the same tissue. In univariate (P=0.0011) and multivariate (P=0.005) analyses, Ets-1 expression showed significant prognostic value for relapse-free survival. Ets-1 is a strong, independent predictor of poor prognosis in breast cancer. This seems – at least in part – to be attributable to its role in transcriptional regulation of factors involved in angiogenesis (VEGF), and extracellular matrix remodeling (PAI-1).

Pretreatment with a Single, Low Dose of Recombinant Human Thyrotropin Allows Dose Reduction of Radioiodine Therapy in Patients with Nodular Goiter

In patients with nodular goiter, radioiodine ((131)I) therapy results in a mean reduction in thyroid volume (TV) of approximately 40% after 1 yr. We have demonstrated that pretreatment with a single, low dose of recombinant human TSH (rhTSH) doubles 24-h radioactive iodine uptake (RAIU) in these patients. We have now studied the safety and efficacy of therapy with a reduced dose of (131)I after pretreatment with rhTSH. Twenty-two patients with nodular goiter received (131)I therapy, 24 h after im administration of 0.01 (n = 12) or 0.03 (n = 10) mg rhTSH. In preceding diagnostic studies using tracer doses of (131)I, 24-h RAIU without and with rhTSH pretreatment (either 0.01 or 0.03 mg) were compared. Therapeutic doses of (131)I were adjusted to the rhTSH-induced increases in 24-h RAIU and were aimed at 100 micro Ci/g thyroid tissue retained at 24 h. Pretreatment with rhTSH allowed dose reduction of (131)I therapy by a factor of 1.9 +/- 0.5 in the 0.01-mg and by a factor of 2.4 +/- 0.4 in the 0.03-mg rhTSH group (P < 0.05, 0.01 vs. 0.03 mg rhTSH). Before and 1 yr after therapy, TV and the smallest cross-sectional area of the tracheal lumen were measured with magnetic resonance imaging. During the year of follow-up, serum TSH, free T(4) (FT(4)), T(3), and TSH receptor antibodies were measured at regular intervals. TV before therapy was 143 +/- 54 ml in the 0.01-mg group and 103 +/- 44 ml in the 0.03-mg rhTSH group. One year after treatment, TV reduction was 35 +/- 14% (0.01 mg rhTSH) and 41 +/- 12% (0.03 mg rhTSH). In both groups, smallest cross-sectional area of the tracheal lumen increased significantly. In the 0.01-mg rhTSH group, serum FT(4) rose, after (131)I treatment, from 15.8 +/- 2.8 to 23.2 +/- 4.4 pM. In the 0.03-mg rhTSH group, serum FT(4) rose from 15.5 +/- 2.5 to 23.5 +/- 5.1 pM. Individual peak FT(4) levels, reached between 1 and 28 d after (131)I treatment, were above the normal range in 12 patients. TSH receptor antibodies were negative in all patients before therapy and became positive in 4 patients. Hyperthyroidism developed in 3 of these 4 patients between 23 and 25 wk after therapy. In conclusion, in patients with nodular goiter pretreatment with a single, low dose of rhTSH allowed approximately 50-60% reduction of the therapeutic dose of radioiodine without compromising the efficacy of TV reduction.

Matrix metalloproteinase inhibitor reversion-inducing cysteine-rich protein with Kazal motifs: a prognostic marker for good clinical outcome in human breast carcinoma.

The recently described reversion‐inducing cysteine‐rich protein with Kazal motifs (RECK) inhibits membrane Type 1 matrix metalloproteinase (MMP‐14), MMP‐2, and MMP‐9 secretion and enzymatic activity. Its expression is essential for normal vasculogenesis. Down‐regulation of RECK has been implicated in tumor angiogenesis and progression.

Cyclin-E is a strong predictor of endocrine therapy failure in human breast cancer.

Recently, cyclin-E was reported to be the most prominent prognostic factor for breast cancer outcome described so far, even surpassing axillary nodal involvement. Earlier studies on the prognostic value of cyclin-E in breast cancer, however, yielded heterogeneous results. Therefore, we set out to confirm and extend these results by quantitative Taqman RT–PCR of cyclin-E levels in 277 resectable breast cancers. Cyclin-E levels were not associated with relapse-free survival (RFS) or overall survival (OS) in the total cohort of patients, or in the subset of patients without involved lymph nodes that were not treated with adjuvant systemic therapy. Besides several classical clinicopathological factors, the interaction between cyclin-E and adjuvant endocrine therapy (P=0.01, HR=3.04, 95% CI: 1.30–7.09) was found to contribute significantly in multivariate analyses. Cyclin-E levels were associated with poor RFS specifically in patients treated with adjuvant endocrine therapy (n=108, P=0.001, HR=4.01, 95% CI: 1.76–9.12), independent of estrogen receptor status. In conclusion, cyclin-E is not a pure prognostic factor in breast cancer, but rather a predictor of failure of endocrine therapy. Differences in literature on the presumed prognostic value of cyclin-E may be due to differences in treatment. Assessment of cyclin-E levels can aid in improving adjuvant treatment selection.

Acute stimulation of the hypothalamic-pituitary-adrenal axis by IL-1ß, TNFα and IL-6: A dose response study

We investigated the effects of iv and intracerebroventricular (icv) administration of increasing doses of recombinant human IL-1ß, TNFα and IL-6 on plasma corticosterone (B) levels in rats. Rats were equipped with a jugular cannula for repeated blood sampling and a subgroup of rats also received an icv implanted cannula. Iv administration of IL-1ß, TNFα or IL-6 and icv administration of IL-1ß and IL-6 induced a significant dose-dependent increase in plasma B levels, whereas icv injection of TNFα in doses up to 1000 ng/rat was not effective. Iv pretreatment of rats with anti-CRH antiserum had no significant overall effect on the plasma B response to iv administered IL-1ß (500 and 3000 ng/rat), whereas the plasma B response to iv TNFα or IL-6 administration (3000 ng/rat) were significantly reduced. Iv pretreatment of the animals with recombinant human IL-1 receptor antagonist (IL-1ra) significantly blocked the plasma B response to iv treatment with IL-1ß, whereas the TNFα- and IL-6-induced increases in plasma B levels were not affected. Our data show that: 1) iv administration of IL-1ß, TNFα or IL-6 and icv administration of IL-1ß or IL-6 dose-dependently stimulate the HPA axis; 2) when given iv or icv, IL-1ß is more powerful than TNFα and IL-6 in activating the HPA axis; 3) endogenous CRH is involved in the activation of the HPA axis by acute iv administration of TNFα and IL-6. It is most likely that in case of iv treatment with IL-1ß a CRH-independent mechanism is involved. This study provides no arguments for the involvement of endogenous IL-1 in TNFα- or IL-6-induced activation of the HPA axis.

Vascular endothelial growth factor is associated with the efficacy of endocrine therapy in patients with advanced breast carcinoma

Vascular endothelial growth factor (VEGF) is a mediator of angiogenesis and is associated with a poor prognosis in patients with primary breast carcinoma. In the current study, the authors investigated whether there was an association between VEGF levels in tumor tissues and response rates to first‐line, systemic therapy in patients with advanced breast carcinoma.

Tissue inhibitors of metalloproteinase expression in human breast cancer: TIMP-3 is associated with adjuvant endocrine therapy success.

Tissue inhibitors of matrix metalloproteinase (TIMPs) may be involved in tumour growth, apoptosis, angiogenesis, invasion, and the development of metastases. This study has evaluated the association of the expression levels of the TIMP forms 1, 2, 3, and 4, measured by quantitative real‐time RT‐PCR, with classical clinicopathological characteristics, ie age, menopausal status, tumour size, histological grade, number of involved lymph nodes, and steroid hormone receptor status, and with disease progression and treatment sensitivity in 273 breast cancer patients. The mRNA levels of TIMP‐1 and TIMP‐2 were not associated with any known clinicopathological tumour feature. TIMP‐3 and TIMP‐4 levels were significantly higher in steroid hormone receptor‐positive samples, although the levels of TIMP‐4 were much lower than those of the other TIMPs. Only TIMP‐3 predicted relapse‐free survival (RFS) time differently depending on post‐surgical treatment as, in particular, the interaction of TIMP‐3 with endocrine therapy (p = 0.008, HR = 0.24, 95% CI = 0.09–0.69) contributed significantly to RFS in multivariate Cox regression analysis. In subgroup analyses, the 107 patients treated with tamoxifen differed greatly in prognosis after dichotomization by the median TIMP‐3 level (p = 0.0003). Thus, high tumour levels of the matrix metalloproteinases inhibitor and pro‐apoptotic factor TIMP‐3 are associated with successful tamoxifen treatment of patients with breast cancer. Copyright

Predictive Impact of Urokinase-Type Plasminogen Activator Plasminogen Activator Inhibitor Type-1 Complex on the Efficacy of Adjuvant Systemic Therapy in Primary Breast Cancer

One of the most thoroughly studied systems in relation to its prognostic relevance in patients with breast cancer, is the plasminogen activation system. This system comprises of, among others, the urokinase-type plasminogen activator (uPA) and its main inhibitor (PAI-1). In this study we investigated whether the uPA:PAI-1 complex is associated with the responsiveness of patients with primary breast cancer to adjuvant systemic therapy. Quantitative enzyme-linked immunosorbent assays were used to assess the levels of uPA, PAI-1, and uPA:PAI-1 complex in 1119 tumors of patients with primary invasive breast cancer. These patients were followed for a median follow-up time of 59 months (range, 2–267 months) after the primary diagnosis. Correlations with well-known clinicopathological factors, and univariate and multivariate survival analyses were performed. High uPA:PAI-1 complex levels were correlated with an adverse histological grade, and inversely associated with negative estrogen and progesterone receptor status. High tumor levels of uPA:PAI-1 complex predicted an early relapse in the univariate relapse-free survival analysis (P < 0.001). The multivariate analysis showed that high uPA:PAI-1 complex levels were associated with a decreased relapse-free survival time (P = 0.033), independently of age, tumor size, number of lymph nodes affected, progesterone receptor status, uPA, adjuvant endocrine, and chemotherapy. More important, it was demonstrated that there is a larger benefit from adjuvant chemotherapy for patients with higher versus lower tumor levels of uPA:PAI-1 complex. The results of this study imply that the expression of uPA:PAI-1 complex independently predicts the efficacy of adjuvant chemotherapy in patients with primary breast cancer.

Intravenous administration of interleukin-1β induces Fos-like immunoreactivity in corticotropin-releasing hormone neurons in the paraventricular hypothalamic nucleus of the rat

It has been shown that acute administration of recombinant human interleukin 1 beta (IL-1) to rats elicits an activation of the pituitary-adrenal axis. In the present study we investigated immunohistochemically the expression of Fos-like immunoreactivity (Fos-LI) in the hypothalamus of rats following intravenous injection of IL-1. One, 2 and 4 h after IL-1 or physiological saline injections, rats were killed and perfused, and the brains processed for Fos-immunohistochemistry. Dense populations of neurons containing Fos-LI-positive nuclei were found in the paraventricular hypothalamic nuclei (PVH) of IL-1-treated rats. In particular, the dorsal medial parvocellular part, but also some of the other parvocellular subdivisions contained many Fos-LI neurons. Maximal induction of staining was found at a dose of 5 micrograms/rat after 1 or 2 h survival, while immunostaining had decreased to almost control levels after 4 h. No Fos-LI was found in the PVH of control animals. Double immunocytochemical staining for Fos and corticotropin-releasing hormone (CRH) revealed that Fos-LI was predominantly present in parvocellular CRH-containing neurons of the PVH. The finding that peripherally injected IL-1 induces Fos-LI in hypothalamic CRH neurons strengthens the hypothesis that these neurons are part of the circuitry mediating IL-1-induced activation of the pituitary-adrenal axis.

Macrophage migration inhibitory factor (MIF) in meningococcal septic shock and experimental human endotoxemia.

Macrophage migration inhibitory factor (MIF) is a mediator of innate immunity and important in the pathogenesis of septic shock. Lipopolysaccharide (LPS) and tumor necrosis factor (TNF) &agr; are reported to be inducers of MIF. We studied MIF and cytokines in vivo in patients with meningococcal disease, in human experimental endotoxemia, and in whole blood cultures using a newly developed sensitive and specific enzyme-linked immunosorbent assay. Twenty patients with meningococcal disease were investigated. For the human endotoxemia model, 8 healthy volunteers were intravenously injected with 2 ng/kg Escherichia coli LPS. Whole blood from healthy volunteers was incubated with LPS or heat-killed meningococci. Macrophage migration inhibitory factor concentration in blood was increased during meningococcal disease and highest in the patients presenting with shock compared with patients without shock. Plasma concentration of MIF correlated with disease severity, the presence of shock and with the cytokines interleukin (IL) 1&bgr;, IL-10, IL-12, and vascular endothelial growth factor, but not with TNF-&agr;. MIF was not detected in blood in experimental endotoxemia, nor after stimulation of whole blood with LPS or meningococci, although high levels of TNF-&agr; were seen in both models. In conclusion, MIF is increased in patients with meningococcal disease and highest in the presence of shock. Macrophage migration inhibitory factor cannot be detected in a human endotoxemia model and is not produced by whole blood cells incubated with LPS or meningococci.