TGF-ß signalling is mediated by two autonomously functioning TßRI:TßRII pairs.

Transforming growth factor (TGF)-ßs are dimeric polypeptides that have vital roles in regulating cell growth and differentiation. They signal by assembling a receptor heterotetramer composed of two TßRI:TßRII heterodimers. To investigate whether the two heterodimers bind and signal autonomously, one of the TGF-ß protomers was substituted to block receptor binding. The substituted dimer, TGF-ß3 WD, bound the TßRII extracellular domain and recruited the TßRI with affinities indistinguishable from TGF-ß3, but with one-half the stoichiometry. TGF-ß3 WD was further shown to retain one-quarter to one-half the signalling activity of TGF-ß3 in three established assays for TGF-ß function. Single-molecule fluorescence imaging with GFP-tagged receptors demonstrated a measurable increase in the proportion of TßRI and TßRII dimers upon treatment with TGF-ß3, but not with TGF-ß3 WD. These results provide evidence that the two TßRI:TßRII heterodimers bind and signal in an autonomous manner. They further underscore how the TGF-ßs diverged from the bone morphogenetic proteins, the ancestral ligands of the TGF-ß superfamily that signal through a RI:RII:RII heterotrimer.

Doxorubicin in combination with a small TGFbeta inhibitor: a potential novel therapy for metastatic breast cancer in mouse models.

BACKGROUND: Recent studies suggested that induction of epithelial-mesenchymal transition (EMT) might confer both metastatic and self-renewal properties to breast tumor cells resulting in drug resistance and tumor recurrence. TGFbeta is a potent inducer of EMT and has been shown to promote tumor progression in various breast cancer cell and animal models. PRINCIPAL FINDINGS: We report that chemotherapeutic drug doxorubicin activates TGFbeta signaling in human and murine breast cancer cells. Doxorubicin induced EMT, promoted invasion and enhanced generation of cells with stem cell phenotype in murine 4T1 breast cancer cells in vitro, which were significantly inhibited by a TGFbeta type I receptor kinase inhibitor (TbetaRI-KI). We investigated the potential synergistic anti-tumor activity of TbetaR1-KI in combination with doxorubicin in animal models of metastatic breast cancer. Combination of Doxorubicin and TbetaRI-KI enhanced the efficacy of doxorubicin in reducing tumor growth and lung metastasis in the 4T1 orthotopic xenograft model in comparison to single treatments. Doxorubicin treatment alone enhanced metastasis to lung in the human breast cancer MDA-MB-231 orthotopic xenograft model and metastasis to bone in the 4T1 orthotopic xenograft model, which was significantly blocked when TbetaR1-KI was administered in combination with doxorubicin. CONCLUSIONS: These observations suggest that the adverse activation of TGFbeta pathway by chemotherapeutics in the cancer cells together with elevated TGFbeta levels in tumor microenvironment may lead to EMT and generation of cancer stem cells resulting in the resistance to the chemotherapy. Our results indicate that the combination treatment of doxorubicin with a TGFbeta inhibitor has the potential to reduce the dose and consequently the toxic side-effects of doxorubicin, and improve its efficacy in the inhibition of breast cancer growth and metastasis.

Cytotoxic and cell vacuolating activity of Vibrio fluvialis isolated from paediatric patients with diarrhoea.

Vibrio fluvialis is a halophilic Vibrio species associated with acute diarrhoeal illness in humans. It has the potential to cause outbreaks and has an association with paediatric diarrhoea. In this study, 11 V. fluvialis strains isolated from hospitalized patients with acute diarrhoea at the Infectious Diseases Hospital, Kolkata were extensively characterized. All the strains showed growth in peptone broth containing 7% NaCl. The strains showed variable results in Voges-Proskauer test and to a vibriostatic agent. There was also variation in their antibiograms, and some of the strains were multidrug resistant. Among the 11 strains, two showed only a single band difference in their PFGE profile and the remaining strains showed nine different PFGE patterns. However, unlike PFGE, the strains exhibited close matches and clustering in their ribotype patterns. The haemolytic effect on sheep red blood cells varied with strains. Partial sequence analysis revealed that the V. fluvialis haemolysin gene has 81% homology with that of the El Tor haemolysin of Vibrio cholerae. A striking finding was the capability of all the strains to evoke distinct cytotoxic and vacuolation effects on HeLa cells.

Molecular characterisation of rough strains of Vibrio cholerae isolated from diarrhoeal cases in India and their comparison to smooth strains.

Sixteen of the 18 Vibrio cholerae rough strains isolated from hospitalised diarrhoea patients were found to contain O1 serotype-specific (wbe) genes and all currently known virulence genes. Expression of the regulatory element ToxR was evident in these strains. Cholera toxin production ability of the rough strains was found to be higher (c. three- to five-fold) as compared to the smooth counterparts and this was transcriptionally regulated. Strains exhibiting the rough phenotype were more amenable to the uptake of CTXphi, which led us to consider that the rough phenotype could play a role in the generation of genetic diversity among V. cholerae strains.

Escalating association of Vibrio cholerae O139 with cholera outbreaks in India.

Between December 1999 and December 2000, teams from the National Institute of Cholera and Enteric Diseases, Calcutta, India, examined eight outbreaks of cholera, which occurred in different parts of the country distant from each other. In two of these outbreaks each, only V. cholerae O1 biotype ElTor or V. cholerae O139 could be isolated, while in the remaining four outbreaks, both O1 and O139 were isolated. The interesting feature is the escalating association of V. cholerae O139 with outbreaks of cholera; two of the most recent outbreaks, one in Calcutta and one in Orissa, were caused exclusively by O139. The O139 strains from the six different outbreaks were genotypically closely related. These trends indicate a shift in the outbreak propensity of V. cholerae O139.