Asthma is a complex airway allergic disease involving the interplay of various cell types, cytokines, and transcriptional factors. Though many factors contribute to disease etiology, the molecular control of disease phenotype and responsiveness is not well understood. Here we report an essential role of the matrix attachment region (MAR)-binding protein SMAR1 in regulating immune response during allergic airway disease. Conditional knockout of SMAR1 in T cells rendered the mice resistant to eosinophilic airway inflammation against ovalbumin (OVA) allergen with low immunoglobulin E (IgE) and interleukin-5 (IL-5) levels. Moreover, a lower IgE/IgG2a ratio and higher interferon-γ (IFN-γ) response suggested aberrant skewing of T-cell differentiation toward type 1 helper T cell (Th1) response. We show that SMAR1 functions as a negative regulator of Th1 and Th17 differentiation by interacting with two potential and similar MAR regions present on the promoters of T-bet and IL-17. Thus, we present SMAR1 as a regulator of T-cell differentiation that favors the establishment of Th2 cells by modulating Th1 and Th17 responses.

Background & objectives:Studies involving animal models of experimental tuberculosis have elucidated the predominant role of cytokines secreted by T cells and macrophages to be an essential component of the immune response against Mycobacterium tuberculosis infection. The immune activities of CD4+ T cells are mediated in part by Th1 cytokine interferon gamma (IFN-γ) which is produced primarily by T cells and natural killer (NK) cells and critical for initiating the immune response against intracellular pathogen such as M. tuberculosis. Nuclear matrix protein SMAR1 plays an important role in V(D)J recombination, T helper cell differentiation and inflammatory diseases. In this study a transgenic mouse model was used to study the role of SMAR1 in M. tuberculosis infection.Methods:Wild type BALB/c, C57BL/6, BALB/c-EGFP-SMAR1 and C57BL/6-SMAR1 transgenic mice were infected with M. tuberculosis (H37Rv). A dose of 100 bacilli was used for infection via respiratory route. Bacterial load in lung and spleen of infected mice was determined at 2, 4, 6 and 8 wk post-infection. Gene expression analysis for Th1 cytokines and inducible nitric oxide synthase (iNOS) was performed in infected lung tissues by quantitative reverse transcription (RT)-PCR.Results:SMAR1 transgenic mice from both BALB/c and C57BL/6 genetic background displayed higher bacillary load and susceptibility to M. tuberculosis infection compared to wild type mice. This susceptibility was attributed due to compromised of Th1 response exhibited by transgenic mice.Interpretation & conclusions:SMAR1 transgenic mice exhibited susceptibility to M. tuberculosis infection in vivo irrespective of genetic background. This susceptibility was attributed to downregulation of Th1 response and its hallmark cytokine IFN-γ. Hence, SMAR1 plays an important role in modulating host immune response after M. tuberculosis infection.

Background: Development of novel agents and drug combinations are urgently needed for the treatment of breast cancer especially triple negative breast cancer (TNBC). P276 is a potent cyclin-dependent kinase inhibitor with excellent antiproliferative activity against various human cancer cell lines. Here, we report the activity of P276, as a single agent and in combination with paclitaxel or gemcitabine/carboplatin in TNBC cell lines. Material and Methods: In vitro effect of P276 on breast cancer cell lines was determined by cytotoxicity assay, flow cytometry, western blotting and immunofluorescence studies. The antiangiogenic potential of P276 was evaluated using HIF-1 a and VEGF inhibition assay, wound healing and tube formation. In vivo efficacy was studied using human xenograft models in SCID and nude mice. Results: Human breast cancer cell lines including TNBC, treated with P276 were found to be highly susceptible with IC50 ranging from 0.3 to 1.0 mM. In MCF-7 (Her2-, BRCA+/−) and MDA-MB-231 (ER-, PR-, Her2-). P276 significantly down regulated cell cycle proteins pRbser780, cyclin D1, Cdk4 and antiapoptotic protein Bcl-2 followed by 80-85% apoptosis. P276 treatment also lead to the inhibition of PARP enzyme activity or PARP cleavage depending on the cell type. Interestingly, P276 inhibited the key angiogenic mediators HIF-1 a and VEGF with IC50 of 0.095mM and 0.31mM respectively in reporter gene based assays. These antiangiogenic effects were confirmed by immunofluorescence and migration studies in TNBC cell lines and endothelial tube formation. In breast cancer patient derived tumor cell line MAXF 401 (basal like) xenografts in nude mice, cyclical dosing of P276 showed growth inhibition of 71%. Moreover, combination studies of P276 with paclitaxel or gemcitabine plus carboplatin showed synergistic effect in TNBC cell lines. Discussion: Our studies provide compelling evidence for the clinical development of P276 for the treatment of triple negative breast cancer either as monotherapy or in combination with paclitaxel or gemcitabine/carboplatin. Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr P6-15-05.