Triple negative breast cancer (TNBC), which includes tumors lacking estrogen receptor, progesterone receptor, and HER-2 proteins, represents one of the most challenging cancers for developing an effective therapy due to lack of a therapeutic target. Here, we tested an immunotherapy approach that is personalized, easy to accomplish, and not requiring establishment of cell lines or gene transfer to treat metastatic TNBC. The defining principle of this cancer immunotherapy technology is the incorporation of glycolipid-anchored forms of immunostimulatory molecules (GPI-ISMs) onto tumor membrane vesicles (TMV) derived from TNBC mice model. We tested our immunotherapy approach in both therapeutic and prophylactic vaccinations. Therapeutic vaccination of mice after tumor resections increased over all survival in mice receiving TNBC tumor membrane-based immunotherapy. Also, prophylactic vaccination of mice with immunotherapy and antibody based combination therapy showed a reduction in lung metastasis by 90% relative to untreated groups in clonogenic assays. To test whether the observed decrease in metastasis in our combination therapy is due to enhanced CD4 or CD8 T cell specific immune response, we depleted CD4 and CD8 T cells separately using their specific antibodies and performed clonogenic assay. Depletion of CD8 (but not CD4) T cells prior to challenge recapitulated full metastasis, suggesting that immunotherapy induced CD8 T cells specific responses. In summary, we have demonstrated the feasibility of this novel immunotherapy in the triple negative breast cancer model when used in combination with antibody therapy. A reduction in metastasis to the lung, prolonged overall survival, and enhanced CD8 T cell responses were observed.

\r\nRishan Singh is a biologist. \r\n

The interactions of compounds on human cell lines are either influenced by the composition of substances present in plant material or alteration of constituents by solvent fractionation. These substances or constituents have an influence on the percentage cytotoxicity readings of compounds in human cell culture. Understanding and correlating the relationship between cytotoxicity and other parameters, such as cell death inducing mechanisms, will assist pharmaceutical chemists to synthesize compounds that can target particular ailments with greater efficiency. This will also allow scientists to understand the interaction of compounds with different cell types for different compound fractions.

Michaela Jirku has graduated in 2011 at the University of Chemistry and Technology, Prague, field of clinical biochemistry. Now she has studied PhD in biochemistry for 4 years. She is a co author of 5 papers in reputed journals

Transient receptor potential melastatin 1 (TRPM1) channel belongs to the superfamily of ion channels that respond to various physiological stimuli like chemosensation, termosensation and mechanosensation. TRP channels have six transmembrane domains with a pore region between the fifth and the sixth segment. Cytosolic N-/C-tails are responsible for regulation of TRPs, which carry binding sites for signal molecules. TRPM1 channel is ubiquitously expressed in most eukaryotic cells and is involved in many cellular processes like transduction of sensory signals and regulation of Ca2+ and Mg2+ homeostasis. Mutations in TRPM1 gene effect on cell cycle in human skin cells. It seems that a loss of TRPM1 in human melanocytes correlates with increased aggressiveness in melanoma and the homozygous loss of TRPM1 in bipolar cells in retina could be a possible explanation for congenital stationary night blindness in humans. \r\nWe studied possible interactions between phosphatidylinositol-4,5-bisphosphate (PIP2) and the N-terminus (NT) of TRPM1. Using bioinformatic approach we identified PIP2 binding site in A451-N566 region. This domain contains several basic amino acids which can interact with anionic phospholipids. Alanine substitution mutagenesis screening revealed the crucial amino acids for these interactions. The equilibrium dissociation constants were estimated using surface plasmon resonance measurement. We identified PIP2-binding site and found mutations that decreased the affinity of TRPM1 NT/PIP2 interaction. Based on this results we concluded that basic residues play crucial role in TRP channels binding to PIP2. Moreover, we have provided the structural insight to TRPM1-NT/PIP2 interaction using computer ligand docking.\r\nSupported by Grants GAUK 238214 and GACR P304/12/G069.\r\n