Clinical impact of high-profile animal-based research reported in the UK national press

Objectives We evaluated animal-based biomedical ‘breakthroughs’ reported in the UK national press in 1995 (25 years prior to the conclusion of this study). Based on evidence of overspeculative reporting of biomedical research in other areas (eg, press releases and scientific papers), we specifically examined animal research in the media, asking, ‘In a given year, what proportion of animal research “breakthroughs”’ published in the UK national press had translated, more than 20 years later, to approved interventions?’ Methods We searched the Nexis media database (LexisNexis.com) for animal-based biomedical reports in the UK national press. The only restrictions were that the intervention should be specific, such as a named drug, gene, biomedical pathway, to facilitate follow-up, and that there should be claims of some clinical promise. Main outcome measures Were any interventions approved for human use? If so, when and by which agency? If not, why, and how far did development proceed? Were any other, directly related interventions approved? Did any of the reports overstate human relevance? Results Overspeculation and exaggeration of human relevance was evident in all the articles examined. Of 27 unique published ‘breakthroughs’, only one had clearly resulted in human benefit. Twenty were classified as failures, three were inconclusive and three were partially successful. Conclusions The results of animal-based preclinical research studies are commonly overstated in media reports, to prematurely imply often-imminent ‘breakthroughs’ relevant to human medicine

Cymbopogon citratus and Camellia sinensis extracts selectively induce apoptosis in cancer cells and reduce growth of lymphoma xenografts in vivo

Cancer is a progressive disease characterized by a biological disharmony in which damaged or mutated cells are able to survive and proliferate by escaping cell death. Moreover, these cells are reported to have elevated levels of reactive oxygen species (ROS) and are highly dependent on cellular defense mechanisms against oxidative stress. A specific cancer of the lymphatic system, lymphoma, affects our body’s infection-fighting cells while posing a great risk to all age groups. Numerous nutraceuticals and natural polyphenolic compounds have a wide range of abilities to alter cellular redox states with potential implications in various diseases. Furthermore, therapeutic options for cancers are mostly nonselective treatments including genotoxic or tubulin-targeting compounds. Some of the natural extracts, containing multiple bioactive compounds, could target multiple pathways in cancer cells to selectively induce cell death. Cymbopogon citratus (lemongrass) and Camellia sinensis (white tea) extracts have been shown to have medicinal properties, however, their activity against lymphoma, as well as mechanistic details, have not been fully characterized. Herein, we report potent anti-cancer properties in dose and time-dependent manners of ethanolic lemongrass and hot water white tea extracts in lymphoma and leukemia models. Both extracts were able to effectively induce apoptosis selectively in these human cancer cell types. Interestingly, ethanolic lemongrass extract induces apoptosis primarily by the extrinsic pathway and was found to be dependent on the generation of ROS. Conversely, apoptotic induction by hot water white tea extract was independent of ROS. Furthermore, both of these extracts caused mitochondrial depolarization and decreased rates of oxygen consumption in lymphoma and leukemia cells, leading to cell death. Most importantly, both these extracts were effective in reducing tumor growth in human lymphoma xenograft models when administered orally. Thus, these natural extracts could have potential for being nontoxic alternatives for the treatment of cancer

Role of tumor architecture in elicitation of effector functions of human cytotoxic T-lymphocytes recognizing melanoma associated antigens

Growth in 3D architectures has been shown to promote the resistance of cancers to treatment with drugs, cytokines, or irradiation, thereby potentially playing an important role in tumor expansion. 3D architectures might also play a role in impairing immunorecognition of cancer cells by cytotoxic T lymphocytes (CTLs) specific for tumor-associated antigens. Culture of HBL, D10 (both HLA-A*0201+, TAA+) and NA8 (HLA-A*0201+, TAA-) melanoma cell lines on poly-Hydroxyethylmethacrylate-coated plates, resulted in generation of 3D multicellular tumor spheroids (MCTS). Kinetics of cell proliferation in MCTS was significantly slower than in monolayer cultures. Following long-term culture (>10-15 days) MCTS showed highly compact and organised cell growth in outer layers, with necrotic cores. To obtain an insight into the role played by tumor architecture in the elicitation of specific gene expression patterns, we addressed gene expression profiles of NA8 melanoma cells cultured in two-dimensional monolayers (2D) or in 3D (MCTS). Oligonucleotide microarray analysis of the expression of over 20,000 genes was performed on cells cultured in standard 2D, in the presence of collagen as model of extracellular matrix (ECM), or in MCTS. Gene expression profiles of cells cultured in 2D in the presence or absence of ECM were highly similar, with more than threefold differences limited to five genes. In contrast, culture in MCTS resulted in the significant, more than threefold, upregulation of the expression of >100 transcripts, while 73 transcripts were more than threefold downregulated. In particular, genes encoding CXCL1, 2, and 3 (GRO-α, -β, and γ), IL-8, CCL20 (MIP-3α), and Angiopoietin-like 4 were significantly upregulated, whereas basic-FGF and CD49d encoding genes were significantly downregulated. Oligonucleotide chip data were validated at the gene and protein level by quantitative real-time PCR, ELISA, and cell surface staining assays. Taken together, our data indicate that structural modifications of the architecture of tumor cell cultures result in a significant upregulation of the expression of a number of genes previously shown to play a role in melanoma progression and metastatic process. Then we investigated the effects of 3D culture on the recognition of melanoma cells by antigenspecific HLA class I-restricted Cytotoxic T-Lymphocytes (CTL). IFN-γ production can be used as a surrogate marker for tumor cell immunorecognition. Co-culture of melanoma spheroids with HLA-A0201 restricted Melan-A/MART-127-35-specific CTL clones resulted in significantly defective TAA recognition by CTL as compared to 2D, as witnessed by decreased IFN-γ production and decreased Fas Ligand, perforin and granzyme B gene expression. Indeed, Melan- A/MART-1 expression, at both gene and protein levels, was significantly decreased in 3D as compared with 2D tumor cell cultures. Concomitantly, a parallel decrease of HLA class I molecule expression was also observed. Differential gene profiling studies on HBL cells showed an increased expression of genes encoding molecules involved in intercellular adhesion, such as junctional adhesion molecule 2 and cadherin-like 1 (>20- and 8-fold up-regulated, respectively) in 3D as compared with 2D cultures. We further identified a multiplicity of mechanisms potentially involved. In particular : 1) MCTS per se limit CTL capacity of recognizing HLA class I restricted antigens by reducing exposed cell surfaces. 2) Expression of melanoma differentiation antigens is down-regulated in tumor cell spheroids as compared to 2D unrelated to hypoxia or increased Oncostatin M gene expression but rather to decreased MITF gene expression. 3) Expression of HLA class I molecules is frequently down-regulated in melanoma MCTS, as compared to 2D, possibly due to decreased IRF-1 gene expression. 4) Lactate production by melanoma cells is increased in MCTS, as compared to 2D and lactate significantly inhibits TAA triggered IFN-γ production by CTL. Taken together, our data suggest that mere growth of melanoma cells in 3D architectures, in the absence of immunoselective pressure, may result in defective recognition by tumor-associated antigen-specific CTL and a constellation of mechanisms are involved in causing this impairment of immunorecognition

Mycobacterium W immunotherapy for treating pulmonary tuberculosis : a systematic review

Includes bibliographical references.Tuberculosis (TB) remains a major health problem in the developing world, accounting for approximately 2 million deaths per year. The search for appropriate and applicable therapies to reduce the burden of TB is essential. Mycobacterium w (M w) is a heat-killed immune-modulating vaccine designed to attenuate the effects of TB infection and reduce the time to sputum negativity, thereby improving cure rates and decreasing transmission rates

Imaging Breast Cancer Progression and Lymph Node Metastases in Murine Models Using MRI and Magnetic Nanoparticles

Most breast cancer related deaths are caused by the spread or metastasis of the primary tumor to distant sites in the body. The lymph nodes are one of the first places where metastases can be detected and are frequently examined for macroscopic metastases to help determine course of treatment for patients. However, little is known about the significance of microscopic metastases and disseminated individual cancer cells within the nodes. The goal of this work was to use MRI to monitor the development of primary tumors and lymphatic metastases in models of breast cancer. In this thesis, we examined the MRI appearance of lymph nodes in several different strains of immune compromised mice (nude, CB -17 SCID, NOD/SCID IL2Rnull) and compared the appearance to immune competent C57/Bl6 strain. We found that immune deficiencies influenced the MRI appearance of nodes and that the nude strain had highly variable lymph node appearance and volume. We also compared orthotopic transplantation models of breast cancer that used both the nude and CB-17 SCID strains using MRI. We found that MRI was most reliable for detecting metastases in the lymph nodes of SCID mice and that the variability of the appearance of nodes in nude mice can lead to their misclassification. We then used the SCID orthotopic breast cancer model to monitor the appearance and retention of iron oxide nanoparticle labeled cancer cells in both the primary tumor and lymph nodes. We found that iron-labeled cells are still detected within the primary tumor after 28 days post-implantation and that these labeled cells almost exclusively migrated to the lymph nodes. The development of improved methods for monitoring the development of the primary tumor and metastases and the roles that different cells populations have in these processes will allow for more accurate knowledge of how cancer cell heterogeneity impacts disease progression. These tools will allow for more effective monitoring of the treatment effect of new drugs on primary tumors and metastatic dissemination

Anticancer Effects and Mechanisms of CFI-400945 and Radiation in Breast Cancer

Breast cancer is a leading cause of death in women and development of new treatments is essential. Polo-like Kinase 4 (PLK4) controls centriole duplication and its inhibition by CFI-400945 induces genomic instability and aneuploidy. Radiation therapy (RT) also induces aneuploidy leading to cell death, although development of radioresistance is common. We hypothesized that CFI-400945 and RT act synergistically in breast cancer. Colony formation assays identified synergistic anticancer effects of CFI-400945 and RT, with combinatorial effects also observed for RT with either siRNA inhibition of PLK4 or with the PLK4 inhibitor Centrinone B. This suggests that the antiproliferative effect of these combinations are, at least partly, mediated through PLK4. Immunocytochemistry for Centrin showed significant overamplification of centrioles in combination compared to single agent treatment, suggesting a possible combined mechanism of action. These results support further translational studies of CFI-400945 and RT as a combination treatment to improve breast cancer outcomes

Triple Positive Microparticles as a “Liquid Biopsy” for Risk Stratification of Prostate Cancer

Prostate cancer remains a substantial contributor to cancer-related mortality worldwide. Current screening methods include obtaining a PSA blood test. However, controversy surrounds its use as it is neither sensitive nor specific. Nanoscale flow cytometry is a type of microfluidics-based technology that allows enumeration of submicron tumor fragments known as microparticles (MPs). In this study, prostate specific microparticles in patient plasma were targeted using fluorophore-conjugated antibodies. Targeted cell surface antigens or biomarkers include: prostate specific membrane antigen (PSMA), six-transmembrane epithelial antigen of the prostate-1 (STEAP1), ghrelin receptor (GHSR1a) and CD151. A statistically significant difference in the level of MP levels was measured with PSMA+STEAP1+GHSR1a and PSMA+STEAP1+CD151 triple-expressing MPs when comparing Gleason score (GS) 6 to GS3+4, GS4+3 and GS≥8 cohorts. In this pilot and exploratory study, I show that MPs have the potential of becoming a “liquid biopsy” that can assist in risk stratification prior to a prostate needle biopsy