A database of whole-body action videos for the study of action, emotion, and untrustworthiness

We present a database of high-definition (HD) videos for the study of traits inferred from whole-body actions. Twenty-nine actors (19 female) were filmed performing different actions—walking, picking up a box, putting down a box, jumping, sitting down, and standing and acting—while conveying different traits, including four emotions (anger, fear, happiness, sadness), untrustworthiness, and neutral, where no specific trait was conveyed. For the actions conveying the four emotions and untrustworthiness, the actions were filmed multiple times, with the actor conveying the traits with different levels of intensity. In total, we made 2,783 action videos (in both two-dimensional and three-dimensional format), each lasting 7 s with a frame rate of 50 fps. All videos were filmed in a green-screen studio in order to isolate the action information from all contextual detail and to provide a flexible stimulus set for future use. In order to validate the traits conveyed by each action, we asked participants to rate each of the actions corresponding to the trait that the actor portrayed in the two-dimensional videos. To provide a useful database of stimuli of multiple actions conveying multiple traits, each video name contains information on the gender of the actor, the action executed, the trait conveyed, and the rating of its perceived intensity. All videos can be downloaded free at the following address: http://www-users.york.ac.uk/~neb506/databases.html. We discuss potential uses for the database in the analysis of the perception of whole-body actions

Genetically Engineering Glycolysis in T Cells Increases Their Antitumor Function

BACKGROUND: T cells play a central role in the antitumor response. However, they often face numerous hurdles in the tumor microenvironment, including the scarcity of available essential metabolites such as glucose and amino acids. Moreover, cancer cells can monopolize these resources to thrive and proliferate by upregulating metabolite transporters and maintaining a high metabolic rate, thereby outcompeting T cells. METHODS: Herein, we sought to improve T-cell antitumor function in the tumor vicinity by enhancing their glycolytic capacity to better compete with tumor cells. To achieve this, we engineered human T cells to express a key glycolysis enzyme, phosphofructokinase, in conjunction with Glucose transporter 3, a glucose transporter. We co-expressed these, along with tumor-specific chimeric antigen or T-cell receptors. RESULTS: Engineered cells demonstrated an increased cytokine secretion and upregulation of T-cell activation markers compared with control cells. Moreover, they displayed superior glycolytic capacity, which translated into an improved in vivo therapeutic potential in a xenograft model of human tumors. CONCLUSION: In summary, these findings support the implementation of T-cell metabolic engineering to enhance the efficacy of cellular immunotherapies for cancer

Preclinical evaluation and structural optimization of anti-BCMA CAR to target multiple myeloma

Chimeric antigen receptor (CAR) T-cell based immunotherapy has become a promising treatment mainly for hematological malignancies. Following the major success of CD19-targeted CAR, new potential targets for other malignancies are required. As such, B-cell maturation antigen (BCMA) is an attractive tumor-associated antigen to be targeted in multiple myeloma (MM). Herein, we aimed at assessing the function and optimal configuration of different BCMA-specific CAR, based on the same targeting moiety but with a different hinge and co-stimulatory domain. We compared their function to that of a previously characterized BCMA-CAR used in clinical trials. All constructs were expressed at high levels by primary human T cells and could trigger cytokine production and cytotoxicity upon co-culture with multiple myeloma targets. Nonetheless, critical differences were observed in off-target activation, exhaustion, and activation marker expression and in vivo antitumoral activity mediated by these different constructs. Interestingly, we noted that CD8-based hinge, combined with a 4-1BB intracellular domain, proved superior compared to IgG4-connecting regions, and/or a CD28-signaling moiety respectively. Overall, this study emphasizes the influence of CAR primary structure on its function and led to the identification of a highly efficient BCMA-specific CAR, namely H8BB, which displayed superior anti-tumoral activity both in vitro and long-term in vivo efficacy

Degradation of HIF-1alpha under Hypoxia Combined with Induction of Hsp90 Polyubiquitination in Cancer Cells by Hypericin: a Unique Cancer Therapy

The perihydroxylated perylene quinone hypericin has been reported to possess potent anti-metastatic and antiangiogenic activities, generated by targeting diverse crossroads of cancer-promoting processes via unique mechanisms. Hypericin is the only known exogenous reagent that can induce forced poly-ubiquitination and accelerated degradation of heat shock protein 90 (Hsp90) in cancer cells. Hsp90 client proteins are thereby destabilized and rapidly degraded. Hsp70 client proteins may potentially be also affected via preventing formation of hsp90-hsp70 intermediate complexes. We show here that hypericin also induces enhanced degradation of hypoxia-inducible factor 1α (HIF-1α) in two human tumor cell lines, U87-MG glioblastoma and RCC-C2VHL−/− renal cell carcinoma and in the non-malignant ARPE19 retinal pigment epithelial cell line. The hypericin-accelerated turnover of HIF-1α, the regulatory precursor of the HIF-1 transcription factor which promotes hypoxic stress and angiogenic responses, overcomes the physiologic HIF-1α protein stabilization which occurs in hypoxic cells. The hypericin effect also eliminates the high HIF-1α levels expressed constitutively in the von-Hippel Lindau protein (pVHL)-deficient RCC-C2VHL−/− renal cell carcinoma cell line. Unlike the normal ubiquitin-proteasome pathway-dependent turnover of HIF-α proteins which occurs in normoxia, the hypericin-induced HIF-1α catabolism can occur independently of cellular oxygen levels or pVHL-promoted ubiquitin ligation of HIF-1α. It is mediated by lysosomal cathepsin-B enzymes with cathepsin-B activity being optimized in the cells through hypericin-mediated reduction in intracellular pH. Our findings suggest that hypericin may potentially be useful in preventing growth of tumors in which HIF-1α plays pivotal roles, and in pVHL ablated tumor cells such as renal cell carcinoma through elimination of elevated HIF-1α contents in these cells, scaling down the excessive angiogenesis which characterizes these tumors

Abstract 2870: Preventive efficacy of curcumin on ENU-induced carcinogenic transformation of ApcMin/+ mammary epithelial (MinMG) cells established from ApcMin/+ mouse

Abstract PURPOSE: ApcMin/+ (Min, multiple intestinal neoplasia) is a point mutation at codon 850 in the murine homolog of human APC tumor suppressor gene. Mutations and hypermethylation of APC gene has recently been reported in up to 45% of sporadic breast cancers. Our recently established ApcMin/+ mammary epithelial cell line [MinMG] from ApcMin/+ Min mouse, exhibit shorter population doubling time and 25% higher proliferation within a week, increased expression of β-catenin and cyclin D1 proteins, compared to normal Apc+/+ C57MG cell line [Apc+/+ C57BL/6J mouse]. In accordance with the two-hit theory of carcinogenesis, MinMG cells are susceptible to a “second hit” and thus prone to malignant transformation. This study was designed to evaluate preventive efficacy of the dietary agent Curcumin [CUR], on N-Ethyl-N-nitrosourea [ENU] induced carcinogenesis of MinMG mammary epithelial cells. EXPERIMENTAL DESIGN: To evaluate the in vitro preventive efficacy of CUR on ENU induced hyperproliferation and transformation, MinMG cells were treated with 20uM CUR for 24hr prior to addition of 20uM ENU for 2hr. The treated cells with parallel controls were further grown for 48hr in regular medium and cell proliferation was measured. Cells were further maintained in regular medium for number of passages to monitor the difference in proliferation rate, at regular weekly interval. Cell proliferation (cell count &amp; MTT assay), Cell cycle analyses, and Anchorage Independent Growth (AIG), served as surrogate endpoint biomarkers to evaluate preventive efficacy of CUR for ENU induced carcinogenic transformation of MinMG cells. RESULTS: MinMG cells exhibit a 3 fold increase in cell proliferation rate induced by 2hr exposure of 20uM ENU compared to untreated cells that persisted for 19 weeks of regular passaging post-treatment. MinMG cells pretreated with 20uM CUR prior to ENU exposure show significant preventive modulation in cell proliferation (12 fold decrease), ratio of quiescent vs proliferative phases of cell cycle (G1:S+G2/M=115% in favor of G1), and number of soft agar colonies (50% reduction), compared to corresponding control. These results show correlation with altered expression of Ki67, β-catenin and cyclin D1 proteins in MinMG cells. CONCLUSIONS: Our results suggest ApcMin/+ mutation in MinMG cells render them at high risk for carcinogenic transformation. This in vitro pre-clinical model validates its use for Apc+/− initiated risk for mammary carcinogenesis. Pretreatment of cells with CUR reduced ENU induced hyperproliferation and transformation in MinMG cells indicating its preventive efficacy for carcinogenic transformation. Thus our data indicates that CUR is efficacious in preventing ENU induced high risk of hyperproliferation and carcinogenic transformation of ApcMin/+ MinMG mammary cells. [Support: NIH-RO1 CA122394] Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 2870.</jats:p