SkiVis: Visual Exploration and Route Planning in Ski Resorts.

Optimal ski route selection is a challenge based on a multitude of factors, such as the steepness, compass direction, or crowdedness. The personal preferences of every skier towards these factors require individual adaptations, which aggravate this task. Current approaches within this domain do not combine automated routing capabilities with user preferences, missing out on the possibility of integrating domain knowledge in the analysis process. We introduce SkiVis, a visual analytics application to interactively explore ski slopes and provide routing recommendations based on user preferences. In collaboration with ski guides and enthusiasts, we elicited requirements and guidelines for such an application and propose different workflows depending on the skiers' familiarity with the resort. In a case study on the resort of Ski Arlberg, we illustrate how to leverage volunteered geographic information to enable a numerical comparison between slopes. We evaluated our approach through a pair-analytics study and demonstrate how it supports skiers in discovering relevant and preference-based ski routes. Besides the tasks investigated in the study, we derive additional use cases from the interviews that showcase the further potential of SkiVis, and contribute directions for further research opportunities.

FS/DS: A Theoretical Framework for the Dual Analysis of Feature Space and Data Space.

With the surge of data-driven analysis techniques, there is a rising demand for enhancing the exploration of large high-dimensional data by enabling interactions for the joint analysis of features (i.e., dimensions). Such a dual analysis of the feature space and data space is characterized by three components, (1) a view visualizing feature summaries, (2) a view that visualizes the data records, and (3) a bidirectional linking of both plots triggered by human interaction in one of both visualizations, e.g., Linking & Brushing. Dual analysis approaches span many domains, e.g., medicine, crime analysis, and biology. The proposed solutions encapsulate various techniques, such as feature selection or statistical analysis. However, each approach establishes a new definition of dual analysis. To address this gap, we systematically reviewed published dual analysis methods to investigate and formalize the key elements, such as the techniques used to visualize the feature space and data space, as well as the interaction between both spaces. From the information elicited during our review, we propose a unified theoretical framework for dual analysis, encompassing all existing approaches extending the field. We apply our proposed formalization describing the interactions between each component and relate them to the addressed tasks. Additionally, we categorize the existing approaches using our framework and derive future research directions to advance dual analysis by including state-of-the-art visual analysis techniques to improve data exploration.

Advanced Dome cellulose/alginate/chitosan composite matrix design with gastric and intestinal co-targeting capacities.

Dome matrix was designed with gastric and intestinal targeting capacities using melatonin and caffeine as model drugs, and alginate, chitosan and cellulose as composite materials. The melatonin, caffeine and intermediate hydroxypropylmethylcelluose-based dispersible modules were prepared through compaction. Caffeine piled module was capped at both ends with melatonin void modules via intermediate dispersible modules into Dome matrix. Dispersion of intermediate module detached melatonin module from Dome matrix and had it floated in stomach providing a more complete melatonin release due to favorable pH-pKa relationship of dissolution medium and drug. With reference to the caffeine module, the detachment of melatonin module facilitated its gastrointestinal transit as a reduced size matrix, with majority of caffeine delivered in colon. The dual site-targeted and -release Dome matrix is applicable as reference oral carrier for pharmaceutical, nutraceutical, functional food and veterinary medicine where a complex formulation and performancein vivoare required.

Radiotherapy orchestrates natural killer cell dependent antitumor immune responses through CXCL8.

Radiotherapy is a mainstay cancer therapy whose antitumor effects partially depend on T cell responses. However, the role of Natural Killer (NK) cells in radiotherapy remains unclear. Here, using a reverse translational approach, we show a central role of NK cells in the radiation-induced immune response involving a CXCL8/IL-8-dependent mechanism. In a randomized controlled pancreatic cancer trial, CXCL8 increased under radiotherapy, and NK cell positively correlated with prolonged overall survival. Accordingly, NK cells preferentially infiltrated irradiated pancreatic tumors and exhibited CD56dim-like cytotoxic transcriptomic states. In experimental models, NF-κB and mTOR orchestrated radiation-induced CXCL8 secretion from tumor cells with senescence features causing directional migration of CD56dim NK cells, thus linking senescence-associated CXCL8 release to innate immune surveillance of human tumors. Moreover, combined high-dose radiotherapy and adoptive NK cell transfer improved tumor control over monotherapies in xenografted mice, suggesting NK cells combined with radiotherapy as a rational cancer treatment strategy.

Toward Mass Video Data Analysis: Interactive and Immersive 4D Scene Reconstruction.

The technical progress in the last decades makes photo and video recording devices omnipresent. This change has a significant impact, among others, on police work. It is no longer unusual that a myriad of digital data accumulates after a criminal act, which must be reviewed by criminal investigators to collect evidence or solve the crime. This paper presents the VICTORIA Interactive 4D Scene Reconstruction and Analysis Framework ("ISRA-4D" 1.0), an approach for the visual consolidation of heterogeneous video and image data in a 3D reconstruction of the corresponding environment. First, by reconstructing the environment in which the materials were created, a shared spatial context of all available materials is established. Second, all footage is spatially and temporally registered within this 3D reconstruction. Third, a visualization of the hereby created 4D reconstruction (3D scene + time) is provided, which can be analyzed interactively. Additional information on video and image content is also extracted and displayed and can be analyzed with supporting visualizations. The presented approach facilitates the process of filtering, annotating, analyzing, and getting an overview of large amounts of multimedia material. The framework is evaluated using four case studies which demonstrate its broad applicability. Furthermore, the framework allows the user to immerse themselves in the analysis by entering the scenario in virtual reality. This feature is qualitatively evaluated by means of interviews of criminal investigators and outlines potential benefits such as improved spatial understanding and the initiation of new fields of application.

Teaching Clustering Algorithms With EduClust: Experience Report and Future Directions.

We share our experiences teaching university students about clustering algorithms using EduClust, an online visualization we developed. EduClust supports professors in preparing teaching material and students in visually and interactively exploring cluster steps and the effects of changing clustering parameters. We used EduClust for two years in our computer science lectures on clustering algorithms and share our experience integrating the online application in a data science curriculum. We also point to opportunities for future development.

Visual Analytics of Volunteered Geographic Information: Detection and Investigation of Urban Heat Islands.

Urban heat islands are local areas where the temperature is much higher than in the vicinity and are a modern phenomenon that occurs mainly in highly developed areas, such as large cities. This effect has a negative impact on energy management in buildings, and also has a direct impact on human health, especially for elderly people. With the advent of volunteered geographic information from private weather station networks, more high-resolution data are now available within cities to better analyze this effect. However, such datasets are large and have heterogeneous characteristics requiring visual-interactive applications to support further analysis. We use machine learning methods to predict urban heat islands occurrences and utilize temporal and spatio-temporal visualizations to contextualize the emergence of urban heat islands to comprehend the influencing causes and their effects. Subsequently, we demonstrate the analysis capabilities of our application by presenting two use cases.

KIR downregulation by IL-12/15/18 unleashes human NK cells from KIR/HLA-I inhibition and enhances killing of tumor cells.

To exploit autologous NK cells for cancer immunotherapy, it is highly relevant to circumvent killer cell immunoglobulin-like receptor (KIR)-mediated self-inhibition of human NK cells by HLA-I-expressing tumor cells. Here, we show that stimulation of NK cells with IL-12/15/18 for two days led to downregulation of surface expression of the inhibitory KIR2DL2/L3, KIR2DL1 and KIR3DL1 receptors on peripheral blood NK cells. Downregulation of KIR expression was attributed to decreased KIR mRNA levels which could be re-induced already 3 days after re-culture in IL-2. Reduced KIR2DL2/L3 expression on IL-12/15/18-activated NK cells resulted in less inhibition upon antibody-mediated KIR engagement and increased CD16-dependent cytotoxicity in redirected lysis assays. Most importantly, downregulated KIR2DL2/L3 expression enabled enhanced cytotoxicity of IL-12/15/18-stimulated NK cells against tumor cells expressing cognate HLA-I molecules. NK cells pre-activated with IL-12/15/18 were previously shown to exert potent anti-tumor activity and memory-like long-lived functionality, mediating remission in a subset of acute myeloid leukemia (AML) patients in a clinical trial. Our study reveals a novel mechanism of IL-12/15/18 in improving the cytotoxicity of NK cells by reducing their sensitivity to inhibition by self-HLA-I due to decreased KIR expression, highlighting the potency of IL-12/15/18-activated NK cells for anti-tumor immunotherapy protocols.

MicroRNA-519a-3p mediates apoptosis resistance in breast cancer cells and their escape from recognition by natural killer cells.

Aggressive breast cancer is associated with poor patient outcome and characterized by the development of tumor cell variants that are able to escape from control of the immune system or are resistant to targeted therapies. The complex molecular mechanisms leading to immune escape and therapy resistance are incompletely understood. We have previously shown that high miR-519a-3p levels are associated with poor survival in breast cancer. Here, we demonstrate that miR-519a-3p confers resistance to apoptosis induced by TRAIL, FasL and granzyme B/perforin by interfering with apoptosis signaling in breast cancer cells. MiR-519a-3p diminished the expression of its direct target genes for TRAIL-R2 (TNFRSF10B) and for caspase-8 (CASP8) and its indirect target gene for caspase-7 (CASP7), resulting in reduced sensitivity and tumor cell apoptosis in response to apoptotic stimuli. Furthermore, miR-519a-3p impaired tumor cell killing by natural killer (NK) cells via downregulation of the NKG2D ligands ULBP2 and MICA on the surface of tumor cells that are crucial for the recognition of these tumor cells by NK cells. We determined that miR-519a-3p was overexpressed in more aggressive mutant TP53 breast cancer that was associated with poor survival. Furthermore, low levels of TRAIL-R2, caspase-7 and caspase-8 correlated with poor survival, suggesting that the inhibitory effect of miR-519a-3p on TRAIL-R2 and caspases may have direct clinical relevance in lowering patient's prognosis. In conclusion, we demonstrate that miR-519a-3p is a critical factor in mediating resistance toward cancer cell apoptosis and impairing tumor cell recognition by NK cells. This joint regulation of apoptosis and immune cell recognition through miR-519a-3p supports the hypothesis that miRNAs are key regulators of cancer cell fate, facilitating cancer progression and evasion from immunosurveillance at multiple and interconnected levels.

Personalized RNA mutanome vaccines mobilize poly-specific therapeutic immunity against cancer.

T cells directed against mutant neo-epitopes drive cancer immunity. However, spontaneous immune recognition of mutations is inefficient. We recently introduced the concept of individualized mutanome vaccines and implemented an RNA-based poly-neo-epitope approach to mobilize immunity against a spectrum of cancer mutations. Here we report the first-in-human application of this concept in melanoma. We set up a process comprising comprehensive identification of individual mutations, computational prediction of neo-epitopes, and design and manufacturing of a vaccine unique for each patient. All patients developed T cell responses against multiple vaccine neo-epitopes at up to high single-digit percentages. Vaccine-induced T cell infiltration and neo-epitope-specific killing of autologous tumour cells were shown in post-vaccination resected metastases from two patients. The cumulative rate of metastatic events was highly significantly reduced after the start of vaccination, resulting in a sustained progression-free survival. Two of the five patients with metastatic disease experienced vaccine-related objective responses. One of these patients had a late relapse owing to outgrowth of ß2-microglobulin-deficient melanoma cells as an acquired resistance mechanism. A third patient developed a complete response to vaccination in combination with PD-1 blockade therapy. Our study demonstrates that individual mutations can be exploited, thereby opening a path to personalized immunotherapy for patients with cancer.

Highly efficient IL-21 and feeder cell-driven ex vivo expansion of human NK cells with therapeutic activity in a xenograft mouse model of melanoma.

Natural killer (NK) cells are promising antitumor effector cells, but the generation of sufficient NK cell numbers for adoptive immunotherapy remains challenging. Therefore, we developed a method for highly efficient ex vivo expansion of human NK cells. Ex vivo expansion of NK cells in medium containing IL-2 and irradiated clinical-grade feeder cells (EBV-LCL) induced a 22-fold NK cell expansion after one week that was significantly increased to 53-fold by IL-21. Repeated stimulation with irradiated EBV-LCL and IL-2 and addition of IL-21 at the initiation of the culture allowed sustained NK cell proliferation with 1011-fold NK cell expansion after 6 weeks. Compared to naive NK cells, expanded NK cells upregulated TRAIL, NKG2D, and DNAM-1, had superior cytotoxicity against tumor cell lines in vitro and produced more IFNγ and TNF-α upon PMA/Iono stimulation. Most importantly, adoptive transfer of NK cells expanded using feeder cells, IL-2 and IL-21 led to significant inhibition of tumor growth in a melanoma xenograft mouse model, which was greater than with NK cells activated with IL-2 alone. Intriguingly, adoptively transferred NK cells maintained their enhanced production of IFNγ and TNF-α upon ex vivo restimulation, although they rapidly lost their capacity to degranulate and mediate tumor cytotoxicity after the in vivo transfer. In conclusion, we developed a protocol for ex vivo NK cell expansion that results in outstanding cell yields. The expanded NK cells possess potent antitumor activity in vitro and in vivo and could be utilized at high numbers for adoptive immunotherapy in the clinic.

Adoptively transferred natural killer cells maintain long-term antitumor activity by epigenetic imprinting and CD4+ T cell help.

Natural killer (NK) cell infusions can induce remissions in subsets of patients with different types of cancer. The optimal strategies for NK cell activation prior to infusion are still under debate. There is recent evidence that NK cells can acquire long-term functional competence by preactivation with the cytokines IL-12/15/18. The mechanisms supporting the maintenance of long-term NK cell antitumor activity are incompletely under-stood. Here, we show that NK cells preactivated in vitro with IL-12/15/18, but not with IL-15 alone, maintained high antitumor activity even 1 mo after transfer into lymphopenic RAG-2-/-γc-/- mice. The NK cell intrinsic ability for IFNγ production coincided with demethylation of the conserved non-coding sequence (CNS) 1 in the Ifng locus, previously shown to enhance transcription of Ifng. In a xenograft melanoma mouse model, human IL-12/15/18-preactivated NK cells rejected tumors more efficiently. In RAG-2-/-γc-/- mice, co-transfer of CD4+ T cells further improved the long-term competence of NK cells for IFNγ production that was dependent on IL-2. CD4+ T cell activation during homeostatic proliferation required macrophages and further promoted the long-term NK cell antitumor activity. Thus, NK cells can "remember" a previous exposure to cytokines by epigenetic imprinting resulting in a remarkable stability of the IFNγ-producing phenotype after adoptive transfer. In addition, our results support combination of cytokine-preactivated NK cells with CD4+ T cell activation upon lymphopenic conditioning to achieve long-term NK cell effector function for cancer immunotherapy.

The HMGB1 protein induces a metabolic type of tumour cell death by blocking aerobic respiration.

The high-mobility group box 1 (HMGB1) protein has a central role in immunological antitumour defense. Here we show that natural killer cell-derived HMGB1 directly eliminates cancer cells by triggering metabolic cell death. HMGB1 allosterically inhibits the tetrameric pyruvate kinase isoform M2, thus blocking glucose-driven aerobic respiration. This results in a rapid metabolic shift forcing cells to rely solely on glycolysis for the maintenance of energy production. Cancer cells can acquire resistance to HMGB1 by increasing glycolysis using the dimeric form of PKM2, and employing glutaminolysis. Consistently, we observe an increase in the expression of a key enzyme of glutaminolysis, malic enzyme 1, in advanced colon cancer. Moreover, pharmaceutical inhibition of glutaminolysis sensitizes tumour cells to HMGB1 providing a basis for a therapeutic strategy for treating cancer.

Exploiting natural killer cells for therapy of melanoma.

During the recent years, immunotherapy has obtained substantial impact on the clinical treatment of melanoma. Besides promising approaches based on T lymphocytes, natural killer (NK) cells have gained more and more attention as anti-melanoma effector cells. NK cell activation is inhibited by HLA class I molecules expressed by target cells, so they preferentially attack tumor cells that express low levels of HLA class I. Partial or complete loss of HLA class I expression is a frequent event during the development of melanoma. In parallel, ligands for activating NK cell receptors become induced upon malignant transformation. Thus, melanoma cells are often efficiently recognized and lysed by NK cells at least in vitro. In vivo, however, melanomas have developed multiple sophisticated strategies to escape from NK cell mediated attack. Several novel approaches aim at harnessing NK cells to treat melanoma patients and to counteract existing tumor escape mechanisms. This review summarizes the most recent advances in the field.

Toward the next generation of NK cell-based adoptive cancer immunotherapy.

The adoptive transfer of interleukin (IL)-2-expanded natural killer (NK) cells has provided unsatisfactory clinical benefits to patients affected by solid tumors. Our study demonstrates that the activation of NK cells with IL-12/IL-15/IL-18 prior to transfer into tumor-bearing mice is critical for obtaining high recovery rates, effector functions in vivo and tumor regression.

Sustained effector function of IL-12/15/18-preactivated NK cells against established tumors.

Natural killer cell (NK cell)-based immunotherapy of cancer is hampered by the transient effector function of NK cells. Recently, mouse IL-12/15/18-preactivated NK cells were shown to persist with sustained effector function in vivo. Our study investigated the antitumor activity of such NK cells. A single injection of syngeneic IL-12/15/18-preactivated NK cells, but neither naive nor IL-15- or IL-2-pretreated NK cells, combined with irradiation substantially reduced growth of established mouse tumors. Radiation therapy (RT) was essential for the antitumor activity of transferred NK cells. IL-12/15/18-preactivated NK cells expressed high levels of IL-2Rα (CD25), and their rapid in vivo proliferation depended on IL-2 produced by CD4+ T cells. IL-12/15/18-preactivated NK cells accumulated in the tumor tissue and persisted at high cell numbers with potent effector function that required the presence of CD4+ T cells. RT greatly increased numbers and function of transferred NK cells. Human IL-12/15/18-preactivated NK cells also displayed sustained effector function in vitro. Our study provides a better understanding for the rational design of immunotherapies of cancer that incorporate NK cells. Moreover, our results reveal an essential role of CD4+ T cell help for sustained antitumor activity by NK cells linking adaptive and innate immunity.

Characterization of MHC ligands for peptide based tumor vaccination.

Short peptides derived from cellular proteins may escape complete destruction during protein catabolism and finally serve as a showcase in the immune system. Exposed at the cell surface to scrutiny by T cells, MHC:peptide complexes mediate a highly specific and immediate information transfer from diseased cells to the cellular immune system. Numerous clinical vaccination trials have been carried out employing MHC-presented peptides for T-cell activation with encouraging results but so far without a final breakthrough. In this review, we briefly highlight the molecular basis of MHC-peptide interactions governed by specificity pockets and anchor residues, as summarized in allele-specific peptide motifs. State-of-the-art technology is comprehensively presented and gives an overview of modern mass spectrometric strategies used for qualitative and quantitative analysis of MHC ligands. We describe the details of the HLA-B*3801 peptide motif by comparing features of natural MHC ligands, resulting in a scoring matrix that enables epitope prediction from any viral or tumor antigen. The pronounced individuality in peptide presentation by MHC molecules, as reflected in the highly specific peptide motifs of different MHC allotypes or the tissue-specific MHC ligandomes, represents a current area of interest within this field. Finally, the identification of post-translational modifications--most important phosphorylations--and the promises this holds will be discussed in this chapter.