1,538 research outputs found

    Converging organoids and extracellular matrix::New insights into liver cancer biology

    Get PDF

    Converging organoids and extracellular matrix::New insights into liver cancer biology

    Get PDF
    Primary liver cancer, consisting primarily of hepatocellular carcinoma (HCC) and cholangiocarcinoma (CCA), is a heterogeneous malignancy with a dismal prognosis, resulting in the third leading cause of cancer mortality worldwide [1, 2]. It is characterized by unique histological features, late-stage diagnosis, a highly variable mutational landscape, and high levels of heterogeneity in biology and etiology [3-5]. Treatment options are limited, with surgical intervention the main curative option, although not available for the majority of patients which are diagnosed in an advanced stage. Major contributing factors to the complexity and limited treatment options are the interactions between primary tumor cells, non-neoplastic stromal and immune cells, and the extracellular matrix (ECM). ECM dysregulation plays a prominent role in multiple facets of liver cancer, including initiation and progression [6, 7]. HCC often develops in already damaged environments containing large areas of inflammation and fibrosis, while CCA is commonly characterized by significant desmoplasia, extensive formation of connective tissue surrounding the tumor [8, 9]. Thus, to gain a better understanding of liver cancer biology, sophisticated in vitro tumor models need to incorporate comprehensively the various aspects that together dictate liver cancer progression. Therefore, the aim of this thesis is to create in vitro liver cancer models through organoid technology approaches, allowing for novel insights into liver cancer biology and, in turn, providing potential avenues for therapeutic testing. To model primary epithelial liver cancer cells, organoid technology is employed in part I. To study and characterize the role of ECM in liver cancer, decellularization of tumor tissue, adjacent liver tissue, and distant metastatic organs (i.e. lung and lymph node) is described, characterized, and combined with organoid technology to create improved tissue engineered models for liver cancer in part II of this thesis. Chapter 1 provides a brief introduction into the concepts of liver cancer, cellular heterogeneity, decellularization and organoid technology. It also explains the rationale behind the work presented in this thesis. In-depth analysis of organoid technology and contrasting it to different in vitro cell culture systems employed for liver cancer modeling is done in chapter 2. Reliable establishment of liver cancer organoids is crucial for advancing translational applications of organoids, such as personalized medicine. Therefore, as described in chapter 3, a multi-center analysis was performed on establishment of liver cancer organoids. This revealed a global establishment efficiency rate of 28.2% (19.3% for hepatocellular carcinoma organoids (HCCO) and 36% for cholangiocarcinoma organoids (CCAO)). Additionally, potential solutions and future perspectives for increasing establishment are provided. Liver cancer organoids consist of solely primary epithelial tumor cells. To engineer an in vitro tumor model with the possibility of immunotherapy testing, CCAO were combined with immune cells in chapter 4. Co-culture of CCAO with peripheral blood mononuclear cells and/or allogenic T cells revealed an effective anti-tumor immune response, with distinct interpatient heterogeneity. These cytotoxic effects were mediated by cell-cell contact and release of soluble factors, albeit indirect killing through soluble factors was only observed in one organoid line. Thus, this model provided a first step towards developing immunotherapy for CCA on an individual patient level. Personalized medicine success is dependent on an organoids ability to recapitulate patient tissue faithfully. Therefore, in chapter 5 a novel organoid system was created in which branching morphogenesis was induced in cholangiocyte and CCA organoids. Branching cholangiocyte organoids self-organized into tubular structures, with high similarity to primary cholangiocytes, based on single-cell sequencing and functionality. Similarly, branching CCAO obtain a different morphology in vitro more similar to primary tumors. Moreover, these branching CCAO have a higher correlation to the transcriptomic profile of patient-paired tumor tissue and an increased drug resistance to gemcitabine and cisplatin, the standard chemotherapy regimen for CCA patients in the clinic. As discussed, CCAO represent the epithelial compartment of CCA. Proliferation, invasion, and metastasis of epithelial tumor cells is highly influenced by the interaction with their cellular and extracellular environment. The remodeling of various properties of the extracellular matrix (ECM), including stiffness, composition, alignment, and integrity, influences tumor progression. In chapter 6 the alterations of the ECM in solid tumors and the translational impact of our increased understanding of these alterations is discussed. The success of ECM-related cancer therapy development requires an intimate understanding of the malignancy-induced changes to the ECM. This principle was applied to liver cancer in chapter 7, whereby through a integrative molecular and mechanical approach the dysregulation of liver cancer ECM was characterized. An optimized agitation-based decellularization protocol was established for primary liver cancer (HCC and CCA) and paired adjacent tissue (HCC-ADJ and CCA-ADJ). Novel malignancy-related ECM protein signatures were found, which were previously overlooked in liver cancer transcriptomic data. Additionally, the mechanical characteristics were probed, which revealed divergent macro- and micro-scale mechanical properties and a higher alignment of collagen in CCA. This study provided a better understanding of ECM alterations during liver cancer as well as a potential scaffold for culture of organoids. This was applied to CCA in chapter 8 by combining decellularized CCA tumor ECM and tumor-free liver ECM with CCAO to study cell-matrix interactions. Culture of CCAO in tumor ECM resulted in a transcriptome closely resembling in vivo patient tumor tissue, and was accompanied by an increase in chemo resistance. In tumor-free liver ECM, devoid of desmoplasia, CCAO initiated a desmoplastic reaction through increased collagen production. If desmoplasia was already present, distinct ECM proteins were produced by the organoids. These were tumor-related proteins associated with poor patient survival. To extend this method of studying cell-matrix interactions to a metastatic setting, lung and lymph node tissue was decellularized and recellularized with CCAO in chapter 9, as these are common locations of metastasis in CCA. Decellularization resulted in removal of cells while preserving ECM structure and protein composition, linked to tissue-specific functioning hallmarks. Recellularization revealed that lung and lymph node ECM induced different gene expression profiles in the organoids, related to cancer stem cell phenotype, cell-ECM integrin binding, and epithelial-to-mesenchymal transition. Furthermore, the metabolic activity of CCAO in lung and lymph node was significantly influenced by the metastatic location, the original characteristics of the patient tumor, and the donor of the target organ. The previously described in vitro tumor models utilized decellularized scaffolds with native structure. Decellularized ECM can also be used for creation of tissue-specific hydrogels through digestion and gelation procedures. These hydrogels were created from both porcine and human livers in chapter 10. The liver ECM-based hydrogels were used to initiate and culture healthy cholangiocyte organoids, which maintained cholangiocyte marker expression, thus providing an alternative for initiation of organoids in BME. Building upon this, in chapter 11 human liver ECM-based extracts were used in combination with a one-step microfluidic encapsulation method to produce size standardized CCAO. The established system can facilitate the reduction of size variability conventionally seen in organoid culture by providing uniform scaffolding. Encapsulated CCAO retained their stem cell phenotype and were amendable to drug screening, showing the feasibility of scalable production of CCAO for throughput drug screening approaches. Lastly, Chapter 12 provides a global discussion and future outlook on tumor tissue engineering strategies for liver cancer, using organoid technology and decellularization. Combining multiple aspects of liver cancer, both cellular and extracellular, with tissue engineering strategies provides advanced tumor models that can delineate fundamental mechanistic insights as well as provide a platform for drug screening approaches.<br/

    Multidisciplinary perspectives on Artificial Intelligence and the law

    Get PDF
    This open access book presents an interdisciplinary, multi-authored, edited collection of chapters on Artificial Intelligence (‘AI’) and the Law. AI technology has come to play a central role in the modern data economy. Through a combination of increased computing power, the growing availability of data and the advancement of algorithms, AI has now become an umbrella term for some of the most transformational technological breakthroughs of this age. The importance of AI stems from both the opportunities that it offers and the challenges that it entails. While AI applications hold the promise of economic growth and efficiency gains, they also create significant risks and uncertainty. The potential and perils of AI have thus come to dominate modern discussions of technology and ethics – and although AI was initially allowed to largely develop without guidelines or rules, few would deny that the law is set to play a fundamental role in shaping the future of AI. As the debate over AI is far from over, the need for rigorous analysis has never been greater. This book thus brings together contributors from different fields and backgrounds to explore how the law might provide answers to some of the most pressing questions raised by AI. An outcome of the Católica Research Centre for the Future of Law and its interdisciplinary working group on Law and Artificial Intelligence, it includes contributions by leading scholars in the fields of technology, ethics and the law.info:eu-repo/semantics/publishedVersio

    Predicting Paid Certification in Massive Open Online Courses

    Get PDF
    Massive open online courses (MOOCs) have been proliferating because of the free or low-cost offering of content for learners, attracting the attention of many stakeholders across the entire educational landscape. Since 2012, coined as “the Year of the MOOCs”, several platforms have gathered millions of learners in just a decade. Nevertheless, the certification rate of both free and paid courses has been low, and only about 4.5–13% and 1–3%, respectively, of the total number of enrolled learners obtain a certificate at the end of their courses. Still, most research concentrates on completion, ignoring the certification problem, and especially its financial aspects. Thus, the research described in the present thesis aimed to investigate paid certification in MOOCs, for the first time, in a comprehensive way, and as early as the first week of the course, by exploring its various levels. First, the latent correlation between learner activities and their paid certification decisions was examined by (1) statistically comparing the activities of non-paying learners with course purchasers and (2) predicting paid certification using different machine learning (ML) techniques. Our temporal (weekly) analysis showed statistical significance at various levels when comparing the activities of non-paying learners with those of the certificate purchasers across the five courses analysed. Furthermore, we used the learner’s activities (number of step accesses, attempts, correct and wrong answers, and time spent on learning steps) to build our paid certification predictor, which achieved promising balanced accuracies (BAs), ranging from 0.77 to 0.95. Having employed simple predictions based on a few clickstream variables, we then analysed more in-depth what other information can be extracted from MOOC interaction (namely discussion forums) for paid certification prediction. However, to better explore the learners’ discussion forums, we built, as an original contribution, MOOCSent, a cross- platform review-based sentiment classifier, using over 1.2 million MOOC sentiment-labelled reviews. MOOCSent addresses various limitations of the current sentiment classifiers including (1) using one single source of data (previous literature on sentiment classification in MOOCs was based on single platforms only, and hence less generalisable, with relatively low number of instances compared to our obtained dataset;) (2) lower model outputs, where most of the current models are based on 2-polar iii iv classifier (positive or negative only); (3) disregarding important sentiment indicators, such as emojis and emoticons, during text embedding; and (4) reporting average performance metrics only, preventing the evaluation of model performance at the level of class (sentiment). Finally, and with the help of MOOCSent, we used the learners’ discussion forums to predict paid certification after annotating learners’ comments and replies with the sentiment using MOOCSent. This multi-input model contains raw data (learner textual inputs), sentiment classification generated by MOOCSent, computed features (number of likes received for each textual input), and several features extracted from the texts (character counts, word counts, and part of speech (POS) tags for each textual instance). This experiment adopted various deep predictive approaches – specifically that allow multi-input architecture - to early (i.e., weekly) investigate if data obtained from MOOC learners’ interaction in discussion forums can predict learners’ purchase decisions (certification). Considering the staggeringly low rate of paid certification in MOOCs, this present thesis contributes to the knowledge and field of MOOC learner analytics with predicting paid certification, for the first time, at such a comprehensive (with data from over 200 thousand learners from 5 different discipline courses), actionable (analysing learners decision from the first week of the course) and longitudinal (with 23 runs from 2013 to 2017) scale. The present thesis contributes with (1) investigating various conventional and deep ML approaches for predicting paid certification in MOOCs using learner clickstreams (Chapter 5) and course discussion forums (Chapter 7), (2) building the largest MOOC sentiment classifier (MOOCSent) based on learners’ reviews of the courses from the leading MOOC platforms, namely Coursera, FutureLearn and Udemy, and handles emojis and emoticons using dedicated lexicons that contain over three thousand corresponding explanatory words/phrases, (3) proposing and developing, for the first time, multi-input model for predicting certification based on the data from discussion forums which synchronously processes the textual (comments and replies) and numerical (number of likes posted and received, sentiments) data from the forums, adapting the suitable classifier for each type of data as explained in detail in Chapter 7

    Minimal information for studies of extracellular vesicles (MISEV2023): From basic to advanced approaches

    Get PDF
    Extracellular vesicles (EVs), through their complex cargo, can reflect the state of their cell of origin and change the functions and phenotypes of other cells. These features indicate strong biomarker and therapeutic potential and have generated broad interest, as evidenced by the steady year-on-year increase in the numbers of scientific publications about EVs. Important advances have been made in EV metrology and in understanding and applying EV biology. However, hurdles remain to realising the potential of EVs in domains ranging from basic biology to clinical applications due to challenges in EV nomenclature, separation from non-vesicular extracellular particles, characterisation and functional studies. To address the challenges and opportunities in this rapidly evolving field, the International Society for Extracellular Vesicles (ISEV) updates its 'Minimal Information for Studies of Extracellular Vesicles', which was first published in 2014 and then in 2018 as MISEV2014 and MISEV2018, respectively. The goal of the current document, MISEV2023, is to provide researchers with an updated snapshot of available approaches and their advantages and limitations for production, separation and characterisation of EVs from multiple sources, including cell culture, body fluids and solid tissues. In addition to presenting the latest state of the art in basic principles of EV research, this document also covers advanced techniques and approaches that are currently expanding the boundaries of the field. MISEV2023 also includes new sections on EV release and uptake and a brief discussion of in vivo approaches to study EVs. Compiling feedback from ISEV expert task forces and more than 1000 researchers, this document conveys the current state of EV research to facilitate robust scientific discoveries and move the field forward even more rapidly

    LIPIcs, Volume 251, ITCS 2023, Complete Volume

    Get PDF
    LIPIcs, Volume 251, ITCS 2023, Complete Volum

    AI: Limits and Prospects of Artificial Intelligence

    Get PDF
    The emergence of artificial intelligence has triggered enthusiasm and promise of boundless opportunities as much as uncertainty about its limits. The contributions to this volume explore the limits of AI, describe the necessary conditions for its functionality, reveal its attendant technical and social problems, and present some existing and potential solutions. At the same time, the contributors highlight the societal and attending economic hopes and fears, utopias and dystopias that are associated with the current and future development of artificial intelligence

    Mining Butterflies in Streaming Graphs

    Get PDF
    This thesis introduces two main-memory systems sGrapp and sGradd for performing the fundamental analytic tasks of biclique counting and concept drift detection over a streaming graph. A data-driven heuristic is used to architect the systems. To this end, initially, the growth patterns of bipartite streaming graphs are mined and the emergence principles of streaming motifs are discovered. Next, the discovered principles are (a) explained by a graph generator called sGrow; and (b) utilized to establish the requirements for efficient, effective, explainable, and interpretable management and processing of streams. sGrow is used to benchmark stream analytics, particularly in the case of concept drift detection. sGrow displays robust realization of streaming growth patterns independent of initial conditions, scale and temporal characteristics, and model configurations. Extensive evaluations confirm the simultaneous effectiveness and efficiency of sGrapp and sGradd. sGrapp achieves mean absolute percentage error up to 0.05/0.14 for the cumulative butterfly count in streaming graphs with uniform/non-uniform temporal distribution and a processing throughput of 1.5 million data records per second. The throughput and estimation error of sGrapp are 160x higher and 0.02x lower than baselines. sGradd demonstrates an improving performance over time, achieves zero false detection rates when there is not any drift and when drift is already detected, and detects sequential drifts in zero to a few seconds after their occurrence regardless of drift intervals

    Efficient Deep Learning for Real-time Classification of Astronomical Transients

    Get PDF
    A new golden age in astronomy is upon us, dominated by data. Large astronomical surveys are broadcasting unprecedented rates of information, demanding machine learning as a critical component in modern scientific pipelines to handle the deluge of data. The upcoming Legacy Survey of Space and Time (LSST) of the Vera C. Rubin Observatory will raise the big-data bar for time- domain astronomy, with an expected 10 million alerts per-night, and generating many petabytes of data over the lifetime of the survey. Fast and efficient classification algorithms that can operate in real-time, yet robustly and accurately, are needed for time-critical events where additional resources can be sought for follow-up analyses. In order to handle such data, state-of-the-art deep learning architectures coupled with tools that leverage modern hardware accelerators are essential. The work contained in this thesis seeks to address the big-data challenges of LSST by proposing novel efficient deep learning architectures for multivariate time-series classification that can provide state-of-the-art classification of astronomical transients at a fraction of the computational costs of other deep learning approaches. This thesis introduces the depthwise-separable convolution and the notion of convolutional embeddings to the task of time-series classification for gains in classification performance that are achieved with far fewer model parameters than similar methods. It also introduces the attention mechanism to time-series classification that improves performance even further still, with significant improvement in computational efficiency, as well as further reduction in model size. Finally, this thesis pioneers the use of modern model compression techniques to the field of photometric classification for efficient deep learning deployment. These insights informed the final architecture which was deployed in a live production machine learning system, demonstrating the capability to operate efficiently and robustly in real-time, at LSST scale and beyond, ready for the new era of data intensive astronomy
    corecore