Perceptions of Cross-Cultural Challenges and Successful Approaches in Facilitating the Improvement of Equine Welfare.

Projects that aim to improve the welfare of equids worldwide usually involve people from different countries and cultures working together. Given that professionals involved with multi-stakeholder projects often work cross-culturally, this study examined their experiences regarding the challenges involved in, and their reflections on, how to work in a culturally sensitive way. Semi-structured interviews were conducted with 14 participants working in a total of 29 countries and analysed using thematic analysis. Key response themes emerged from the responses to questions covering the areas of perceptions of animal welfare, challenges working cross-culturally and embracing cultural sensitivity. The overriding theme regarding perceptions of animal welfare was that of barriers to animal welfare, under which emerged the subthemes of limited financial and material resources, limited understanding of the tenets of animal welfare, and attachment to traditional medicines and practices. Exploring the key challenges resulted in two themes: challenges regarding the local context and etiquette, and those regarding working with different stakeholders. Considering cultural sensitivity, again, two themes emerged: the importance of trust and respect, and of working with local partners. Previous works have highlighted the importance of shared linguistic knowledge, interpersonal skills and cultural knowledge, and these elements also emerged in this research. As well as providing insights into the challenges of working cross-culturally, the findings of this study have enabled the development of suggestions for how this work could be taken forward in a practical way to be of use to professionals in this sector.

Development of college student focused care in a psychiatric hospital: Preliminary clinical outcomes and future directions.

OBJECTIVE: The unique features of college experience make it essential to address escalating mental-health challenges beyond college campuses. In 2010, we launched college-student focused (CSF) care nested within an adult day treatment program in a psychiatric hospital. The CSF care consists of student group therapies, individual consultation services for hospital staff and student-patients, and student-focused mental health guides for patients and families. This study preliminarily examined the clinical impact of CSF care on post-treatment symptoms reduction. PARTICIPANTS AND METHODS: In 235 college student-patients admitted to the day program between 2011 July to 2013 January, we assessed the targeted outcomes of the CSF care, using a newly developed CSF questionnaire. RESULTS: Higher levels of CSF care-related outcomes predicted reduced post-treatment depression and anxiety, even after controlling for baseline clinical symptoms and post-treatment skills usage. CONCLUSIONS: These results highlighted the benefits and need for CSF care on a healthcare system level.

Perceptions of Fear and Anxiety in Horses as Reported in Interviews with Equine Behaviourists.

One of the key welfare concerns for horses in the United Kingdom is lack of recognition of fear in horses. This study aimed to gain an understanding of how well horse care givers recognise fear and/or anxiety in horses by interviewing equine behaviourists (who interact with large numbers of horse care givers and talk to them about this topic routinely). The experiences of Animal Behaviour and Training Council (ABTC)-registered equine behaviourists working with horse caregivers were examined, including the ability of clients to recognise fear and/or anxiety in horses, how clients respond when discussing fear as the reason for their horse's behaviour, and what explanations the participants use to explain fear and anxiety. Semi-structured interviews were conducted with nine participants and analysed using thematic analysis before being written up to reflect the discussion points. When asked how well horse caregivers recognise fear and/or anxiety in horses, three key response themes emerged: caregivers are extremely poor at recognizing fear and anxiety in horses; some clients do recognise behavioural signs indicating fear and/or anxiety but only the overt signs (e.g., rearing, running away) rather than the more subtle signs (e.g., tension in face, subtle avoidance behaviours such as a hesitant gait); and fear and/or anxiety behaviour is often misinterpreted or mislabelled. These key themes recurred throughout several other interview questions. This study has provided initial insights into the lack of recognition of fear and anxiety of horses by their caregivers in the United Kingdom as well as tried and tested approaches to conversations to change this. Such synthesis of experience and techniques across the equine behaviour sector, together with the information gained regarding perception of equine caregivers, could be a valuable approach to improve the effectiveness of behaviour consultations and welfare initiatives.

Dynamics of Metabolic Pathways and Stress Response Patterns during Human Neural Stem Cell Proliferation and Differentiation.

Understanding early nervous system stress response mechanisms is crucial for studying developmental neurotoxicity and devising neuroprotective treatments. We used hiPSC-derived long-term self-renewing neuroepithelial stem (lt-NES) cells differentiated for up to 12 weeks as an in vitro model of human neural development. Following a transcriptome analysis to identify pathway alterations, we induced acute oxidative stress (OS) using tert-butyl hydroperoxide (TBHP) and assessed cell viability at different stages of neural differentiation. We studied NRF2 activation, autophagy, and proteasomal function to explore the contribution and interplay of these pathways in the acute stress response. With increasing differentiation, lt-NES cells showed changes in the expression of metabolic pathway-associated genes with engagement of the pentose phosphate pathway after 6 weeks, this was accompanied by a decreased susceptibility to TBHP-induced stress. Microarray analysis revealed upregulation of target genes of the antioxidant response KEAP1-NRF2-ARE pathway after 6 weeks of differentiation. Pharmacological inhibition of NRF2 confirmed its vital role in the increased resistance to stress. While autophagy was upregulated alongside differentiation, it was not further increased upon oxidative stress and had no effect on stress-induced cell loss and the activation of NRF2 downstream genes. In contrast, proteasome inhibition led to the aggravation of the stress response resulting in decreased cell viability, derangement of NRF2 and KEAP1 protein levels, and lacking NRF2-pathway activation. Our data provide detailed insight into the dynamic regulation and interaction of pathways involved in modulating stress responses across defined time points of neural differentiation.

Birth plans: A systematic, integrative review into their purpose, process, and impact.

BACKGROUND: The birth plan was introduced in the 1980s to facilitate communication between maternity care providers and women and increase agency for childbearing women in the face of medicalised birth. Forty years on, the birth plan is a heterogeneous document with uncertainty surrounding its purpose, process, and impact. The aim of this review was to synthesise the evidence and improve understanding of the purpose, process and impact of the birth plan on childbearing women's experiences and outcomes. METHODS: This systematic review followed the PRISMA guidelines. A comprehensive search strategy was designed and applied to electronic databases CINAHL, MEDLINE, PsychINFO, Cochrane Library, Scopus, and ClinicalTrials.gov. Articles were appraised using the Crowe Critical Appraisal Tool and a five-step integrative approach to analysis followed. FINDINGS: Eleven articles were identified, all quantitative in nature. It is clear that the general purpose of birth plans is communication, with decision making a key factor. Even though the processes of birth planning were varied, having a birth plan was associated with generally positive birth outcomes. CONCLUSIONS: Despite the heterogeneity of birth plans, birth plans were associated with positive outcomes for childbearing women when developed in collaboration with care providers. The act of collaboratively creating a birth plan may improve obstetric outcomes, aid realistic expectations, and improve satisfaction and the sense of control.

Systems analysis of miRNA biomarkers to inform drug safety.

microRNAs (miRNAs or miRs) are short non-coding RNA molecules which have been shown to be dysregulated and released into the extracellular milieu as a result of many drug and non-drug-induced pathologies in different organ systems. Consequently, circulating miRs have been proposed as useful biomarkers of many disease states, including drug-induced tissue injury. miRs have shown potential to support or even replace the existing traditional biomarkers of drug-induced toxicity in terms of sensitivity and specificity, and there is some evidence for their improved diagnostic and prognostic value. However, several pre-analytical and analytical challenges, mainly associated with assay standardization, require solutions before circulating miRs can be successfully translated into the clinic. This review will consider the value and potential for the use of circulating miRs in drug-safety assessment and describe a systems approach to the analysis of the miRNAome in the discovery setting, as well as highlighting standardization issues that at this stage prevent their clinical use as biomarkers. Highlighting these challenges will hopefully drive future research into finding appropriate solutions, and eventually circulating miRs may be translated to the clinic where their undoubted biomarker potential can be used to benefit patients in rapid, easy to use, point-of-care test systems.

Attitudes of the Equestrian Public towards Equine End-of-Life Decisions.

A key welfare concern for the equine population in the U.K. has been identified as delayed death, leading to prolonged suffering of horses. Reasons why some horse owners fail to have their horses euthanised include financial cost, emotional attachment, peer pressure, negative attitudes towards killing and poor recognition of behavioural indicators of equine pain and stress. The Five Freedoms framework of welfare was used to build a Likert-style survey to investigate the factors underlying attitudes of horse owners towards welfare measures in an end-of-life decision. Participants were asked to respond to hypothetical welfare scenarios and to give details of any horses they had had euthanised. The survey was conducted predominantly via equestrian Facebook groups and obtained 160 participant responses. Reliability of the scale was acceptable, with Cronbach's α=0.89. Principal Component Analysis was used to load the hypothetical scenarios onto seven factors containing 62.2% of the variance. The first four factors could be categorized according to "Ethology-informed Management", "Traditional Horse Management", "Emotional Issues" and "Physical Issues". Participants were more likely to consider euthanasia for physical issues, compared with issues relating to affective state and/or ethology, although it was not clear whether this was due to disregard for welfare issues relating to mental health or failure to recognise them as such. A large number of responses stated that the scenario had no bearing on whether a horse should be euthanised, again suggesting a lack of recognition of welfare issues and their implications. When asked to state their reasons for euthanising their horses, participants cited almost exclusively physical reasons, with the exception of those citing dangerous behaviour. Only a small number of responses also included consideration of affective and/or ethological factors, suggesting that welfare issues concerning affective state and/or behaviour are at risk of omission from end-of-life decisions.

Response of UK Horse, Pony and Donkey Owners to the Early Stages of the COVID-19 Pandemic.

In the UK, March 2020 was a time of great uncertainty as COVID-19 became increasingly widespread. The government responded by making suggestions about how people could reduce the risk of spread on 16 March, moved swiftly into closing schools on the 18 March before announcing a mandatory lockdown on the 23rd March. This was a challenging time for UK equestrians who had to balance maintaining their equine's routine and daily care alongside the increasing biosecurity measures. A cross-sectional survey was distributed to UK equestrians via social media over two days (28 and 29 March 2020) to better understand the decisions made by UK horse, pony and donkey owners during this time. Data from 452 respondents were generated across all four countries comprising the UK, although there were no significant differences in owner response to the pandemic between locations. The changes respondents made differed between the 16th and the 18th of March 2020, with an early emphasis on improving yard biosecurity and opting to stop riding, as well as reducing the time spent at the yard. After the 18 March, respondents placed more emphasis on risk reduction by changing the activities they did with their horse, including riding, with common examples including avoiding "high risk" activities such as riding on busy roads, jumping, riding young or nervous horses. Few respondents reported having an emergency plan in place should they become ill or otherwise unable to care for their equine. The findings highlight areas that would significantly benefit from in-depth investigation in future research. Equestrian behaviour and mindset around risk-taking and risk perception have already been researched in relation to equestrian activities and sport but have received little attention in the context of wider health challenges. Understanding the uptake of emergency planning and preparation in the UK equestrian community also warrants consideration. Using this information effectively to promote forward planning is likely to be of great benefit in equestrian responses to future health or climate-related crises.

Escalating Mean Arterial Pressure in Severe Traumatic Brain Injury: A Prospective, Observational Study.

To investigate cerebral autoregulatory status in patients with severe traumatic brain injury (TBI), guidelines now suggest active manipulation of mean arterial pressure (MAP). There is a paucity of data, however, describing the effect on intracranial pressure (ICP) when MAP is raised. Consecutive patients with TBI requiring ICP monitoring were enrolled from November 2019 to April 2020. The MAP and ICP were recorded continuously, and clinical annotations were made whenever intravenous vasopressors were commenced or adjusted to defend cerebral perfusion pressure (CPP) targets. A significant change in MAP burden was defined as MAP >100min.mm Hg over 15 min. The primary outcome was the change in ICP burden over the same 15-min period. Bedside and clinical parameters were then compared between these groups. Twenty-eight patients were enrolled, providing 212 clinical events, of which 60 were deemed significant. Over the first 15 min, 65% were associated with a net negative ICP burden. A greater reduction in ICP burden was observed with events occurring in patients without a history of hypotension at scene (p = 0.016), after three days post-injury (p = 0.0018), and where the pressure-reactivity index (PRx) was <0.25 (p = 0.0005) or the ICP amplitude to CPP correlation coefficient (RAC) was <-0.10 (p = 0.0036) at the initiation of vasopressor changes. The ICP burden in the first 15 min was highly correlated with the next 15-min period. In patients with severe TBI requiring ICP monitoring, increasing MAP to pursue a CPP target was followed by a net negative ICP burden in approximately two-thirds of events. These data suggest a MAP challenge may be a useful adjunct in managing intracranial hypertension.

Functionality of primary hepatic non-parenchymal cells in a 3D spheroid model and contribution to acetaminophen hepatotoxicity.

In addition to hepatocytes, the liver comprises a host of specialised non-parenchymal cells which are important to consider in the development of in vitro models which are both physiologically and toxicologically relevant. We have characterized a 3D co-culture system comprising primary human hepatocytes (PHH) and non-parenchymal cells (NPC) and applied it to the investigation of acetaminophen-induced toxicity. Firstly, we titrated ratios of PHH:NPC and confirmed the presence of functional NPCs via both immunohistochemistry and activation with both LPS and TGF-ß. Based on these data we selected a ratio of 2:1 PHH:NPC for further studies. We observed that spheroids supplemented with NPCs were protected against acetaminophen (APAP) toxicity as determined by ATP (up to threefold difference in EC50 at day 14 compared to hepatocytes alone) and glutathione depletion, as well as miR-122 release. APAP metabolism was also altered in the presence of NPCs, with significantly lower levels of APAP-GSH detected. Expression of several CYP450 enzymes involved in the bioactivation of APAP was also lower in NPC-containing spheroids. Spheroids containing NPCs also expressed higher levels of miRNAs which have been implicated in APAP-induced hepatotoxicity, including miR-382 and miR-155 which have potential roles in liver regeneration and inflammation, respectively. These data indicate that the interaction between hepatocytes and NPCs can have significant metabolic and toxicological consequences important for the correct elucidation of hepatic safety mechanisms.

Modification of the cyclopropyl moiety of abacavir provides insight into the structure activity relationship between HLA-B*57:01 binding and T-cell activation.

BACKGROUND: Abacavir is associated with hypersensitivity reactions in individuals positive for the HLA-B*57:01 allele. The drug binds within the peptide binding groove of HLA-B*57:01 altering peptides displayed on the cell surface. Presentation of these HLA-abacavir-peptide complexes to T-cells is hypothesized to trigger a CD8+ T-cell response underpinning the hypersensitivity. Thus, the aim of this study was to explore the relationship between the structure of abacavir with HLA-B*57:01 binding and the CD8+ T-cell activation. METHODS: Seventeen abacavir analogues were synthesized and cytokine secretion from abacavir/abacavir analogue-responsive CD8+ T-cell clones was measured using IFN-γ ELIspot. In silico docking studies were undertaken to assess the predicted binding poses of the abacavir analogues within the HLA-B*57:01 peptide binding groove. In parallel, the effect of selected abacavir analogues on the repertoire of self-peptides presented by cellular HLA-B*57:01 was characterized using mass spectrometry. RESULTS: Abacavir and ten analogues stimulated CD8+ T-cell IFN-γ release. Molecular docking of analogues that retained antiviral activity demonstrated a relationship between predicted HLA-B*57:01 binding orientations and the ability to induce a T-cell response. Analogues that stimulated T-cells displayed a perturbation of the natural peptides displayed by HLA-B*57:01. The antigen-specific CD8+ T-cell response was dependent on the enantiomeric form of abacavir at both cyclopropyl and cyclopentyl regions. CONCLUSION: Alteration of the chemical constitution of abacavir generates analogues that retain a degree of pharmacological activity, but have variable ability to activate T-cells. Modelling and immunopeptidome analysis delineate how drug HLA-B*57:01 binding and peptide display by antigen presenting cells relate to the activation of CD8+ T-cells.

Regulation of Peritoneal Inflammatory Response to Implant Material Using an Ex Vivo Model System.

BACKGROUND: Implants used in abdominal wall reconstruction are associated with intra-abdominal inflammation that can cause complications such as adhesions, fistulae, or failure of the implant. This study analyzed the inflammatory response of human peritoneum explants when exposed to different implant materials including synthetic and biological (cross-linked and non-cross-linked). MATERIALS AND METHODS: Human peritoneum explants (parietal and visceral) were incubated in culture with implants used for abdominal wall reconstruction. Implants included Permacol (biological implant with chemical cross-linking); Biodesign and Strattice (biological implants without chemical cross-linking); Prolene (synthetic nonabsorbable); and Vicryl (synthetic absorbable). Control peritoneum samples were incubated without implant. Cytokine concentrations and corresponding gene expression were measured by enzyme-linked immunosorbent assay and quantitative polymerase chain reaction, respectively. Further evaluation included assessment of tissue viability and implant-cytokine adsorption. RESULTS: Incubation of human peritoneal explants with Biodesign or Strattice was associated with a significant reduction in interleukin-6, interleukin-1ß, and tumour necrosis factor alpha protein and gene expression compared with control. These could not be explained by reduced cell viability or implant-cytokine adsorption. Incubation of explants in Biodesign-conditioned media displayed a similar effect to incubation of explants with Biodesign itself. CONCLUSIONS: Human peritoneal explants cultured with different mesh implant materials show an altered inflammatory cytokine response suggesting a tissue-specific response. Downregulation of key inflammatory cytokines by the peritoneum exposed to non-cross-linked biological implants may be mediated by the release of soluble factors from these implants inhibiting cytokine gene expression. This ex vivo human peritoneal system provides a novel preclinical model to investigate peritoneum-implant interactions.

Improving the Recognition of Equine Affective States.

A key welfare problem for horses is that people commonly fail to recognise, and consequently neglect to resolve, equine behavioural signs of distress, worsening the welfare of the horse and potentially putting the safety of the handler at risk as a result. Members of equestrian Facebook groups were asked to view six videos and assess the horse's behaviour in each; the authors selected the videos and considered each video to demonstrate behaviour associated with negative affective states. An additional six equine behaviourists also completed the survey as an "expert comparison group" from whom we could define "correct" answers; their responses were consistent with each other and the views of the authors. Although the majority of respondents successfully recognised behaviour indicative of distress in some instances, behaviour associated with negative affective states was commonly missed; videos featuring natural horsemanship and bridle-less riding were particularly interpreted incorrectly to be positive experiences for the horses. Binary logistic regression analysis (72.1% success rate) confirmed that the different video types (ridden dressage, natural horsemanship, in-hand dressage, bridle-less riding, Western reining and behavioural rehabilitation) were strong predictors for obtaining a correct answer (p < 0.01) but that experience of equine-ownership was not. Of the equestrian activities preferred by participants, only proponents of clicker training showed an increased likelihood of obtaining the correct answer (p = 0.05). Even when behavioural signs suggestive of negative affective states were recognised, a minority of respondents stated that they would be happy for their horse to be treated similarly. In conclusion, behavioural signs of equine distress are poorly recognised; they therefore warrant an increased prominence in education and the outreach activity of welfare organisations, in order to reduce equine suffering.

PIK3CA Cooperates with KRAS to Promote MYC Activity and Tumorigenesis via the Bromodomain Protein BRD9.

Tumor formation is generally linked to the acquisition of two or more driver genes that cause normal cells to progress from proliferation to abnormal expansion and malignancy. In order to understand genetic alterations involved in this process, we compared the transcriptomes of an isogenic set of breast epithelial cell lines that are non-transformed or contain a single or double knock-in (DKI) of PIK3CA (H1047R) or KRAS (G12V). Gene set enrichment analysis revealed that DKI cells were enriched over single mutant cells for genes that characterize a MYC target gene signature. This gene signature was mediated in part by the bromodomain-containing protein 9 (BRD9) that was found in the SWI-SNF chromatin-remodeling complex, bound to the MYC super-enhancer locus. Small molecule inhibition of BRD9 reduced MYC transcript levels. Critically, only DKI cells had the capacity for anchorage-independent growth in semi-solid medium, and CRISPR-Cas9 manipulations showed that PIK3CA and BRD9 expression were essential for this phenotype. In contrast, KRAS was necessary for DKI cell migration, and BRD9 overexpression induced the growth of KRAS single mutant cells in semi-solid medium. These results provide new insight into the earliest transforming events driven by oncoprotein cooperation and suggest BRD9 is an important mediator of mutant PIK3CA/KRAS-driven oncogenic transformation.

Communication strategies for rare cancers: a systematic review protocol.

BACKGROUND: Rare cancers comprise almost a quarter of all cancers in Europe, and patients generally have poorer outcomes than those suffering from more common cancers. This is attributed in part to a general lack of knowledge and awareness of rare cancers. This review aims to examine the communication strategies being used throughout the world to inform on rare cancers and to highlight any opportunities for improvement. METHODS: A systematic review of literature published in English prior to November 2018 will be conducted, screening articles from the electronic databases MEDLINE, PubMed, EMBASE, Web of Science, PsycINFO, CINAHL Plus and the Cochrane Database of Systematic Reviews. Grey literature databases (GreyLit, OpenGrey) will also be searched in order to screen for any unpublished works. As well as primary literature, reference lists will be examined via forward and reverse citation screening. The review will be reported using the Preferred Reporting Items for Systematic Reviews and Meta-Analysis (PRISMA). Titles and abstracts will first be examined for eligibility, with remaining studies undergoing a full-text screening before being included in the final review. Individual studies will be screened for bias, and a meta-analysis performed provided there is enough data. If insufficient homogenous literature exists, a narrative summary of the literature will be produced. DISCUSSION: Despite the broad topic and width of study type that will be considered, this review hopes to provide a reflective summary of the communication strategies available for people living with and working with rare cancer. It aims to reveal any gaps in the resources available, to contribute to the long-term improvement of diagnosis and management of rare cancers. SYSTEMATIC REVIEW REGISTRATION: PROSPERO CRD42018099784.

Comparison of Hepatic 2D Sandwich Cultures and 3D Spheroids for Long-term Toxicity Applications: A Multicenter Study.

Primary human hepatocytes (PHHs) are commonly used for in vitro studies of drug-induced liver injury. However, when cultured as 2D monolayers, PHH lose crucial hepatic functions within hours. This dedifferentiation can be ameliorated when PHHs are cultured in sandwich configuration (2Dsw), particularly when cultures are regularly re-overlaid with extracellular matrix, or as 3D spheroids. In this study, the 6 participating laboratories evaluated the robustness of these 2 model systems made from cryopreserved PHH from the same donors considering both inter-donor and inter-laboratory variability and compared their suitability for use in repeated-dose toxicity studies using 5 different hepatotoxins with different toxicity mechanisms. We found that expression levels of proteins involved in drug absorption, distribution, metabolism, and excretion, as well as catalytic activities of 5 different CYPs, were significantly higher in 3D spheroid cultures, potentially affecting the exposure of the cells to drugs and their metabolites. Furthermore, global proteomic analyses revealed that PHH in 3D spheroid configuration were temporally stable whereas proteomes from the same donors in 2Dsw cultures showed substantial alterations in protein expression patterns over the 14 days in culture. Overall, spheroid cultures were more sensitive to the hepatotoxic compounds investigated, particularly upon long-term exposures, across testing sites with little inter-laboratory or inter-donor variability. The data presented here suggest that repeated-dosing regimens improve the predictivity of in vitro toxicity assays, and that PHH spheroids provide a sensitive and robust system for long-term mechanistic studies of drug-induced hepatotoxicity, whereas the 2Dsw system has a more dedifferentiated phenotype and lower sensitivity to detect hepatotoxicity.

tsRNA signatures in cancer.

Small, noncoding RNAs are short untranslated RNA molecules, some of which have been associated with cancer development. Recently we showed that a class of small RNAs generated during the maturation process of tRNAs (tRNA-derived small RNAs, hereafter "tsRNAs") is dysregulated in cancer. Specifically, we uncovered tsRNA signatures in chronic lymphocytic leukemia and lung cancer and demonstrated that the ts-4521/3676 cluster (now called "ts-101" and "ts-53," respectively), ts-46, and ts-47 are down-regulated in these malignancies. Furthermore, we showed that tsRNAs are similar to Piwi-interacting RNAs (piRNAs) and demonstrated that ts-101 and ts-53 can associate with PiwiL2, a protein involved in the silencing of transposons. In this study, we extended our investigation on tsRNA signatures to samples collected from patients with colon, breast, or ovarian cancer and cell lines harboring specific oncogenic mutations and representing different stages of cancer progression. We detected tsRNA signatures in all patient samples and determined that tsRNA expression is altered upon oncogene activation and during cancer staging. In addition, we generated a knocked-out cell model for ts-101 and ts-46 in HEK-293 cells and found significant differences in gene-expression patterns, with activation of genes involved in cell survival and down-regulation of genes involved in apoptosis and chromatin structure. Finally, we overexpressed ts-46 and ts-47 in two lung cancer cell lines and performed a clonogenic assay to examine their role in cell proliferation. We observed a strong inhibition of colony formation in cells overexpressing these tsRNAs compared with untreated cells, confirming that tsRNAs affect cell growth and survival.

Transcriptional, Functional, and Mechanistic Comparisons of Stem Cell-Derived Hepatocytes, HepaRG Cells, and Three-Dimensional Human Hepatocyte Spheroids as Predictive In Vitro Systems for Drug-Induced Liver Injury.

Reliable and versatile hepatic in vitro systems for the prediction of drug pharmacokinetics and toxicity are essential constituents of preclinical safety assessment pipelines for new medicines. Here, we compared three emerging cell systems-hepatocytes derived from induced pluripotent stem cells, HepaRG cells, and three-dimensional primary human hepatocyte (PHH) spheroids-at transcriptional and functional levels in a multicenter study to evaluate their potential as predictive models for drug-induced hepatotoxicity. Transcriptomic analyses revealed widespread gene expression differences between the three cell models, with 8148 of 17,462 analyzed genes (47%) being differentially expressed. Expression levels of genes involved in the metabolism of endogenous as well as xenobiotic compounds were significantly elevated in PHH spheroids, whereas genes involved in cell division and endocytosis were significantly upregulated in HepaRG cells and hepatocytes derived from induced pluripotent stem cells, respectively. Consequently, PHH spheroids were more sensitive to a panel of drugs with distinctly different toxicity mechanisms, an effect that was amplified by long-term exposure using repeated treatments. Importantly, toxicogenomic analyses revealed that transcriptomic changes in PHH spheroids were in compliance with cholestatic, carcinogenic, or steatogenic in vivo toxicity mechanisms at clinically relevant drug concentrations. Combined, the data reveal important phenotypic differences between the three cell systems and suggest that PHH spheroids can be used for functional investigations of drug-induced liver injury in vivo in humans.