An intersectional approach to exploring lived realities and harnessing the creativity of ethnic minority youth for health gains: protocol for a multiphase mixed method study.

BACKGROUND: Understanding the diversity and multiplicity of identities experienced by youth in Aotearoa (Te reo Maori name of the country) New Zealand (NZ) is vital to promoting their wellbeing. Ethnic minority youth (EMY) in NZ (defined as those identifying with Asian, Middle Eastern, Latin American and African ethnic origins) have been historically under-studied and under-counted, despite reporting high levels of discrimination, a major determinant of mental health and wellbeing and potentially a proxy for other inequities. In this paper, we describe the protocol for a multi-year study that examines, using an intersectional approach, how multiple marginalised identities impact mental and emotional wellbeing of EMY. METHODS: This is a multiphase, multi-method study designed to capture the diversity of lived realities of EMY who self-identify with one or more additional marginalised intersecting identity (the population referred here as EMYi). Phase 1 (Descriptive study) will involve secondary analyses of national surveys to examine the prevalence and relationships between discrimination and wellbeing of EMYi. Phase 2 (Study on public discourse) will analyse data from media narratives, complemented by interviews with stakeholders to explore discourses around EMYi. Phase 3 (Study on lived experience) will examine lived experiences of EMYi to discuss challenges and sources of resilience, and how these are influenced by public discourse. Phase 4 (Co-design phase) will use a creative approach that is youth-centered and participatory, and will involve EMYi, creative mentors and health service, policy and community stakeholders as research partners and advisors. It will employ participatory generative creative methods to explore strengths-based solutions to discriminatory experiences. DISCUSSION: This study will explore the implications of public discourse, racism and multiple forms of marginalisation on the wellbeing of EMYi. It is expected to provide evidence on the impacts of marginalisation on their mental and emotional wellbeing and inform responsive health practice and policy. Using established research tools and innovative creative means, it will enable EMYi to propose their own strength-based solutions. Further, population-based empirical research on intersectionality and health is still nascent, and even more scarce in relation to youth. This study will present the possibility of expanding its applicability in public health research focused on under-served communities.

Equity and the financial costs of informal caregiving in palliative care: a critical debate.

BACKGROUND: Informal caregivers represent the foundation of the palliative care workforce and are the main providers of end of life care. Financial pressures are among the most serious concerns for many carers and the financial burden of end of life caregiving can be substantial. METHODS: The aim of this critical debate paper was to review and critique some of the key evidence on the financial costs of informal caregiving and describe how these costs represent an equity issue in palliative care. RESULTS: The financial costs of informal caregiving at the end of life can be significant and include carer time costs, out of pocket costs and employment related costs. Financial burden is associated with a range of negative outcomes for both patient and carer. Evidence suggests that the financial costs of caring are not distributed equitably. Sources of inequity are reflective of those influencing access to specialist palliative care and include diagnosis (cancer vs non-cancer), socio-economic status, gender, cultural and ethnic identity, and employment status. Effects of intersectionality and the cumulative effect of multiple risk factors are also a consideration. CONCLUSIONS: Various groups of informal end of life carers are systematically disadvantaged financially. Addressing these, and other, determinants of end of life care is central to a public health approach to palliative care that fully recognises the value of carers. Further research exploring these areas of inequity in more depth and gaining a more detailed understanding of what influences financial burden is required to take the next steps towards meeting this aspiration. We will address the conclusions and recommendations we have made in this paper through the work of our recently established European Association of Palliative Care (EAPC) Taskforce on the financial costs of family caregiving.

A cellular taxonomy of the adult human spinal cord.

The mammalian spinal cord functions as a community of cell types for sensory processing, autonomic control, and movement. While animal models have advanced our understanding of spinal cellular diversity, characterizing human biology directly is important to uncover specialized features of basic function and human pathology. Here, we present a cellular taxonomy of the adult human spinal cord using single-nucleus RNA sequencing with spatial transcriptomics and antibody validation. We identified 29 glial clusters and 35 neuronal clusters, organized principally by anatomical location. To demonstrate the relevance of this resource to human disease, we analyzed spinal motoneurons, which degenerate in amyotrophic lateral sclerosis (ALS) and other diseases. We found that compared with other spinal neurons, human motoneurons are defined by genes related to cell size, cytoskeletal structure, and ALS, suggesting a specialized molecular repertoire underlying their selective vulnerability. We include a web resource to facilitate further investigations into human spinal cord biology.

Factors predicting forgone healthcare among Asian adolescents in New Zealand: unmasking variations in aggregate data.

AIM: To examine the relationship between social support, safety, healthcare experience and forgone healthcare for Asian secondary school students in New Zealand by unmasking variations in aggregate Asian data. METHODS: The study population included 1,911 Asians (1,272 East Asians and 604 South Asians) from the Youth19 survey. The reference group included 3,053 Pakeha. RESULTS: We found disparities in family socioeconomic status (SES), social support, safety in school and neighbourhood, healthcare experience and forgone healthcare between East Asians and South Asians compared to Pakeha. One in five Asians (20%) reported forgone healthcare. Compared to their Pakeha peers (18%), Asian students (AOR=1.18, CI=1.04-1.33) and East Asian students (AOR=1.24, CI=1.06-1.45) were more likely to experience forgone healthcare, but South Asian students were not (AOR=1.05, CI=0.86-1.28). Important unique predictors of forgone healthcare for both East and South Asian students were: being discriminated against by health professionals due to ethnicity, not having a family member to talk about their worries with, and unfair treatment by teachers. Other unique predictors varied: lower community and family SES, not getting enough quality time with family, and being bullied at school were significant predictors for East Asian students; low perceived neighbourhood safety was a predictor for South Asian students. CONCLUSIONS: A complicated picture underlies the seemingly positive findings for the overall Asian group. We highlight the importance of disaggregating Asian youth data into East Asian and South Asian, to identify disparities in risk/protective factors and better inform targeted interventions.

Transcriptional and Translational Dynamics of Zika and Dengue Virus Infection.

Zika virus (ZIKV) and dengue virus (DENV) are members of the Flaviviridae family of RNA viruses and cause severe disease in humans. ZIKV and DENV share over 90% of their genome sequences, however, the clinical features of Zika and dengue infections are very different reflecting tropism and cellular effects. Here, we used simultaneous RNA sequencing and ribosome footprinting to define the transcriptional and translational dynamics of ZIKV and DENV infection in human neuronal progenitor cells (hNPCs). The gene expression data showed induction of aminoacyl tRNA synthetases (ARS) and the translation activating PIM1 kinase, indicating an increase in RNA translation capacity. The data also reveal activation of different cell stress responses, with ZIKV triggering a BACH1/2 redox program, and DENV activating the ATF/CHOP endoplasmic reticulum (ER) stress program. The RNA translation data highlight activation of polyamine metabolism through changes in key enzymes and their regulators. This pathway is needed for eIF5A hypusination and has been implicated in viral translation and replication. Concerning the viral RNA genomes, ribosome occupancy readily identified highly translated open reading frames and a novel upstream ORF (uORF) in the DENV genome. Together, our data highlight both the cellular stress response and the activation of RNA translation and polyamine metabolism during DENV and ZIKV infection.

Co-creating a large-scale adolescent health survey integrated with access to digital health interventions.

AIM: This paper describes how we engaged with adolescents and health providers to integrate access to digital health interventions as part of a large-scale secondary school health and wellbeing survey in New Zealand. METHODS: We conducted nine participatory, iterative co-design sessions involving 29 adolescents, and two workshops with young people (n = 11), digital and health service providers (n = 11) and researchers (n = 9) to gain insights into end-user perspectives on the concept and how best to integrate digital interventions in to the survey. RESULTS: Students' perceived integrating access to digital health interventions into a large-scale youth health survey as acceptable and highly beneficial. They did not want personalized/normative feedback, but thought that every student should be offered all the help options. Participants identified key principles: assurance of confidentiality, usability, participant choice and control, and language. They highlighted wording as important for ease and comfort, and emphasised the importance of user control. Participants expressed that it would be useful and acceptable for survey respondents to receive information about digital help options addressing a range of health and wellbeing topics. CONCLUSION: The methodology of adolescent-practitioner-researcher collaboration and partnership was central to this research and provided useful insights for the development and delivery of adolescent health surveys integrated with digital help options. The results from the ongoing study will provide useful data on the impact of digital health interventions integrated in large-scale surveys, as a novel methodology. Future research on engaging with adolescents once interventions are delivered will be useful to explore benefits over time.

Cell Type-Specific In Vitro Gene Expression Profiling of Stem Cell-Derived Neural Models.

Genetic and genomic studies of brain disease increasingly demonstrate disease-associated interactions between the cell types of the brain. Increasingly complex and more physiologically relevant human-induced pluripotent stem cell (hiPSC)-based models better explore the molecular mechanisms underlying disease but also challenge our ability to resolve cell type-specific perturbations. Here, we report an extension of the RiboTag system, first developed to achieve cell type-restricted expression of epitope-tagged ribosomal protein (RPL22) in mouse tissue, to a variety of in vitro applications, including immortalized cell lines, primary mouse astrocytes, and hiPSC-derived neurons. RiboTag expression enables depletion of up to 87 percent of off-target RNA in mixed species co-cultures. Nonetheless, depletion efficiency varies across independent experimental replicates, particularly for hiPSC-derived motor neurons. The challenges and potential of implementing RiboTags in complex in vitro cultures are discussed.

Spatiotemporal dynamics of molecular pathology in amyotrophic lateral sclerosis.

Paralysis occurring in amyotrophic lateral sclerosis (ALS) results from denervation of skeletal muscle as a consequence of motor neuron degeneration. Interactions between motor neurons and glia contribute to motor neuron loss, but the spatiotemporal ordering of molecular events that drive these processes in intact spinal tissue remains poorly understood. Here, we use spatial transcriptomics to obtain gene expression measurements of mouse spinal cords over the course of disease, as well as of postmortem tissue from ALS patients, to characterize the underlying molecular mechanisms in ALS. We identify pathway dynamics, distinguish regional differences between microglia and astrocyte populations at early time points, and discern perturbations in several transcriptional pathways shared between murine models of ALS and human postmortem spinal cords.

Unexpected similarities between C9ORF72 and sporadic forms of ALS/FTD suggest a common disease mechanism.

Amyotrophic lateral sclerosis (ALS) and frontotemporal dementia (FTD) represent two ends of a disease spectrum with shared clinical, genetic and pathological features. These include near ubiquitous pathological inclusions of the RNA-binding protein (RBP) TDP-43, and often the presence of a GGGGCC expansion in the C9ORF72 (C9) gene. Previously, we reported that the sequestration of hnRNP H altered the splicing of target transcripts in C9ALS patients (Conlon et al., 2016). Here, we show that this signature also occurs in half of 50 postmortem sporadic, non-C9 ALS/FTD brains. Furthermore, and equally surprisingly, these 'like-C9' brains also contained correspondingly high amounts of insoluble TDP-43, as well as several other disease-related RBPs, and this correlates with widespread global splicing defects. Finally, we show that the like-C9 sporadic patients, like actual C9ALS patients, were much more likely to have developed FTD. We propose that these unexpected links between C9 and sporadic ALS/FTD define a common mechanism in this disease spectrum.

The PLK1 inhibitor GSK461364A is effective in poorly differentiated and anaplastic thyroid carcinoma cells, independent of the nature of their driver mutations.

BACKGROUND: Poorly differentiated thyroid carcinoma (PDTC) and anaplastic thyroid carcinoma (ATC) are the most aggressive forms of thyroid cancer. Despite their low incidence, they account for a disproportionate number of thyroid cancer-related deaths because of their resistance to most therapeutic approaches. We have generated mouse models that develop ATC ([Pten, p53](thyr-/-) mice) and follicular thyroid cancer with areas of poor differentiation (Pten(thyr-/-),Kras(G12D) mice). Comparative gene expression profiling of human and mouse ATCs reveals a common "mitotic signature" in which mitotic kinases, including Polo-like kinase-1 (PLK1), are found deregulated in both species. Most genes from this signature are also upregulated in poorly differentiated tumors developing in Pten(thyr-/-),Kras(G12D) mice. PLK1 is a crucial driving force for normal mitotic spindle formation, centrosome maturation, and separation, and its overexpression has been demonstrated in a wide range of tumors. METHODS: Human and mouse ATC and PDTC cell lines were treated with the PLK1 inhibitor GSK461364A, and proliferation, apoptosis, and mitotic spindle alterations were analyzed. Furthermore, immunocompetent mice were injected in the flank with mouse ATC cells, and treated with placebo or GSK461364A. RESULTS: We show that the PLK1 inhibitor GSK461364A inhibits cell proliferation and induces cell death in both mouse ATC- and PDTC-derived cell lines and in several human ATC cell lines carrying different driver mutations. Dose-dependent changes in chromosome alignment and spindle assembly during mitosis are observed after treatment, together with changes in the mitotic index. FACS analysis reveals a G2/M phase arrest, followed by apoptosis, and mitotic slippage in cells with PI3K activation. GSK461364A is also effective in vivo, in an allograft model of ATC. CONCLUSIONS: Taken together, these data suggest that PLK1 targeting is a promising and effective therapeutic approach against PDTC cells and undifferentiated thyroid carcinoma cells.

Inhibition of AMPK and Krebs cycle gene expression drives metabolic remodeling of Pten-deficient preneoplastic thyroid cells.

Rapidly proliferating and neoplastically transformed cells generate the energy required to support rapid cell division by increasing glycolysis and decreasing flux through the oxidative phosphorylation (OXPHOS) pathway, usually without alterations in mitochondrial function. In contrast, little is known of the metabolic alterations, if any, which occur in cells harboring mutations that prime their neoplastic transformation. To address this question, we used a Pten-deficient mouse model to examine thyroid cells where a mild hyperplasia progresses slowly to follicular thyroid carcinoma. Using this model, we report that constitutive phosphoinositide 3-kinase (PI3K) activation caused by PTEN deficiency in nontransformed thyrocytes results in a global downregulation of Krebs cycle and OXPHOS gene expression, defective mitochondria, reduced respiration, and an enhancement in compensatory glycolysis. We found that this process does not involve any of the pathways classically associated with the Warburg effect. Moreover, this process was independent of proliferation but contributed directly to thyroid hyperplasia. Our findings define a novel metabolic switch to glycolysis driven by PI3K-dependent AMPK inactivation with a consequent repression in the expression of key metabolic transcription regulators.

Thyrocyte-specific inactivation of p53 and Pten results in anaplastic thyroid carcinomas faithfully recapitulating human tumors.

Anaplastic thyroid carcinoma (ATC) is the most aggressive form of thyroid cancer, and often derives from pre-existing well-differentiated tumors. Despite a relatively low prevalence, it accounts for a disproportionate number of thyroid cancer-related deaths, due to its resistance to any therapeutic approach. Here we describe the first mouse model of ATC, obtained by combining in the mouse thyroid follicular cells two molecular hallmarks of human ATC: activation of PI3K (via Pten deletion) and inactivation of p53. By 9 months of age, over 75% of the compound mutant mice develop aggressive, undifferentiated thyroid tumors that evolve from pre-existing follicular hyperplasia and carcinoma. These tumors display all the features of their human counterpart, including pleomorphism, epithelial-mesenchymal transition, aneuploidy, local invasion, and distant metastases. Expression profiling of the murine ATCs reveals a significant overlap with genes found deregulated in human ATC, including genes involved in mitosis control. Furthermore, similar to the human tumors, [Pten, p53]thyr-/- tumors and cells are highly glycolytic and remarkably sensitive to glycolysis inhibitors, which synergize with standard chemotherapy. Taken together, our results show that combined PI3K activation and p53 loss faithfully reproduce the development of thyroid anaplastic carcinomas, and provide a compelling rationale for targeting glycolysis to increase chemotherapy response in ATC patients.