Although high-throughput RNA sequencing (RNA-seq) has greatly advanced small non-coding RNA (sncRNA) discovery, the currently widely used complementary DNA library construction protocol generates ...biased sequencing results. This is partially due to RNA modifications that interfere with adapter ligation and reverse transcription processes, which prevent the detection of sncRNAs bearing these modifications. Here, we present PANDORA-seq (panoramic RNA display by overcoming RNA modification aborted sequencing), employing a combinatorial enzymatic treatment to remove key RNA modifications that block adapter ligation and reverse transcription. PANDORA-seq identified abundant modified sncRNAs-mostly transfer RNA-derived small RNAs (tsRNAs) and ribosomal RNA-derived small RNAs (rsRNAs)-that were previously undetected, exhibiting tissue-specific expression across mouse brain, liver, spleen and sperm, as well as cell-specific expression across embryonic stem cells (ESCs) and HeLa cells. Using PANDORA-seq, we revealed unprecedented landscapes of microRNA, tsRNA and rsRNA dynamics during the generation of induced pluripotent stem cells. Importantly, tsRNAs and rsRNAs that are downregulated during somatic cell reprogramming impact cellular translation in ESCs, suggesting a role in lineage differentiation.
Human embryogenesis entails complex signalling interactions between embryonic and extra-embryonic cells. However, how extra-embryonic cells direct morphogenesis within the human embryo remains ...largely unknown due to a lack of relevant stem cell models. Here, we have established conditions to differentiate human pluripotent stem cells (hPSCs) into yolk sac-like cells (YSLCs) that resemble the post-implantation human hypoblast molecularly and functionally. YSLCs induce the expression of pluripotency and anterior ectoderm markers in human embryonic stem cells (hESCs) at the expense of mesoderm and endoderm markers. This activity is mediated by the release of BMP and WNT signalling pathway inhibitors, and, therefore, resembles the functioning of the anterior visceral endoderm signalling centre of the mouse embryo, which establishes the anterior-posterior axis. Our results implicate the yolk sac in epiblast cell fate specification in the human embryo and propose YSLCs as a tool for studying post-implantation human embryo development
Muscle satellite stem cells (MuSCs) are responsible for skeletal muscle growth and regeneration. Despite their differentiation potential, human MuSCs have limited in vitro expansion and in vivo ...migration capacity, limiting their use in cell therapies for diseases affecting multiple skeletal muscles. Several protocols have been developed to derive MuSC‐like progenitors from human induced pluripotent stem (iPS) cells (hiPSCs) to establish a source of myogenic cells with controllable proliferation and differentiation. However, current hiPSC myogenic derivatives also suffer from limitations of cell migration, ultimately delaying their clinical translation. Here we use a multi‐disciplinary approach including bioinformatics and tissue engineering to show that DLL4 and PDGF‐BB improve migration of hiPSC‐derived myogenic progenitors. Transcriptomic analyses demonstrate that this property is conserved across species and multiple hiPSC lines, consistent with results from single cell motility profiling. Treated cells showed enhanced trans‐endothelial migration in transwell assays. Finally, increased motility was detected in a novel humanised assay to study cell migration using 3D artificial muscles, harnessing advanced tissue modelling to move hiPSCs closer to future muscle gene and cell therapies.
Synopsis
This study describes an advanced framework to assess, model and enhance migration of human skeletal myogenic progenitor cells using platforms including induced pluripotent stem (iPS) cells, RNAseq, single cell profiling and tissue engineering.
Activation of NOTCH and platelet‐derived growth factor (PDGF) signalling pathways induces conserved transcriptional changes in mouse and human tissue‐ and iPS cell‐derived myogenic progenitors (hiMPs).
Combined activation of NOTCH and PDGF signalling via Delta‐like 4 (DLL4) and PDGF‐BB treatment assessed via single cell migratory profiling results in enhanced motility of hiMPs.
Organ‐on‐chip and transwell assays show increased trans‐endothelial migration of treated hiMPs not mediated by enhanced adhesion.
A novel 3D assay modelling intramuscular migration using engineered muscles shows increased motility of treated hiMPs in a biomimetic environment.
This study describes an advanced framework to assess, model and enhance migration of human skeletal myogenic progenitor cells using platforms including induced pluripotent stem (iPS) cells, RNAseq, single cell profiling and tissue engineering.
Poverty and inequality are both the starting point, and the ultimate outcome, for most neglected tropical diseases (NTDs). As a group of 20 diseases, NTDs are typified by their prevalence among the ...poor, excluded, and marginalised within society. In the absence of quality healthcare provision, many NTDs lead to long term disability, disfigurement, and stigma, which in turn act to reinforce the exclusion and poverty experienced by the afflicted. The path leading to illness is often determined by a widespread lack of access to formal education, timely healthcare, adequate living conditions, employment and nutrition. The reasons for such deprivation are complicated, but ultimately reduce to a persistence of inequalities in affected regions. These inequalities can manifest differently depending on the demographic being studied: gender, ethnicity, geographic location, level of formal education, can all determine the ease with which NTDs are transmitted, diagnosed or treated. Yet, regardless of contextual differences, NTDs continue to persist because individuals within endemic regions experience a healthcare system that is, either directly or indirectly, inequitable. The healthcare system referred to in this context is not confined to simply formal healthcare settings. Rather, it encompasses local healers, community healthcare professionals or volunteers, drug administration programmes, community mental health provision, the formal education system, and, in extension, even the infrastructure that exists to physically connect individuals to their healthcare provider. Failing to ensure parity across each of these components for every citizen equates to a discriminatory healthcare system which fails to recognise the individual’s basic human right to “a standard of living adequate for the health and well-being of themselves and of their family, including food, clothing, housing and medical care and necessary social services” (Article 25- Universal Declaration of Human Rights)
The health of our population is one of our nation’s most important assets. Optimal health not only forms a central component of our happiness, but it is also vital for a strong economy. Despite this, ...reports suggest that population health is declining, with the average adult expected to spend 20% of their life in ill-health. Given the ever-increasing burden of non- communicable disease, such as cardiovascular disease and obesity-related conditions, alongside our growing and ageing population, the need for adequate strategies to prevent ill- health has never been greater. However, as our healthcare model is relatively centralised, the development, coordination and delivery of comprehensive prevention strategies is incredibly difficult. This is not only because a central system cannot make strategies that are flexible enough to cater for every demographic, but also because the average person spends very little time engaged directly with formal healthcare services. To circumvent these issues, focus has turned to the communities in which people live, work and play as an asset to prevent ill-health and promote wellbeing. By combining rapid literature reviews with surveys of Cambridgeshire-based community groups, this report aims to investigate the role that community-led initiatives play in improving the health and wellbeing of the communities they serve, and to further identify policies that can be updated or implemented in order to support communities in this pursuit.
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