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Disease Models & Mechanisms 07/08/2020 05:17
ABSTRACT. Advances in sequencing technology have led to an explosion in the number of known genetic variants of human genes. A major challenge is to now determine which of these variants contribute to diseases as a result of their effect on gene function. Here, we describe a generic approach using the yeast Saccharomyces cerevisiae to quickly develop gene-specific in vivo assays that can be used to quantify the level of function of a genetic variant. Using synthetic dosage lethality screening, ‘sentinel’ yeast strains are identified that are sensitive to overexpression of a human disease gene. Variants of the gene can then be functionalized in a high-throughput fashion through simple growth assays using solid or liquid media. Sentinel intera.
Disease Models & Mechanisms 07/07/2020 03:51
ABSTRACT. Organs-on-chips are broadly defined as microfabricated surfaces or devices designed to engineer cells into microscale tissues with native-like features and then extract physiologically relevant readouts at scale. Because they are generally compatible with patient-derived cells, these technologies can address many of the human relevance limitations of animal models. As a result, organs-on-chips have emerged as a promising new paradigm for patient-specific disease modeling and drug development. Because neuromuscular diseases span a broad range of rare conditions with diverse etiology and complex pathophysiology, they have been especially challenging to model in animals and thus are well suited for organ-on-chip approaches. In this Re.
Disease Models & Mechanisms 06/26/2020 07:34
ABSTRACT. First Person is a series of interviews with the first authors of a selection of papers published in Disease Models & Mechanisms, helping early-career researchers promote themselves alongside their papers. Alec Nickolls is first author on ‘Human embryoid bodies as a 3D tissue model of the extracellular matrix and α-dystroglycanopathies’, published in DMM. Alec conducted the research described in this article while a PhD student in Carsten Bönnemann's lab at National Institute of Neurological Disorders and Stroke, National Institutes of Health, Bethesda, MD, USA. He is now a postdoctoral fellow in the lab of Alexander Chesler at National Center for Complementary and Integrative Health, National Institutes of Health, Bethesda, MD, US.
Disease Models & Mechanisms 06/26/2020 07:34
ABSTRACT. The basal lamina is a specialized sheet of dense extracellular matrix (ECM) linked to the plasma membrane of specific cell types in their tissue context, which serves as a structural scaffold for organ genesis and maintenance. Disruption of the basal lamina and its functions is central to many disease processes, including cancer metastasis, kidney disease, eye disease, muscular dystrophies and specific types of brain malformation. The latter three pathologies occur in the α-dystroglycanopathies, which are caused by dysfunction of the ECM receptor α-dystroglycan. However, opportunities to study the basal lamina in various human disease tissues are restricted owing to its limited accessibility. Here, we report the generation of embry.
Disease Models & Mechanisms 06/26/2020 07:22
ABSTRACT. Maladaptive signaling by pro-inflammatory cytokines (PICs), such as TNFα, IL1β and IFN, can activate downstream signaling cascades that are implicated in the development and progression of multiple inflammatory diseases. Despite playing critical roles in pathogenesis, the availability of in vivo models in which to model tissue-specific induction of PICs is limited. To bridge this gap, we have developed a novel multi-gene expression system dubbed Cre-enabled and tetracycline-inducible transgenic system for conditional, tissue-specific expression of pro-inflammatory cytokines (CETI-PIC3). This binary transgenic system permits the stoichiometric co-expression of proteins Tumor necrosis factor a (Tnfa), Interleukin-1 beta (Il1b) and In.
Disease Models & Mechanisms 06/26/2020 07:22
ABSTRACT. First Person is a series of interviews with the first authors of a selection of papers published in Disease Models & Mechanisms, helping early-career researchers promote themselves alongside their papers. Sara Ibrahim is first author on ‘A novel Cre-enabled tetracycline-inducible transgenic system for tissue-specific cytokine expression in the zebrafish: CETI-PIC3’, published in DMM. Sara is an MD-PhD student in the lab of Emily K. Sims and Ryan M. Anderson at Indiana University School of Medicine, Indianapolis, IN, USA, developing and utilizing molecular biology techniques to study β cell dysfunction.
Disease Models & Mechanisms 06/26/2020 04:41
ABSTRACT. Mammalian organs consist of diverse, intermixed cell types that signal to each other via ligand-receptor interactions – an interactome – to ensure development, homeostasis and injury-repair. Dissecting such intercellular interactions is facilitated by rapidly growing single-cell RNA sequencing (scRNA-seq) data; however, existing computational methods are often not readily adaptable by bench scientists without advanced programming skills. Here, we describe a quantitative intuitive algorithm, coupled with an optimized experimental protocol, to construct and compare interactomes in control and Sendai virus-infected mouse lungs. A minimum of 90 cells per cell type compensates for the known gene dropout issue in scRNA-seq and achieves c.
Disease Models & Mechanisms 06/26/2020 04:41
ABSTRACT. First Person is a series of interviews with the first authors of a selection of papers published in Disease Models & Mechanisms, helping early-career researchers promote themselves alongside their papers. Dimitrije Stanković is first author on ‘A Drosophila model to study retinitis pigmentosa pathology associated with mutations in the core splicing factor Prp8’, published in DMM. Dimitrije is a PhD student in the lab of Mirka Uhlirova at CECAD Research Center in Cologne, Germany, investigating the functional consequences of aberrant pre-mRNA splicing in the context of regulation of gene expression, activation of stress signaling pathways and DNA damage.
Disease Models & Mechanisms 06/26/2020 04:41
ABSTRACT. Retinitis pigmentosa (RP) represents genetically heterogeneous and clinically variable disease characterized by progressive degeneration of photoreceptors resulting in a gradual loss of vision. The autosomal dominant RP type 13 (RP13) has been linked to the malfunction of PRPF8, an essential component of the spliceosome. Over 20 different RP-associated PRPF8 mutations have been identified in human patients. However, the cellular and molecular consequences of their expression in vivo in specific tissue contexts remain largely unknown. Here, we establish a Drosophila melanogaster model for RP13 by introducing the nine distinct RP mutations into the fly PRPF8 ortholog prp8 and express the mutant proteins in precise spatiotemporal patt.
Disease Models & Mechanisms 06/24/2020 04:09
ABSTRACT. Necrotizing enterocolitis (NEC) is the leading cause of death from gastrointestinal disease in premature infants and remains stubbornly difficult to treat in many cases. Much of our understanding of NEC pathogenesis has been gained through the study of highly translational animal models. However, most models of NEC are limited by their overall complexity and by the fact that they do not incorporate human tissue. To address these limitations, investigators have recently developed precision-based ex vivo models of NEC, also termed ‘NEC-in-a-dish’ models, which provide the opportunity to increase our understanding of this disease and for drug discovery. These approaches involve exposing intestinal cells from either humans or animals w.
Disease Models & Mechanisms 06/24/2020 04:09
ABSTRACT. Human disorders of the post-squalene cholesterol biosynthesis pathway frequently result in skeletal abnormalities, yet our understanding of the mechanisms involved is limited. In a forward-genetic approach, we have found that a late-onset skeletal mutant, named koliber nu7 , is the result of a cis -acting regulatory mutation leading to loss of methylsterol monooxygenase 1 ( msmo1 ) expression within pre-hypertrophic chondrocytes. Generated msmo1 nu81 knockdown mutation resulted in lethality at larval stage. We demonstrated that this is a result of both cholesterol deprivation and sterol intermediate accumulation by creating a mutation eliminating activity of Lanosterol synthase (Lss). Our results indicate that double lss nu60 ;msmo.
Disease Models & Mechanisms 06/23/2020 03:37
ABSTRACT. The outbreak of COVID-19 has stalled both the basic, clinical and non-COVID medical research. The scientific community has shown extraordinary flexibility and resilience in responding to the pandemic. However, funding restructuring, risk of infection, cancelation of scientific conferences and delayed experiments have already proven detrimental to the career opportunities of early-career scientists. Moreover, school closures and a lack of systematic support for childcare have been additional challenges for early- and mid-career researchers who have young children. This Editorial describes an early-career researcher's experience and highlights how after efficiently contributing to ‘flattening the curve’ of COVID-19 infections, the re.
Disease Models & Mechanisms 06/17/2020 11:00
ABSTRACT. Metastasis is facilitated by the formation of pre-metastatic niches through the remodelling of the extracellular matrix (ECM) promoted by haematopoietic and stromal cells. The impact of these primed sites is pronounced for intraperitoneal metastases, where the cavity-exposed ECM supports the attachment of the disseminating tumour cells. Likewise, implantation of biomaterial scaffolds influences metastatic progression systemically through a foreign body reaction (FBR). In this study, we integrated the concept of creating an artificial niche to capture tumour cells actively disseminating in the peritoneal cavity with a therapeutic strategy modulating the interactions of metastatic cells with the ECM. The aim was to transform a dissem.
Disease Models & Mechanisms 06/15/2020 09:37
ABSTRACT. Tissue fibrosis is the deposition of excessive extracellular matrix and can occur as part of the body's natural wound healing process upon injury, or as a consequence of diseases such as systemic sclerosis. Skin fibrosis contributes to significant morbidity due to the prevalence of injuries resulting from trauma and burn. Fibroblasts, the principal cells of the dermis, synthesize extracellular matrix to maintain the skin during homeostasis and also play a pivotal role in all stages of wound healing. Although it was previously believed that fibroblasts are homogeneous and mostly quiescent cells, it has become increasingly recognized that numerous fibroblast subtypes with unique functions and morphologies exist. This Review provides
Disease Models & Mechanisms 06/15/2020 09:37
ABSTRACT. First Person is a series of interviews with the first authors of a selection of papers published in Disease Models & Mechanisms, helping early-career researchers promote themselves alongside their papers. Amélie Bacle is first author on ‘A comprehensive study of phospholipid fatty acid rearrangements in metabolic syndrome: correlations with organ dysfunction’, published in DMM. Amélie conducted the research described in this article while a postdoc in Thierry Ferreira’s lab at Université de Poitiers, Poitiers, France, where she investigated lipid membranes, their composition, and the link between their properties, cellular processes and diseases, using computational techniques such as molecular modelization. She is now working in a.
Disease Models & Mechanisms 06/03/2020 08:16
ABSTRACT. Mature human erythrocytes contain a rich pool of microRNAs (miRNAs), which result from differentiation of the erythrocytes during the course of haematopoiesis. Recent studies have described the effect of erythrocytic miRNAs on the invasion and growth of the malaria parasite Plasmodium falciparum during the asexual blood stage of its life cycle. In this work, we have identified two erythrocytic miRNAs, miR-150-3p and miR-197-5p, that show favourable in silico hybridization with Plasmodium apicortin, a protein with putative microtubule-stabilizing properties. Co-expression of P. falciparum apicortin and these two miRNAs in a cell line model resulted in downregulation of apicortin at both the RNA and protein level. To create a disease.
Disease Models & Mechanisms 06/03/2020 08:16
ABSTRACT. First Person is a series of interviews with the first authors of a selection of papers published in Disease Models & Mechanisms, helping early-career researchers promote themselves alongside their papers. Malabika Chakrabarti is first author on ‘Targeted repression of Plasmodium apicortin by host microRNA impairs malaria parasite growth and invasion’, published in DMM. Malabika is a PhD student in the lab of Dr Shailja Singh at Jawaharlal Nehru University, New Delhi, India, investigating the functional roles of human microRNA in the pathogenicity of the malaria parasite.
Disease Models & Mechanisms 06/01/2020 08:08
ABSTRACT. Netherton syndrome is a monogenic autosomal recessive disorder primarily characterized by the detachment of the uppermost layer of the epidermis, the stratum corneum . It results from mutations in the SPINK5 gene, which codes for a kallikrein inhibitor. Uncontrolled kallikrein activity leads to premature desquamation, resulting in a severe epidermal barrier defect and subsequent life-threatening systemic infections and chronic cutaneous inflammation. Here, we show that genetic activation of the transcription factor nuclear factor (erythroid-derived 2)-like 2 (Nfe2l2/Nrf2) in keratinocytes of Spink5 knockout mice, a model for Netherton syndrome, significantly alleviates their cutaneous phenotype. Nrf2 activation promoted attachment
Disease Models & Mechanisms 05/29/2020 03:35
ABSTRACT. Glaucoma is a leading cause of blindness, affecting up to 70 million people worldwide. High intraocular pressure (IOP) is a major risk factor for glaucoma. It is well established that inefficient aqueous humor (AqH) outflow resulting from structural or functional alterations in ocular drainage tissues causes high IOP, but the genes and pathways involved are poorly understood. We previously demonstrated that mutations in the gene encoding the serine protease PRSS56 induces ocular angle closure and high IOP in mice and identified reduced ocular axial length as a potential contributing factor. Here, we show that Prss56 –/– mice also exhibit an abnormal iridocorneal angle configuration characterized by a posterior shift of ocular drain.
Disease Models & Mechanisms 05/29/2020 03:35
ABSTRACT. First Person is a series of interviews with the first authors of a selection of papers published in Disease Models & Mechanisms, helping early-career researchers promote themselves alongside their papers. Sukalp Muzumdar is first author on ‘Genetic activation of Nrf2 reduces cutaneous symptoms in a murine model of Netherton syndrome’, published in DMM. Sukalp conducted the research described in this article while a PhD student in the lab of Prof. Dr Sabine Werner and PD Dr Matthias Schäfer at ETH Zurich, Zurich, Switzerland. He is now a postdoctoral fellow in the lab of Jesse Gillis at Cold Spring Harbor Laboratory, Cold Spring Harbor, NY, USA, investigating the mechanisms behind rare diseases and exploring avenues for novel treat.

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