154 results on '"Quinlan RA"'
Search Results
2. αB-crystallin, vimentin and increased p53 expression levels in breast cancer is associated with poor prognosis.
- Author
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Quinlan, PR, primary, Sreseli, R, additional, Quinlan, RA, additional, Hadad, S, additional, Bray, SE, additional, Kernohan, N, additional, Kellock, D, additional, Baker, L, additional, Purdie, C, additional, Jordan, L, additional, and Thompson, AM, additional
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- 2009
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3. A novel frameshift variant in BCOR causes congenital nuclear cataract.
- Author
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Berry V, Ponnekanti MB, Pontikos N, Quinlan RA, and Michaelides M
- Abstract
Background: BCL6 co-repressor ( BCOR ) gene variants are involved in oculofaciocardiodental (OFCD) syndrome, acute myeloid leukaemia, renal tumours, and photoreceptor degenerative diseases. Here, we describe a British family with a pathogenic heterozygous variant in the BCOR gene causing congenital nuclear cataract., Methods: Whole-exome sequencing was conducted on an individual affected by X-linked dominant congenital cataract in a three-generation family to establish the underlying genetic basis. Bioinformatics analysis confirmed the variants with damaging pathogenicity scores., Results: A novel likely pathogenic frameshift variant BCOR NM_001123385.1: c.3621del; p.Lys1207AsnfsTer31, was identified and found to co-segregate with the disease in this family., Conclusions: This is apparently the first report of a variant in BCOR causing X-linked dominant congenital cataract which is potentially isolated or presenting with a remarkably mild systemic phenotype. Our findings extend the genetic basis for congenital cataract and add to the phenotypic spectrum of BCOR variants.
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- 2024
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4. Inhibition of PDIs Downregulates Core LINC Complex Proteins, Promoting the Invasiveness of MDA-MB-231 Breast Cancer Cells in Confined Spaces In Vitro.
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Young N, Gui Z, Mustafa S, Papa K, Jessop E, Ruddell E, Bevington L, Quinlan RA, Benham AM, Goldberg MW, Obara B, and Karakesisoglou I
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- Humans, Cell Line, Tumor, Female, Down-Regulation drug effects, Breast Neoplasms metabolism, Breast Neoplasms pathology, Membrane Proteins metabolism, Nuclear Proteins metabolism, Nuclear Envelope metabolism, Triple Negative Breast Neoplasms metabolism, Triple Negative Breast Neoplasms pathology, Intracellular Signaling Peptides and Proteins, Neoplasm Invasiveness, Protein Disulfide-Isomerases metabolism
- Abstract
Eukaryotic cells tether the nucleoskeleton to the cytoskeleton via a conserved molecular bridge, called the LINC complex. The core of the LINC complex comprises SUN-domain and KASH-domain proteins that directly associate within the nuclear envelope lumen. Intra- and inter-chain disulphide bonds, along with KASH-domain protein interactions, both contribute to the tertiary and quaternary structure of vertebrate SUN-domain proteins. The significance of these bonds and the role of PDIs (protein disulphide isomerases) in LINC complex biology remains unclear. Reducing and non-reducing SDS-PAGE analyses revealed a prevalence of SUN2 homodimers in non-tumorigenic breast epithelia MCF10A cells, but not in the invasive triple-negative breast cancer MDA-MB-231 cell line. Furthermore, super-resolution microscopy revealed SUN2 staining alterations in MCF10A, but not in MDA-MB-231 nuclei, upon reducing agent exposure. While PDIA1 levels were similar in both cell lines, pharmacological inhibition of PDI activity in MDA-MB-231 cells led to SUN-domain protein down-regulation, as well as Nesprin-2 displacement from the nucleus. This inhibition also caused changes in perinuclear cytoskeletal architecture and lamin downregulation, and increased the invasiveness of PDI-inhibited MDA-MB-231 cells in space-restrictive in vitro environments, compared to untreated cells. These results emphasise the key roles of PDIs in regulating LINC complex biology, cellular architecture, biomechanics, and invasion.
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- 2024
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5. Pregnancy Management in HIV Viral Controllers: Twenty Years of Experience.
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Short CS, Byrne L, Hagan-Bezgin A, Quinlan RA, Anderson J, Brook G, De Alwis O, de Ruiter A, Farrugia P, Fidler S, Hamlyn E, Hartley A, Murphy S, Noble H, Oomeer S, Roedling S, Rosenvinge M, Rubinstein L, Shah R, Singh S, Thorne E, Toby M, Wait B, Sarner L, and Taylor GP
- Abstract
(1) Background: The evidence base for the management of spontaneous viral controllers in pregnancy is lacking. We describe the management outcomes of pregnancies in a series of UK women with spontaneous HIV viral control (<100 copies/mL 2 occasions before or after pregnancy off ART). (2) Methods: A multi-centre, retrospective case series (1999-2021) comparing pre- and post-2012 when guidelines departed from zidovudine-monotherapy (ZDVm) as a first-line option. Demographic, virologic, obstetric and neonatal information were anonymised, collated and analysed in SPSS. (3) Results: A total of 49 live births were recorded in 29 women, 35 pre-2012 and 14 post. HIV infection was more commonly diagnosed in first reported pregnancy pre-2012 (15/35) compared to post (2/14), p = 0.10. Pre-2012 pregnancies were predominantly managed with ZDVm (28/35) with pre-labour caesarean section (PLCS) (24/35). Post-2012 4/14 received ZDVm and 10/14 triple ART, p = 0.002. Post-2012 mode of delivery was varied (5 vaginal, 6 PLCS and 3 emergency CS). No intrapartum ZDV infusions were given post-2012 compared to 11/35 deliveries pre-2012. During pregnancy, HIV was detected (> 50 copies/mL) in 14/49 pregnancies (29%) (median 92, range 51-6084). Neonatal ZDV post-exposure prophylaxis was recorded for 45/49 infants. No transmissions were reported. (4) Conclusion: UK practice has been influenced by the change in guidelines, but this has had little impact on CS rates.
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- 2024
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6. The redox-responsive roles of intermediate filaments in cellular stress detection, integration and mitigation.
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Pérez-Sala D and Quinlan RA
- Subjects
- Vimentin analysis, Vimentin metabolism, Oxidation-Reduction, Intermediate Filaments chemistry
- Abstract
Intermediate filaments are critical for cell and tissue homeostasis and for stress responses. Cytoplasmic intermediate filaments form versatile and dynamic assemblies that interconnect cellular organelles, participate in signaling and protect cells and tissues against stress. Here we have focused on their involvement in redox signaling and oxidative stress, which arises in numerous pathophysiological situations. We pay special attention to type III intermediate filaments, mainly vimentin, because it provides a physical interface for redox signaling, stress responses and mechanosensing. Vimentin possesses a single cysteine residue that is a target for multiple oxidants and electrophiles. This conserved residue fine tunes vimentin assembly, response to oxidative stress and crosstalk with other cellular structures. Here we integrate evidence from the intermediate filament and redox biology fields to propose intermediate filaments as redox sentinel networks of the cell. To support this, we appraise how vimentin detects and orchestrates cellular responses to oxidative and electrophilic stress., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 The Author(s). Published by Elsevier Ltd.. All rights reserved.)
- Published
- 2024
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7. The major inducible small heat shock protein HSP20-3 in the tardigrade Ramazzottius varieornatus forms filament-like structures and is an active chaperone.
- Author
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Al-Ansari M, Fitzsimons T, Wei W, Goldberg MW, Kunieda T, and Quinlan RA
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- Humans, Amino Acid Sequence, HSP20 Heat-Shock Proteins genetics, HSP20 Heat-Shock Proteins metabolism, Molecular Chaperones metabolism, Heat-Shock Response, Heat-Shock Proteins, Small metabolism
- Abstract
The tardigrade Ramazzottius varieornatus has remarkable resilience to a range of environmental stresses. In this study, we have characterised two members of the small heat shock protein (sHSP) family in R. varieornatus, HSP20-3 and HSP20-6. These are the most highly upregulated sHSPs in response to a 24 h heat shock at 35
0 C of adult tardigrades with HSP20-3 being one of the most highly upregulated gene in the whole transcriptome. Both R. varieornatus sHSPs and the human sHSP, CRYAB (HSPB5), were produced recombinantly for comparative structure-function studies. HSP20-3 exhibited a superior chaperone activity than human CRYAB in a heat-induced protein aggregation assay. Both tardigrade sHSPs also formed larger oligomers than CRYAB as assessed by size exclusion chromatography and transmission electron microscopy of negatively stained samples. Whilst both HSP20-3 and HSP20-6 formed particles that were variable in size and larger than the particles formed by CRYAB, only HSP20-3 formed filament-like structures. The particles and filament-like structures formed by HSP20-3 appear inter-related as the filament-like structures often had particles located at their ends. Sequence analyses identified two unique features; an insertion in the middle region of the N-terminal domain (NTD) and preceding the critical-sequence identified in CRYAB, as well as a repeated QNTN-motif located in the C-terminal domain of HSP20-3. The NTD insertion is expected to affect protein-protein interactions and subunit oligomerisation. Removal of the repeated QNTN-motif abolished HSP20-3 chaperone activity and also affected the assembly of the filament-like structures. We discuss the potential contribution of HSP20-3 to protein condensate formation., Competing Interests: Declaration of Competing interest No competing interests., (Copyright © 2023 The Author(s). Published by Elsevier Inc. All rights reserved.)- Published
- 2024
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8. Evaluation of cataract formation in fish exposed to environmental radiation at Chernobyl and Fukushima.
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Lerebours A, Regini J, Quinlan RA, Wada T, Pierscionek B, Devonshire M, Kalligeraki AA, Uwineza A, Young L, Girkin JM, Warwick P, Smith K, Hoshino M, Uesugi K, Yagi N, Terrill N, Shebanova O, Snow T, and Smith JT
- Subjects
- Animals, Scattering, Small Angle, X-Ray Diffraction, Chernobyl Nuclear Accident, Cataract etiology, Cataract veterinary, Cataract metabolism, Crystallins
- Abstract
Recent studies apparently finding deleterious effects of radiation exposure on cataract formation in birds and voles living near Chernobyl represent a major challenge to current radiation protection regulations. This study conducted an integrated assessment of radiation exposure on cataractogenesis using the most advanced technologies available to assess the cataract status of lenses extracted from fish caught at both Chernobyl in Ukraine and Fukushima in Japan. It was hypothesised that these novel data would reveal positive correlations between radiation dose and early indicators of cataract formation. The structure, function and optical properties of lenses were analysed from atomic to millimetre length scales. We measured the short-range order of the lens crystallin proteins using Small Angle X-Ray Scattering (SAXS) at both the SPring-8 and DIAMOND synchrotrons, the profile of the graded refractive index generated by these proteins, the epithelial cell density and organisation and finally the focal length of each lens. The results showed no evidence of a difference between the focal length, the epithelial cell densities, the refractive indices, the interference functions and the short-range order of crystallin proteins (X-ray diffraction patterns) in lens from fish exposed to different radiation doses. It could be argued that animals in the natural environment which developed cataract would be more likely, for example, to suffer predation leading to survivor bias. But the cross-length scale study presented here, by evaluating small scale molecular and cellular changes in the lens (pre-cataract formation) significantly mitigates against this issue., Competing Interests: Declaration of competing interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Jim T. Smith reports financial support was provided by Natural Environment Research Council. Adelaide Lerebours reports financial support was provided by Natural Environment Research Council. Jim T. Smith reports financial support was provided by Science and Technology Funding Council. Adelaide Lerebours reports financial support was provided by Science and Technology Funding Council. Roy Quinlan reports financial support was provided by EU Framework Programme for Research and Innovation Euratom. Alexia Kalligeraki reports financial support was provided by National Eye Research Foundation. Roy Quinlan reports financial support was provided by Fight for Sight. Roy Quinlan reports was provided by Leverhulme Trust. John Girkin, Laura Young reports financial support was provided by Engineering and Physical Sciences Research Council. Jim T. Smith reports a relationship with The Chernobyl Spirit Community Interest Company that includes: board membership and equity or stocks., (Copyright © 2023 The Authors. Published by Elsevier B.V. All rights reserved.)
- Published
- 2023
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9. Multimorbidity due to novel pathogenic variants in the WFS1/RP1/NOD2 genes: autosomal dominant congenital lamellar cataract, retinitis pigmentosa and Crohn's disease in a British family.
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Berry V, Ionides A, Georgiou M, Quinlan RA, and Michaelides M
- Subjects
- Humans, Multimorbidity, Eye Proteins genetics, Nod2 Signaling Adaptor Protein genetics, Microtubule-Associated Proteins genetics, Crohn Disease genetics, Retinitis Pigmentosa epidemiology, Cataract epidemiology
- Abstract
Background: A five generation family has been analysed by whole exome sequencing (WES) for genetic associations with the multimorbidities of congenital cataract (CC), retinitis pigmentosa (RP) and Crohn's disease (CD)., Methods: WES was performed for unaffected and affected individuals within the family pedigree followed by bioinformatic analyses of these data to identify disease-causing variants with damaging pathogenicity scores., Results: A novel pathogenic missense variant in WFS1 : c.1897G>C; p.V633L, a novel pathogenic nonsense variant in RP1 : c.6344T>G; p.L2115* and a predicted pathogenic missense variant in NOD 2: c.2104C>T; p.R702W are reported. The three variants cosegregated with the phenotypic combinations of autosomal dominant CC, RP and CD within individual family members., Conclusions: Here, we report multimorbidity in a family pedigree listed on a CC register, which broadens the spectrum of potential cataract associated genes to include both RP1 and NOD2 ., Competing Interests: Competing interests: None declared., (© Author(s) (or their employer(s)) 2023. Re-use permitted under CC BY. Published by BMJ.)
- Published
- 2023
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10. Independent Membrane Binding Properties of the Caspase Generated Fragments of the Beaded Filament Structural Protein 1 (BFSP1) Involves an Amphipathic Helix.
- Author
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Jarrin M, Kalligeraki AA, Uwineza A, Cawood CS, Brown AP, Ward EN, Le K, Freitag-Pohl S, Pohl E, Kiss B, Tapodi A, and Quinlan RA
- Subjects
- Humans, Cell Membrane metabolism, Intermediate Filament Proteins metabolism, Membranes metabolism, Caspases metabolism, Lens, Crystalline metabolism
- Abstract
Background: BFSP1 (beaded filament structural protein 1) is a plasma membrane, Aquaporin 0 (AQP0/MIP)-associated intermediate filament protein expressed in the eye lens. BFSP1 is myristoylated, a post-translation modification that requires caspase cleavage at D433. Bioinformatic analyses suggested that the sequences 434-452 were α-helical and amphipathic., Methods and Results: By CD spectroscopy, we show that the addition of trifluoroethanol induced a switch from an intrinsically disordered to a more α-helical conformation for the residues 434-467. Recombinantly produced BFSP1 fragments containing this amphipathic helix bind to lens lipid bilayers as determined by surface plasmon resonance (SPR). Lastly, we demonstrate by transient transfection of non-lens MCF7 cells that these same BFSP1 C-terminal sequences localise to plasma membranes and to cytoplasmic vesicles. These can be co-labelled with the vital dye, lysotracker, but other cell compartments, such as the nuclear and mitochondrial membranes, were negative. The N-terminal myristoylation of the amphipathic helix appeared not to change either the lipid affinity or membrane localisation of the BFSP1 polypeptides or fragments we assessed by SPR and transient transfection, but it did appear to enhance its helical content., Conclusions: These data support the conclusion that C-terminal sequences of human BFSP1 distal to the caspase site at G433 have independent membrane binding properties via an adjacent amphipathic helix.
- Published
- 2023
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11. Identification and quantification of ionising radiation-induced oxysterol formation in membranes of lens fibre cells.
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Uwineza A, Cummins I, Jarrin M, Kalligeraki AA, Barnard S, Mol M, Degani G, Altomare AA, Aldini G, Schreurs A, Balschun D, Ainsbury EA, Dias IH, and Quinlan RA
- Abstract
Ionising radiation (IR) is a cause of lipid peroxidation, and epidemiological data have revealed a correlation between exposure to IR and the development of eye lens cataracts. Cataracts remain the leading cause of blindness around the world. The plasma membranes of lens fibre cells are one of the most cholesterolrich membranes in the human body, forming lipid rafts and contributing to the biophysical properties of lens fibre plasma membrane. Liquid chromatography followed by mass spectrometry was used to analyse bovine eye lens lipid membrane fractions after exposure to 5 and 50 Gy and eye lenses taken from wholebody 2 Gy-irradiated mice. Although cholesterol levels do not change significantly, IR dose-dependant formation of the oxysterols 7β-hydroxycholesterol, 7-ketocholesterol and 5, 6-epoxycholesterol in bovine lens nucleus membrane extracts was observed. Whole-body X-ray exposure (2 Gy) of 12-week old mice resulted in an increase in 7β-hydroxycholesterol and 7-ketocholesterol in their eye lenses. Their increase regressed over 24 h in the living lens cortex after IR exposure. This study also demonstrated that the IR-induced fold increase in oxysterols was greater in the mouse lens cortex than the nucleus. Further work is required to elucidate the mechanistic link(s) between oxysterols and IR-induced cataract, but these data evidence for the first time that IR exposure of mice results in oxysterol formation in their eye lenses., Competing Interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (© 2022 The Authors. Published by Elsevier B.V.)
- Published
- 2023
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12. Editorial: The wetware credentials of intermediate filaments involves coordinating, organising and networking in cells and tissues.
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Leube RE and Quinlan RA
- Abstract
Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.
- Published
- 2023
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13. Insights into the biochemical and biophysical mechanisms mediating the longevity of the transparent optics of the eye lens.
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Quinlan RA and Clark JI
- Subjects
- Humans, Protein Aggregates, Collagen metabolism, Aging, Cataract metabolism, Crystallins metabolism, Lens, Crystalline metabolism
- Abstract
In the human eye, a transparent cornea and lens combine to form the "refracton" to focus images on the retina. This requires the refracton to have a high refractive index "n," mediated largely by extracellular collagen fibrils in the corneal stroma and the highly concentrated crystallin proteins in the cytoplasm of the lens fiber cells. Transparency is a result of short-range order in the spatial arrangement of corneal collagen fibrils and lens crystallins, generated in part by post-translational modifications (PTMs). However, while corneal collagen is remodeled continuously and replaced, lens crystallins are very long-lived and are not replaced and so accumulate PTMs over a lifetime. Eventually, a tipping point is reached when protein aggregation results in increased light scatter, inevitably leading to the iconic protein condensation-based disease, age-related cataract (ARC). Cataracts account for 50% of vision impairment worldwide, affecting far more people than other well-known protein aggregation-based diseases. However, because accumulation of crystallin PTMs begins before birth and long before ARC presents, we postulate that the lens protein PTMs contribute to a "cataractogenic load" that not only increases with age but also has protective effects on optical function by stabilizing lens crystallins until a tipping point is reached. In this review, we highlight decades of experimental findings that support the potential for PTMs to be protective during normal development. We hypothesize that ARC is preventable by protecting the biochemical and biophysical properties of lens proteins needed to maintain transparency, refraction, and optical function., Competing Interests: Conflict of interest The authors declare that they have no conflicts of interest with the contents of this article., (Copyright © 2022 The Authors. Published by Elsevier Inc. All rights reserved.)
- Published
- 2022
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14. A recurrent variant in LIM2 causes an isolated congenital sutural/lamellar cataract in a Japanese family.
- Author
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Berry V, Fujinami K, Mochizuki K, Iwata T, Pontikos N, Quinlan RA, and Michaelides M
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- Eye Proteins, Humans, Japan, Membrane Proteins genetics, Mutation, Pedigree, Cataract congenital, Cataract genetics
- Abstract
Background: Genetically determined cataract is both clinically and molecularly highly heterogeneous. Here, we have identified a heterozygous variant in the lens integral membrane protein LIM2, the second most abundant protein in the lens, responsible for congenital sutural/lamellar cataract in a three-generation Japanese family., Methods: Whole exome sequencing (WES) was undertaken in one affected and one unaffected individual from a family with autosomal dominant congenital cataract to establish the underlying genetic basis., Results: A recurrent missense variant LIM2: c.388C>T; p.R130C was identified and found to co-segregate with disease. In addition, one variant COL11A1:c.3788C>T of unknown significance (VUS) was also identified., Conclusions: We report a variant in LIM2 causing an isolated autosomal-dominant congenital sutural/lamellar cataract in a Japanese family. This is the first report of a LIM2 variant in the Japanese population. Hence, we expand the mutation spectrum of LIM2 variants in different ethnic groups.
- Published
- 2022
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15. Variants in PAX6, PITX3 and HSF4 causing autosomal dominant congenital cataracts.
- Author
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Berry V, Ionides A, Pontikos N, Moore AT, Quinlan RA, and Michaelides M
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- Humans, Mutation, Pedigree, Cataract congenital, Heat Shock Transcription Factors genetics, Homeodomain Proteins genetics, PAX6 Transcription Factor genetics, Transcription Factors genetics
- Abstract
Background: Lens development is orchestrated by transcription factors. Disease-causing variants in transcription factors and their developmental target genes are associated with congenital cataracts and other eye anomalies., Methods: Using whole exome sequencing, we identified disease-causing variants in two large British families and one isolated case with autosomal dominant congenital cataract. Bioinformatics analysis confirmed these disease-causing mutations as rare or novel variants, with a moderate to damaging pathogenicity score, with testing for segregation within the families using direct Sanger sequencing., Results: Family A had a missense variant (c.184 G>A; p.V62M) in PAX6 and affected individuals presented with nuclear cataract. Family B had a frameshift variant (c.470-477dup; p.A160R*) in PITX3 that was also associated with nuclear cataract. A recurrent missense variant in HSF4 (c.341 T>C; p.L114P) was associated with congenital cataract in a single isolated case., Conclusions: We have therefore identified novel variants in PAX6 and PITX3 that cause autosomal dominant congenital cataract., (© 2021. Crown.)
- Published
- 2022
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16. The eye lens as an aging paradigm par excellence.
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Quinlan RA and Giblin F
- Subjects
- Lens, Crystalline
- Published
- 2022
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17. Pathogenic variants in the CYP21A2 gene cause isolated autosomal dominant congenital posterior polar cataracts.
- Author
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Berry V, Pontikos N, Ionides A, Kalitzeos A, Quinlan RA, and Michaelides M
- Subjects
- Female, Humans, Male, Mutation, Mutation, Missense, Pedigree, Phenotype, Steroid 21-Hydroxylase genetics, Adrenal Hyperplasia, Congenital genetics, Cataract congenital, Cataract genetics, Lens, Crystalline
- Abstract
Background: Congenital cataracts are the most common cause of visual impairment worldwide. Inherited cataract is a clinically and genetically heterogeneous disease. Here we report disease-causing variants in a novel gene, CYP21A2 , causing autosomal dominant posterior polar cataract. Variants in this gene are known to cause autosomal recessive congenital adrenal hyperplasia (CAH)., Methods: Using whole-exome sequencing (WES), we have identified disease-causing sequence variants in two families of British and Irish origin, and in two isolated cases of Asian-Indian and British origin. Bioinformatics analysis confirmed these variants as rare with damaging pathogenicity scores. Segregation was tested within the families using direct Sanger sequencing., Results: A nonsense variant NM_000500.9 c.955 C > T; p.Q319* was identified in CYP21A2 in two families with posterior polar cataract and in an isolated case with unspecified congenital cataract phenotype. This is the same variant previously linked to CAH and identified as Q318* in the literature. We have also identified a rare missense variant NM_000500.9 c.770 T > C; p.M257T in an isolated case with unspecified congenital cataract phenotype., Conclusion: This is the first report of separate sequence variants in CYP21A2 associated with congenital cataract. Our findings extend the genetic basis for congenital cataract and add to the phenotypic spectrum of CYP21A2 variants and particularly the CAH associated Q318* variant. CYP21A2 has a significant role in mineralo- and gluco-corticoid biosynthesis. These findings suggest that CYP21A2 may be important for extra-adrenal biosynthesis of aldosterone and cortisol in the eye lens.
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- 2022
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18. Lens Epithelial Cell Proliferation in Response to Ionizing Radiation.
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Barnard S, Uwineza A, Kalligeraki A, McCarron R, Kruse F, Ainsbury EA, and Quinlan RA
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- Animals, Cell Differentiation radiation effects, Cell Proliferation radiation effects, Dose-Response Relationship, Radiation, Epithelial Cells, Female, Humans, Mice, Inbred C57BL, Radiation Dosage, Radiation Exposure, Radiation, Ionizing, Mice, Lens, Crystalline radiation effects
- Abstract
Lens epithelial cell proliferation and differentiation are naturally well regulated and controlled, a characteristic essential for lens structure, symmetry and function. The effect of ionizing radiation on lens epithelial cell proliferation has been demonstrated in previous studies at high acute doses, but the effect of dose and dose rate on proliferation has not yet been considered. In this work, mice received single acute doses of 0.5, 1 and 2 Gy of radiation, at dose rates of 0.063 and 0.3 Gy/min. Eye lenses were isolated postirradiation at 30 min up until 14 days and flat-mounted. Then, cell proliferation rates were determined using biomarker Ki67. As expected, radiation increased cell proliferation 2 and 24 h postirradiation transiently (undetectable 14 days postirradiation) and was dose dependent (changes were very significant at 2 Gy; P = 0.008). A dose-rate effect did not reach significance in this study (P = 0.054). However, dose rate and lens epithelial cell region showed significant interactions (P < 0.001). These observations further our mechanistic understanding of how the lens responds to radiation., (©2022 by Radiation Research Society. All rights of reproduction in any form reserved.)
- Published
- 2022
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19. Introduction to the Special LDLensRad Focus Issue.
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Ainsbury EA, Dalke C, Mancuso M, Kadhim M, Quinlan RA, Azizova T, Dauer LT, Dynlacht JR, Tanner R, and Hamada N
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- Animals, Dose-Response Relationship, Radiation, Europe, Humans, Japan, Mice, Inbred C57BL, Occupational Exposure, Radiation Dosage, Radiation Tolerance, Mice, Cataract ethnology, Lens, Crystalline radiation effects
- Abstract
Recent epidemiological and experimental animal data, as well as reanalyses of data previously accumulated, indicate that the lens of the eye is more radiosensitive than was previously thought. This has resulted in a reduction of the occupational lens dose limit within the European Union countries, Japan and elsewhere. This Commentary introduces the work done by the LDLensRad Consortium contained within this Focus Issue, towards advancement of understanding of the mechanisms of low dose radiation cataract., (©2022 by Radiation Research Society. All rights of reproduction in any form reserved.)
- Published
- 2022
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20. The importance of the epithelial fibre cell interface to lens regeneration in an in vivo rat model and in a human bag-in-the-lens (BiL) sample.
- Author
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Wu W, Lois N, Prescott AR, Brown AP, Van Gerwen V, Tassignon MJ, Richards SA, Saunter CD, Jarrin M, and Quinlan RA
- Subjects
- Actins metabolism, Aged, Animals, Aquaporins metabolism, Cadherins metabolism, Cell Proliferation physiology, Epithelial Cells ultrastructure, Epithelial-Mesenchymal Transition physiology, Eye Proteins metabolism, Female, Fibronectins metabolism, Humans, In Situ Nick-End Labeling, Lens Capsule, Crystalline cytology, Lens Capsule, Crystalline surgery, Lens, Crystalline ultrastructure, Male, Microscopy, Electron, Microscopy, Fluorescence, Models, Animal, Nerve Tissue Proteins metabolism, Proteomics, Rats, Rats, Sprague-Dawley, Tandem Mass Spectrometry, Capsule Opacification metabolism, Epithelial Cells physiology, Lens Implantation, Intraocular, Lens, Crystalline physiology, Regeneration physiology
- Abstract
Human lens regeneration and the Bag-in-the-Lens (BIL) surgical treatment for cataract both depend upon lens capsule closure for their success. Our studies suggest that the first three days after surgery are critical to their long-term outcomes. Using a rat model of lens regeneration, we evidenced lens epithelial cell (LEC) proliferation increased some 50 fold in the first day before rapidly declining to rates observed in the germinative zone of the contra-lateral, un-operated lens. Cell multi-layering at the lens equator occurred on days 1 and 2, but then reorganised into two discrete layers by day 3. E- and N-cadherin expression preceded cell polarity being re-established during the first week. Aquaporin 0 (AQP0) was first detected in the elongated cells at the lens equator at day 7. Cells at the capsulotomy site, however, behaved very differently expressing the epithelial mesenchymal transition (EMT) markers fibronectin and alpha-smooth muscle actin (SMA) from day 3 onwards. The physical interaction between the apical surfaces of the anterior and posterior LECs from day 3 after surgery preceded cell elongation. In the human BIL sample fibre cell formation was confirmed by both histological and proteome analyses, but the cellular response is less ordered and variable culminating in Soemmerring's ring (SR) formation and sometimes Elschnig's pearls. This we evidence for lenses from a single patient. No bow region or recognisable epithelial-fibre cell interface (EFI) was evident and consequently the fibre cells were disorganised. We conclude that lens cells require spatial and cellular cues to initiate, sustain and produce an optically functional tissue in addition to capsule integrity and the EFI., (Copyright © 2021. Published by Elsevier Ltd.)
- Published
- 2021
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21. Vaginal Microbiota, Genital Inflammation and Extracellular Matrix Remodelling Collagenase: MMP-9 in Pregnant Women With HIV, a Potential Preterm Birth Mechanism Warranting Further Exploration.
- Author
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Short CS, Quinlan RA, Wang X, Preda VG, Smith A, Marchesi JR, Lee YS, MacIntyre DA, Bennett PR, and Taylor GP
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- Collagenases, Extracellular Matrix, Female, Humans, Infant, Newborn, Inflammation, Matrix Metalloproteinase 9, Pregnancy, Pregnant Women, RNA, Ribosomal, 16S genetics, Vagina, HIV Infections complications, Microbiota, Premature Birth
- Abstract
Background: Pregnant women living with HIV infection (PWLWH) have elevated rates of preterm birth (PTB) in which HIV and cART are implicated. PWLWH also have a high prevalence of adverse vaginal microbiota, which associate with genital tract inflammation. The mechanism underlying PTB in PWLWH is unknown. We present the first data in PWLWH on genital-tract matrix-metalloproteinase-9(MMP-9), an important collagenase implicated in labour onset, and tissue inhibitor of metalloproteinases-1(TIMP-1) and explore correlations with local inflammation and vaginal bacteria., Material and Methods: Cervical vaginal fluid (CVF) collected by a soft cup and high vaginal swabs (HVS) were obtained from PWLWH and HIV uninfected pregnant women (HUPW) at three antenatal time points. Maternal characteristics, combination antiretroviral therapy (cART) exposure, and pregnancy outcome were recorded. Concentrations of MMP-9, TIMP-1 and ten cytokines were measured by immunoassays. Vaginal microbiota composition was determined through 16S rRNA amplicon sequencing. MMP-9, TIMP-1 and cytokine concentrations were compared by HIV status, cART, and prematurity and in PWLWH correlations with polymorphonuclear leucocytes, cytokines and bacterial genera were explored., Results: CVF was available for 50 PWLWH (108 samples) and 12 HUPW (20 samples) between gestation weeks 14-38. Thirty-six PWLWH conceived on cART and 14 initiated post-conception. There were five and one PTB outcomes in PWLWH and HUPW respectively. PWLWH had higher mean CVF concentrations of MMP-9 (p<0.001) and TIMP-1 (p=0.035) in the second trimester compared with HUPW with a similar trend in the third trimester. PWLWH also had higher CVF values of cytokines: IL-1β, IL-8, IL-12 and TNF-α in both trimesters compared to HUPW (p ≤ 0.003). In PWLWH, MMP-9 positively correlated with TIMP-1 (r=0.31, p=0.002) and CVF polymorphonuclear leucocytes (r=0.57, p=0.02). Correlations were observed between MMP-9 and three cytokines: IL-1β (r=0.61), IL-8 (r=0.57) and TNF-α (r=0.64), p<0.001, similarly for TIMP-1. Abundance of anaerobic pathobionts correlated with MMP-9: Gardnerella (r=0.44, p<0.001), Atopobium (r=0.33, p=0.005), and Prevotella genera (r=0.39, p<0.001). Conversely proportion of Lactobacillus genera negatively correlated with MMP-9 (rho=-0.46, p<0.001). MMP-9/TIMP-1 ratio increased with gestational age at sampling in PWLWH, but this was no longer significant after adjusting for confounders and no difference by prematurity was observed in this sub-study., Conclusions: Here we show strong correlations of MMP-9 to genital tract inflammation and sub-optimal bacterial genera in PWLWH indicating the ascending genital tract infection pathway may be a contributory mechanism to the high risk of PTB., Competing Interests: PB reports personal fees and shares and stock ownership in ObsEva Pharmaceuticals, personal fees from GlaxoSmithKline that are both outside the submitted work. PB and DM have a patent for microRNA markers to predict cervical shortening and preterm birth issued again outside of the submitted work. The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Short, Quinlan, Wang, Preda, Smith, Marchesi, Lee, MacIntyre, Bennett and Taylor.)
- Published
- 2021
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22. Cluster analyses of the TCGA and a TMA dataset using the coexpression of HSP27 and CRYAB improves alignment with clinical-pathological parameters of breast cancer and suggests different epichaperome influences for each sHSP.
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Quinlan PR, Figeuredo G, Mongan N, Jordan LB, Bray SE, Sreseli R, Ashfield A, Mitsch J, van den Ijssel P, Thompson AM, and Quinlan RA
- Subjects
- Cluster Analysis, Female, HSP27 Heat-Shock Proteins genetics, HSP27 Heat-Shock Proteins metabolism, Heat-Shock Proteins analysis, Humans, Molecular Chaperones analysis, alpha-Crystallin B Chain metabolism, Breast Neoplasms genetics, Heat-Shock Proteins metabolism, Heat-Shock Proteins, Small, Molecular Chaperones metabolism
- Abstract
Our cluster analysis of the Cancer Genome Atlas for co-expression of HSP27 and CRYAB in breast cancer patients identified three patient groups based on their expression level combination (high HSP27 + low CRYAB; low HSP27 + high CRYAB; similar HSP27 + CRYAB). Our analyses also suggest that there is a statistically significant inverse relationship between HSP27 and CRYAB and known clinicopathological markers in breast cancer. Screening an unbiased 248 breast cancer patient tissue microarray (TMA) for the protein expression of HSP27 and phosphorylated HSP27 (HSP27-82pS) with CRYAB also identified three patient groups based on HSP27 and CRYAB expression levels. TMA24 also had recorded clinical-pathological parameters, such as ER and PR receptor status, patient survival, and TP53 mutation status. High HSP27 protein levels were significant with ER and PR expression. HSP27-82pS associated with the best patient survival (Log Rank test). High CRYAB expression in combination with wild-type TP53 was significant for patient survival, but a different patient outcome was observed when mutant TP53 was combined with high CRYAB expression. Our data suggest that HSP27 and CRYAB have different epichaperome influences in breast cancer, but more importantly evidence the value of a cluster analysis that considers their coexpression. Our approach can deliver convergence for archival datasets as well as those from recent treatment and patient cohorts and can align HSP27 and CRYAB expression to important clinical-pathological features of breast cancer., (© 2022. The Author(s).)
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- 2021
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23. Joining European Scientific Forces to Face Pandemics.
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Vasconcelos MH, Alcaro S, Arechavala-Gomeza V, Baumbach J, Borges F, Brevini TAL, Rivas JL, Devaux Y, Hozak P, Keinänen-Toivola MM, Lattanzi G, Mohr T, Murovska M, Prusty BK, Quinlan RA, Pérez-Sala D, Scheibenbogen C, Schmidt HHHW, Silveira I, Tieri P, Tolios A, and Riganti C
- Subjects
- Communication, Europe, Humans, Laboratory Personnel, Pandemics, SARS-CoV-2 genetics, Biomedical Research organization & administration, COVID-19 virology, SARS-CoV-2 physiology
- Abstract
Despite the international guidelines on the containment of the coronavirus disease 2019 (COVID-19) pandemic, the European scientific community was not sufficiently prepared to coordinate scientific efforts. To improve preparedness for future pandemics, we have initiated a network of nine European-funded Cooperation in Science and Technology (COST) Actions that can help facilitate inter-, multi-, and trans-disciplinary communication and collaboration., (Copyright © 2020 The Authors. Published by Elsevier Ltd.. All rights reserved.)
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- 2021
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24. The genetic landscape of crystallins in congenital cataract.
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Berry V, Ionides A, Pontikos N, Georgiou M, Yu J, Ocaka LA, Moore AT, Quinlan RA, and Michaelides M
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- DNA Mutational Analysis, Humans, Mutation, Missense genetics, Pedigree, Cataract genetics, Crystallins genetics, Lens, Crystalline
- Abstract
Background: The crystalline lens is mainly composed of a large family of soluble proteins called the crystallins, which are responsible for its development, growth, transparency and refractive index. Disease-causing sequence variants in the crystallins are responsible for nearly 50% of all non-syndromic inherited congenital cataracts, as well as causing cataract associated with other diseases, including myopathies. To date, more than 300 crystallin sequence variants causing cataract have been identified., Methods: Here we aimed to identify the genetic basis of disease in five multi-generation British families and five sporadic cases with autosomal dominant congenital cataract using whole exome sequencing, with identified variants validated using Sanger sequencing. Following bioinformatics analysis, rare or novel variants with a moderate to damaging pathogenicity score, were filtered out and tested for segregation within the families., Results: We have identified 10 different heterozygous crystallin variants. Five recurrent variants were found: family-A, with a missense variant (c.145C>T; p.R49C) in CRYAA associated with nuclear cataract; family-B, with a deletion in CRYBA1 (c.272delGAG; p.G91del) associated with nuclear cataract; and family-C, with a truncating variant in CRYGD (c.470G>A; W157*) causing a lamellar phenotype; individuals I and J had variants in CRYGC (c.13A>C; T5P) and in CRYGD (c.418C>T; R140*) causing unspecified congenital cataract and nuclear cataract, respectively. Five novel disease-causing variants were also identified: family D harboured a variant in CRYGC (c.179delG; R60Qfs*) responsible for a nuclear phenotype; family E, harboured a variant in CRYBB1 (c.656G>A; W219*) associated with lamellar cataract; individual F had a variant in CRYGD (c.392G>A; W131*) associated with nuclear cataract; and individuals G and H had variants in CRYAA (c.454delGCC; A152del) and in CRYBB1 (c.618C>A; Y206*) respectively, associated with unspecified congenital cataract. All novel variants were predicted to be pathogenic and to be moderately or highly damaging., Conclusions: We report five novel variants and five known variants. Some are rare variants that have been reported previously in small ethnic groups but here we extend this to the wider population and record a broader phenotypic spectrum for these variants.
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- 2020
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25. Three-dimensional data capture and analysis of intact eye lenses evidences emmetropia-associated changes in epithelial cell organization.
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Kalligeraki AA, Isted A, Jarrin M, Uwineza A, Pal R, Saunter CD, Girkin JM, Obara B, and Quinlan RA
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- Animals, Epithelium physiology, Mice, Mice, Inbred C57BL, Microscopy, Confocal methods, Emmetropia physiology, Epithelial Cells physiology, Lens, Crystalline physiology
- Abstract
Organ and tissue development are highly coordinated processes; lens growth and functional integration into the eye (emmetropia) is a robust example. An epithelial monolayer covers the anterior hemisphere of the lens, and its organization is the key to lens formation and its optical properties throughout all life stages. To better understand how the epithelium supports lens function, we have developed a novel whole tissue imaging system using conventional confocal light microscopy and a specialized analysis software to produce three-dimensional maps for the epithelium of intact mouse lenses. The open source software package geometrically determines the anterior pole position, the equatorial diameter, and three-dimensional coordinates for each detected cell in the epithelium. The user-friendly cell maps, which retain global lens geometry, allow us to document age-dependent changes in the C57/BL6J mouse lens cell distribution characteristics. We evidence changes in epithelial cell density and distribution in C57/BL6J mice during the establishment of emmetropia between postnatal weeks 4-6. These epithelial changes accompany a previously unknown spheroid to lentoid shape transition of the lens as detected by our analyses. When combined with key findings from previous mouse genetic and cell biological studies, we suggest a cytoskeleton-based mechanism likely underpins these observations.
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- 2020
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26. Whole Exome Sequencing Reveals Novel and Recurrent Disease-Causing Variants in Lens Specific Gap Junctional Protein Encoding Genes Causing Congenital Cataract.
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Berry V, Ionides A, Pontikos N, Moghul I, Moore AT, Quinlan RA, and Michaelides M
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- Amino Acid Sequence, Animals, Base Sequence, Cataract genetics, Connexins chemistry, Exome, Female, Genes, Dominant, Genetic Variation, High-Throughput Nucleotide Sequencing, Male, Models, Molecular, Pedigree, Phenotype, Protein Conformation, Sequence Alignment, Sequence Homology, Amino Acid, United Kingdom, Vertebrates genetics, Cataract congenital, Connexins genetics, Lens, Crystalline metabolism, Mutation, Missense, Exome Sequencing
- Abstract
Pediatric cataract is clinically and genetically heterogeneous and is the most common cause of childhood blindness worldwide. In this study, we aimed to identify disease-causing variants in three large British families and one isolated case with autosomal dominant congenital cataract, using whole exome sequencing. We identified four different heterozygous variants, three in the large families and one in the isolated case. Family A, with a novel missense variant (c.178G>C, p.Gly60Arg) in GJA8 with lamellar cataract; family B, with a recurrent variant in GJA8 (c.262C>T, p.Pro88Ser) associated with nuclear cataract; and family C, with a novel variant in GJA3 (c.771dupC, p.Ser258GlnfsTer68) causing a lamellar phenotype. Individual D had a novel variant in GJA3 (c.82G>T, p.Val28Leu) associated with congenital cataract. Each sequence variant was found to co-segregate with disease. Here, we report three novel and one recurrent disease-causing sequence variant in the gap junctional protein encoding genes causing autosomal dominant congenital cataract. Our study further extends the mutation spectrum of these genes and further facilitates clinical diagnosis. A recurrent p.P88S variant in GJA8 causing isolated nuclear cataract provides evidence of further phenotypic heterogeneity associated with this variant.
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- 2020
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27. Site-specific phosphorylation and caspase cleavage of GFAP are new markers of Alexander disease severity.
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Battaglia RA, Beltran AS, Delic S, Dumitru R, Robinson JA, Kabiraj P, Herring LE, Madden VJ, Ravinder N, Willems E, Newman RA, Quinlan RA, Goldman JE, Perng MD, Inagaki M, and Snider NT
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- Adult, Alexander Disease diagnosis, Alexander Disease genetics, Astrocytes metabolism, Binding Sites genetics, Brain metabolism, Brain pathology, Cell Line, Glial Fibrillary Acidic Protein genetics, Humans, Induced Pluripotent Stem Cells metabolism, Infant, Intermediate Filaments metabolism, Mutation, Phosphorylation, Proteolysis, Severity of Illness Index, Alexander Disease metabolism, Biomarkers metabolism, Caspases metabolism, Glial Fibrillary Acidic Protein metabolism
- Abstract
Alexander disease (AxD) is a fatal neurodegenerative disorder caused by mutations in glial fibrillary acidic protein (GFAP), which supports the structural integrity of astrocytes. Over 70 GFAP missense mutations cause AxD, but the mechanism linking different mutations to disease-relevant phenotypes remains unknown. We used AxD patient brain tissue and induced pluripotent stem cell (iPSC)-derived astrocytes to investigate the hypothesis that AxD-causing mutations perturb key post-translational modifications (PTMs) on GFAP. Our findings reveal selective phosphorylation of GFAP-Ser13 in patients who died young, independently of the mutation they carried. AxD iPSC-astrocytes accumulated pSer13-GFAP in cytoplasmic aggregates within deep nuclear invaginations, resembling the hallmark Rosenthal fibers observed in vivo. Ser13 phosphorylation facilitated GFAP aggregation and was associated with increased GFAP proteolysis by caspase-6. Furthermore, caspase-6 was selectively expressed in young AxD patients, and correlated with the presence of cleaved GFAP. We reveal a novel PTM signature linking different GFAP mutations in infantile AxD., Competing Interests: RB, AB, SD, RD, JR, PK, LH, VM, RQ, JG, MP, MI No competing interests declared, NR, EW, RN are paid employees of ThermoFisher Scientific, whose products were used to complete parts of the study. ThermoFisher Scientific had no role in the study design, data analysis, decision to publish, or preparation of the manuscript. NS is a member of the Scientific Advisory Board for Elise's Corner Fund, which supported part of this work, (© 2019, Battaglia et al.)
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- 2019
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28. BFSP1 C-terminal domains released by post-translational processing events can alter significantly the calcium regulation of AQP0 water permeability.
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Tapodi A, Clemens DM, Uwineza A, Jarrin M, Goldberg MW, Thinon E, Heal WP, Tate EW, Nemeth-Cahalan K, Vorontsova I, Hall JE, and Quinlan RA
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- Adolescent, Adult, Aged, Amino Acid Sequence, Animals, Blotting, Western, Caspases metabolism, Cell Membrane Permeability, Cells, Cultured, Epithelial Cells metabolism, Humans, Immunohistochemistry, Lens, Crystalline cytology, MCF-7 Cells metabolism, Microscopy, Electron, Scanning, Middle Aged, Molecular Sequence Data, Myristates metabolism, Oocytes, Protein Domains, Transfection, Xenopus laevis, Young Adult, Aquaporins metabolism, Body Water metabolism, Calcium metabolism, Eye Proteins metabolism, Intermediate Filament Proteins metabolism, Protein Processing, Post-Translational
- Abstract
BFSP1 (beaded filament structural protein 1, filensin) is a cytoskeletal protein expressed in the eye lens. It binds AQP0 in vitro and its C-terminal sequences have been suggested to regulate the water channel activity of AQP0. A myristoylated fragment from the C-terminus of BFSP1 was found in AQP0 enriched fractions. Here we identify BFSP1 as a substrate for caspase-mediated cleavage at several C-terminal sites including D433. Cleavage at D433 exposes a cryptic myristoylation sequence (434-440). We confirm that this sequence is an excellent substrate for both NMT1 and 2 (N-myristoyl transferase). Thus caspase cleavage may promote formation of myristoylated fragments derived from the BFSP1 C-terminus (G434-S665). Myristoylation at G434 is not required for membrane association. Biochemical fractionation and immunogold labeling confirmed that C-terminal BFSP1 fragments containing the myristoylation sequence colocalized with AQP0 in the same plasma membrane compartments of lens fibre cells. To determine the functional significance of the association of BFSP1 G434-S665 sequences with AQP0, we measured AQP0 water permeability in Xenopus oocytes co-transfected with transcripts expressing both AQP0 and various C-terminal domain fragments of BFSP1 generated by caspase cleavage. We found that different fragments dramatically alter the response of AQP0 to different concentrations of Ca
2+ . The complete C-terminal fragment (G434-S665) eliminates calcium regulation altogether. Shorter fragments can enhance regulation by elevated calcium or reverse the response, indicative of the regulatory potential of BFSP1 with respect to AQP0. In particular, elimination of the myristoylation site by the mutation G434A reverses the order of water permeability sensitivity to different Ca2+ concentrations., (Copyright © 2019 The Authors. Published by Elsevier Ltd.. All rights reserved.)- Published
- 2019
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29. Cataractogenic load - A concept to study the contribution of ionizing radiation to accelerated aging in the eye lens.
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Uwineza A, Kalligeraki AA, Hamada N, Jarrin M, and Quinlan RA
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- Animals, Humans, Oxidative Stress radiation effects, Radiation, Ionizing, Aging radiation effects, Cataract etiology, Lens, Crystalline radiation effects
- Abstract
Ionizing radiation (IR) damages DNA and other macromolecules, including proteins and lipids. Most cell types can repair DNA damage and cycle continuously their macromolecules as a mechanism to remove defective proteins and lipids. In those cells that lack nuclei and other organelles, such as lens fiber cells and mammalian erythrocytes, IR-induced damage to macromolecules is retained because they cannot be easily replenished. Whilst the life span for an erythrocyte is several months, the life span of a human lens is decades. There is very limited turnover in lens macromolecules, therefore the aging process greatly impacts lens structure and function over its lifetime. The lens is a tissue where biomolecular longevity, lifelong retention of its components and continued growth are integral to its homeostasis. These characteristics make the lens an excellent model to study the contribution of retained macromolecular damage over time. Epidemiological data have revealed a significant association between exposure to IR, the loss of lens optical function and the formation of cataracts (cataractogenesis) later in life. Lifestyle, genetic and environmental factors all contribute to cataractogenesis due to their effect on the aging process. Cataract is an iconic age-related disease in humans. IR is a recognised cause of cataract and the occupational lens dose limit is reduced from 150 to 20 mGy / year averaged over 5 years (ICRP Publication 118). Understanding the effects of low dose IR on the lens and its role in cataractogenesis is therefore very important. So we redefine "cataractogenic load" as a term to account for the combined lifestyle, genetic and environmental processes that increase biomolecular damage to lens macromolecules leading to cataract formation. These processes weaken metabolic defenses, increase post-translational protein modifications, and alter the lipid structure and content of the lens. IR exposure is a significant insult to the lens because of free radical generation and the ensuing oxidative stress. We support the concept that damage caused by IR compounds the aging process by increasing the cataractogenic load, hereby accelerating lens aging and its loss of function., (Copyright © 2019 The Authors. Published by Elsevier B.V. All rights reserved.)
- Published
- 2019
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30. Dotting the eyes: mouse strain dependency of the lens epithelium to low dose radiation-induced DNA damage.
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Barnard SGR, Moquet J, Lloyd S, Ellender M, Ainsbury EA, and Quinlan RA
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- Animals, DNA Repair radiation effects, Dose-Response Relationship, Radiation, Epithelium metabolism, Epithelium radiation effects, Mice, Mice, Inbred BALB C, Mice, Inbred C57BL, Radiation Tolerance genetics, Species Specificity, Time Factors, DNA Damage, Lens, Crystalline metabolism, Lens, Crystalline radiation effects
- Abstract
Purpose: Epidemiological evidence regarding the radiosensitivity of the lens of the eye and radiation cataract development has led to changes in the EU Basic Safety Standards for protection of the lens against ionizing radiation. However, mechanistic details of lens radiation response pathways and their significance for cataractogenesis remain unclear. Radiation-induced DNA damage and the potential impairment of repair pathways within the lens epithelium, a cell monolayer that covers the anterior hemisphere of the lens, are likely to be involved., Materials and Methods: In this work, the lens epithelium has been analyzed for its DNA double-strand break (DSB) repair response to ionizing radiation. The responses of epithelial cells located at the anterior pole (central region) have been compared to at the very periphery of the monolayer (germinative and transitional zones). Described here are the different responses in the two regions and across four strains (C57BL/6, 129S2, BALB/c and CBA/Ca) over a low dose (0-25 mGy) in-vivo whole body X-irradiation range up to 24 hours post exposure., Results: DNA damage and repair as visualized through 53BP1 staining was present across the lens epithelium, although repair kinetics appeared non-uniform. Epithelial cells in the central region have significantly more 53BP1 foci. The sensitivities of different mouse strains have also been compared., Conclusions: 129S2 and BALB/c showed higher levels of DNA damage, with BALB/c showing significantly less inter-individual variability and appearing to be a more robust model for future DNA damage and repair studies. As a result of this study, BALB/c was identified as a suitable radiosensitive lens strain to detect and quantify early low dose ionizing radiation DNA damage effects in the mouse eye lens specifically, as an indicator of cataract formation.
- Published
- 2018
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31. γ-Crystallin redox-detox in the lens.
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Quinlan RA and Hogg PJ
- Subjects
- Amino Acid Substitution, Cataract physiopathology, Cysteine chemistry, Humans, Mutation, Oxidation-Reduction, Oxidoreductases Acting on Sulfur Group Donors genetics, gamma-Crystallins genetics, Disulfides metabolism, Lens, Crystalline metabolism, Oxidoreductases Acting on Sulfur Group Donors metabolism, gamma-Crystallins metabolism
- Abstract
In the vertebrate eye, limiting oxidation of proteins and lipids is key to maintaining lens function and avoiding cataract formation. A study by Serebryany et al. identifies a surprising contributor to the eye's oxidative defense in their demonstration that γD-crystallin (HγD) functions as an oxidoreductase and uses disulfide exchange to initiate aggregation of mutant crystallins that mimic oxidative damage. These insights suggest a mechanism by which a dynamic pool of closely packed proteins might avoid oxidation-driven protein-folding traps, providing new avenues to understand the basis of a human disease with global impact., (© 2018 Quinlan and Hogg.)
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- 2018
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32. Heat shock proteins are differentially expressed in brain and spinal cord: implications for multiple sclerosis.
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Gorter RP, Nutma E, Jahrei MC, de Jonge JC, Quinlan RA, van der Valk P, van Noort JM, Baron W, and Amor S
- Subjects
- Adult, Aged, Aged, 80 and over, Demyelinating Diseases, Female, Gray Matter pathology, Humans, Immunohistochemistry, Male, Middle Aged, White Matter pathology, Astrocytes metabolism, Brain pathology, Gray Matter metabolism, Heat-Shock Proteins metabolism, Multiple Sclerosis metabolism, Neurons metabolism, Spinal Cord pathology, White Matter metabolism
- Abstract
Multiple sclerosis (MS) is a chronic neurodegenerative disease characterized by demyelination, inflammation and neurodegeneration throughout the central nervous system. Although spinal cord pathology is an important factor contributing to disease progression, few studies have examined MS lesions in the spinal cord and how they differ from brain lesions. In this study we have compared brain and spinal cord white (WM) and grey (GM) matter from MS and control tissues, focusing on small heat shock proteins (HSPB) and HSP16.2. Western blotting was used to examine protein levels of HSPB1, HSPB5, HSPB6, HSPB8 and HSP16.2 in brain and spinal cord from MS and age-matched non-neurological controls. Immunohistochemistry was used to examine expression of the HSPs in MS spinal cord lesions and controls. Expression levels were quantified using ImageJ. Western blotting revealed significantly higher levels of HSPB1, HSPB6 and HSPB8 in MS and control spinal cord compared to brain tissues. No differences in HSPB5 and HSP16.2 protein levels were observed, although HSPB5 protein levels were higher in brain WM versus GM. In MS spinal cord lesions, increased HSPB1 and HSPB5 expression was observed in astrocytes, and increased neuronal expression of HSP16.2 was observed in normal-appearing GM and type 1 GM lesions. The high constitutive expression of several HSPBs in spinal cord and increased expression of HSPBs and HSP16.2 in MS illustrate differences between brain and spinal cord in health and upon demyelination. Regional differences in HSP expression may reflect differences in astrocyte cytoskeleton composition and influence inflammation, possibly affecting the effectiveness of pharmacological agents., (© 2018 The Authors. Clinical & Experimental Immunology published by John Wiley & Sons Ltd on behalf of British Society for Immunology.)
- Published
- 2018
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33. Non-invasive in vivo quantification of the developing optical properties and graded index of the embryonic eye lens using SPIM.
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Young LK, Jarrin M, Saunter CD, Quinlan RA, and Girkin JM
- Abstract
Graded refractive index lenses are inherent to advanced visual systems in animals. By understanding their formation and local optical properties, significant potential for improved ocular healthcare may be realized. We report a novel technique measuring the developing optical power of the eye lens, in a living animal, by exploiting the orthogonal imaging modality of a selective plane illumination microscope (SPIM). We have quantified the maturation of the lenticular refractive index at three different visible wavelengths using a combined imaging and ray tracing approach. We demonstrate that the method can be used with transgenic and vital dye labeling as well as with both fixed and living animals. Using a key eye lens morphogen and its inhibitor, we have measured their effects both on lens size and on refractive index. Our technique provides insights into the mechanisms involved in the development of this natural graded index micro-lens and its associated optical properties., Competing Interests: The authors declare that there are no conflicts of interest related to this article.
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- 2018
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34. A rim-and-spoke hypothesis to explain the biomechanical roles for cytoplasmic intermediate filament networks.
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Quinlan RA, Schwarz N, Windoffer R, Richardson C, Hawkins T, Broussard JA, Green KJ, and Leube RE
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- Animals, Cell Membrane physiology, Cell Membrane ultrastructure, Cytoplasm physiology, Epithelial Cells physiology, Epithelial Cells ultrastructure, Humans, Intermediate Filaments physiology, Mechanotransduction, Cellular, Models, Molecular, Skin ultrastructure, Intermediate Filaments ultrastructure
- Abstract
Textbook images of keratin intermediate filament (IF) networks in epithelial cells and the functional compromization of the epidermis by keratin mutations promulgate a mechanical role for this important cytoskeletal component. In stratified epithelia, keratin filaments form prominent radial spokes that are focused onto cell-cell contact sites, i.e. the desmosomes. In this Hypothesis, we draw attention to a subset of keratin filaments that are apposed to the plasma membrane. They form a rim of filaments interconnecting the desmosomes in a circumferential network. We hypothesize that they are part of a rim-and-spoke arrangement of IFs in epithelia. From our review of the literature, we extend this functional role for the subplasmalemmal rim of IFs to any cell, in which plasma membrane support is required, provided these filaments connect directly or indirectly to the plasma membrane. Furthermore, cytoplasmic IF networks physically link the outer nuclear and plasma membranes, but their participation in mechanotransduction processes remain largely unconsidered. Therefore, we also discuss the potential biomechanical and mechanosensory role(s) of the cytoplasmic IF network in terms of such a rim (i.e. subplasmalemmal)-and-spoke arrangement for cytoplasmic IF networks., Competing Interests: Competing interestsThe authors declare no competing or financial interests., (© 2017. Published by The Company of Biologists Ltd.)
- Published
- 2017
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35. αB-crystallin is a sensor for assembly intermediates and for the subunit topology of desmin intermediate filaments.
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Sharma S, Conover GM, Elliott JL, Der Perng M, Herrmann H, and Quinlan RA
- Subjects
- Amino Acid Sequence, Binding Sites, Desmin chemistry, Desmin genetics, Desmin ultrastructure, Humans, Intermediate Filaments chemistry, Intermediate Filaments genetics, Intermediate Filaments ultrastructure, Point Mutation, Protein Binding, Protein Domains, Desmin metabolism, Intermediate Filaments metabolism, alpha-Crystallin B Chain metabolism
- Abstract
Mutations in the small heat shock protein chaperone CRYAB (αB-crystallin/HSPB5) and the intermediate filament protein desmin, phenocopy each other causing cardiomyopathies. Whilst the binding sites for desmin on CRYAB have been determined, desmin epitopes responsible for CRYAB binding and also the parameters that determine CRYAB binding to desmin filaments are unknown. Using a combination of co-sedimentation centrifugation, viscometric assays and electron microscopy of negatively stained filaments to analyse the in vitro assembly of desmin filaments, we show that the binding of CRYAB to desmin is subject to its assembly status, to the subunit organization within filaments formed and to the integrity of the C-terminal tail domain of desmin. Our in vitro studies using a rapid assembly protocol, C-terminally truncated desmin and two disease-causing mutants (I451M and R454W) suggest that CRYAB is a sensor for the surface topology of the desmin filament. Our data also suggest that CRYAB performs an assembly chaperone role because the assembling filaments have different CRYAB-binding properties during the maturation process. We suggest that the capability of CRYAB to distinguish between filaments with different surface topologies due either to mutation (R454W) or assembly protocol is important to understanding the pathomechanism(s) of desmin-CRYAB myopathies.
- Published
- 2017
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36. The functional roles of the unstructured N- and C-terminal regions in αB-crystallin and other mammalian small heat-shock proteins.
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Carver JA, Grosas AB, Ecroyd H, and Quinlan RA
- Subjects
- Animals, Heat-Shock Proteins, Small metabolism, Humans, Models, Molecular, Protein Aggregates, Protein Binding, Protein Conformation, Protein Interaction Domains and Motifs, Protein Interaction Maps, Protein Multimerization, alpha-Crystallin B Chain metabolism, Heat-Shock Proteins, Small chemistry, alpha-Crystallin B Chain chemistry
- Abstract
Small heat-shock proteins (sHsps), such as αB-crystallin, are one of the major classes of molecular chaperone proteins. In vivo, under conditions of cellular stress, sHsps are the principal defence proteins that prevent large-scale protein aggregation. Progress in determining the structure of sHsps has been significant recently, particularly in relation to the conserved, central and β-sheet structured α-crystallin domain (ACD). However, an understanding of the structure and functional roles of the N- and C-terminal flanking regions has proved elusive mainly because of their unstructured and dynamic nature. In this paper, we propose functional roles for both flanking regions, based around three properties: (i) they act in a localised crowding manner to regulate interactions with target proteins during chaperone action, (ii) they protect the ACD from deleterious amyloid fibril formation and (iii) the flexibility of these regions, particularly at the extreme C-terminus in mammalian sHsps, provides solubility for sHsps under chaperone and non-chaperone conditions. In the eye lens, these properties are highly relevant as the crystallin proteins, in particular the two sHsps αA- and αB-crystallin, are present at very high concentrations.
- Published
- 2017
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37. THE IMPACT OF CIRCADIAN RHYTHMS ON MEDICAL IMAGING AND RADIOTHERAPY REGIMES FOR THE PAEDIATRIC PATIENT.
- Author
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Forssell-Aronsson E and Quinlan RA
- Subjects
- Apoptosis, Cell Cycle, Child, Humans, Neoplasms diagnostic imaging, Neoplasms radiotherapy, Radiation, Ionizing, Circadian Rhythm, Diagnostic Imaging, Radiation Exposure
- Abstract
Daily rhythmic changes are found in cellular events in cell cycle, DNA repair, apoptosis and angiogenesis in both normal and tumour tissue, as well as in enzymatic activity and drug metabolism. In this paper, we hypothesize that circadian rhythms need to be considered in radiation protection and optimization in personalized medicine, especially for paediatric care. The sensitivity of the eye lens to ionizing radiation makes the case for limiting damage to the lens epithelium by planning medical radio-imaging procedures for the afternoon, rather than the morning. Equally, the tumour and normal tissue response to radiotherapy is also subject to diurnal variation enabling optimization of time of treatment., (© The Author 2016. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oup.com.)
- Published
- 2017
- Full Text
- View/download PDF
38. The eye lens - a paradigm for healthy living.
- Author
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Quinlan RA
- Subjects
- History, 20th Century, History, 21st Century, Lens Diseases genetics, Lens Diseases metabolism, United States, Lens Diseases history, Lens, Crystalline, Molecular Biology history, Ophthalmology history
- Published
- 2017
- Full Text
- View/download PDF
39. Small molecules, both dietary and endogenous, influence the onset of lens cataracts.
- Author
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Barnes S and Quinlan RA
- Subjects
- Aging physiology, Animals, Antioxidants physiology, Glucocorticoids adverse effects, Humans, Metabolic Networks and Pathways, Vitamins physiology, Cataract etiology, Cataract prevention & control, Crystallins physiology, Diet, Glutathione physiology, Lens, Crystalline metabolism, Sphingolipids physiology
- Abstract
How the lens ages successfully is a lesson in biological adaption and the emergent properties of its complement of cells and proteins. This living tissue contains some of the oldest proteins in our bodies and yet they remain functional for decades, despite exposure to UV light, to reactive oxygen species and all the other hazards to protein function. This remarkable feat is achieved by a shrewd investment in very stable proteins as lens crystallins, by providing a reservoir of ATP-independent protein chaperones unequalled by any other tissue and by an oxidation-resistant environment. In addition, glutathione, a free radical scavenger, is present in mM concentrations and the plasma membranes contain oxidation-resistant sphingolipids what compromises lens function as it ages? In this review, we examine the role of small molecules in the prevention or causation of cataracts, including those associated with diet, metabolic pathways and drug therapy (steroids)., (Copyright © 2016 Elsevier Ltd. All rights reserved.)
- Published
- 2017
- Full Text
- View/download PDF
40. Sub-nanometre mapping of the aquaporin-water interface using multifrequency atomic force microscopy.
- Author
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Ricci M, Quinlan RA, and Voïtchovsky K
- Abstract
Aquaporins are integral membrane proteins that regulate the transport of water and small molecules in and out of the cell. In eye lens tissue, circulation of water, ions and metabolites is ensured by a microcirculation system in which aquaporin-0 (AQP0) plays a central role. AQP0 allows water to flow beyond the diffusion limit through lens membranes. AQP0 naturally arranges in a square lattice. The malfunction of AQP0 is related to numerous diseases such as cataracts. Despite considerable research into its structure, function and dynamics, the interface between the protein and the surrounding liquid and the effect of the lattice arrangement on the behaviour of water at the interface with the membrane are still not fully understood. Here we use a multifrequency atomic force microscopy (AFM) approach to map both the liquid at the interface with AQP0 and the protein itself with sub-nanometer resolution. Imaging using the fundamental eigenmode of the AFM cantilever probes mainly the interfacial water at the surface of the membrane. The results highlight a well-defined region that surrounds AQP0 tetramers and where water exhibits a higher affinity for the protein. Imaging in the second eigenmode is dominated by the mechanical response of the protein and provides sub-molecular details of the protein surface and the sub-surface structure. The relationship between modes and harmonics is also examined.
- Published
- 2016
- Full Text
- View/download PDF
41. The lipidation profile of aquaporin-0 correlates with the acyl composition of phosphoethanolamine lipids in lens membranes.
- Author
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Ismail VS, Mosely JA, Tapodi A, Quinlan RA, and Sanderson JM
- Subjects
- Animals, Aquaporins genetics, Cattle, Ethanolamines chemistry, Eye Proteins genetics, Gene Expression, Humans, Lens, Crystalline chemistry, Lipoylation, Membranes, Young Adult, Aquaporins metabolism, Arachidonic Acids metabolism, Ethanolamines metabolism, Eye Proteins metabolism, Lens, Crystalline metabolism, Protein Processing, Post-Translational
- Abstract
The lens fiber major intrinsic protein (otherwise known as aquaporin-0 (AQP0), MIP26 and MP26) has been examined by mass spectrometry (MS) in order to determine the speciation of acyl modifications to the side chains of lysine residues and the N-terminal amino group. The speciation of acyl modifications to the side chain of one specific, highly conserved lysine residue (K238) and the N-terminal amino group of human and bovine AQP0 revealed, in decreasing order of abundance, oleoyl, palmitoyl, stearoyl, eicosenoyl, dihomo-γ-linolenoyl, palmitoleoyl and eicosadienoyl modifications. In the case of human AQP0, an arachidonoyl modification was also found at the N-terminus. The relative abundances of these modifications mirror the fatty acid composition of lens phosphatidylethanolamine lipids. This lipid class would be expected to be concentrated in the inner leaflet of the lens fiber membrane to which each of the potential AQP0 lipidation sites is proximal. Our data evidence a broad lipidation profile that is both species and site independent, suggesting a chemical-based ester aminolysis mechanism to explain such modifications., (Copyright © 2016. Published by Elsevier B.V.)
- Published
- 2016
- Full Text
- View/download PDF
42. Ionizing radiation induced cataracts: Recent biological and mechanistic developments and perspectives for future research.
- Author
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Ainsbury EA, Barnard S, Bright S, Dalke C, Jarrin M, Kunze S, Tanner R, Dynlacht JR, Quinlan RA, Graw J, Kadhim M, and Hamada N
- Subjects
- Animals, Cataract pathology, Corneal Opacity etiology, Humans, Lens, Crystalline metabolism, Lens, Crystalline physiology, Lens, Crystalline radiation effects, Models, Animal, Cataract etiology, Radiation, Ionizing
- Abstract
The lens of the eye has long been considered as a radiosensitive tissue, but recent research has suggested that the radiosensitivity is even greater than previously thought. The 2012 recommendation of the International Commission on Radiological Protection (ICRP) to substantially reduce the annual occupational equivalent dose limit for the ocular lens has now been adopted in the European Union and is under consideration around the rest of the world. However, ICRP clearly states that the recommendations are chiefly based on epidemiological evidence because there are a very small number of studies that provide explicit biological, mechanistic evidence at doses <2Gy. This paper aims to present a review of recently published information on the biological and mechanistic aspects of cataracts induced by exposure to ionizing radiation (IR). The data were compiled by assessing the pertinent literature in several distinct areas which contribute to the understanding of IR induced cataracts, information regarding lens biology and general processes of cataractogenesis. Results from cellular and tissue level studies and animal models, and relevant human studies, were examined. The main focus was the biological effects of low linear energy transfer IR, but dosimetry issues and a number of other confounding factors were also considered. The results of this review clearly highlight a number of gaps in current knowledge. Overall, while there have been a number of recent advances in understanding, it remains unknown exactly how IR exposure contributes to opacification. A fuller understanding of how exposure to relatively low doses of IR promotes induction and/or progression of IR-induced cataracts will have important implications for prevention and treatment of this disease, as well as for the field of radiation protection., (Crown Copyright © 2016. Published by Elsevier B.V. All rights reserved.)
- Published
- 2016
- Full Text
- View/download PDF
43. Purification of Protein Chaperones and Their Functional Assays with Intermediate Filaments.
- Author
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Perng MD, Huang YS, and Quinlan RA
- Subjects
- Chromatography, Gel, Chromatography, Ion Exchange, Cytoskeletal Proteins isolation & purification, Escherichia coli, Heat-Shock Proteins chemistry, Humans, Recombinant Proteins chemistry, Recombinant Proteins isolation & purification, Cytoskeletal Proteins chemistry, Heat-Shock Proteins isolation & purification, Intermediate Filaments chemistry
- Abstract
Intermediate filament (IF) scaffolds facilitate small heat shock protein (sHSP) function, while IF function is sHSP dependent. sHSPs interact with IFs and the importance of this interaction is to maintain the individuality of the IFs and to modulate interfilament interactions both in networks and in assembly intermediates. Mutations in both sHSPs and their interacting IF proteins phenocopy each other in the human diseases they cause. This establishes a key functional relationship between these two very distinct protein families, and it also evidences the role of this cytoskeleton-chaperone complex in the cellular stress response. In this chapter, we describe the detailed experimental protocols for the preparation of purified IF proteins and sHSPs to facilitate the study in vitro of their functional interactions. In addition, we describe the detailed biochemical procedures to assess the effect of sHSP on the assembly of IFs, the binding to IFs, and the prevention of noncovalent filament-filament interactions using in vitro cosedimentation, electron microscopy, and viscosity assays. These assays are valuable research tools to study and manipulate sHSP-IF complexes in vitro and therefore to determine the structure-function detail of this complex, and how it contributes to cellular, tissue, and organismal homeostasis and the in vivo stress response., (© 2016 Elsevier Inc. All rights reserved.)
- Published
- 2016
- Full Text
- View/download PDF
44. In vivo, Ex Vivo, and In Vitro Approaches to Study Intermediate Filaments in the Eye Lens.
- Author
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Jarrin M, Young L, Wu W, Girkin JM, and Quinlan RA
- Subjects
- Animals, Cells, Cultured, Humans, In Vitro Techniques, Intermediate Filament Proteins metabolism, Intermediate Filaments metabolism, Lens, Crystalline metabolism
- Abstract
The role of the eye lens is to focus light into the retina. To perform this unique function, the ocular lens must be transparent. Previous studies have demonstrated the expression of vimentin, BFSP1, and BFSP2 in the eye lens. These intermediate filament (IF) proteins are essential to the optical properties of the lens. They are also important to its biomechanical properties, to the shape of the lens fiber cells, and to the organization and function of the plasma membrane. The eye lens is an iconic model in developmental studies, as a result different vertebrate models, including zebrafish, have been developed to study lens formation. In the present chapter, we have summarized the new approaches and the more breakthrough models (e.g., iPSc) that can be used to study the function of IFs in the ocular lens. We have presented three different groups of models. The first group includes in vitro models, where IFs can be studied and manipulated in lens cell cultures. The second includes ex vivo models. These replicate better the complex lens cell differentiation processes and the role(s) played by IFs. The third class is the in vivo models, and here, we have focused on Zebrafish and new imaging approaches using selective plane illumination microscopy. Finally, we present protocols on how to use these lens models to study IFs., (Copyright © 2016 Elsevier Inc. All rights reserved.)
- Published
- 2016
- Full Text
- View/download PDF
45. Radiation protection of the eye lens in medical workers--basis and impact of the ICRP recommendations.
- Author
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Barnard SG, Ainsbury EA, Quinlan RA, and Bouffler SD
- Subjects
- Humans, Radiation Dosage, Radiation Monitoring methods, Health Personnel, Lens, Crystalline radiation effects, Occupational Diseases prevention & control, Radiation Injuries prevention & control, Radiation Protection methods
- Abstract
The aim of this article was to explore the evidence for the revised European Union basic safety standard (BSS) radiation dose limits to the lens of the eye, in the context of medical occupational radiation exposures. Publications in the open literature have been reviewed in order to draw conclusions on the exposure profiles and doses received by medical radiation workers and to bring together the limited evidence for cataract development in medical occupationally exposed populations. The current status of relevant radiation-protection and monitoring practices and procedures is also considered. In conclusion, medical radiation workers do receive high doses in some circumstances, and thus working practices will be impacted by the new BSS. However, there is strong evidence to suggest that compliance with the new lower dose limits will be possible, although education and training of staff alongside effective use of personal protective equipment will be paramount. A number of suggested actions are given with the aim of assisting medical and associated radiation-protection professionals in understanding the requirements.
- Published
- 2016
- Full Text
- View/download PDF
46. DRUG DISCOVERY. A new dawn for cataracts.
- Author
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Quinlan RA
- Subjects
- Animals, Female, Humans, Male, Cataract drug therapy, Cataract metabolism, Lanosterol pharmacology, Lanosterol therapeutic use, Protein Aggregates drug effects, Protein Aggregation, Pathological drug therapy
- Published
- 2015
- Full Text
- View/download PDF
47. A dimensionless ordered pull-through model of the mammalian lens epithelium evidences scaling across species and explains the age-dependent changes in cell density in the human lens.
- Author
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Wu JJ, Wu W, Tholozan FM, Saunter CD, Girkin JM, and Quinlan RA
- Subjects
- Adult, Aged, Aged, 80 and over, Aging pathology, Animals, Cattle, Cell Survival, Epithelium metabolism, Epithelium pathology, Female, Humans, Lens, Crystalline pathology, Male, Mice, Middle Aged, Rabbits, Rats, Species Specificity, Aging metabolism, Cell Differentiation, Cell Proliferation, Lens, Crystalline metabolism, Models, Biological
- Abstract
We present a mathematical (ordered pull-through; OPT) model of the cell-density profile for the mammalian lens epithelium together with new experimental data. The model is based upon dimensionless parameters, an important criterion for inter-species comparisons where lens sizes can vary greatly (e.g. bovine (approx. 18 mm); mouse (approx. 2 mm)) and confirms that mammalian lenses scale with size. The validated model includes two parameters: β/α, which is the ratio of the proliferation rate in the peripheral and in the central region of the lens; and γ(GZ), a dimensionless pull-through parameter that accounts for the cell transition and exit from the epithelium into the lens body. Best-fit values were determined for mouse, rat, rabbit, bovine and human lens epithelia. The OPT model accounts for the peak in cell density at the periphery of the lens epithelium, a region where cell proliferation is concentrated and reaches a maximum coincident with the germinative zone. The β/α ratio correlates with the measured FGF-2 gradient, a morphogen critical to lens cell survival, proliferation and differentiation. As proliferation declines with age, the OPT model predicted age-dependent changes in cell-density profiles, which we observed in mouse and human lenses.
- Published
- 2015
- Full Text
- View/download PDF
48. The lens of the eye: exposures in the UK medical sector and mechanistic studies of radiation effects.
- Author
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Bouffler SD, Peters S, Gilvin P, Slack K, Markiewicz E, Quinlan RA, Gillan J, Coster M, Barnard S, Rothkamm K, and Ainsbury E
- Subjects
- Animals, Eye Diseases pathology, Eye Diseases physiopathology, Humans, Mice, Radiation Dosage, Radiation Injuries pathology, Radiation Injuries physiopathology, Risk, United Kingdom, Eye Diseases etiology, Lens, Crystalline radiation effects, Occupational Exposure, Ophthalmology, Optometry, Radiation Injuries etiology
- Abstract
The recommendation from the International Commission on Radiological Protection that the occupational equivalent dose limit for the lens of the eye should be reduced to 20 mSv year(-1), averaged over 5 years with no year exceeding 50 mSv, has stimulated a discussion on the practicalities of implementation of this revised dose limit, and the most appropriate risk and protection framework to adopt. This brief paper provides an overview of some of the drivers behind the move to a lower recommended dose limit. The issue of implementation in the medical sector in the UK has been addressed through a small-scale survey of doses to the lens of the eye amongst interventional cardiologists and radiologists. In addition, a mechanistic study of early and late post-irradiation changes in the lens of the eye in in-vivo-exposed mice is outlined. Surveys and studies such as those described can contribute to a deeper understanding of fundamental and practical issues, and therefore contribute to a robust evidence base for ensuring adequate protection of the eye while avoiding undesirable restrictions to working practices., (© The International Society for Prosthetics and Orthotics Reprints and permissions: sagepub.co.uk/journalsPermissions.nav.)
- Published
- 2015
- Full Text
- View/download PDF
49. Nonlinear ionizing radiation-induced changes in eye lens cell proliferation, cyclin D1 expression and lens shape.
- Author
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Markiewicz E, Barnard S, Haines J, Coster M, van Geel O, Wu W, Richards S, Ainsbury E, Rothkamm K, Bouffler S, and Quinlan RA
- Subjects
- Algorithms, Animals, Cell Line, DNA Breaks, Double-Stranded radiation effects, DNA Repair, Dose-Response Relationship, Radiation, Epithelial Cells metabolism, Epithelial Cells radiation effects, Female, Histones metabolism, Humans, Immunoblotting, Lens, Crystalline cytology, Lens, Crystalline metabolism, Lymphocytes drug effects, Lymphocytes metabolism, Male, Mice, Inbred C57BL, Microscopy, Fluorescence, Models, Biological, Cell Proliferation radiation effects, Cell Shape radiation effects, Cyclin D1 metabolism, Lens, Crystalline radiation effects, X-Rays
- Abstract
Elevated cataract risk after radiation exposure was established soon after the discovery of X-rays in 1895. Today, increased cataract incidence among medical imaging practitioners and after nuclear incidents has highlighted how little is still understood about the biological responses of the lens to low-dose ionizing radiation (IR). Here, we show for the first time that in mice, lens epithelial cells (LECs) in the peripheral region repair DNA double strand breaks (DSB) after exposure to 20 and 100 mGy more slowly compared with circulating blood lymphocytes, as demonstrated by counts of γH2AX foci in cell nuclei. LECs in the central region repaired DSBs faster than either LECs in the lens periphery or lymphocytes. Although DSB markers (γH2AX, 53BP1 and RAD51) in both lens regions showed linear dose responses at the 1 h timepoint, nonlinear responses were observed in lenses for EdU (5-ethynyl-2'-deoxy-uridine) incorporation, cyclin D1 staining and cell density after 24 h at 100 and 250 mGy. After 10 months, the lens aspect ratio was also altered, an indicator of the consequences of the altered cell proliferation and cell density changes. A best-fit model demonstrated a dose-response peak at 500 mGy. These data identify specific nonlinear biological responses to low (less than 1000 mGy) dose IR-induced DNA damage in the lens epithelium.
- Published
- 2015
- Full Text
- View/download PDF
50. A silk purse from a sow's ear-bioinspired materials based on α-helical coiled coils.
- Author
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Quinlan RA, Bromley EH, and Pohl E
- Subjects
- Animals, Biomechanical Phenomena, Hair chemistry, Humans, Keratins chemistry, Keratins metabolism, Protein Structure, Secondary, Silk chemistry, Silk metabolism, Biomimetics, Intermediate Filaments chemistry
- Abstract
This past few years have heralded remarkable times for intermediate filaments with new revelations of their structural properties that has included the first crystallographic-based model of vimentin to build on the experimental data of intra-filament interactions determined by chemical cross-linking. Now with these and other advances on their assembly, their biomechanical and their cell biological properties outlined in this review, the exploitation of the biomechanical and structural properties of intermediate filaments, their nanocomposites and biomimetic derivatives in the biomedical and private sectors has started., (Copyright © 2015 The Authors. Published by Elsevier Ltd.. All rights reserved.)
- Published
- 2015
- Full Text
- View/download PDF
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