Genetic and cellular basis of functional cilia assembly
Lead Research Organisation:
University of Edinburgh
Department Name: UNLISTED
Abstract
Cilia are specialized structures found on the surface of most mammalian cells, playing key sensory and sometimes movement functions. Defects in cilia function result in a broad range of genetic diseases termed ciliopathies. These can have devastating effects on human development (birth defects) or postnatal health, including blindness, obesity and kidney failure. Hundreds of genes are involved in building and maintaining this highly conserved structure, which is highly dynamic and biochemically complex. However, we know very little about why some cilia types are more affected in human disease than others. In order to understand how different cell types have configured cilia as specialized signaling ‘antennae’ to interpret environmental signals, we have engineered a series of mice expressing the latest molecular tags and markers to allow us to exquisitely profile cilia subtypes, in both healthy and diseased states.
Ciliopathies are rare genetic diseases, for which there are no effective treatments. For a subset of these diseases affecting accessible tissues, like the airways of primary ciliary dyskinesia (PCD) patients, we have been developing genome editing strategies to determine can we fix the right cells types efficiently and whether this is treatment well-tolerated.
We hope to better understand the disease mechanisms underlying ciliopathies in order to develop effective diagnostic as well as therapeutic strategies to benefit patients and their families.
Ciliopathies are rare genetic diseases, for which there are no effective treatments. For a subset of these diseases affecting accessible tissues, like the airways of primary ciliary dyskinesia (PCD) patients, we have been developing genome editing strategies to determine can we fix the right cells types efficiently and whether this is treatment well-tolerated.
We hope to better understand the disease mechanisms underlying ciliopathies in order to develop effective diagnostic as well as therapeutic strategies to benefit patients and their families.
Technical Summary
Ciliopathies are a diverse class of human genetic diseases, with over 20 recognized syndromes caused by mutations at ~100 different loci. While mammalian cilia are ubiquitous and highly conserved structures, the clinical features associated with their dysfunction are highly pleiotropic with varying degrees of severity and penetrance among tissues. While the causative genes are now being rapidly identified, the cellular and developmental basis for this phenotypic complexity remains poorly understood and controversial. In this programme we aim to:
1. Investigate the cellular pleiotropy of ciliopathies by determining why different cell types vary in their vulnerability to mutations affecting cilia, despite their ubiquity. To better understand cilial diversity and how this results in variable phenotypic consequences upon ciliary dysfunction, we are building tools to interrogate clinically relevant cilial types at a molecular, structural and physiological level. We are engineering a series of precisely-targeted in vivo proximity labelling tracers to common cilial transport machineries and novel cilial biosensors to track cilial dynamics in vivo, in healthy and diseased states.
2. Develop therapeutic strategies for somatic gene editing in ciliopathies, focusing on improving delivery to the correct cell type, and biasing editing of intragenic mutations to therapeutically beneficial events. The recent therapeutic game-changer for rare human genetic disease is the possibility of gene correction using targeted CRISPR technology for precise and efficient corrective genome-editing. If targeted to the most relevant cell type(s), this offers the possibility of a somatic cure for some genetic diseases in which affected cell types are accessible, such as the postnatal airways (affected in primary ciliary dyskinesia: PCD) and the eye (affected in retinitis pigmentosa: RP). Importantly both PCD and RP are characterized by their staggering polygenic natures: over 40 and 100 genes are implicated in PCD and RP respectively, which complicates a conventional “one-size-fits-all” gene therapy for all patients. In contrast, therapeutic gene editing could be personalized based on an individual’s genetic mutation(s). However, there remain at least two sizeable obstacles to making gene editing a therapeutic reality, namely (a) targeted delivery of the editing machinery and (b) improving repair efficiency. To address these problems, we have developed fluorescent reporter mouse models that allow a sensitive read-out of gene-editing events in time and space, which we will use to investigate how best to target cells of interest safely and in a therapeutically effective manner.
Our aims fully align with the Unit’s mission to understand genetic disease and disease mechanisms, as well as its emphasis on the genome and brain biology: our whole organism approach is essential for understanding why different tissues or organs are particularly susceptible to perturbation across the wide phenotypic spectrum of human ciliopathies.
1. Investigate the cellular pleiotropy of ciliopathies by determining why different cell types vary in their vulnerability to mutations affecting cilia, despite their ubiquity. To better understand cilial diversity and how this results in variable phenotypic consequences upon ciliary dysfunction, we are building tools to interrogate clinically relevant cilial types at a molecular, structural and physiological level. We are engineering a series of precisely-targeted in vivo proximity labelling tracers to common cilial transport machineries and novel cilial biosensors to track cilial dynamics in vivo, in healthy and diseased states.
2. Develop therapeutic strategies for somatic gene editing in ciliopathies, focusing on improving delivery to the correct cell type, and biasing editing of intragenic mutations to therapeutically beneficial events. The recent therapeutic game-changer for rare human genetic disease is the possibility of gene correction using targeted CRISPR technology for precise and efficient corrective genome-editing. If targeted to the most relevant cell type(s), this offers the possibility of a somatic cure for some genetic diseases in which affected cell types are accessible, such as the postnatal airways (affected in primary ciliary dyskinesia: PCD) and the eye (affected in retinitis pigmentosa: RP). Importantly both PCD and RP are characterized by their staggering polygenic natures: over 40 and 100 genes are implicated in PCD and RP respectively, which complicates a conventional “one-size-fits-all” gene therapy for all patients. In contrast, therapeutic gene editing could be personalized based on an individual’s genetic mutation(s). However, there remain at least two sizeable obstacles to making gene editing a therapeutic reality, namely (a) targeted delivery of the editing machinery and (b) improving repair efficiency. To address these problems, we have developed fluorescent reporter mouse models that allow a sensitive read-out of gene-editing events in time and space, which we will use to investigate how best to target cells of interest safely and in a therapeutically effective manner.
Our aims fully align with the Unit’s mission to understand genetic disease and disease mechanisms, as well as its emphasis on the genome and brain biology: our whole organism approach is essential for understanding why different tissues or organs are particularly susceptible to perturbation across the wide phenotypic spectrum of human ciliopathies.
Organisations
- University of Edinburgh (Lead Research Organisation)
- UNIVERSITY OF EDINBURGH (Collaboration)
- Beat-PCD (Collaboration)
- Aarhus University (Collaboration)
- University of San Francisco (Collaboration)
- Human Technopole (Collaboration)
- University of Toronto (Collaboration)
- Scottish Genome Partnership (Collaboration)
People |
ORCID iD |
Pleasantine Mill (Principal Investigator) |
Publications
Ford MJ
(2018)
A Cell/Cilia Cycle Biosensor for Single-Cell Kinetics Reveals Persistence of Cilia after G1/S Transition Is a General Property in Cells and Mice.
in Developmental cell
Waddell SH
(2023)
A TGFß-ECM-integrin signaling axis drives structural reconfiguration of the bile duct to promote polycystic liver disease.
in Science translational medicine
Quidwai T
(2021)
A WDR35-dependent coat protein complex transports ciliary membrane cargo vesicles to cilia
in eLife
Kumar D
(2023)
Centriolar satellites expedite mother centriole remodeling to promote ciliogenesis
in eLife
Zur Lage P
(2018)
Ciliary dynein motor preassembly is regulated by Wdr92 in association with HSP90 co-chaperone, R2TP.
in The Journal of cell biology
Van Kerckvoorde M
(2021)
Correction to: Live Imaging and Analysis of Cilia and Cell Cycle Dynamics with the Arl13bCerulean-Fucci2a Biosensor and Fucci Tools.
in Methods in molecular biology (Clifton, N.J.)
Younger NT
(2022)
In Vivo Modeling of Patient Genetic Heterogeneity Identifies New Ways to Target Cholangiocarcinoma.
in Cancer research
Van Kerckvoorde M
(2021)
Live Imaging and Analysis of Cilia and Cell Cycle Dynamics with the Arl13bCerulean-Fucci2a Biosensor and Fucci Tools.
in Methods in molecular biology (Clifton, N.J.)
Related Projects
Project Reference | Relationship | Related To | Start | End | Award Value |
---|---|---|---|---|---|
MC_UU_00007/1 | 01/04/2018 | 31/03/2023 | £662,000 | ||
MC_UU_00007/2 | Transfer | MC_UU_00007/1 | 01/04/2018 | 31/03/2023 | £3,730,000 |
MC_UU_00007/3 | Transfer | MC_UU_00007/2 | 01/04/2018 | 31/05/2022 | £3,053,000 |
MC_UU_00007/4 | Transfer | MC_UU_00007/3 | 01/04/2018 | 31/03/2023 | £1,772,000 |
MC_UU_00007/5 | Transfer | MC_UU_00007/4 | 01/04/2018 | 31/03/2023 | £4,524,000 |
MC_UU_00007/6 | Transfer | MC_UU_00007/5 | 01/04/2018 | 31/03/2023 | £2,878,000 |
MC_UU_00007/7 | Transfer | MC_UU_00007/6 | 01/04/2018 | 31/03/2023 | £2,829,000 |
MC_UU_00007/8 | Transfer | MC_UU_00007/7 | 01/04/2018 | 31/12/2022 | £4,072,000 |
MC_UU_00007/9 | Transfer | MC_UU_00007/8 | 01/04/2018 | 31/03/2023 | £3,137,000 |
MC_UU_00007/10 | Transfer | MC_UU_00007/9 | 01/04/2018 | 31/03/2023 | £6,948,000 |
MC_UU_00007/11 | Transfer | MC_UU_00007/10 | 01/04/2018 | 31/03/2023 | £2,421,000 |
MC_UU_00007/12 | Transfer | MC_UU_00007/11 | 01/04/2018 | 31/03/2023 | £1,205,000 |
MC_UU_00007/13 | Transfer | MC_UU_00007/12 | 01/04/2018 | 31/03/2023 | £1,174,000 |
MC_UU_00007/14 | Transfer | MC_UU_00007/13 | 01/04/2018 | 31/03/2023 | £1,838,000 |
MC_UU_00007/15 | Transfer | MC_UU_00007/14 | 01/04/2018 | 31/03/2023 | £2,551,000 |
MC_UU_00007/16 | Transfer | MC_UU_00007/15 | 01/04/2018 | 31/03/2023 | £1,496,000 |
MC_UU_00007/17 | Transfer | MC_UU_00007/16 | 01/04/2018 | 31/03/2023 | £1,886,000 |
Description | Attended and co-authored a paper as an invited genome editing expert 'Genome Editing for Human Health: Roundtable to explore future public engagement priorities' organized by the Department for Business, Energy and Industrial Strategy and Sciencewise |
Geographic Reach | National |
Policy Influence Type | Participation in a guidance/advisory committee |
URL | https://sciencewise.org.uk/wp-content/uploads/2018/07/Genome-Editing-for-Human-Health-Briefing-Paper... |
Description | Congenital anomalies: patient-led functional genomics. |
Amount | £3,600,000 (GBP) |
Funding ID | MC_PC_21044 |
Organisation | Medical Research Council (MRC) |
Sector | Public |
Country | United Kingdom |
Start | 04/2022 |
End | 04/2022 |
Description | EMBO Short-term Fellowship, Awardee: Tooba Quidwai, Title: WDR35/ IFT121 functions like COPI vesicles to transport cilia specific cargo to cilia. |
Amount | € 10,000 (EUR) |
Organisation | European Molecular Biology Organisation |
Sector | Charity/Non Profit |
Country | Germany |
Start | 01/2019 |
End | 06/2019 |
Description | ERC Consolidator Grant 2019 |
Amount | € 1,965,459 (EUR) |
Funding ID | 866355 - CiliaCircuits |
Organisation | European Research Council (ERC) |
Sector | Public |
Country | Belgium |
Start | 04/2020 |
End | 05/2025 |
Description | EUROPEAN CONSORTIUM FOR COMMUNICATING GENE AND CELL THERAPY RESEARCH (EuroCGT) |
Amount | £2,000,000 (GBP) |
Funding ID | 965241 |
Organisation | European Research Council (ERC) |
Sector | Public |
Country | Belgium |
Start | 07/2021 |
End | 07/2026 |
Description | Edinburgh and Lothian Health Fund, Awardee: Roly Megaw |
Amount | £400,000 (GBP) |
Organisation | NHS Lothian |
Sector | Public |
Country | United Kingdom |
Start | 10/2018 |
End | 04/2019 |
Description | IDENTIFYING THERAPEUTIC TARGETS FOR RETINITIS PIGMENTOSA BY DEFINING PHOTORECEPTOR CELL DEATH -Megaw, Vendrell, Mill & Forrester |
Amount | £60,000 (GBP) |
Organisation | The Royal College of Surgeons of Edinburgh |
Sector | Charity/Non Profit |
Country | United Kingdom |
Start | 07/2020 |
End | 07/2021 |
Description | Identification of novel therapies for inherited eye disease- Megaw, Mill Vallejos |
Amount | £24,357,742 (GBP) |
Organisation | Fight for Sight |
Sector | Charity/Non Profit |
Country | United Kingdom |
Start | 05/2022 |
End | 05/2025 |
Description | National Productivity Investment Fund (NPIF) PhD Innovation Placement Fund (Awardee:Daniel Dodd) |
Amount | £25,000 (GBP) |
Organisation | Medical Research Council (MRC) |
Sector | Public |
Country | United Kingdom |
Start | 09/2019 |
End | 03/2020 |
Description | Wellcome Clinical Research Career Development Fellowship (Dr Roly Megaw) |
Amount | £697,263 (GBP) |
Funding ID | Building a photoreceptor: Defining RPGR's role in disc morphogenesis |
Organisation | Wellcome Trust |
Sector | Charity/Non Profit |
Country | United Kingdom |
Start | 02/2020 |
End | 02/2023 |
Description | Wellcome Trust Institutional Strategic Support Funds 3 (ISSF3), Awardee: Roly Megaw, Support for strengthening application for a Wellcome Trust Clinical Research Career Development Fellowship |
Amount | £52,000 (GBP) |
Organisation | Wellcome Trust |
Sector | Charity/Non Profit |
Country | United Kingdom |
Start | 02/2019 |
End | 02/2021 |
Description | Wellcome Trust Seed Award in Science: Towards in vivo genome editing of post-mitotic mammalian photoreceptors for treatment of Inherited Retinal Dystrophies Awardee: Roly Megaw |
Amount | £100,000 (GBP) |
Organisation | Wellcome Trust |
Sector | Charity/Non Profit |
Country | United Kingdom |
Start | 04/2019 |
End | 04/2021 |
Title | In vivo gene editing reporter |
Description | We have generated a mouse model for rapid and scaleable monitoring gene editing events in situ, allowing tracking of potentially damaging non-homologous end-joining (NHEJ) versus therapeutically desirable homology-driven repair (HDR) events in a whole organism, tissue or field of primary cells. This model will be essential for optimizing gene editing reagents and vehicles to establish robust protocol for therapeutic gene editing in target tissues of interest. |
Type Of Material | Model of mechanisms or symptoms - mammalian in vivo |
Year Produced | 2015 |
Provided To Others? | No |
Impact | MRC Confidence in Concept award. |
Title | PCD Mouse models |
Description | We have generated several mouse models of PCD disease genes with a range of human disease-like mutations (indels, duplications, insertions and inversions) which we will use for testing therapeutic benefit of somatic gene editing. In parallel, these models will also be used in collaboration with the UK CF Gene Therapy Consortium looking at repurposing reagents for gene replacement therapies for PCD. These animals are excellent models of human PCD disease, from pathology to molecular mechanism. |
Type Of Material | Model of mechanisms or symptoms - mammalian in vivo |
Year Produced | 2014 |
Provided To Others? | No |
Impact | Carnegie Collaborative Research Grant Mali, GR, Yeyati, PL, Mizuno, S, Dodd, DO, Tennant, PA, Keighren, MA, Zur Lage, P, Shoemark, A, Garcia-Munoz, A, Shimada, A, Takeda, H, Edlich, F, Takahashi, S, von Kreisheim, A, Jarman, AP & Mill, P 2018, 'ZMYND10 functions in a chaperone relay during axonemal dynein assembly' eLIFE, vol. 7. DOI: 10.7554/eLife.34389 |
URL | https://elifesciences.org/articles/34389 |
Description | COST Action BEAT-PCD |
Organisation | Beat-PCD |
Country | Belgium |
Sector | Private |
PI Contribution | As member of this European-based consortium, we are part of the Basic Science work group, that is intended to encourage communication between basic and clinical scientists working on cilia and PCD. In particular, we will be providing models of PCD (animal and cellular) with the aim of developing them to evaluate pre-clinical studies for novel therapies for PCD. |
Collaborator Contribution | COST Actions provide funds for cooperation, collaboration and the development of ideas through meetings, workshops, and short term lab visits. It aims to help seek more direct forms of funding down the way. |
Impact | Funding for meetings, workshops and travel. I presented a podium talk at the Gordon Research Conference (February 2017, Galveston Texas) Cilia, Mucus and Mucociliary Interactions meeting. I faciliated a PowerHour Session on diversity in science and chaired a session at the Gordon Research Conference (February 2019, Barga Italy) Cilia Mucus and Mucociliary Interactions meeting. I was selected to be the vice-chair for the 2021 and co-chair for the 2023 GRC Cilia, Mucus and Mucociliary Interactions meetings. I spoke at the 5th BEAT-PCD meeting in Poznan, Poland March 2019. Gardner LE, Horton KL, Shoemark A, Lucas JS, Nielsen KG, Kobbernagel H, Rubbo B, Hirst RA, Kouis P, Ullmann N, Reula A, Rumman N, Mitchison HM, Pinto A, Richardson C, Schmidt A, Thompson J, Gaupmann R, Dabrowski M, Mill P, Carr SB, Norris DP, Kuehni CE, Goutaki M, Hogg C. Proceedings of the 4th BEAT-PCD Conference and 5th PCD Training School. BMC Proc. 2020 Jun 19;14(Suppl 8):7. doi: 10.1186/s12919-020-00191-3. PMID: 32577127; PMCID: PMC7304082. |
Start Year | 2015 |
Description | Functional Conservation in Ciliary Trafficking |
Organisation | University of Edinburgh |
Department | Centre for Discovery Brain Sciences |
Country | United Kingdom |
Sector | Academic/University |
PI Contribution | Characterization of function of ciliary candidate genes in mammalian cell culture and mouse mutant models. |
Collaborator Contribution | Identification and characterization of putative ciliary candidate genes in Drosophila cilia. |
Impact | PAPERS: Hall,E.A., Keighren, M., Ford, M., Davey, T., Jarman, A.P., Smith, L.B.S, Jackson I.J. and Mill, P. (2013). Acute Versus Chronic Loss of Mammalian Azi1/Cep131 Results in Distinct Ciliary Phenotypes. PLoS Genetics. 9(12): e1003928. doi:10.1371/journal.pgen.1003928. Diggle CP, Moore DJ, Mali G, zur Lage P, Ait-Lounis A, Schmidts M, Shoemark A, Garcia Munoz A, Halachev MR, Gautier P, Yeyati PL, Bonthron DT, Carr IM, Hayward B, Markham AF, Hope JE, von Kriegsheim A, Mitchison HM, Jackson IJ, Durand B, Reith W, Sheridan E, Jarman AP, Mill P. HEATR2 plays a conserved role in assembly of the ciliary motile apparatus. PLoS Genet. 2014 Sep 18;10(9):e1004577. doi: 10.1371/journal.pgen.1004577. eCollection 2014 Sep. zur Lage, P, Stefanopoulou, P, Styczynska, K, Quinn, N, Mali, G, Von Kriegsheim, A, Mill, P & Jarman, A 2018, 'Ciliary dynein motor preassembly is regulated by Wdr92 in association with HSP90 co-chaperone, R2TP' The Journal of Cell Biology. DOI: 10.1083/jcb.201709026 Mali, GR, Yeyati, PL, Mizuno, S, Dodd, DO, Tennant, PA, Keighren, MA, Zur Lage, P, Shoemark, A, Garcia-Munoz, A, Shimada, A, Takeda, H, Edlich, F, Takahashi, S, von Kreisheim, A, Jarman, AP & Mill, P 2018, 'ZMYND10 functions in a chaperone relay during axonemal dynein assembly' eLIFE, vol. 7. DOI: 10.7554/eLife.34389 PhD candidates: Girish Mail (graduated), Daniel Dodd |
Start Year | 2007 |
Description | Role of Centriolar Satellites in Mammalian Development |
Organisation | University of San Francisco |
Country | United States |
Sector | Academic/University |
PI Contribution | In collaboration with Jeremy Reiter and Laurence Pelletier, we have generated null alleles as well as SNAP-tagged alleles of the key centriolar satellite gene PCM-1 in mouse by CRISPR gene editing. If PCM1 was essential for centriolar satellites in vivo as it is in vitro, we would expect mutant embryos to be embryonic lethal, with profound defects in ciliogenesis, centriole duplication and DNA damage response. However, this is not the case in mice. Null mutants are perinatal lethal, with escapers showing key features of human ciliopathies including hydrocephaly, infertility, cystic kidneys and cerebellar malformations. |
Collaborator Contribution | The Reiter and Pelletier labs have contributed key complementary cell biology experiments in human cell lines using advanced imaging and genetics. |
Impact | This collaboration has resulted in a manuscript in preparation, we would expect to be pre-printed and submitted before the end of March 2022. |
Start Year | 2019 |
Description | Role of Centriolar Satellites in Mammalian Development |
Organisation | University of Toronto |
Country | Canada |
Sector | Academic/University |
PI Contribution | In collaboration with Jeremy Reiter and Laurence Pelletier, we have generated null alleles as well as SNAP-tagged alleles of the key centriolar satellite gene PCM-1 in mouse by CRISPR gene editing. If PCM1 was essential for centriolar satellites in vivo as it is in vitro, we would expect mutant embryos to be embryonic lethal, with profound defects in ciliogenesis, centriole duplication and DNA damage response. However, this is not the case in mice. Null mutants are perinatal lethal, with escapers showing key features of human ciliopathies including hydrocephaly, infertility, cystic kidneys and cerebellar malformations. |
Collaborator Contribution | The Reiter and Pelletier labs have contributed key complementary cell biology experiments in human cell lines using advanced imaging and genetics. |
Impact | This collaboration has resulted in a manuscript in preparation, we would expect to be pre-printed and submitted before the end of March 2022. |
Start Year | 2019 |
Description | Role of IFT-A in ciliary trafficking |
Organisation | Aarhus University |
Country | Denmark |
Sector | Academic/University |
PI Contribution | WDR35 is a non-core member of the highly conserved retrograde IFT-A complex and is mutated in human ciliopathies including Short-rib polydactyly, Sensenbrenner's syndrome and Ellis van Creuzfeld disease, suggesting a spectrum of syndromic skeletal dysplasias. We had previously generated null mouse mutants (Mill et al 2011 American Journal of Human Genetics) which we used for these studies. Here we showed that the absence of WDR35, small mutant cilia form but fail to enrich in diverse classes of ciliary membrane proteins. In Wdr35 mouse mutants, the non-core IFT-A components are degraded and core components accumulate at the ciliary base. We reveal deep sequence homology of WDR35 and other IFT-A subunits to a and ß' COPI coatomer subunits. |
Collaborator Contribution | With the Pigino lab, using electron tomography techiniques we were able to demonstrate an accumulation of 'coat-less' vesicles that fail to fuse with Wdr35 mutant cilia in vivo. With the Lorentzen lab, we were able to demonstratethat recombinant non-core IFT-As can bind directly to specific lipids, including those involved in membrane curvature. Our collaborative studies provided the first in situ evidence of a novel coat function for WDR35, likely with other IFT-A proteins, in delivering ciliary membrane cargo necessary for cilia elongation |
Impact | Quidwai T, Wang J, Hall EA, Petriman NA, Leng W, Kiesel P, Wells JN, Murphy LC, Keighren MA, Marsh JA, Lorentzen E, Pigino G, Mill P. A WDR35-dependent coat protein complex transports ciliary membrane cargo vesicles to cilia. Elife. 2021 Nov 4;10:e69786. doi: 10.7554/eLife.69786. PMID: 34734804; PMCID: PMC8754431. EMBO Short-Term Fellowship: Tooba Quidwai, to lab of Gaia Pigino |
Start Year | 2019 |
Description | Role of IFT-A in ciliary trafficking |
Organisation | Human Technopole |
Country | Italy |
Sector | Charity/Non Profit |
PI Contribution | WDR35 is a non-core member of the highly conserved retrograde IFT-A complex and is mutated in human ciliopathies including Short-rib polydactyly, Sensenbrenner's syndrome and Ellis van Creuzfeld disease, suggesting a spectrum of syndromic skeletal dysplasias. We had previously generated null mouse mutants (Mill et al 2011 American Journal of Human Genetics) which we used for these studies. Here we showed that the absence of WDR35, small mutant cilia form but fail to enrich in diverse classes of ciliary membrane proteins. In Wdr35 mouse mutants, the non-core IFT-A components are degraded and core components accumulate at the ciliary base. We reveal deep sequence homology of WDR35 and other IFT-A subunits to a and ß' COPI coatomer subunits. |
Collaborator Contribution | With the Pigino lab, using electron tomography techiniques we were able to demonstrate an accumulation of 'coat-less' vesicles that fail to fuse with Wdr35 mutant cilia in vivo. With the Lorentzen lab, we were able to demonstratethat recombinant non-core IFT-As can bind directly to specific lipids, including those involved in membrane curvature. Our collaborative studies provided the first in situ evidence of a novel coat function for WDR35, likely with other IFT-A proteins, in delivering ciliary membrane cargo necessary for cilia elongation |
Impact | Quidwai T, Wang J, Hall EA, Petriman NA, Leng W, Kiesel P, Wells JN, Murphy LC, Keighren MA, Marsh JA, Lorentzen E, Pigino G, Mill P. A WDR35-dependent coat protein complex transports ciliary membrane cargo vesicles to cilia. Elife. 2021 Nov 4;10:e69786. doi: 10.7554/eLife.69786. PMID: 34734804; PMCID: PMC8754431. EMBO Short-Term Fellowship: Tooba Quidwai, to lab of Gaia Pigino |
Start Year | 2019 |
Description | Whole genome sequencing for novel PCD candidates |
Organisation | Scottish Genome Partnership |
Country | United Kingdom |
Sector | Learned Society |
PI Contribution | Along with the pediatric respiratory team at the Hospital for Sick Children Edinburgh, we have molecularly diagnosed 9/9 cases of primary ciliary dyskinesia (PCD) in their care. This involves identification of a novel PCD candidate gene and mode of inheritance for the disease, which has spurred on an international collaboration for additional patient mutations and my lab is generating novel mouse models as well as in vitro experiments to get at disease mechanisms. |
Collaborator Contribution | The cost of sequencing 9 trios of Scottish PCD patients. |
Impact | We will have a draft manuscript to be submitted on the WGS diagnosis of the 9 patients, followed by a large mechanistic paper with international collaborators in the next year. I will be speaking at the UK PCD foundation as a keynote in May on this work and our genome editing project. Black, HA, Marion de Proce, S, Campos, JL, Meynert, A, Halachev, MR, Marsh, J, Callaghan, C, Hirst, RA, Murray, J, Macleod, K, Urqhart, D, Unger, S, Aitman, T*, Mill, P*. (2021) Whole genome sequencing identifies pathogenic single nucleotide and structural variants in a series of primary ciliary dyskinesia patients. (Under revision ERS Open ERJ-02769-2020) |
Start Year | 2018 |
Description | 'Genome Editing for Human Health: Roundtable to explore future public engagement priorities' |
Form Of Engagement Activity | A formal working group, expert panel or dialogue |
Part Of Official Scheme? | No |
Geographic Reach | National |
Primary Audience | Policymakers/politicians |
Results and Impact | I sat on a high-level round table and was involved in organizing a paper as an invited genome editing expert entitled 'Genome Editing for Human Health: Roundtable to explore future public engagement priorities' organized by the Department for Business, Energy and Industrial Strategy and Sciencewise, March 22,2018. Researchers, ethicists, policy makers, public engagement groups and more were in attendance. |
Year(s) Of Engagement Activity | 2018 |
URL | https://sciencewise.org.uk/wp-content/uploads/2018/07/Genome-Editing-for-Human-Health-event-report-F... |
Description | BBC Alba Interview on our Genome Editing research for PCD in Gaelic |
Form Of Engagement Activity | A broadcast e.g. TV/radio/film/podcast (other than news/press) |
Part Of Official Scheme? | No |
Geographic Reach | National |
Primary Audience | Public/other audiences |
Results and Impact | We were interviewed answering questions about our research on 'genome surgery' for the ciliopathies and taking a tour of the facilities from the tissue culture suites to the advanced imaging set-ups. |
Year(s) Of Engagement Activity | 2019 |
URL | https://www.cilialab.co.uk/genome-editing-for-pcd-bbc-alba-segment-15112019 |
Description | Featured in the 'Humans In STEM' instagram campaign to highlight diversity in Science (BEIS initiative) |
Form Of Engagement Activity | Engagement focused website, blog or social media channel |
Part Of Official Scheme? | No |
Geographic Reach | National |
Primary Audience | Public/other audiences |
Results and Impact | Multimedia campaign by BEIS to highlight diversity of STEM with pictures and personal narratives. |
Year(s) Of Engagement Activity | 2019 |
URL | https://www.instagram.com/p/Bzh7BxVhRx1/ |
Description | Invited 'role model' and 'exemplar communicator' as an invited expert speaker in the 'GCSE Science in Action', the Training Partnership virtual event (11/2020) |
Form Of Engagement Activity | A talk or presentation |
Part Of Official Scheme? | No |
Geographic Reach | National |
Primary Audience | Schools |
Results and Impact | I spoke briefly about my background, my training and my research as a cell and developmental biologist to encourage students to go into STEM disciplines ahead of their high school subject choices. |
Year(s) Of Engagement Activity | 2020 |
Description | Invited expert panellist for the PCD Support UK Annual General Meeting (06/2021)- virtual |
Form Of Engagement Activity | Participation in an activity, workshop or similar |
Part Of Official Scheme? | No |
Geographic Reach | National |
Primary Audience | Patients, carers and/or patient groups |
Results and Impact | An annual meeting of PCD patients, carers and families from across the 4 nations which was virtual this year due to COVID, and likely reached a live audience of about 60 people as well as those watching the recording at a later date. |
Year(s) Of Engagement Activity | 2021 |
Description | Invited keynote talk at the US PCD Foundation 'PCD on the Move' Scientific Meeting, Minneapolis, MN, USA |
Form Of Engagement Activity | A formal working group, expert panel or dialogue |
Part Of Official Scheme? | No |
Geographic Reach | International |
Primary Audience | Professional Practitioners |
Results and Impact | I delivered the keynote address for the meeting on our work to develop 'genome surgery' to correct PCD mutations in patients- a rare disease with huge genetic heterogeneity and currently no treatment to an audience of ~120 invitees of patients, patient advocates, physicians, physiotherapists and clinical specialists from mostly the US, some Canada, Japan and Europe. |
Year(s) Of Engagement Activity | 2019 |
Description | Invited panellist in Chan-Zuckerburg Science Initiative Workshop: CZI Advancing Diagnosis in Rare Disease Through Patient-Researcher Collaborations Using Single-Cell Methods to discuss challenges and opportunities related to building research partnerships and diagnosing rare diseases on (06/ 2021). |
Form Of Engagement Activity | A formal working group, expert panel or dialogue |
Part Of Official Scheme? | No |
Geographic Reach | International |
Primary Audience | Supporters |
Results and Impact | A closed invitation-only workshop facilitated by CZI of patient advocacy groups and researchers in rare disease space using single cell technologies to work towards improving diagnosis, management and treatments for rare disease by improving partnerships- 10 patient groups, 10 top international researchers and 12 CZI team. |
Year(s) Of Engagement Activity | 2021 |
Description | Invited panellist to 14th Annual Pfizer Frontiers in Human Disease Symposium 'The Role of Cilia in Biology and Medicine' |
Form Of Engagement Activity | A talk or presentation |
Part Of Official Scheme? | No |
Geographic Reach | International |
Primary Audience | Industry/Business |
Results and Impact | A two day virtual meeting of presentations and round-table discussions open to all Pfizer employees across the world in a topic they consider of broad interest and importance to their mission. Open also by registration to other interested parties including academics, clinicians, patient advocacy groups and more. |
Year(s) Of Engagement Activity | 2022 |
URL | https://www.pfizer.com/frontiers-human-disease-symposium |
Description | Invited speaker to Cold Spring Harbor Asia Cilia and Centrosomes Meeting, Awaji, Japan |
Form Of Engagement Activity | A talk or presentation |
Part Of Official Scheme? | No |
Geographic Reach | International |
Primary Audience | Professional Practitioners |
Results and Impact | A biennial international meeting schedule for 2021 but COVID delayed to Feb 28-Mar 3 2023, the leading Asian meeting for cilia and centrosome science. ~100 attendees were present, with hybrid viewing options to recorded talks also available. |
Year(s) Of Engagement Activity | 2023 |
URL | https://www.csh-asia.org/?content/1177 |
Description | Invited talk and outreach events at the 2019 Scottish PCD Family Support Day (Glasgow) |
Form Of Engagement Activity | A formal working group, expert panel or dialogue |
Part Of Official Scheme? | No |
Geographic Reach | National |
Primary Audience | Patients, carers and/or patient groups |
Results and Impact | I gave a lay talk on our 'genome surgery' work for ciliopathies which was recorded and uploaded on YouTube for those who could not attend. We also ran a series of outreach activities like 'Color Your Lungs' aprons and 'Strawberry DNA Extraction' for the kids and teenagers. My PhD student Fraser McPhie and postdoc Peter Tennant also facilitated a discussion group with the kids and teens on 'Living with PCD' as a satellite event. |
Year(s) Of Engagement Activity | 2019 |
URL | https://www.youtube.com/watch?v=HFjYue3KvaE |
Description | Organizer and scientific chair of the US PCD Foundation Scientific Meeting 'PCD on The Move' 08/2021 |
Form Of Engagement Activity | Participation in an activity, workshop or similar |
Part Of Official Scheme? | No |
Geographic Reach | International |
Primary Audience | Professional Practitioners |
Results and Impact | I was part of the organizing committee and host of the virtual two day event (due to COVID) which brought together top international primary ciliary dyskinesia basic science researchers as well as clinical research and care teams from across the Americas and Europe. |
Year(s) Of Engagement Activity | 2021 |
Description | Primary Ciliary Dyskinesia (PCD) Awareness Day 2018 |
Form Of Engagement Activity | Participation in an activity, workshop or similar |
Part Of Official Scheme? | No |
Geographic Reach | Regional |
Primary Audience | Patients, carers and/or patient groups |
Results and Impact | To mark PCD awareness month, we convened Scotland-based cilia researchers, clinicians, families and advocates to talk together about emerging research, diagnostics and therapeutics for PCD, as well as the significant challenges that still exist for patients and families with PCD at the MRC Human Genetics Unit. Primary ciliary dyskinesia (PCD) is a relatively rare, genetic disorder resulting from abnormalities in cilia - the microscopic hair-like projections on the cell surface that have important functions, including beating to clear secretions from the lungs. People with PCD can have problems affecting their lungs, nose, sinuses, ears and fertility. It is a life-long condition for which there is currently no cure. A clinical presentation from Dr Kenneth MacLeod, Royal Hospital for Sick Children, highlighted the challenges in diagnosing and treating PCD. Talks from researchers across the University of Edinburgh highlighted some of the genes and molecular pathways that can lead to PCD, emphasising the importance of basic research using fruit flies, chicks and mice to better understand this rare and genetically diverse disease. Researchers outlined the potential impact of research on early genetic diagnosis and development of bespoke therapies, including genome editing. In the future, patients could have their genome sequenced and learn they have PCD, but go beyond just telling them they need to live with that genetic disorder, use this information to choose which technology to treat their mutation, possibly cure them. Dr Susanne Shanks of the PCD Family Support Group Scotland, gave a clear and frank presentation outlining her family story, including the long journey to diagnosis and the impact of PCD on daily and family life. There was great discussion and exchanges throughout the day. |
Year(s) Of Engagement Activity | 2018 |
URL | https://www.cilialab.co.uk/pcd2018overview |
Description | Shedding light on genetic disorders: the importance of the patient voice |
Form Of Engagement Activity | A talk or presentation |
Part Of Official Scheme? | No |
Geographic Reach | Local |
Primary Audience | Other audiences |
Results and Impact | This was an inaugural seminar for all IGMM staff and students, prior to Rare Disease Day, to recognise that patients and their families are at the heart of IGMM research. The quality of MRC IGMM research and the potential to maximise its impact depends upon building and maintaining successful partnerships with multiple partners and stakeholders, including patients, NHS clinicians and policy makers. Families affected by genetic disorders rely on research to understand disease and improve their health and quality of life. Professor Margaret Frame, IGMM Director, hosted this event attended by patient advocates for the rare genetic disorders primary ciliary dyskinesia (PCD) and Annie's syndrome, as well the Genetic Alliance UK. My group provided updates on their research on PCD and the FitzPatrick's lab presented overviews of their work on rare genetic conditions. This was then followed by an institute-wide Seminar, where the patient advocates for genetic disorders shared the diagnostic journeys that their families have been on and how dialogue with clinical researchers at the IGMM has driven biomedical research and clinical impact. |
Year(s) Of Engagement Activity | 2019 |
URL | https://www.ed.ac.uk/igmm/news-and-events/news-2019/shining-a-light-genetic-disorders |