Rhomboids and cellular aspects of development

Lead Research Organisation: MRC Laboratory of Molecular Biology

Abstract

Epithelial sheets of cells are one of the major tissue types in all animals. They also form the basic building blocks of all tubular organs, such as lungs, kidney, vasculature, in both vertebrates such as us humans, but also in invertebrates such as fruit flies. Many severe developmental defects are the result of a failure of tubes forming correctly, such as neural tube closure defects, and furthermore 80% of cancers originate from epithelial tissues. We want to dissect the basic mechanisms of how simple epithelial sheets deform in a highly coordinated manner to turn into complex tissue shapes. We use a process of tube formation in the fruit fly embryo as our main model: the formation of the tubes of the salivary glands. The macroscopic process of this tube formation is conserved and very similar to for instance the early formation of tubes in the development of lungs in mouse or humans. In contrast to vertebrates, fruit flies have the advantage of excellent genetic tools and allow easy imaging and manipulation. We use a combination of genetic methods, live imaging and computational tools to understand how healthy organs form, and his will allow us in the future to better understand the causes of diseases or malformation of tubular epithelia.

Technical Summary

Epithelia constitute one of the major tissue types in all animals and form the basic building blocks of tubular organs in both vertebrates and invertebrates. Simple epithelial sheets are also the basic starting point in the development of most animals. We want to dissect how simple epithelial sheets deform in a highly concerted manner to turn into complex tubular tissue shapes. We use the formation of the tubes of the salivary glands in the Drosophila embryos as our main model system, but have also begun to compare our findings to the related process of budding morphogenesis in embryonic mouse lungs. Coordinated changes in the shape of many neighbouring cells, as well as cell rearrangements are the major drivers of tissue formation. Cell shape changes are driven by the intracellular cytoskeleton. We have in the past shown how crosstalk between the actomyosin cytoskeleton and microtubules is crucial to maintain a pool of actomyosin that drives apical constriction of cells during tissue bending. Coordination between cytoskeletal responses between neighbouring cells allows concerted tissue-scale changes in shape, often mediated through cell-cell adhesion receptors. We discovered that the apical polarity protein Crumbs, through newly identified homophilic interactions of its extracellular domain, can determine where a multicellular actomyosin cable is formed.
The study of epithelial morphogenesis over the last decade has mostly focussed on events within the most apical portion of the cells. Our previous analysis of microtubule-actomyosin interactions and interdependence in the placode have clearly shown that structures that span the whole length of the cells, the longitudinal microtubules bundles, can have profound effects on events within the apical surface, the medial actomyosin contractions, and thus the whole cell volume and events within need to be studied. We now want to follow on from this analysis to address what happens in the remaining 90% volume of these epithelial cells during tube formation.
We plan to dissect the control of cell shape changes during tissue morphogenesis by addressing questions at different scales and integrating the information gathered at each scale:
1) At the molecular level, we will analyse how responses of different cytoskeletal systems are coordinated during cell shape changes. In particular we will analyse how the microtubule cytoskeleton influences the actomyosin cytoskeleton and what the role of cytolinker proteins is in this process.
2) At the cellular level, we will dissect how cell-cell adhesion systems, in particular Crumbs homophilic interactions, impact on the intracellular actomyosin cytoskeleton and how cytoskeletal responses are coupled between cells, such as for instance during the assembly of multicellular actomyosin cables.
3) At the tissue level, we will segment and track epithelial cells in 3D over time, to understand the contribution of different cell behaviours to morphogenesis and also to uncover novel cell behaviours that drive shape changes at the tissue scale.
We propose that the combination of approaches detailed above will give us a very detailed understanding of the cellular events that drive tube formation at the tissue level. The mutant analysis as well as the search for molecular factors driving these cellular behaviours will allow us to extend the understanding of epithelial morphogenesis from its current apical-centric view to a true 4D appreciation.

Publications

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Adrain C (2012) New lives for old: evolution of pseudoenzyme function illustrated by iRhoms. in Nature reviews. Molecular cell biology

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Adrain C (2014) Regulation of receptor tyrosine kinase ligand processing. in Cold Spring Harbor perspectives in biology

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Aldaz S (2010) Live imaging of Drosophila imaginal disc development. in Proceedings of the National Academy of Sciences of the United States of America

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Blobel CP (2009) The role of protease activity in ErbB biology. in Experimental cell research

 
Description Austrian Fellowship
Amount £54,000 (GBP)
Organisation Austrian Academy of Sciences 
Sector Academic/University
Country Austria
Start 01/2006 
End 01/2009
 
Description CRUK Project Grant
Amount £156,800 (GBP)
Organisation Cancer Research UK 
Sector Charity/Non Profit
Country United Kingdom
Start 01/2011 
End 06/2014
 
Description Characterization of the cytoskeletal "pigs" as a direct target of Notch signalling
Amount £121,718 (GBP)
Funding ID 090279 
Organisation Wellcome Trust 
Sector Charity/Non Profit
Country United Kingdom
Start 01/2011 
 
Description EMBO Long-term Fellowship
Amount £54,000 (GBP)
Organisation European Molecular Biology Organisation 
Sector Charity/Non Profit
Country Germany
Start 01/2006 
End 01/2008
 
Description EMBO Long-term Fellowship
Amount £54,000 (GBP)
Organisation European Molecular Biology Organisation 
Sector Charity/Non Profit
Country Germany
Start 01/2007 
End 01/2009
 
Description EMBO Long-term Fellowship
Amount £27,000 (GBP)
Organisation European Molecular Biology Organisation 
Sector Charity/Non Profit
Country Germany
Start 01/2008 
End 01/2009
 
Description HFSP Postdoctoral Fellowship
Amount £54,000 (GBP)
Organisation Human Frontier Science Program (HFSP) 
Sector Charity/Non Profit
Country France
Start 01/2007 
End 01/2009
 
Description MRCT Development Gap Funding
Amount £121,534 (GBP)
Organisation MRC-Technology 
Sector Private
Country United Kingdom
Start 01/2008 
End 01/2010
 
Description Marie-Curie Fellowship
Amount £140,700 (GBP)
Organisation Marie Sklodowska-Curie Actions 
Sector Charity/Non Profit
Country Global
Start 01/2006 
End 01/2008
 
Description Marie-Curie Fellowship
Amount £150,000 (GBP)
Organisation Marie Sklodowska-Curie Actions 
Sector Charity/Non Profit
Country Global
Start 01/2008 
End 01/2010
 
Description Novartis iRhom2 
Organisation Novartis
Country Global 
Sector Private 
PI Contribution We have provided intellectual insight and knockout mice to allow Novartis to assess whether iRhom2 is a potential drug target.
Collaborator Contribution Platform technology for assessing the phenotype of our knockout mice. Experience in inflammation research.
Impact Unpublished data about iRhom2 KO mice.
Start Year 2012
 
Description Stephen Verhelst 
Organisation Technical University of Munich
Country Germany 
Sector Academic/University 
PI Contribution We led this work and did essentially all the work that was published.
Collaborator Contribution Intellectual input and material, leading to a joint publication.
Impact Publication. PMID 20890268
Start Year 2007
 
Description LMB Open Day 
Form Of Engagement Activity Participation in an open day or visit at my research institution
Part Of Official Scheme? No
Geographic Reach Regional
Primary Audience Public/other audiences
Results and Impact After moving into its new building, the LMB held an Open Day on 22 June 2013 to showcase our research in its new surroundings and with its new facilities.
I was involved in running tours of the Cell Biology Division, introducing our research model, the fruit fly Drosophila melanogaster, to interested audiences.
The day was attended by over 700 people.
Year(s) Of Engagement Activity 2013
 
Description Pint of Science research talk 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach Local
Primary Audience Public/other audiences
Results and Impact This was a research talk as part of the country-wide Pint of Science festival, where scientists are invited to give talks to a lay audience about their research in a pub and engage ins discussion afterwards.

Increased public interest and understanding of basic biological research.
Year(s) Of Engagement Activity 2014
URL http://www.pintofscience.com/#!cam-mon-body/c1xy1
 
Description Postdoc (Clara Sidor) Pint of Science Festival 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach Local
Primary Audience Public/other audiences
Results and Impact Clara joined the organising committee for the Pint of Science Festival 2014 and took on organisation of on of the evening events.
Year(s) Of Engagement Activity 2014
 
Description Postdoc (Dr Yara Sanchez -Corrales) demonstrated experiments as part of Cambridge Science Festival 2015 
Form Of Engagement Activity Participation in an open day or visit at my research institution
Part Of Official Scheme? No
Geographic Reach Regional
Primary Audience Public/other audiences
Results and Impact Volunteer in Cambridge Science Festival 2015:
Yara participated as a demonstrator during the Cambridge Science Festival.
Together with other colleagues from the LMB, we set up two activities:
1) See your cells: people could take some cells from the mount, stain them
and look at them under the microscope
2) Quiz of microscopy: people could look at different organisms under the
microscope and match them.
Year(s) Of Engagement Activity 2015
 
Description Postdoc (Dr Yara Sanchez-Corrales) became STEM ambassador 
Form Of Engagement Activity A formal working group, expert panel or dialogue
Part Of Official Scheme? No
Geographic Reach Local
Primary Audience Undergraduate students
Results and Impact STEM Ambassadors use their enthusiasm and commitment to encourage young people to enjoy STEM subjects. They open the doors to a world of opportunities and possibilities which come from pursuing STEM subjects and careers.
Year(s) Of Engagement Activity 2015
URL http://www.stemnet.org.uk/ambassadors/
 
Description Postdoc (Gemma Girdler) LMB Open Day 
Form Of Engagement Activity Participation in an open day or visit at my research institution
Part Of Official Scheme? No
Geographic Reach Regional
Primary Audience Public/other audiences
Results and Impact After moving into its new building, the LMB held an Open Day on 22 June 2013 to showcase our research in its new surroundings and with its new facilities.
Gemma was involved in running outreach activities and experiments, as well as tours of the Cell Biology Division.
The day was attended by over 700 people.
Year(s) Of Engagement Activity 2013
 
Description Postdoc (Gemma Girdler) Microscopes for Schools Visit 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach Local
Primary Audience Schools
Results and Impact Microscopes for Schools was established at the LMB as a way of getting primary school children interested in Biology. Microscopes and specimen to look at and identify are brought into schools on request.
Year(s) Of Engagement Activity 2012
 
Description Postdoc (Gemma Girdler) Science on a Saturday, Cambridge Science Festival 
Form Of Engagement Activity Participation in an open day or visit at my research institution
Part Of Official Scheme? No
Geographic Reach Regional
Primary Audience Public/other audiences
Results and Impact Volunteer in Cambridge Science Festival 2012:
Gemma participated as a demonstrator during the Cambridge Science Festival.
Together with other colleagues from the LMB, she set up two activities:
1) See your cells: people could take some cells from the mount, stain them
and look at them under the microscope
2) Quiz of microscopy: people could look at different organisms under the
microscope and match them.
Year(s) Of Engagement Activity 2012
 
Description Science on a Saturday during Cambridge Science Week 
Form Of Engagement Activity Participation in an open day or visit at my research institution
Part Of Official Scheme? No
Geographic Reach Local
Primary Audience Public/other audiences
Results and Impact Many families and local people attend the Open Days during Science Week and get engaged in the activity spur on by Departments and Research Institutes. During a 3-4 hour session more than a hundred people usually attended the different displays put up and used the microscopes provided.

n.a.
Year(s) Of Engagement Activity 2012,2013,2014