Optical Imaging of Fly Brains for Neurodegeneration Research

Lead Research Organisation: University of Cambridge
Department Name: Chemical Engineering and Biotechnology

Abstract

Approximately 1 million people in the UK alone are suffering from neurodegenerative diseases, and the number is rising. They pose a huge burden on individuals suffering from these devastating diseases, their relatives, and the economy as a whole, and despite all research efforts there are currently no cures. A key phenomenon linking diseases such as Parkinsons and Alzheimers is that proteins, which are normally harmless, start to change shape, to aggregate and to misbehave so as to become toxic to the neurones in the patients brains. Nobody knows what mechanisms leads to this dysfunction nor how to prevent it. A huge problem is that one cannot with current tools follow how the proteins form into toxic structures in living individuals. What we propose here is to develop, by drawing on the expertise of physicists and geneticists, a novel platform that will permit us to study the formation of such toxic aggregates directly in living brains, not human brains of course, but the brains of fruit flies, which are amenable to optical microscopes and can serve as models of what might be going on in humans. If we succeed in building such a technology, we would have a tool with which to understand the disease and with which to test potential drugs which prevent the toxic structures from forming.

Technical Summary

The aim of this proposal, submitted under the discipline hopping initiative, is for a geneticist to cross the boundary into optical engineering to develop technology with which it is possible to image the formation of toxic protein aggregates in living fly (Drosophila melanogaster) brains. We seek funds for Claire Michel from the department of Genetics to join the Laser Analytics group of Dr. Kaminski in the Department of Chemical Engineering, where novel optical microscopy platforms are being developed. Ms. Michel will be given thorough training in the design and operation of multimodal imaging technologies, co-ordinated through a combination of practical research in instrumentation design and data processing and workshops (which are to be funded and organised through the CamBRIDGESens network (http://sensors.cam.ac.uk), of which Kaminski is the director and which is funded by the EPSRC Bridging the Gaps initiative. We thus ask for Ms Michel, whose background is in Drosophila genetics, to be funded for 12 months to ?discipline hop? into an optical engineering lab. Simultaneous with this activity two other discipline hops will take place: Kaminski, the PI, is a physicist with no background in biology, and he has timed this project to coincide with a research sabbatical which he will use to learn about genetic manipulation of flies, dissection of fly brains, and fundamental aspects of neuronal disease research in fly models, practical aspects of which he will learn from Claire Michel. The fly genetics laboratory of Dr. Damian Crowther, where Michel performed her PhD work, will also benefit from the ?discipline hop?: By obtaining a fundamental understanding of the potential and limitation of modern imaging techniques it will be possible to guide future high impact experiments with which it will be possible to elucidate functional aspects of potential drugs directly in flies to serve as a flexible and high level model for neurodegenerative disease.

Publications

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Arnon ZA (2018) Opal-like Multicolor Appearance of Self-Assembled Photonic Array. in ACS applied materials & interfaces

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Blacker TS (2017) Investigating State Restriction in Fluorescent Protein FRET Using Time-Resolved Fluorescence and Anisotropy. in The journal of physical chemistry. C, Nanomaterials and interfaces

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Chakrabortee S (2012) Intrinsically disordered proteins as molecular shields. in Molecular bioSystems

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Chan FT (2011) HomoFRET fluorescence anisotropy imaging as a tool to study molecular self-assembly in live cells. in Chemphyschem : a European journal of chemical physics and physical chemistry

 
Description 2019 exercise
Amount £400 (GBP)
Organisation European Biophysical Societies' Association 
Sector Charity/Non Profit
Country Germany
Start 02/2018 
End 02/2018
 
Description ARUK emergency grant
Amount £18,000 (GBP)
Organisation Alzheimer's Research UK 
Sector Charity/Non Profit
Country United Kingdom
Start 12/2011 
End 05/2012
 
Description British Biophysical Society Travel Grant
Amount £309 (GBP)
Organisation British Biophysical Society 
Start 05/2018 
End 05/2018
 
Description Cambridge ARUK Travel Grant
Amount £488 (GBP)
Organisation Alzheimer's Research UK 
Sector Charity/Non Profit
Country United Kingdom
Start 05/2018 
End 05/2018
 
Description Cambridge ARUK travel grant
Amount £300 (GBP)
Organisation Alzheimer's Research UK 
Sector Charity/Non Profit
Country United Kingdom
Start 03/2019 
End 03/2019
 
Description EBSA bursary
Amount £330 (GBP)
Organisation European Biophysical Societies' Association 
Sector Charity/Non Profit
Country Germany
Start 02/2018 
End 02/2018
 
Description Equipment grant
Amount £96,864 (GBP)
Funding ID ARUK-EG2012A-1 
Organisation Alzheimer's Research UK 
Sector Charity/Non Profit
Country United Kingdom
Start 01/2012 
 
Description Newnham College Travel Grant
Amount £731 (GBP)
Organisation University of Cambridge 
Department Newnham College
Sector Academic/University
Country United Kingdom
Start 06/2018 
End 09/2018
 
Description Research Fellowship Deutsche Forschungsgemeinschaft (DFG)
Amount € 140,400 (EUR)
Funding ID LA 3609/2-1 
Organisation German Research Foundation 
Sector Public
Country Germany
Start 04/2015 
End 03/2018
 
Title Microscopic assays for quantification of in situ protein aggregation 
Description We have developed a microscopy platform with which it is possible to measure the degree of protein aggregation in live cells and organisms. This permits other researchers to assess the mode of action and efficacy of potential aggregation inhibiting drugs. We make all instruments we develop available to other researchers via CAIC, Cambridge Advanced Imaging Centre, http://www.pdn.cam.ac.uk/caic/index.shtml, where they are available for booking. 
Type Of Material Technology assay or reagent 
Year Produced 2011 
Provided To Others? Yes  
Impact We have been able to monitor the effect of mutations of FUS and its aggregation in C elegans and were able to correlate the degree of aggregation with a toxic phenotype. Preliminary outcomes were published in Human Molecular Genetics in 2011 (10.1093/hmg/ddr417) and a further publication is currently in preparation with the Tanz Center for research into Neurodegeneration. 
URL http://www.pdn.cam.ac.uk/caic/index.shtml
 
Description Analysis of the Native Structure, Stability and Aggregation of Biotinylated Human Lysozyme 
Organisation University of Cambridge
Department Department of Chemistry
Country United Kingdom 
Sector Academic/University 
PI Contribution We have performed superresolution imaging of human lysozyme fibrils in situ
Collaborator Contribution Collaborators produced labelled human lysozyme fibrils for analysis
Impact Ahn M, De Genst E, Kaminski Schierle GS, Erdelyi M, Kaminski CF, Dobson CM, Kumita JR, "Analysis of the Native Structure, Stability and Aggregation of Biotinylated Human Lysozyme", PLoS One (2012), PLoS ONE 7(11): e50192. doi:10.1371
Start Year 2011
 
Description Droplet Cages as a Platform for Monitoring Amyloid Aggregation with Florian Hollfelder 
Organisation University of Cambridge
Department Department of Biochemistry
Country United Kingdom 
Sector Academic/University 
PI Contribution We combined our FRET aggregation assay with their droplet cages to develop a new platform to study amyloid aggregation.
Collaborator Contribution Collaborators developed droplet cage assay.
Impact Droplet Cages as a Platform for Monitoring Amyloid Aggregation Publications Shared PhD student from Oct 2014
Start Year 2011
 
Description Imaging of protein aggregation in C. elegans with Peter St George Hyslop 
Organisation University of Toronto
Department Tanz Centre for Research in Neurodegenerative Diseases
Country Canada 
Sector Academic/University 
PI Contribution We performed fluorescence lifetime imaging microscopy to monitor FUS aggregation in C.Elegans and effects and to test a toxic gain of function hypothesis of FUS aggregation.
Collaborator Contribution Reagents, Samples.
Impact Research Paper: ALS mutations in FUS cause neuronal dysfunction and death in Caenorhabditis elegans by a dominant gain-of-function mechanism. Murakami T, Yang SP, Xie L, Kawano T, Fu D, Mukai A, Bohm C, Chen F, Robertson J, Suzuki H, Tartaglia GG, Vendruscolo M, Kaminski Schierle GS, Chan FT, Moloney A, Crowther D, Kaminski CF, Zhen M, St George-Hyslop P. Hum Mol Genet. 2011 Oct 11. [Epub ahead of print] PMID: 21949354 Update in 2016: paper published in Neuron.
Start Year 2011
 
Description In vivo fluorescent lifetime imaging of a redox-sensitive green fluorescent protein tracks disulfide bond formation in the endoplasmic reticulum with David Ron and Edward Avezov 
Organisation University of Cambridge
Department Institute of Metabolic Science (IMS)
Country United Kingdom 
Sector Academic/University 
PI Contribution We provided TCSPC imaging tools for sensitive redox measurements in live cells. Update in 2016: we have now set up Structure Illumination Microscopy imaging of ER peristalsis
Collaborator Contribution Provided and designed sensors.
Impact In vivo fluorescent lifetime imaging of a redox-sensitive green fluorescent protein tracks disulfide bond formation in the endoplasmic reticulum Edward Avezov, Mikael Winters, Gabriele S Kaminski Schierle, Clemens Kaminski, David Ron
Start Year 2011
 
Description Integrated Microfluidic- Nanosensor System for Neuro- Functional Imaging and Neurotransmission Detection (Dr. E. Nugent) 
Organisation University of Cambridge
Department Department of Physics
Country United Kingdom 
Sector Academic/University 
PI Contribution Jane is currently setting up a brain on chip platform to study amyloid propagation in live neurons.
Collaborator Contribution Dr. Eileen Nugent has been co-supervising Jane.
Impact Eileen Nugent, Clemens F. Kaminski and Gabriele S. Kaminski Schierle: Super-resolution imaging of alpha-synuclein polymorphisms and their potential role in neurodegeneration, Integr. Biol., 2017, Advance Article, doi:10.1039/C6IB00206D
Start Year 2016
 
Description Interactions between aSynuclein and synaptic vesicles 
Organisation Swiss Federal Institute of Technology in Lausanne (EPFL)
Country Switzerland 
Sector Public 
PI Contribution Prepared recombinant protein and purified synaptic vesicles for interaction studies
Collaborator Contribution Microfluidics-integrated nanophotonics-enhanced IR spectroscopy platform
Impact Paper in preparation. Molecular/structural biology and bionanophotonics
Start Year 2018
 
Description Intrinsically Disordered Proteins As molecular shields with Prof Tunnacliffe 
Organisation University of Cambridge
Department Department of Chemical Engineering and Biotechnology
Country United Kingdom 
Sector Academic/University 
PI Contribution We applied the techniques developed to study amyloidogenesis in vivo to a new problem of a similar nature: chaperone activity of intrinsically disordered proteins to prevent aggregation.
Collaborator Contribution Reagents, infrastructure, and people
Impact Research Paper: Sohini Chakrabortee, Rashmi Tripathi, Matthew Watson, Gabriele S. Kaminski Schierle, Davy P. Kurniawan, Clemens F. Kaminski, Michael J. Wise and Alan Tunnacliffe Mol. BioSyst., 2012, Advance Article DOI: 10.1039/C1MB05263B
Start Year 2011
 
Description Investigating the role of water in protein structure and aggregation 
Organisation Swiss Federal Institute of Technology in Lausanne (EPFL)
Country Switzerland 
Sector Public 
PI Contribution Prepared recombinant protein alpha synuclein for aggregation studied in the presence of different salts. Morphology studies
Collaborator Contribution Used second harmonic scattering to determine mobility of water around fibrils
Impact Paper in preparation. Molecular/structural biology and physics/biophotonics
Start Year 2018
 
Description Investigating the role of water in protein structure and aggregation 
Organisation University of Cambridge
Country United Kingdom 
Sector Academic/University 
PI Contribution Prepared recombinant protein alpha synuclein for aggregation studied in the presence of different salts. Morphology studies
Collaborator Contribution Runing and analysis of small angle neuron scattering data
Impact Paper in preparation, application for SANS beam time. Molecular biology and structural biology
Start Year 2018
 
Description Investigating the role of water in protein structure and aggregation 
Organisation University of Vermont
Country United States 
Sector Academic/University 
PI Contribution Prepared recombinant protein alpha synuclein for aggregation studied in the presence of different salts. Morphology studies
Collaborator Contribution Molecular dynamic simulations of peptide fragments in different salts
Impact Paper in preparation. Molecular/structural biology and physics/chemistry
Start Year 2018
 
Description Setting up a brain on chip platform for the study of prion like propagation in amyloid-related diseases (Prof. Knowles) 
Organisation University of Cambridge
Department Department of Physics
Country United Kingdom 
Sector Academic/University 
PI Contribution Jane has been designing a new microfluidic based platform to sort and trap single neurons and to grow and differentiate them for live cell imaging. For this she has produced several PDMS devices, optimised flow conditions to guarantee fluidic isolation and is currently testing cell viability in these devices. She has further designed and implemented a device incubation chamber that can easily be adapted to fit on various microscope stages.
Collaborator Contribution Prof. Knowles PhD student Lianne Roode has been involved in this project and has helped with the design and evaluation of the original microfluidic design.
Impact We are currently writing up a manuscript on this.
Start Year 2016
 
Description Small molecule inhibition of protein aggregation 
Organisation Perrigo
Department Elan Pharmaceuticals
Country Ireland 
Sector Private 
PI Contribution We will test their novel small molecule drugs in our cellular models of amyloid protein aggregation
Collaborator Contribution Provide and design drugs
Impact Under preparation
Start Year 2012
 
Description Structure of monomeric aSynuclein 
Organisation University of Antwerp
Country Belgium 
Sector Academic/University 
PI Contribution Prepared recombinant protein alpha synuclein for aggregation studied in the presence of different salts. Morphology studies
Collaborator Contribution native mass spec and ion mobility mass spec
Impact Two papers in preparation. Molecular biology and structural biology
Start Year 2018
 
Description Structure of monomeric aSynuclein 
Organisation University of Exeter
Country United Kingdom 
Sector Academic/University 
PI Contribution Prepared recombinant protein and purified synaptic vesicles for interaction studies
Collaborator Contribution Hydrogen-Deuterium Exchange Mass Spectroscopy
Impact Paper in preparation. Molecular biology and structural biology
Start Year 2016
 
Description Structure of monomeric aSynuclein 
Organisation University of Leeds
Country United Kingdom 
Sector Academic/University 
PI Contribution Prepared recombinant protein alpha synuclein for aggregation studied in the presence of different salts. Morphology studies
Collaborator Contribution native mass spec and ion mobility mass spec
Impact Two papers in preparation. Molecular biology and structural biology
Start Year 2018
 
Description Study of monomeric Tau propagation from cell to cell with Prof Eckard Mandelkow 
Organisation German Centre for Neurodegenerative Diseases
Country Germany 
Sector Public 
PI Contribution We have applied multi parametric and super resolution imaging techniques to the study of Tau propagation from cell to cell
Collaborator Contribution Prof Eckard Mandelkow provided us with purified recombinant Tau protein, both unlabelled and fluorescently labelled.
Impact Michel CH, Kumar S, Pinotsi D, Tunnacliffe A, St George-Hyslop P, Mandelkow E, Mandelkow E-M, Kaminski CF, Kaminski Schierle GS, "Extracellular Monomeric Tau is Sufficient to Initiate the Spread of Tau Pathology", J. Biol. Chem. (2014), 289: 956-967. This collaboration is multi-disciplinary and involved biochemistry and physics
Start Year 2010
 
Description Super-resolution imaging of nanotubes involved in the transfer of Tau from neuron to neuron (Luc Buee) 
Organisation National Institute of Health and Medical Research (INSERM)
Department Lille (INSERM)
Country France 
Sector Public 
PI Contribution We would carry out super-resolution imaging on samples
Collaborator Contribution Will provide cells, plasmids for expression of Tau and expertise in nanotubes
Impact Invited talk given by Dr Gabriele Kaminski Schierle in Lille for a seminar on New technologies and Neurosciences
Start Year 2015
 
Description Synthetic 3D Neuronal Networks (Prof. Heutnik, Dr. Cesare) 
Organisation Eberhard Karls University of Tubingen
Department Natural and Medical Sciences Institute
Country Germany 
Sector Academic/University 
PI Contribution We have written a proposal that we submitted to the VW Foundation.
Collaborator Contribution Joint proposal to VW Foundation.
Impact Joint proposal submitted
Start Year 2016
 
Description Synthetic 3D Neuronal Networks (Prof. Heutnik, Dr. Cesare) 
Organisation German Centre for Neurodegenerative Diseases
Country Germany 
Sector Public 
PI Contribution We have written a proposal that we submitted to the VW Foundation.
Collaborator Contribution Joint proposal to VW Foundation.
Impact Joint proposal submitted
Start Year 2016
 
Description Synthetic 3D networks 
Organisation German Centre for Neurodegenerative Diseases
Department Tübingen Research Campus
Country Germany 
Sector Private 
PI Contribution Written joint proposal to VW Foundation.
Collaborator Contribution Written joint proposal to VW Foundation.
Impact Written joint proposal to VW Foundation.
Start Year 2016
 
Description aSynuclein-lipid interaction 
Organisation University of Cambridge
Country United Kingdom 
Sector Academic/University 
PI Contribution Prepared recombinant alpha synuclein and brain extract lipid samples for aSyn/membrane interactions studies. Performed AFM analysis.
Collaborator Contribution Access to FastScan AFM in Pharmacology
Impact Paper in preparation. Molecular biology and structural biology
Start Year 2018
 
Description CamBRAIN : Teaching children about neurons 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach Regional
Primary Audience Schools
Results and Impact Teaching children about neurons
Year(s) Of Engagement Activity 2018
 
Description Episode of Elemental Ideas on Cambridge TV 25 Minute TV show about super-resolution microscopy 
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 Clemens Kaminski, Gabriele Kaminski Schierle, Colin Hockings, Florian Ströhl, Nathan Curry

Episode of Elemental Ideas on Cambridge TV 25 Minute TV show about super-resolution microscopy
01/07/2016 Explaining research to the public through a video of our group's lab and research work, with interviews of the two group leaders and several postdoctoral research associates and PhD students.
The videos can be accessed for free online.

Elemental Ideas is Cambridge TV's science magazine programme. They've produced more than 50 programmes since August 2015, covering research topics including string theory, diabetes, exoplanets, vulcanology, epigenetics and the wildlife of New Zealand.
Year(s) Of Engagement Activity 2016
URL http://www.cambridge-tv.co.uk/super-resolution-microscopy/
 
Description Pint of Science Festival Cambridge 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach Regional
Primary Audience Public/other audiences
Results and Impact Public lecture at Pint of Science Festival.
Year(s) Of Engagement Activity 2016
 
Description PreLighter 
Form Of Engagement Activity A formal working group, expert panel or dialogue
Part Of Official Scheme? No
Geographic Reach National
Primary Audience Public/other audiences
Results and Impact Writing highlight sections on interesting preprints for the PreLight website and twitter feed
Year(s) Of Engagement Activity 2018,2019
 
Description Research Photography Exhibition 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach Regional
Primary Audience Public/other audiences
Results and Impact Finalist in STEM photography competition at Hughes Hall College, Cambridge
Year(s) Of Engagement Activity 2019
 
Description STEM in Song 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach National
Primary Audience Schools
Results and Impact Encouraging school-aged girls to engage with the STEM subjects through music. Awards event with science demos, song launch at the LMB, music video at https://www.youtube.com/watch?v=7c4KMOWoZW4 . Reach: 15300 (according to Facebook insights)
Year(s) Of Engagement Activity 2018
URL https://www.youtube.com/watch?v=7c4KMOWoZW4
 
Description Seminar on Science and Brewing 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach Regional
Primary Audience Public/other audiences
Results and Impact Explaining science to the public
Year(s) Of Engagement Activity 2018
 
Description So you want to be a scientist?' experiment and speaking presentation 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach Regional
Primary Audience Schools
Results and Impact Talking to school children about life as a scientist and GCSE options to take
Year(s) Of Engagement Activity 2018
 
Description Talk and lab tour to funders of ARUK, Feb 2016 
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 Supporters
Results and Impact Engage with funders on the research carried out thanks to their donations
Year(s) Of Engagement Activity 2016
 
Description Talk and lab tour to funders of ARUK, May 2015 
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 Supporters
Results and Impact Engage with funders on the research carried out thanks to their donations
Year(s) Of Engagement Activity 2015
 
Description Talk on A short history of Microscopy, Cambridge 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach Local
Primary Audience Public/other audiences
Results and Impact Public lecture on A short history of Microscopy
Year(s) Of Engagement Activity 2017
 
Description Tech Me Out team volunteer 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach Regional
Primary Audience Public/other audiences
Results and Impact Pint of Science Festival Cambridge
Year(s) Of Engagement Activity 2018