Chemical probes for lysosomal biology
Theo Murphy meeting organised by Dr Simon Wheeler and Professor Elizabeth New.
Lysosomal function underpins both cellular health and disease. Despite this our understanding of how these organelles work, and how they contribute to important pathologies, is far from complete. The use of chemical sensors to elucidate these mechanisms provides an exciting but largely unexploited opportunity to facilitate fundamental research and ultimately to develop therapies.
The schedule of talks and speaker biographies are available below. Speaker abstracts will be available closer to the meeting date.
Poster session
There will be a poster session on Monday 09 September. If you would like to present a poster, please submit your proposed title, abstract (up to 200 words), author list, and the name of the proposed presenter and institution to the Scientific Programmes team no later than Friday 26 July 2024.
Attending this event
This event is intended for researchers in relevant fields, and is a residential meeting taking place at Hilton York, 1 Tower Street, York, YO1 9WD.
- Free to attend
- Advance registration essential (more information about registration will be available soon)
- This is an in person meeting
- Catering options are available to purchase during registration. Participants are responsible for their own accommodation booking.
Enquiries: contact the Scientific Programmes team
Organisers
Schedule
Chair
Professor Frances Platt FMedSci FRS, University of Oxford, UK
Professor Frances Platt FMedSci FRS, University of Oxford, UK
Professor Frances Platt obtained her BSc from Imperial College London (Zoology) and her PhD from the University of Bath, UK. She was a post-doctoral fellow at Washington University Medical School in St. Louis, USA. She was a Lister Institute Senior Research Fellow and is currently Professor of Biochemistry and Pharmacology at the University of Oxford. Her main research interests include the biology and pathobiology of glycosphingolipids and lysosomal disorders. Her research led to the development of miglustat for the treatment of glycosphingolipid lysosomal storage diseases. Professor Platt was awarded the Alan Gordon Memorial Award and the Horst Bickel Award for advances in metabolic disease therapy. She was elected a fellow of the Academy of Medical Sciences in 2011 and was the recipient of a Royal Society Wolfson Merit Award in 2013. In 2016 she became a Wellcome Trust Investigator in Science. She was appointed Head of the Department of Pharmacology in 2020 and was elected a Fellow of the Royal Society in 2021.
| 09:00-09:05 |
Welcome and introduction
Dr Simon Wheeler, De Montfort University, UK
Dr Simon Wheeler, De Montfort University, UKSimon Wheeler studied chemistry at the University of Bristol before working in the pharma industry for 14 years as a synthetic chemist. Redundancy prompted him to retrain as a pharmacist at De Montfort University but he quickly found that this profession offered insufficient opportunity for his curiosity. He returned to De Montfort to study for a PhD in cell biology (under the supervision of Dr Dan Sillence) where he examined the cellular mechanisms of lysosomal storage disorders especially Niemann-Pick type C. After a period of post-doctoral work on fluorescent europium-based probes with Dr Steve Butler at Loughborough University he returned to De Montfort University in August 2022 as a lecturer in pharmaceutical chemistry. His research interests are in fragment based drug discovery and crown ether-based fluorescent probes for Ca2+. |
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| 09:05-09:45 |
Controlling ion flux across the lysosome through two-pore channels
Much evidence points to lysosomes as non-canonical stores of Ca2+. This extends the role of lysosomes to signalling processes. A number of Ca2+-permeable lysosomal ion channels have been identified and shown to regulate numerous functions including many aspects of membrane trafficking. Malfunction of these channels is linked to a growing number of diseases. In this talk, I will cover recent advances in understanding how two-pore channels (TPCs) control flux of Ca2+ and other ions across the lysosomal membrane. The picture that emerges is an unusual one in which TPCs are able to switch their ion selectivity in an agonist dependent manner to differentially control lysosomal function. I will discuss molecular mechanisms underpinning their activation by natural and newly identified synthetic mimics. And how the local Ca2+ release events that TPCs generate are converted into global Ca2+ signals through cross-talk with other Ca2+ channels on neighbouring canonical Ca2+ stores. 
Professor Sandip Patel, UCL, UK
Professor Sandip Patel, UCL, UKSandip Patel is Professor of Cell Signalling at University College London. He obtained a BSc in Medical Biochemistry from Birmingham and a PhD in Pharmacology from Cambridge. Sandip held Research Fellowships funded by the Wellcome Trust and Hayward Foundation at several institutions in the USA and UK including Oxford prior to tenure. He was elected to the membership of Academia Europaea in 2020 and awarded the GL Brown Prize Lecture from the Physiological Society in 2023. Sandip sits on panels of various funding bodies including the Medical Research Council and he is currently the Deputy Head of the Department of Cell and Developmental Biology at UCL. His research has demonstrated the widespread physiological importance of so-called ‘acidic calcium stores’ within our cells and helped define the molecular mechanisms underpinning their actions. |
| 09:45-10:30 |
Lysosomes at the nexus of cellular homeostasis and cancer
Transmembrane proteins represent ~20-30% of the human proteome and are subjected to turnover by endocytic uptake and delivery to the lysosome, where their degradation poses a unique challenge due to their hydrophobic, phospholipid-embedded a-helical domains that are inaccessible to endopeptidases. Here, we answer the long-standing question of how lysosomes degrade transmembrane proteins: a key missing piece in the life cycle of this important class of proteins that mediate signal transduction, nutrient import, adhesion, and migration. Combining lysosomal proteomics with functional genomic data mining, untargeted metabolomics, and biochemical reconstitution, we find that a previously mischaracterised enzyme that we rename Lysosomal Leucine Aminopeptidase (LyLAP) is most tightly associated with elevated endocytic activity and enables transmembrane protein degradation. LyLAP performs a unique function not found among known lysosomal hydrolases, namely, processive disassembly of hydrophobic a-helices triggered by their N-terminal hydrophobic (often Leucine) residues. Importantly, LyLAP is upregulated in pancreatic ductal adenocarcinoma (PDA), an aggressive cancer that relies on macropinocytosis for nutrient uptake. Strikingly, loss of LyLAP activity has catastrophic consequences for lysosomes, ultimately leading to PDA cell death. Thus, LyLAP enables lysosomal degradation of membrane proteins, and may represent a targetable vulnerability in highly endocytic cancer cells. 
Dr Aakriti Jain, University of California, Berkeley, USA
Dr Aakriti Jain, University of California, Berkeley, USAAakriti obtained her BS in Chemical and Biomolecular Engineering from the University of California, Berkeley, and her MS in Biological Sciences from Université Diderot, Paris and the Center for Research and Interdisciplinary studies. During her PhD at the Francis Crick Institute in London, funded by the Marie-Sklowdoska-Curie doctoral fellowship, she studied dysregulation of mitochondrial metabolism in liver cancer. Following her doctoral work, she has been a postdoctoral researcher in Roberto Zoncu’s lab at UC Berkeley. In the Zoncu lab, she is not only continuing her study of metabolic perturbations in disease, but also overall organelle homeostasis pathways. By combining biochemical, cell biological and large-scale omics techniques, she is dissecting various aspects of lysosomal biology in both cancer and neurodegenerative disorders. |
| 10:30-11:00 |
Break
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| 11:00-11:45 |
Lysosomes as targets for cancer therapy
Being originally discovered as cellular recycling bins, lysosomes are today recognised as versatile signalling organelles that control a wide range of cellular functions that are essential not only for the well-being of normal cells but also for malignant transformation and cancer progression. In addition to their core functions in waste disposal and recycling of macromolecules and energy, lysosomes serve as an indispensable support system for malignant phenotype by promoting cell growth, cytoprotective autophagy, drug resistance, pH homeostasis, invasion, metastasis and genomic integrity. On the other hand, malignant transformation reduces the stability of lysosomal membranes rendering cancer cells sensitive to lysosome-dependent cell death. Notably, many clinically approved cationic amphiphilic drugs widely used for the treatment of other diseases accumulate in lysosomes, interfere with their cancer-promoting and cancer-supporting functions, and destabilise their membranes thereby opening intriguing possibilities for cancer therapy. Here, I will discuss the emerging evidence that supports the supplementation of current cancer therapies with lysosome-targeting cationic amphiphilic drugs and the molecular mechanisms by which they disturb lysosomal function and permeabilise lysosomal membranes. Funding: Danish Cancer Society, Danish National Research Foundation, European Research Council, and Novo Nordisk Foundation. 
Dr Marja Jäättelä, Danish Cancer Institute, Denmark
Dr Marja Jäättelä, Danish Cancer Institute, DenmarkMarja Jäättelä received MD and DMSc degrees from the University of Helsinki in 1989 and 1990, respectively. Following postdoctoral training in Copenhagen, New York and Ann Arbor, she was appointed the head of the Apoptosis Laboratory at the Danish Cancer Institute in 1997 and promoted to the Head of Cell Death and Metabolism Unit in 2012. Additionally, she holds a professorship in Cell Death and Metabolism at the University of Copenhagen and is the director of the Center for Autophagy, Recycling and Disease. Her research focuses on cellular metabolism and lysosomal biology in cancer, and her work is best characterized as a successful combination of uncompromised and innovative basic science with strong translational interest. She is an internationally respected scientist as evidenced by over 50,000 citations, frequent lectures in major international meetings, ample funding and memberships in numerous learned societies, advisory boards, editorial boards and evaluation panels. |
| 11:45-12:30 |
GBA1 and parkinsonism: new tools for new times
To explore the pathogenesis of GBA1-associated synucleinopathies, we genetically engineered GBA1 in an iPSC line derived from a patient with both GD (GBA1: N370S/N370S) and PD using CRISPR/Cas9 and piggyBac systems and generated a panel of isogenic lines (GBA1: KO/KO, and WT/WT) to model the impact of enzyme and lipid substrate levels. DANs differentiated from these isogenic lines demonstrated descending levels of GCase protein: the WT/WT line exhibited the highest, followed by the N370S/N370S, whereas the KO/KO line showed only a trace. Glycosylation analysis demonstrated endoplasmic reticulum (ER) retention of a modest fraction of GCase N370S. Immunofluorescence with the sensitive GCase antibody hGCase-1/23 revealed targeting of GCase N370S to lysosomes. Both in vitro GCase activity assay using the fluorogenic substrate 4MU and live-cell GCase assay with LysoFQ-GBA demonstrated reduced GCase activity in the N370S/N370S and no activity in the KO/KO line. Surprisingly, lipid analysis using supercritical fluid chromatography-mass spectrometry revealed substantial glucosylceramide and glucosylsphingosine accumulation only in the KO/KO line, despite reduced GCase activity in the N370S/N370S. Proteomic profiling of lysosomes purified via the LysoIP approach revealed that the KO/KO lysosomes had substantial lipid accumulation and reduced levels of Cathepsin F. Ongoing experiments aim to further elucidate the mechanistic implications of these findings. 
Yu Chen, National Human Genome Research Institute, USA
Yu Chen, National Human Genome Research Institute, USADr Yu Chen, a Staff Scientist of the Medical Genetics Branch, is a cell biologist in the National Human Genome Research Institute at National Institutes of Health (NIH). He received his PhD from Institute of Biophysics, Chinese Academy of Sciences, trained in cell biology at the NIH, and in translational science at the National Centre for Advancing Translational Sciences (NCATS). Dr Chen studies the association between glucocerebrosidase and parkinsonism, focusing on the role of lysosomal pathways in parkinsonism and the development of small molecule chaperone therapy for Gaucher disease and for parkinsonism. |
Chair
Professor Elizabeth New, University of Sydney, Australia
Professor Elizabeth New, University of Sydney, Australia
Dr Elizabeth New undertook her undergraduate and Masters studies at the University of Sydney. She completed her PhD studies in 2010 at the University of Durham with Professor David Parker. Liz was then a Royal Commission for the Exhibition of 1851 Research Fellow at the University of California, Berkeley, working with Professor Chris Chang. In 2012, she returned to the University of Sydney, holding a Discovery Early Career Research Fellowship from the Australian Research Council from 2012-2014, and a Westpac Research Fellowship from 2016. Liz’s research interests lie in the development of small molecule sensors for the study of oxidative stress and metal ions in biology.
| 13:30-14:15 |
Photo-activatable and clickable lipid analogs as tools to investigate the transport of lysosomal sphingosine
Sphingolipids are major eukaryotic lipids defined by a common sphingoid backbone. They are particularly enriched at the plasma membrane and they undergo constitutive turnover along the endocytic pathway. The bulk of sphingolipid recycling takes place in late endosomes and lysosomes. Here, catabolism of complex sphingolipids produces sphingosine and long chain sphingoid bases, which are then exported to the ER for synthesis of new sphingolipids. This process, called sphingosine recycling, is critical for cellular lipid homeostasis, yet its mechanisms are largely unknown. To address this, we utilised a recently developed functionalised sphingosine probe. The probe pre-localises into the lysosome and is activated by light. Its other functionalities allow the detection of interaction partners as well as following its subcellular distribution and metabolic conversions. Using this tool, we investigated the role of the sterol transporter STARD3 in sphingosine recycling. Overexpression of STARD3 in cells increased the probe’s metabolic conversion into higher sphingolipids, whereas STARD3 depletion led to delayed export from the lysosome. In vitro approaches showed that the lipid binding domain of STARD3 can bind sphingosine, supporting the hypothesis that STARD3 is a sphingosine transporter operating at the lysosome – ER contact sites. This work paves the way for further research on protein mediated sphingosine transport and on mechanisms that regulate the recycling of cellular sphingolipids. 
Dr Denisa Jamecna, University of Osnabrück, Germany
Dr Denisa Jamecna, University of Osnabrück, GermanyDr Jamecna studied Biology at the Masaryk University in Brno and at the University College London. She received her PhD from the Université Côte d’Azur in Nice in 2018. During her PhD in the group of Dr Bruno Antonny, she studied how intrinsically disordered N-terminal regions in OSBP and related proteins affect membrane tethering geometry and dynamics of membrane contact sites. Until recently, she worked as a Walter Benjamin postdoctoral fellow in the lab of Dr Doris Höglinger at the Biochemistry Center Heidelberg. Her research focused on the mechanism of sphingosine transport at the lysosome - ER contact sites. Since June 2024, she has started as a junior group leader at the University of Osnabrück. The work of her group combines cell biology, biochemistry, chemical biology and structural biology. They specialize in using synthetic photoactivatable and clickable (pac) lipid probes and their organelle-targeted derivatives to investigate subcellular lipid distribution, metabolism and protein-lipid interactions. |
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| 14:15-15:00 |
Visualisation of endosome/ lysosomal labile iron by ferrous ion-selective fluorescent probes
Iron is the most abundant transition metal in the human body and plays numerous essential roles. The homeostasis of iron is tightly regulated, as both deficiency and overload can lead to various dysfunctions. However, much of intracellular iron homeostasis and trafficking remains poorly understood, largely due to the lack of effective fluorescent probes for detecting the labile iron pool. Lysosomes and endosomes are key organelles in intracellular iron trafficking, as they are involved in endocytic iron uptake, storage, and subcellular iron recycling processes. We have developed a series of ferrous ion-selective fluorescent probes using N-oxide chemistry, some of which have been applied to study iron status in endosomes and lysosomes under both physiological and pathological conditions. Among these, a lysosome-specific fluorescent probe, HMRhoNox, was designed and employed to monitor transferrin-mediated iron uptake and the lysosomal accumulation of ferrous ions during ferroptosis. Additionally, we developed a plasma membrane-anchoring fluorescent probe to enable real-time imaging of iron uptake. This presentation will detail the development of these probes, their imaging capabilities, and recent findings from their application. 
Dr Takusu Hirayama, Gifu Pharmaceutical University, Japan
Dr Takusu Hirayama, Gifu Pharmaceutical University, JapanDr Tasuku Hirayama received his BS degree from Kyoto University in 2004. He also obtained his MS (2006) and PhD (2009) in chemical biology from Kyoto University under the direction of Professor Yukio Yamamoto. He joined Professor Christopher Chang’s group at University of California, Berkeley as a postdoctoral fellow. In 2010, he joined Professor Hideko Nagasawa’s group in Gifu Pharmaceutical University as an assistant professor. He has been an associate professor in Gifu Pharmaceutical University since 2016. His research interests include development of unique chemical tools to understand biological functions of metal ions. |
| 15:00-15:30 |
Break
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| 15:30-16:15 |
Organelle-targeted sensors and multimodal probes for cellular imaging
Understanding the biochemical composition of cells and organelles is crucial for comprehending physiological and pathological processes. This requires tools that can identify and monitor the chemistry of organelles within cells. We are interested in developing fluorogenic labels, and responsive fluorescent sensors for cellular redox state and redox-active metal ions. To this end, we are developing lysosome-targeted redox sensors and investigating more robust lysosomal-targeting groups. Multimodal imaging, which integrates the benefits of two or more imaging techniques, is becoming increasingly popular. Conventional probes are typically limited to a single analyte; by combining multiple imaging modalities, more comprehensive information can be obtained from a single sample. Developing multimodal probes compatible with several imaging techniques is therefore highly desirable. We have prepared a set of lysosomally-targeted multimodal imaging agents that can be used for fluorescence microscopy and additional modalities, including phosphorescence, Raman, and X-ray fluorescence microscopies. 
Dr Liam Adair, The University of Sydney, Australia
Dr Liam Adair, The University of Sydney, AustraliaLiam Adair completed an MSci Chemistry with Medicinal Chemistry at the University of Glasgow, with a year in industry at AstraZeneca. He remained at Glasgow for PhD studies in total synthesis with Professor Rudi Marquez and Dr Joëlle Prunet. Since 2018 he has been a postdoctoral researcher at the University of Sydney, where he is currently a research fellow in the ARC Centre of Excellence for Innovations in Peptide and Protein Science. His research involves the design, synthesis, and application of targeted and responsive fluorophores and multimodal probes for bioimaging. |
| 16:15-17:00 |
Towards fluorescent sensors for lysosomal Ca 2+ and K+
Dr Simon Wheeler, De Montfort University, UK
Dr Simon Wheeler, De Montfort University, UKSimon Wheeler studied chemistry at the University of Bristol before working in the pharma industry for 14 years as a synthetic chemist. Redundancy prompted him to retrain as a pharmacist at De Montfort University but he quickly found that this profession offered insufficient opportunity for his curiosity. He returned to De Montfort to study for a PhD in cell biology (under the supervision of Dr Dan Sillence) where he examined the cellular mechanisms of lysosomal storage disorders especially Niemann-Pick type C. After a period of post-doctoral work on fluorescent europium-based probes with Dr Steve Butler at Loughborough University he returned to De Montfort University in August 2022 as a lecturer in pharmaceutical chemistry. His research interests are in fragment based drug discovery and crown ether-based fluorescent probes for Ca2+. |
Chair
Dr Simon Wheeler, De Montfort University, UK
Dr Simon Wheeler, De Montfort University, UK
Simon Wheeler studied chemistry at the University of Bristol before working in the pharma industry for 14 years as a synthetic chemist. Redundancy prompted him to retrain as a pharmacist at De Montfort University but he quickly found that this profession offered insufficient opportunity for his curiosity. He returned to De Montfort to study for a PhD in cell biology (under the supervision of Dr Dan Sillence) where he examined the cellular mechanisms of lysosomal storage disorders especially Niemann-Pick type C. After a period of post-doctoral work on fluorescent europium-based probes with Dr Steve Butler at Loughborough University he returned to De Montfort University in August 2022 as a lecturer in pharmaceutical chemistry. His research interests are in fragment based drug discovery and crown ether-based fluorescent probes for Ca2+.
| 09:00-09:45 |
A genetically encoded lysosomal pH sensor for in vivo applications in aging and neurodegenerative disease
A highly acidic pH is required for the optimal functioning of lysosomal enzymes. Loss of lysosomal intraluminal acidity can disrupt cellular protein and lipid homeostasis and is linked to age-related diseases such as neurodegeneration. To understand how lysosomal pH changes with age and disease, we have developed a robust new lysosomal pH biosensor that we call FIRE-pHLy for Fluorescent Indicator REporting on pH of the Lysosome. Using FIRE-pHLy, we have performed cell-based high-throughput fluorescence screening assays for modifiers of lysosomal pH in undifferentiated and differentiated SH-SY5Y neuroblastoma cells. We have also performed a whole genome screen for genetic modifiers of lysosomal pH in iPSC-derived neurons (induced neurons or iNeurons.) Finally, we have expressed FIRE-pHLy in C. elegans and M. musculus. In C. elegans, lysosomal pH changes during development and aging, and manipulation of lysosomal pH can directly impact lifespan. Thus, lysosomal pH serves as both a biomarker and therapeutic target in aging and neurodegeneration. 
Dr Aimee Kao, University of California San Francisco, USA
Dr Aimee Kao, University of California San Francisco, USADr Aimee Kao, MD, PhD, is the John Douglas French Foundation Endowed Professor of Neurology at the University of California, San Francisco. She studied Neuroscience at Brown University and received MD/PhD degrees from the University of Iowa. She completed Neurology residency at UCSF, followed by fellowships in Behavioral Neurology and molecular genetics of ageing. She is an expert in managing Alzheimer's disease and age-related cognitive disorders. Dr Kao’s laboratory investigates the basic mechanisms underlying neurodegenerative disorders, with a focus on how ageing, stress and gene mutations disrupt lysosomal function and protein homeostasis. She currently directs the UCSF Medical Scientist Training Program and is principle investigator of an NIH-sponsored Tau Center without Walls. Dr Kao has been awarded the Paul G. Allen Family Foundation Distinguished Investigator Award in Neurodegenerative Diseases, the Glenn Award for Research in the Biological Mechanisms of Aging and the American Neurological Society’s Derek Denny Brown Award. |
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| 09:45-10:30 |
Interrogation of lysosome biology by pH sensitive nanoparticles
Engineered and tailor-made nanomaterials (NM) are of increasing relevance for current and future developments in the life and material sciences for applications, eg as drug carriers, fluorescent sensors, and multimodal labels in bioanalytical assays, and reporters for imaging applications. For instance, NM-based reporters and sensors, that are labelled or stained with a multitude of conventional or sensor dyes, have several advantages as compared to molecular probes like enhanced brightness, ie amplified signals, ease of designing ratiometric systems by combining analyte-sensitive and inert reference dyes, and increased photostability. Moreover, this can enable the use of hydrophobic dyes in aqueous environments. For rational NM design, choosing and tailoring the intrinsic physicochemical properties, such as particle size, size distribution, morphology, and surface chemistry of the NM application-specific considerations like biocompatibility, ease and low cost of preparation, and colloidal stability and performance in the targeted environment must be considered. In this lecture, different design concepts of inorganic, organic, and hybrid NM and microparticles with hydrophilic surface chemistries and different functionalities are presented that can be used for the targeting of lysosomes; and to monitor functional parameters of endo-lysosomal compartments, like pH or enable oxygen sensing. 
Dr Isabella Tavernaro, Federal Institute for Materials Research and Testing, Germany
Dr Isabella Tavernaro, Federal Institute for Materials Research and Testing, GermanyBeing a chemist by training, Dr Isabella Tavernaro specialized in surface-modified organic and inorganic nanomaterials for biomedical applications during her doctoral studies at Justus-Liebig University Giessen. After graduating in 2014, she joined the Nano Cell Interactions group at the INM-Leibniz Institute for New Materials in Saarbrücken as a postdoctoral fellow. Her research work focused on nanosafety and the development of tailored nanomaterials for safe(r)-by-design approaches. In 2020, she joined the Federal Institute for Materials Research and Testing (BAM) in Berlin. In division Biophotonics, led by Dr Ute Resch-Genger, she develops nanoscale test materials and multi-method characterization strategies for quantifying surface functional groups, as well as molecular and nanoscale optical sensors. |
| 10:30-11:00 |
Break
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| 11:00-11:45 |
Progress and challenges for protein-based probes of metal ions
An ever-growing selection of high performance fluorescent protein-based biosensors is revolutionizing our ability to visualize metal ion concentrations in cells. In this seminar I will describe our most recent efforts to use protein engineering to make a new generation of genetically encoded biosensors, and chemigenetic biosensors augmented with synthetic molecules, for calcium ion, potassium ion, and sodium ion. Key to this effort is our reliance on the use of directed protein evolution to iteratively and reliably improve the properties of biosensors. By creating biosensors with different colours, specificities, and affinities, we are opening up new opportunities for researchers to use multiplexed imaging to investigate dynamics changes in metal ion concentrations in organelles, cells, and whole tissues in model organisms. 
Professor Robert E Campbell, The University of Tokyo, Japan
Professor Robert E Campbell, The University of Tokyo, JapanDr Robert E Campbell is a Professor in the Department of Chemistry, School of Science, at The University of Tokyo (2018 - present). He earned his PhD in Chemistry at the University of British Columbia (1994-2000) and undertook postdoctoral research at the University of California, San Diego (2000-2003). From 2003 to 2023 he was a Professor at the University of Alberta. He is a leading developer of optogenetic tools, including a complete spectral palette of fluorescent protein-based calcium ion indicators. He has distributed >8000 samples of fluorescent protein-based tools to labs around the world. Recognitions include a Stanford Neurosciences Institute Visiting Scholar Award (2017), the Teva Canada Limited Biological and Medicinal Chemistry Award (2016), the Rutherford Memorial Medal from the Royal Society of Canada (2015), and the Boehringer Ingelheim Research Excellence Award (2014), and a Canada Research Chair (2004-2014). |
| 11:45-12:30 |
Understanding the role of solute carriers in lysosomal homeostasis and signalling
Solute carriers (SLCs) play key roles in lysosomal homeostasis and intracellular signalling as essential regulators of metabolite and ion exchange with the cytoplasm. Recent advances in structural biology have revealed an unexpected evolutionary links between SLCs and intracellular receptors, opening a new and exciting time in transporter biology. In this talk I will discuss our recent results in understanding the concepts and mechanisms through which SLC proteins can both transport and signal across membranes and the unique role of lysosomal lipids in regulating SLC function. 
Professor Simon Newstead, University of Oxford, UK
Professor Simon Newstead, University of Oxford, UKSimon Newstead holds the David Phillips Chair of Molecular Biophysics in the Department of Biochemistry, is a member of the Kavli Institute for Nanoscience Discovery in Oxford and is a Professorial Fellow at Corpus Christi College, Oxford. Simon received his MBiochem (Hons) degree from the University of Bath in 2001 and his PhD in protein crystallography at St Andrews in 2004. Simon then joined the membrane protein laboratory of Professor So Iwata at Imperial College London, where he worked on structural studies of secondary active transporters and methods development in membrane protein structural biology. In 2009, he was awarded an MRC career development award to establish a research group in Oxford focused on cellular nutrient uptake and drug transport. In 2013, he became an Associate Professor in the Department of Biochemistry and senior subject tutor (Ordinary Student) in Biochemistry at Christ Church, Oxford. In 2015, he was promoted to Professor and in 2019 elected to the Royal Society of Biology. In 2022, Simon was appointed to the David Phillips Chair in Molecular Biophysics and Professorial Fellow at Corpus Christi College. Simon currently leads the Structural Biology and Molecular Biophysics research theme in the Department of Biochemistry. |
Chair
Dr Aimee Kao, University of California San Francisco, USA
Dr Aimee Kao, University of California San Francisco, USA
Dr Aimee Kao, MD, PhD, is the John Douglas French Foundation Endowed Professor of Neurology at the University of California, San Francisco. She studied Neuroscience at Brown University and received MD/PhD degrees from the University of Iowa. She completed Neurology residency at UCSF, followed by fellowships in Behavioral Neurology and molecular genetics of ageing. She is an expert in managing Alzheimer's disease and age-related cognitive disorders. Dr Kao’s laboratory investigates the basic mechanisms underlying neurodegenerative disorders, with a focus on how ageing, stress and gene mutations disrupt lysosomal function and protein homeostasis. She currently directs the UCSF Medical Scientist Training Program and is principle investigator of an NIH-sponsored Tau Center without Walls. Dr Kao has been awarded the Paul G. Allen Family Foundation Distinguished Investigator Award in Neurodegenerative Diseases, the Glenn Award for Research in the Biological Mechanisms of Aging and the American Neurological Society’s Derek Denny Brown Award.
| 13:30-14:15 |
Super-resolution imaging of sub-cellular dynamics of endolysosomes
As a process of cellular uptake, endocytosis, with gradient acidity in different endocytic vesicles, is vital for the homeostasis of intracellular nutrients and other functions. To study the dynamics of endolysosomes, we developed pH-sensitive molecular probes imaging the sub-cellular dynamics of endocytic vesicles using a super-resolution fluorescence technique, structured illumination microscopy (SIM). Firstly, a membrane-anchored pH probe, ECGreen was synthesised to visualise endocytic vesicles under SIM. Being sensitive to acidity with increasing fluorescence at low pH, ECGreen, can differentiate early and late endosomes as well as endolysosomes. Meanwhile, membrane-anchoring not only improves the durability of ECGreen, but also provides an excellent anti-photobleaching property for long-time imaging with SIM, which makes ECGreen a good probe for elucidating the interaction between mitochondria and endocytic vesicles in autophagy. Based on ECGreen, a multi-dimensional model containing spatial, temporal, and pH information was successfully developed to analyse endocytic dynamics for its unclear roles in cellular functions. Secondly, by taking advantages of intramolecular charge transfer and a pH-sensitive pyridine unit, we developed a cationic quinolinium-based fluorescent probe, PyQPMe, which exhibits excellent ratiometric fluorescent response to endolysosomes at different stage of interest. By applying PyQPMe as a small molecular probe, we were able to reveal a constant conversion rate from early endosome to late endosome during autophagy using the super-resolution imaging. 
Dr Jiajie Diao, University of Cincinnati College of Medicine, USA
Dr Jiajie Diao, University of Cincinnati College of Medicine, USADr Jiajie Diao is Associate Professor at University of University of Cincinnati College of Medicine, and a co-director of the Center for Chemical Imaging in Biomedicine. Dr Diao received his PhD in physics from University of Illinois at Urbana-Champaign and conducted his postdoc research at Stanford University. His research is using advanced biophysical tools to study subcellular dynamics of organells. Dr Diao has published more than 130 papers in top-tier journals including Nature, PNAS, Nat Struct Mol Biol, Nat Commun, Nat Protoc, Cell Chem Biol, Cell Rep, eLife, JACS, Angew Chem, ACS Nano, Biomaterials. In 2017, he was awarded the Young Scientist Prize in Biological Physics by the International Union of Pure and Applied Physics. He is an editorial board member of BMC Biology and a guest editor of PNAS, and has been serving as a reviewer for more than 80 journals. |
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| 14:15-15:00 |
Morpho-functional characterisation of lysosomes by correlative light and electron microscopy
Cells reprogram and adapt their endo-lysosomal systems to their metabolic requirements in physiology and pathology. Yet, molecular and ultrastructural adaptations of endo-lysosomal organelles to metabolic stimuli at nanoscale, and how this is exploited particularly by cancer cells remain notoriously obscure. Addressing this, we have developed endo-lysosomal biology tailored correlative microscopy approaches to combine molecular specificity and live-cell capabilities of fluorescence microscopy (FM) with high-resolution ultrastructure of electron microscopy (EM). We label and visualise endo-lysosomal organelles in (live) cells with fluorescent biosensors after which they are processed for EM and imaged using a (volume) EM technique. We correlate hundreds of fluorescently labelled organelles enabling quantitative analysis of the molecular-functional-ultrastructural data within a single dataset. We have employed our methods to map the ultrastructural distribution of endo-lysosomal proteins; to link single organelle dynamics to their 3D ultrastructure; to image transient contact sites between the endoplasmic reticulum and lysosomes; and to monitor functional parameters of endo-lysosomal compartments, ie enzyme activities, pH, and calcium content. The presented CLEM workflows are compatible with a large repertoire of probes and can integrate information on protein localisation, functional status, dynamics and 3D ultrastructure, all in a single sample and at the level of single organelles. 
Dr Nalan Liv, University Medical Center Utrecht, The Netherlands
Dr Nalan Liv, University Medical Center Utrecht, The NetherlandsNalan Liv is Assistant Professor of Cell Biology, at Center for Molecular Medicine, University Medical Center Utrecht (UMCU). She leads a cellular nanoimaging and endo-lysosomal biology research team, and coordinates the Cell Microscopy Core. Nalan’s current research is dedicated to high-resolution understanding of intracellular structure-function relations in cancer cell biology, and she is awarded with multiple grants to study the role of lysosomes in carcinogenesis. Her team develops innovative high-resolution microscopy approaches and utilizes a combination of molecular biology, live-cell fluorescence microscopy, and ultrastructural electron microscopy tools to resolve how the endo-lysosomal system is rewired in cancer. |
| 15:00-15:30 |
Break
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| 15:30-16:15 |
New sub-cellular chemical imaging techniques with potential to probe lysosomal biology
Vibrational spectroscopy has found many applications in the biological sciences. Its capabilities include direct, label-free and in situ characterisation of protein secondary structures (including protein aggregates), lipid profiling, and investigation of metabolism (eg lactate, glucose, glycogen, organic phosphates). Unfortunately, a common form of vibrational spectroscopy, infrared spectroscopy, suffers from relatively poor spatial resolution due to the longer wavelengths of infrared light (compared to bright field or fluorescence microscopy). Other modalities of vibrational spectroscopy, such as Raman spectroscopy offer improved diffraction-limited spatial resolution, but can suffer substantial limitations from cell and tissue autofluorescence. Substantial efforts have been undertaken for both infrared and Raman spectroscopy to develop improved instrumentation, light sources, and imaging protocols to enable direct sub-cellular chemical imaging with these two techniques. In my talk I will present the recent developments in the field of vibrational spectroscopy, along with approaches to integrate these techniques in multi-modal or correlative workflows, which could open new possibilities to probe the biology of lysosomes and other organelles. 
Dr Mark Hackett, Curtin University, Australia
Dr Mark Hackett, Curtin University, AustraliaDr Hackett is a mid-career researcher in the field of analytical chemistry and applied spectroscopy, currently completing an Australian Research Council Future Fellowship at Curtin University. He completed his PhD at The University of Sydney (2011, Chemistry), which was followed by two-postdoctoral fellowships at the University of Saskatchewan, Canada (2011 – 2016). His primary research interest is the development and optimization of novel elemental and bio-molecular microscopy techniques for application to the field of neuroscience to study the mechanisms of brain ageing and neurodegenerative disease. The main spectroscopic techniques involved in his research program include Fourier transform infrared spectroscopy (FTIR), Raman spectroscopy, X-ray fluorescence microscopy (XFM) and X-ray absorption spectroscopy (XAS). |
| 16:15-16:55 |
Rational design of fluorescent nano probes for long term tracking and super-resolution imaging of lysosomal dynamics
Lysosomes are membrane-enclosed small spherical cytoplasmic organelles. Malfunctioning and abnormalities in lysosomes can cause a plethora of neurodegenerative diseases. Consequently, understanding the structural information on lysosomes down to a subnanometer level is essential. Recently, super-resolution imaging enables us to visualise dynamical processes in living cells down to subnanometer accuracy by breaking the diffraction limit. Herein, the advancement of various fluorescent nanoprobes that have been utilised in our lab recently for super-resolution imaging of lysosomal dynamics are presented. Dr Nandi will discuss the detail single molecule level photophysical of coinage metal nanoclusters for their potential use in the super-resolution imaging of lysosomes. He will also highlight the rational design of fluorescent superparamagnetic iron oxide nanoparticles (SPIONs) to enhance their photon budget as well as the magnetic resonance imaging contrast. The SPIONs were highly specific in staining and imaging the lysosomes of HeLa cells with high contrast. 
Dr Chayan Nandi, Indian Institute of Technology Mandi, India
Dr Chayan Nandi, Indian Institute of Technology Mandi, IndiaDr Chayan Kanti Nandi, is a Professor at the Indian Institute of Technology Mandi (IIT Mandi). He completed his PhD in 2006 in physical chemistry at the Indian Institute of Technology Kanpur India. Professor Nandi was a post doctoral fellow in Geothe University Germany (2006-2009) and in Princeton University (2009-2010). He received the prestigious Alexander Von Humboldt (AvH) Fellowship during his stay in Germany. After completing his post doctoral fellowship, he joined IIT Mandi as an Assistant Professor in 2010. He has several awards in his name. He has been awarded the Chemical Research Society of India (CRSI) bronze medal in 2020 for his outstanding research in nanomaterials chemistry. His main research focus is to design new fluorescent nanomaterials such as carbon nanodots, metal nanoclusters, quantum dots, super paramagnetics iron oxide nanoparticles for their application in single molecule localization based super resolution microscopy. |
| 16:55-17:00 |
Closing remarks
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