The Nanoscale Phenotype of Immune Responses in Health and Disease

Lead Research Organisation: Imperial College London
Department Name: Life Sciences

Abstract

Familiarity with the body's response to a cut or an infection - redness, tenderness and inflammation - belies the wonders taking place, where swarms of different cells move in to fight off germs, as well as repair the damage and deal with the debris. Far from conscious control, this reflex is essential for our survival. A simple view of this is that the immune system attacks germs which invade the body opportunistically. But over the last few decades, a painstaking, game-changing scientific adventure unfolded in which the world of immunity has opened up for what it really is: not simply a few types of immune cells which attack germs, but a multi-layered, dynamic lattice of interlocking sub-systems, one of the most complex and important frontiers of scientific enquiry we know of.
Advances in technology are helping us understand the immune system as never before, and to develop medicines which boost the system to fight cancer better, to dampen it to combat the symptoms of auto-immune disease, and to help develop better vaccines. Activating and inhibitory receptors on the surface of immune cells are critical determinants of immune activity. The level of each receptor and its ligand, and how well they bind, are primary determinants of disease outcomes. However, advances in microscopy are now revealing a host of other factors which control immune responses. This includes protrusions from immune cells which contact other cells, a complex nanoscale organisation of activating and inhibitory receptors, clusters of proteins secreted by immune cells to kill diseased cells, and novel mechanisms by which immune cells can detach from one target cell to attack again. Understanding immunity on a nanoscale is a major new frontier and will lead to completely new ideas for medicine.
My own research laboratory has decades of experience studying human immune cells called Natural Killer (NK) cells. These immune cells are able to directly kill cancer cells, and are a hot topic in developing new cancer therapies. NK cells are also important in viral defence, microbial pathogens, autoimmune diseases, reproductive complications and transplantation. Their activation is regulated by many activating and inhibitory receptors at their surface. However, the central tenet of this proposal is that their activity is also influenced by nanoscale processes, beyond simple ligation of receptors. For example, the presence of a receptor may remain similar in health and disease, but its nanoscale organisation can be altered to affect its activity. Also, it is entirely unexplored whether or not disease impacts NK cell protrusion density or the capacity of NK cells for serial killing, and so on. Indeed, nanoscale processes which regulate NK cells (and other immune cells) may be a major factor missing in our understanding of health and disease.
Here, we will compare NK cells from healthy donors and cancer patients, assessing every stage of their interaction with a cancer cell on a nanoscale - from an initial cell-cell contact to the assembly of a synapse, release of effector particles, subsequent detachment and serial engagement. Activating and inhibitory receptors will be mapped to understand signal integration and immune response thresholds, the structure and function of immune cell secretions will be analysed, and determinants of cell detachment and serial killing will be determined, and then compared in health and disease. Single cell secretions, visualised by super-resolution microscopy, will lead to a new approach to characterising immune responses. We will also compare types of NK cell. For example, it is entirely unexplored if memory-like NK cells exhibit faster interaction dynamics and greater serial killing. A large consortium of collaborators will facilitate this complex interdisciplinary endeavour, from using new instrumentation, developing image analysis and access to clinical samples. Strong links to industry will translate these new ideas to medicines.

Technical Summary

Cell-contact dependent immune responses are vital to our health. We now know that an immune cell interaction with a target cell involves several stages: initial contact, assembly of an immune synapse, release of effector proteins, subsequent detachment and finally, engagement with another target. However at each stage, there is a paucity of understanding what happens on a nanoscale. Evidence is accumulating that initial contacts occurs via cell protrusions, such that their density and dynamics may control immune cell sensitivity. Also, the nanoscale organisation of activating and inhibitory receptors has a huge impact on activation thresholds as, for example, discrete receptor clusters can segregate or coalesce to impact membrane proximal signal integration. Then upon activation, proteins secreted to kill infected or cancerous cells are organised in nanoscale supramolecular attack particles which are little understood. More broadly, secretions, including exosomes, are heterogeneous and our understanding of their functions and variability is still in its infancy. Mechanisms for immune cell detachment are also not clear, but involve nanoscale processes for removing receptor interactions. Thus, focussing on human Natural Killer (NK) cells as cells vital for defence against infections and cancer, and with NK cell-based therapies currently a hot topic, we will study each of these processes using super-resolution microscopy in combination with molecular and cell biology techniques. We will compare different subsets of NK cells isolated from blood, lung tissue, bone marrow, tumours and leukaemia. We will also compare NK cell behaviour in 3D matrix, within model tumour spheroids, and when cells are kept under hypoxic conditions. This will lead to understanding NK cell responses on a nanoscale and how this impacts health and disease, in turn leading to novel ideas for medicine. Translational outcomes will be pursued with GSK, Bristol Myers Squibb and Continuum Life Sciences.

Publications

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Ambrose AR (2023) Analyzing Single Cell Secretions by "Shadow Imaging". in Methods in molecular biology (Clifton, N.J.)

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Sheppard S (2024) Fatty acid oxidation fuels natural killer cell responses against infection and cancer. in Proceedings of the National Academy of Sciences of the United States of America

 
Description Cell biology of the TIGIT/CD96 regulatory axis
Amount £65,000 (GBP)
Organisation GlaxoSmithKline (GSK) 
Sector Private
Country Global
Start 09/2022 
End 09/2025
 
Description Optimisation of CAR T cell therapies
Amount £1,170,000 (GBP)
Organisation Bristol-Myers Squibb 
Sector Private
Country United States
Start 02/2023 
End 01/2027
 
Title Shadow Imaging 
Description Generation of cell shadows, visible by microscopy, to enable the identification and quantification of immune cell secretions at the single-cell level. In this method we activate cells on ligand coated surfaces to trigger the secretion of cellular components such as lytic granules or extracellular vesicles. We then rapidly pulse-stain the slide with fluorescently conjugated antibodies against the surface ligands, which stains the slide everywhere where cells are not attached. The cells are then removed and secreted components stained and imaged with each individual cells secretions located within a single cells shadow. 
Type Of Material Technology assay or reagent 
Year Produced 2023 
Provided To Others? Yes  
Impact This enables the identification of secretions from individual cells and importantly to visualise and quantify cells which are not secreting anything at all. This enables more robust and accurate quantification of the heterogeneity of secretions form cell populations. 
URL https://link.springer.com/protocol/10.1007/978-1-0716-3135-5_26
 
Title Mass cytometry analysis of peripheral blood and dissociated tumour cell (DTC) samples from renal and lung cancer patients 
Description Mass cytometry analysis of peripheral blood and dissociated tumour cell (DTC) samples from renal and lung cancer patients 
Type Of Material Database/Collection of data 
Year Produced 2023 
Provided To Others? Yes  
Impact Used in our publication, TIGIT can inhibit T cell activation via ligation-induced nanoclusters, independent of CD226 co-stimulation, Nature Communications, 2023 
URL https://www.nature.com/articles/s41467-023-40755-3#Sec37
 
Description Partnership with Bristol Myers Squibb 
Organisation Bristol-Myers Squibb
Country United States 
Sector Private 
PI Contribution Optimisation of CAR T cell therapies
Collaborator Contribution To provide deeper insights into the mechanisms of action for tandem CAR T cells, to inform functional outcomes, translational development, and optimize future tandem CAR T design.
Impact Providing deep insights into the mechanisms of action for tandem CAR T cells, to inform functional outcomes, translational development, and optimize future tandem CAR T design.
Start Year 2023
 
Description Partnership with GSK 
Organisation GlaxoSmithKline (GSK)
Country Global 
Sector Private 
PI Contribution Invetsigtaion of the TIGIT axis with GSK
Collaborator Contribution This is a PhD studentship funded by GSK to study the nanoscale organisatiuon of proteins involved in the TIGIT/CD96 axis
Impact This a PhD studentship and the student is working on this project until 2026
Start Year 2023
 
Description Partnership with UCL, Kwee Young 
Organisation University College London
Country United Kingdom 
Sector Academic/University 
PI Contribution We are imaging immune recognition by cells from patients.
Collaborator Contribution From UCLH, we obtain samples of bone marrow and peripheral blood lymphocytes from myeloma patients
Impact We aim to learn what happens to patients in the course of myeloma
Start Year 2024
 
Description Article in THE SUNDAY TIMES about the naming of diseases 
Form Of Engagement Activity A magazine, newsletter or online publication
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Public/other audiences
Results and Impact I published an article in THE SUNDAY TIMES about how we name diseases.
Year(s) Of Engagement Activity 2022
URL https://www.thetimes.co.uk/article/why-we-should-give-a-monkeys-about-the-names-we-give-diseases-p57...
 
Description Article in The Times Literary Supplement 
Form Of Engagement Activity A magazine, newsletter or online publication
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Media (as a channel to the public)
Results and Impact Trigger warning, by Daniel M. Davis, in The Times Literary Supplement. Available online here: https://www.the-tls.co.uk/articles/allergic-theresa-macphail-book-review-daniel-m-davis/
Year(s) Of Engagement Activity 2024
URL https://www.the-tls.co.uk/articles/allergic-theresa-macphail-book-review-daniel-m-davis/
 
Description Article published in The Times about the history of vaccination 
Form Of Engagement Activity A magazine, newsletter or online publication
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Public/other audiences
Results and Impact Feature article published in THE SUNDAY TIMES: A little more vaccination, Elvis and the race to beat polio, June 26th 2022
Year(s) Of Engagement Activity 2022
URL https://www.thetimes.co.uk/article/a-little-more-vaccination-elvis-presley-and-the-race-to-beat-poli...
 
Description Article published in WIRED magazine 
Form Of Engagement Activity A magazine, newsletter or online publication
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Public/other audiences
Results and Impact Article published in Wired magazine about microbiomes
Year(s) Of Engagement Activity 2023
URL https://www.wired.co.uk/article/microbes-health-science
 
Description BBC Radio 4, Best Medicine 
Form Of Engagement Activity A broadcast e.g. TV/radio/film/podcast (other than news/press)
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Media (as a channel to the public)
Results and Impact I was a guest on a live panel show, Best Medicine, in which I discussed how the immune system works, and why it is important
Year(s) Of Engagement Activity 2023
URL https://www.bbc.co.uk/programmes/m001ryf0
 
Description BBC Radio 4, The Infinite Monkey Cage 
Form Of Engagement Activity A broadcast e.g. TV/radio/film/podcast (other than news/press)
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Media (as a channel to the public)
Results and Impact I was on a live panel show for BBC Radio 4, with Prof. Brian Cox, Robin Ince, Sarah Gilbert and Chris can Tullekan, discussing what science has learnt from Covid, and what Covid has taught us about science.
Year(s) Of Engagement Activity 2023
URL https://www.bbc.co.uk/sounds/play/m001f4lp
 
Description Interview for The Times on Cancer treatments 
Form Of Engagement Activity A magazine, newsletter or online publication
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Public/other audiences
Results and Impact Article in the Times about cancer treatments features interview with me
Year(s) Of Engagement Activity 2022
URL https://www.thetimes.co.uk/article/advances-show-tide-is-turning-at-last-in-the-war-on-cancer-3fp57n...
 
Description Particiaption in 'Science Busking' events 
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 Two sessions with year-7 pupils from one of the Harris Academy schools in London. It's called "science busking" or "exciting science". They come to Imperial College for the day and we teach them "magic tricks" that have basic science principles behind them (friction, combustion, surface tension etc), and we explain that too. The goal is to get them to be excited about science and maybe learn something that will come up in their classroom later in the year, make them feel empowered etc.
Year(s) Of Engagement Activity 2023,2024
URL https://www.imperial.ac.uk/be-inspired/schools-outreach/wohl-reach-out-lab/our-activities/
 
Description Participation in The Brilliant Club 
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 A series of tutorials to high school students, from disadvantaged backgrounds, with the aim to widen participation in higher education.
Year(s) Of Engagement Activity 2023,2024
URL https://thebrilliantclub.org/