DIAD-Dual Imaging and Diffraction Beamline
I13-2-Diamond Manchester Imaging
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Sarah
Davidson
,
Davide
Simone
,
Kathrin
Jansen
,
Max
Cowan
,
Caio
Machado
,
Ian
Reekie
,
Ananya
Bhalla
,
Rowie
Borst
,
Cesar
Prada Medina
,
Joshua
Bull
,
Zhi Yi
Wong
,
Sarah
Hill
,
Micon
Garvilles
,
Sam
Pledger
,
Patricia Reis
Nisa
,
Nora Rebecca
Schwingen
,
Dylan
Windell
,
Moustafa
Attar
,
Catherine
Disney
,
Andrew J.
Bodey
,
Alissa
Parmenter
,
Helen
Byrne
,
Sharif
Ahmed
,
Shashidhara
Marathe
,
Peter
Lee
,
Chris
Mahony
,
Adam P.
Croft
,
Stephen
Sansom
,
Mark C.
Coles
,
Christopher D.
Buckley
Diamond Proposal Number(s):
[30542, 34348]
Open Access
Abstract: The cellular basis for site-specific inflammation remains unclear. In human fingers, proximal interphalangeal (PIP) joints are preferentially affected by inflammatory arthritis, whereas distal interphalangeal joints are spared, providing a model to investigate the predilection of inflammation to distinct sites. Here we combine single-cell RNA sequencing, imaging and X-ray tomography to examine cellular composition, spatial organization and structure of finger joints during fetal development. PIP joints had a larger synovial volume and were enriched for PI16+ ‘universal’ fibroblasts. These cells were located in perivascular regions and at developing tendon–ligament interfaces. PI16+ fibroblasts exhibited both a shared inflammatory and cell-type-specific response to cytokine stimulation, suggesting that the combination of their spatial location and transcriptional responses promote inflammation. We suggest that differences in the stoichiometry of mesenchymal cells established in utero, including the key role of PI16+ fibroblasts, is a general principle that drives inflammation susceptibility across tissues.
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Jun 2026
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I03-Macromolecular Crystallography
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Christopher P.
Tinworth
,
Justyna
Wojno-Picon
,
Michael
Adam
,
Stephan
Gade
,
Ashley P.
Hancock
,
David J.
Hirst
,
Jonathan P.
Hutchinson
,
Semra
Kitchen
,
Despoina
Koumantou
,
Jonathan
Lea
,
Stephanie
Lehmann
,
John
Liddle
,
Richard
Lonsdale
,
Margarete
Neu
,
Leng
Nickels
,
Alex
Phillipou
,
James E.
Rowedder
,
Paul
Rowland
,
Jessica L.
Schneck
,
Paul
Scott-Stevens
,
Hester
Sheehan
,
Michael
Steidel
,
Chloe L.
Tayler
,
Ioannis
Temponeras
,
Kevin
Thang
,
David F.
Tough
,
Giovanni
Vitulli
,
Ian D.
Wall
,
Robert J.
Young
,
Nico
Zinn
,
Simon
Peace
,
Efstratios
Stratikos
Diamond Proposal Number(s):
[37907]
Abstract: Endoplasmic reticulum aminopeptidase 1 (ERAP1) regulates immune responses by proteolytically processing peptides presented by major histocompatibility class I molecules (MHC-I). ERAP1 can reduce the immunogenicity of cancer cells by destroying cancer-associated antigenic peptides or contribute to autoimmunity by generating self-antigenic peptides. ERAP1 inhibition has emerged as a tractable approach for cancer immunotherapy and specific classes of autoimmune diseases. We describe the discovery of a potent and selective ERAP1 inhibitor that targets its regulatory allosteric site. The compound has favorable in vivo pharmacokinetics, oral bioavailability, can regulate the immunopeptidome of cancer cells, and enhance tumor antigenicity in vivo controlling growth. When administered in the murine collagen-induced arthritis model, we observed no exacerbation of autoimmune responses but rather a dose-dependent therapeutic benefit. Our results demonstrate that ERAP1 inhibition is a tractable approach to modulating immune responses, provide mechanistic insight, and are valuable in vivo tools for interrogating ERAP1 biology and further drug development.
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Jun 2026
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I03-Macromolecular Crystallography
I04-1-Macromolecular Crystallography (fixed wavelength)
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Diamond Proposal Number(s):
[39203]
Abstract: Tissue inhibitor of metalloproteinases-1 (TIMP-1) is a critical regulator of extracellular matrix remodelling and an important mediator of remyelination in demyelinating disorders such as multiple sclerosis. In addition, TIMP-1 has emerged as a promising therapeutic target in cancer due to its interaction with CD63, which promotes tumorigenic signalling and carcinogenesis. Although several structures of TIMP-1 bound to matrix metalloproteinases have been reported, no unbound structure with all druggable sites available has previously been reported. Here, we present the first unbound crystal structure of human TIMP-1, resolved at 1.95 Å resolution. Comparison with the MMP-bound complex reveals localized conformational changes and altered intramolecular hydrogen bonding in the unbound structure, indicating increased structural plasticity in the absence of the protease. Crystals were obtained in multiple conditions, but only two diffracted to high resolution. Although optimization and seeding did not significantly improve the morphology, the additive screen enhanced both the morphology and reproducibility and provided intrinsic cryoprotection. The resulting crystal form proved compatible with soaking-based screening campaigns, providing a robust structural basis for the discovery of TIMP-1 ligands with clinical potential.
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Apr 2026
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I03-Macromolecular Crystallography
I23-Long wavelength MX
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Céline
Zheng-Gérard
,
Jana
Joha
,
Maria
Carrasquero
,
Kamel
El Omari
,
Edward
Lowe
,
Shirish
Dubey
,
Simon J.
Draper
,
Yu-Chi
Chang
,
Hsi-Hsien
Lin
,
Alan D.
Salama
,
Kirsty
Mchugh
,
Elena
Seiradake
Diamond Proposal Number(s):
[18069]
Open Access
Abstract: Granulomatosis with polyangiitis is a life-threatening systemic vasculitis, characterised by anti-neutrophil cytoplasmic autoantibodies (ANCA) most commonly against proteinase 3 (PR3), a protease expressed intracellularly and on the surface of neutrophils. Most cell surface PR3 is bound to the receptor CD177; however, the molecular mechanism of the interactions is not well understood. Here, we present crystal structures of CD177 in complex with PR3 and unliganded CD177. We describe a mainly hydrophobic binding interface between PR3 and CD177, involving the first two Ly6/uPAR (LU) domains of CD177. These form a globular structure which is connected to downstream domains via a flexible linker. Using a panel of PR3-ANCA-positive patient samples, we show that a significant proportion of ANCAs target the CD177-binding site of PR3 in these samples. Structure-guided mutation of the CD177-binding site on PR3 is effective in reducing PR3-ANCA binding. The results demonstrate that the CD177-binding surface of PR3 harbours a major PR3-ANCA epitope, and that the extent of binding to this surface varies between different patients.
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Feb 2026
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I04-Macromolecular Crystallography
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Thembaninkosi
Gaule
,
Katie J.
Simmons
,
Kieran
Walker
,
Francesco
Del Galdo
,
Rebecca L.
Ross
,
Hema
Viswambharan
,
Jahnavi
Krishnappa
,
Jack
Pacey
,
Martin
Mcphillie
,
Darren C.
Tomlinson
,
Azhar
Maqbool
Open Access
Abstract: Tissue fibrosis is a hallmark of systemic sclerosis (SSc) and results from the persistent activation of fibroblasts and excessive accumulation of extracellular matrix component such as collagen. Recent evidence implicates the matricellular protein Tenascin-C (TNC) in promoting self-sustaining fibroblast activation and fibrosis via its interaction with Toll-like receptor 4 (TLR4). In this study, we utilized Adhiron-guided ligand discovery to identify small molecule inhibitors targeting the fibrinogen-like globe domain of TNC, a key mediator of TLR4 activation. Two lead compounds (464 and 830) demonstrated structural similarity, favourable ADME profiles, and robust anti-fibrotic activity in vitro. Treatment of dermal fibroblasts derived from SSc patients with either compound significantly reduced Transforming growth factor-β-induced expression of fibrotic genes, ACTA2, COL1A1, COL1A2, and CCN2, and inhibited myofibroblast differentiation. These studies may facilitate the development of effective targeted therapy for fibrosis in SSc and support this novel strategy for small molecule development.
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Dec 2025
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I24-Microfocus Macromolecular Crystallography
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Tosca
Holtrop
,
Arianne M.
Brandsma
,
Louris J.
Feitsma
,
Steffen
Krohn
,
Petra
Moerer
,
Frederique
Van Den Haak
,
Anouk
Versnel
,
Leonie
Voss
,
Elsemieke M.
Passchier
,
Maaike
Nederend
,
J. H. Marco
Jansen
,
Anouk G.
Van Mourik
,
Rolf T.
Urbanus
,
Diane
Van Der Woude
,
Roger E. G.
Schutgens
,
Rene E. M.
Toes
,
Bert J. C.
Janssen
,
Anja
Lux
,
Kevin
Budding
,
Matthias
Peipp
,
Jeanette H. W.
Leusen
Open Access
Abstract: Overactivation of FcγRI by immune complexes (IC) is implicated in various autoimmune disorders and neuropathies. Currently, no effective FcγRI-specific blocking antibodies are available. Here we report preclinical data revealing two anti-FcγRI antibodies, C01 and C04, with high affinity, Fab-mediated binding within the IgG binding site on extracellular domain 2 of FcγRI. Both C01 and C04 block 90% of IgG and IC binding, and displace ~60% of pre-bound ICs without activating FcγRI, thereby minimizing the risk of aggravating inflammation. In the context of autoimmunity, C01 and C04 inhibit RA patient-derived autoantibody-IC binding to monocytes, macrophages and activated neutrophils, meanwhile they also inhibit the binding of opsonized platelets to monocytes from patients with immune thrombocytopenia. In vivo, C01 and C04 reduce IgG-dependent platelet depletion in humanized immunodeficient FcRγ-/- mice. Structural studies confirm that C01 and C04 achieve their blocking effects through Fab-mediated binding to FcγRI. Our data thus suggest that C01 and C04 may offer therapeutic potential for autoimmune disorders.
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Nov 2025
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VMXi-Versatile Macromolecular Crystallography in situ
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Martina
Steglich
,
Nicole
Larrieux
,
Ari
Zeida
,
Joaquín
Dalla Rizza
,
Sonia R.
Salvatore
,
Mariana
Bonilla
,
Matías N.
Möller
,
Alejandro
Buschiazzo
,
Beatriz
Alvarez
,
Francisco J.
Schopfer
,
Lucía
Turell
Diamond Proposal Number(s):
[33300]
Open Access
Abstract: Nitroalkene fatty acids (NO2-FAs) are formed endogenously. They regulate cell signaling pathways and are being developed clinically to treat inflammatory diseases. NO2-FAs are electrophilic and form thioether adducts with glutathione (GSH), which are exported from cells. Glutathione transferases (GSTs), a superfamily of enzymes, contribute to the cellular detoxification of hydrophobic electrophiles by catalyzing their conjugation to GSH. Herein, we evaluated the capacity of five human GSTs (M1-1, M2-2, M4-4, A4-4, and P1-1) to catalyze the reaction between nitrooleic acid (NO2-OA) and GSH. The reaction was monitored by HPLC-ESI-MS/MS and catalytic activity was detected with hGSTs M1-1 and A4-4. Using stopped-flow spectrophotometry, a 1400 and 7500-fold increase in the apparent second-order rate constant was observed for hGST M1-1 and hGST A4-4, respectively, compared to the uncatalyzed reaction (pH 7.4, 25 °C), in part due to a higher availability of the thiolate. The crystal structure of hGST M1-1 in complex with the adduct was solved at 2.55 Å resolution, revealing that the ligand was bound within the reaction center, and establishing a foundation to build a model of hGST A4-4 in complex with the adduct. A larger number of interactions between the enzyme and the fatty acid were observed for hGST A4-4 compared to hGST M1-1, probably contributing to the increased catalysis. Altogether, these results show, for the first time, that hGSTs can catalyze the reaction between GSH and NO2-FAs, likely affecting the signaling actions of these metabolites and expanding the repertoire of GST reactions.
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Feb 2025
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I03-Macromolecular Crystallography
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Simon
Nicolle
,
Mike
Barker
,
John
Barrett
,
Matthew
Campbell
,
Justyna
Wojno-Picon
,
Stephen J.
Atkinson
,
Helen
Aylott
,
Hripsimee
Kessedjian
,
Yanan
He
,
Cassie
Messenger
,
Emma
Roberts
,
Claus
Spitzfaden
,
Joelle
Le
,
Nico
Zinn
,
Thilo
Werner
,
Birgit
Dümpelfeld
,
Marcus
Bantscheff
,
Don O.
Somers
,
Heather
Reid
,
Kevin
Thang
,
Thomas
Gobbetti
,
Huw D.
Lewis
Diamond Proposal Number(s):
[29352]
Abstract: Therapeutics promoting the endogenous production of IL-10 have the potential to restore homeostasis in inflammatory disorders such as inflammatory bowel disease (IBD). Here we describe the identification of a series of IL-10 upregulators based on a pyrimidyl-piperidine scaffold through a high throughput phenotypic CD4+ T-cell multiplex assay. In vitro optimization of the initial hit yielded a lead with good potency and an in vitro clearance profile, compound 3–7, which additionally demonstrated efficacy in a murine endotoxin challenge PK–PD mechanistic model. Target deconvolution efforts identified compound 3–7 as a highly selective CDK8/19 inhibitor, and crystallographic studies unveiled its binding mode to the CDK8/Cyclin-C complex, characterized by an unusual water-mediated hydrogen bond to the kinase hinge region.
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Jan 2025
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I04-Macromolecular Crystallography
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Oscar
Mammoliti
,
Christel
Menet
,
Céline
Cottereaux
,
Javier
Blanc
,
Ann
De Blieck
,
Ghjuvanni
Coti
,
Raphaël
Geney
,
Line
Oste
,
Koen
Ostyn
,
Adeline
Palisse
,
Evelyne
Quinton
,
Benoit
Schmitt
,
Monica
Borgonovi
,
Isabelle
Parent
,
Catherine
Jagerschmidt
,
Steve
De Vos
,
Béatrice
Vayssiere
,
Miriam
López-Ramos
,
Kenji
Shoji
,
Reginald
Brys
,
David
Amantini
,
René
Galien
,
Caroline
Joannesse
Open Access
Abstract: Janus kinase (JAK) inhibitors have gathered interest as treatments for several inflammatory and autoimmune diseases. The four first marketed inhibitors target JAK1, with varying selectivity towards other JAK family members, but none inhibit tyrosine kinase-2 (TYK2) at clinically relevant doses. TYK2 is required for the signaling of the interleukin (IL)-12 and IL-23 cytokines, which are key to the polarization of TH1 and TH17 cells, respectively; two cell subtypes that play major roles in inflammatory diseases. Herein, we report our effort towards the optimization of a potent and selective dual JAK1/TYK2 inhibitor series starting from a HTS hit. Structural information revealed vectors required to improve both JAK1 and TYK2 potency as well as selectivity towards JAK2. The potent inhibition of both JAK1 (3.5 nM) and TYK2 (5.7 nM) in biochemical assays by our optimized lead compound, as well as its notable selectivity against JAK2, were confirmed in cellular and whole blood assays. Inhibition of TYK2 by the lead compound was demonstrated by dose-dependent efficacy in an IL-23-induced psoriasis-like inflammation mouse model.
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Nov 2024
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I03-Macromolecular Crystallography
I04-Macromolecular Crystallography
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Garry
Dolton
,
Anna
Bulek
,
Aaron
Wall
,
Hannah
Thomas
,
Jade R.
Hopkins
,
Cristina
Rius
,
Sarah A. E.
Galloway
,
Thomas
Whalley
,
Li Rong
Tan
,
Théo
Morin
,
Nader
Omidvar
,
Anna
Fuller
,
Katie
Topley
,
Md Samiul
Hasan
,
Shikha
Jain
,
Nirupa
D’souza
,
Thomas
Hodges-Hoyland
,
Owen B.
Spiller
,
Deborah
Kronenberg-Versteeg
,
Barbara
Szomolay
,
Hugo A.
Van Den Berg
,
Lucy C.
Jones
,
Mark
Peakman
,
David K.
Cole
,
Pierre J.
Rizkallah
,
Andrew K.
Sewell
Diamond Proposal Number(s):
[10462, 18812]
Open Access
Abstract: CD8+ T cells destroy insulin-producing pancreatic β cells in type 1 diabetes through HLA class I–restricted presentation of self-antigens. Combinatorial peptide library screening was used to produce a preferred peptide recognition landscape for a patient-derived T cell receptor (TCR) that recognized the preproinsulin-derived (PPI-derived) peptide sequence LWMRLLPLL in the context of disease risk allele HLA A*24:02. Data were used to generate a strong superagonist peptide, enabling production of an autoimmune HLA A*24:02–peptide–TCR structure by crystal seeding. TCR binding to the PPI epitope was strongly focused on peptide residues Arg4 and Leu5, with more flexibility at other positions, allowing the TCR to strongly engage many peptides derived from pathogenic bacteria. We confirmed an epitope from Klebsiella that was recognized by PPI-reactive T cells from 3 of 3 HLA A*24:02+ patients. Remarkably, the same epitope selected T cells from 7 of 8 HLA A*24+ healthy donors that cross-reacted with PPI, leading to recognition and killing of HLA A*24:02+ cells expressing PPI. These data provide a mechanism by which molecular mimicry between pathogen and self-antigens could have resulted in the breaking of self-tolerance to initiate disease.
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Sep 2024
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