I04-1-Macromolecular Crystallography (fixed wavelength)
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Michael
Berlin
,
Jennifer
Cantley
,
Fabio
Broccatelli
,
Lin
Cao
,
Huifen
Chen
,
Tommy K.
Cheung
,
Andrew P.
Crew
,
Dean
Dinicola
,
Hanqing
Dong
,
Matthew
Grimmer
,
Brian D.
Hamman
,
Alicia
Harbin
,
Mingtao
He
,
Xiuxian
Hu
,
Alison J.
Hole
,
Thomas
Januario
,
Philip S.
Kerry
,
Xiangjia
Liu
,
Connor
Quinn
,
Christopher M.
Rose
,
Emma
Rousseau
,
Lawrence B.
Snyder
,
Leanna R.
Staben
,
Gan
Wang
,
Jing
Wang
,
Xiaofen
Ye
,
Robert L.
Yauch
,
Peter S.
Dragovich
Diamond Proposal Number(s):
[23279]
Open Access
Abstract: Modification of the VHL-binding fragments contained in proteolysis targeting chimeras (PROTACs) that potently degrade the BRM protein (also known as SMARCA2) improved degradation selectivity over the closely-related paralog protein BRG1 (SMARCA4). In particular, replacement of the phenyl-thiazole entity commonly employed in the generation of VHL-dependent PROTACs with pyridyl-thiazole, phenyl imidazole, and phenyl-nitrile moieties consistently improved the BRG1/BRM degradation selectivity ratios of multiple, structurally-diverse degrader compounds. Crystal structures of these new VHL-binding fragments in complex with the VHL protein were obtained to better understand their interactions. Some of these VHL alterations, the phenyl-nitrile substitution in particular, afforded molecules that displayed strong antiproliferative activities against BRM-dependent (BRG1-mutant) cancers but minimal potency toward wild-type cell lines. One such compound (21, G-9293) was profiled in detailed broad proteomics and chromatin accessibility experiments, and its biological properties were clearly differentiated from a less-selective BRM-degrader (5, A947) in the latter assessment. The highly selective molecule (21, G-9293) was also extensively profiled in vitro using a panel of lung cancer cell lines (defined by BRG1 or BRM status) along with several prostate cancer lines. It exhibited similar antiproliferation activity relative to the less-selective BRM-degrader (5, A947) against the lung lines but significantly diminished potency toward the prostate cancer cells.
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May 2026
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I04-Macromolecular Crystallography
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Philipp
Münick
,
Dimitrios-Ilias
Balourdas
,
Julianne S.
Funk
,
Büşra
Yüksel
,
Danai
Mavridi
,
Justin
Heftel
,
Birgit
Dreier
,
Jonas V.
Schaefer
,
Birgit
Schäfer
,
Stefan
Knapp
,
Tümay
Telatar
,
Baki
Akgül
,
Andreas
Plückthun
,
Thorsten
Stiewe
,
Andreas C.
Joerger
,
Volker
Dötsch
Abstract: The tumor suppressor p53 is the most frequently mutated protein in tumors and a target for drug development. More than 2000 cancer-associated p53 missense mutations have been reported, most of them located in the DNA-binding domain (DBD). Due to the low intrinsic thermostability of the latter, they often lead to unfolding at physiological temperature. Stabilizing the DBD with small molecules has been shown to be effective in reactivating the cavity-creating cancer mutant Y220C. Unfortunately, the majority of p53 mutants seem to lack druggable binding pockets for small molecules. Here we show that a designed ankyrin repeat protein (DARPin) that binds to the p53 DBD stabilizes temperature-sensitive (TS) p53 cancer mutants, thereby compensating for mutation-induced loss of stability. We determined high-resolution crystal structures of multiple DARPin–mutant p53 complexes, providing mechanistic insights into this mode of stabilization. Reporter gene assays across a comprehensive panel of cancer-associated mutants revealed reactivation of the majority of TS mutants, whereas DNA-contact mutants and those with local misfolding of the DNA-binding surface remained inactive, as expected. We demonstrate that this reactivation induces the transcription of canonical p53 target genes and elicits antiproliferative effects in cancer cell lines. A combination of this DARPin with an mRNA/lipid nanoparticle-based transfection approach may have the potential to reactivate most TS p53 mutants and resensitize cancer cells to chemotherapy.
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May 2026
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I13-2-Diamond Manchester Imaging
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Karo
De Rycke
,
Marina
Horvat
,
Lisa
Caboor
,
Petra
Vermassen
,
Griet
De Smet
,
Sophie
Lobbestael
,
Marta
Santana Silva
,
Wouter
Steyaert
,
Matthias
Van Impe
,
Patrick
Segers
,
Julie
De Backer
,
Patrick
Sips
Diamond Proposal Number(s):
[32919]
Open Access
Abstract: Fibrillin defects lead to severe cardiovascular complications in Marfan syndrome (MFS), including aortic dilation, dissection, and rupture. To model MFS, zebrafish mutants lacking various fibrillin genes were generated. Among these mutant lines, only fibrillin-3–deficient zebrafish exhibited cardiovascular phenotypes mimicking human disease. Multimodal imaging revealed early cardiac defects, bulbus arteriosus dilation, and valve abnormalities. Transcriptomic analysis identified altered regulation of pathways related to extracellular matrix homeostasis and immune system activation. This zebrafish model, recapitulating key cardiovascular features of MFS, provides a valuable platform to investigate disease mechanisms and identify novel treatment strategies.
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May 2026
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Holger
Greschik
,
Florian
Friedrich
,
Ludwig
Seifert
,
Farnoush
Mousavizadeh
,
Francesco
Fiorentino
,
Johannes
Walz
,
Lin
Zhang
,
Jianyu
Li
,
Emanuele
Fabbrizi
,
Stefano
Tomassi
,
Farhad
Panahi
,
Niklas
Papenkordt
,
Silas L.
Wurnig
,
Johannes
Osterroth
,
Anna M.
Strasser
,
Jan
Ruprecht
,
Aurélien F. A.
Moumbock
,
Martin
Hügle
,
Manuela
Sum
,
Ling
Peng
,
Sheng
Wang
,
Adina A.
Baniahmad
,
Laura
Pulido-Cortés
,
H. Th. Marc
Timmers
,
Ralf
Flaig
,
Eric
Metzger
,
Bernhard
Breit
,
Oliver
Einsle
,
Stefan
Günther
,
Dante
Rotili
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Antonello
Mai
,
Roland
Schüle
,
Manfred
Jung
Open Access
Abstract: The chromatin remodeler CHD1, a regulator of gene activity and potential drug target in prostate cancer (PCa), contains a tandem chromodomain (tCD) binding histone H3 trimethylated at lysine 4 (H3K4me3). We developed the first submicromolar inhibitors (2n and 2s) that target the H3K4me3 binding site of the CHD1 tCD with Kd values of 0.15 μM and 0.14 μM, respectively. Co-crystal structures of these quinoline-based compounds revealed aromatic cage interactions and extended ligand contacts in other parts of the H3K4me3 peptide pocket as the main determinants of high-affinity ligand binding. 2n and 2s engage endogenous CHD1 in cell lysates or the exogenous CHD1 tCD in cells. Furthermore, we provide evidence for selectivity against a panel of methyl-lysine readers and epigenetic enzymes as well as impairment of PCa cell viability. Due to their high potency and defined binding mode, our ligands offer new directions for further optimization.
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May 2026
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Krios IV-Titan Krios IV at Diamond
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Rosie M.
Mundy
,
Kasim
Waraich
,
Emily A.
Bates
,
Pierre J.
Rizkallah
,
Alexander T.
Baker
,
Mark T.
Young
,
Edward
Morris
,
Paula C. A.
Da Fonseca
,
Carly M.
Bliss
,
David
Matthews
,
David
Bhella
,
Alan L.
Parker
Diamond Proposal Number(s):
[31827]
Open Access
Abstract: Adenoviruses are widely used as vectors for subunit vaccines and oncolytic therapies. Efficient vectors must infect target cells and deliver therapeutic transgenes at high levels. Species D adenoviruses, such as human adenovirus type 10 (HAdV-D10), are promising candidates due to low seroprevalence in humans. Here, we present the cryo-electron microscopy structure of the HAdV-D10 capsid alongside transcriptomic profiling of infected cells to inform vector development. The fiber shaft, essential for cell entry, was resolved at 10 Å, revealing a previously uncharacterized ‘umbrella’ motif. Viral transcript analysis using an ORF-centric pipeline uncovered key differences from HAdV-C5, including abundant expression of a transcript encoding a protein equivalent to mature protein VII. These findings highlight the importance of detailed vector characterization prior to clinical translation and support the advancement of HAdV-D10 as a next-generation platform for gene delivery and vaccine development.
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Apr 2026
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Krios I-Titan Krios I at Diamond
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Brajabandhu
Pradhan
,
Senthil T.
Kumar
,
Jessica
Wagner
,
Rodrigo
Gallardo
,
Gabriele
Orlando
,
Matthias
De Vleeschouwer
,
Valentina
Zorzini
,
Jillian
Madine
,
Nikolaos
Louros
,
Jonas J.
Neher
,
Frederic
Rousseau
,
Joost
Schymkowitz
Diamond Proposal Number(s):
[41018]
Open Access
Abstract: Amyloid assembly in vivo occurs in complex environments where multiple aggregation-prone species coexist. Aβ and medin are prevalent amyloids in ageing humans that co-localize in cerebral amyloid angiopathy (CAA), yet their structural interactions remain poorly understood. Here, using cryo-electron microscopy, we determine high-resolution fibril structures from in vitro mixtures of Aβ40 and medin. From the same reaction, we resolve three distinct fibril populations: (i) a previously characterized Aβ40 polymorph that also forms in isolation, (ii) a Aβ40 polymorph with Aβ42-like features, including ordered N- and C-terminal regions, and (iii) the atomic structure of full-length medin fibrils. Biochemical and immunogold analyses demonstrate Aβ–medin association within mixed assemblies, though medin is not resolved within the ordered Aβ core. These findings support two non-exclusive mechanisms: transient heterotypic interactions redirecting Aβ folding, or partial medin incorporation into fibril architecture. Our data reveal how coexisting amyloids remodel each other’s polymorphic landscapes.
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Apr 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
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Robert L.
Hudkins
,
Eric
Allen
,
Samhita
Iyer
,
Alexandra
Balcer
,
Melissa
Neal
,
Qing
Ye
,
Marc
Rideout
,
Caleb B.
Frye
,
Kirk J.
Nelson
,
Isaac D.
Hoffman
,
Jacqueline H.
Starrett
,
Todd
Harris
,
Ronald V.
Swanson
,
Daniel C.
Bensen
Diamond Proposal Number(s):
[41212]
Abstract: Genetic alterations in FGFR2 drive multiple malignancies, most notably intrahepatic cholangiocarcinoma, where they occur in ∼10–15% of patients. While approved pan-FGFR inhibitors provide clinical benefit, their durability is limited by acquired, often polyclonal, on-target resistance mutations affecting key regions of the FGFR2 kinase domain, including the gatekeeper residue (V565), molecular brake residues (N550, E566, K642), and other key variants. These liabilities motivate the development of next-generation inhibitors. Given FGFR2-associated toxicities and the need for subtype selectivity, FGFR4 inhibition was prioritized as a selectivity determinant, while sparing FGFR1 was considered less critical. Guided by structure-based drug design, a reversible aminopyrimidine screening hit was optimized into a novel covalent inhibitor series active against FGFR2 wild-type and clinically relevant resistance mutations. An advanced lead 13 showed favorable potency, ADME properties, and demonstrated proof-of-concept in vivo efficacy in an FGFR2-amplified xenograft model comparable with the standard of care.
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Apr 2026
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I04-Macromolecular Crystallography
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Abstract: The design of irreversible drugs has resulted, over the past decade, in several new therapeutics in oncology that present improved pharmacodynamic and pharmacoKInetic properties compared to reversible ligands. Nevertheless, most ligands to date are designed to target a cysteine (Cys) residue, which is not a very common amino acid and only rarely occurs in protein target binding sites, thereby limiting the applicability of this covalent targeting approach. Recent work from our laboratory and others suggests that, after Cys, histidine (His) residues can be particularly suitable for covalent substitution with proper electrophiles. Using a ligand-first, structure-based approach, we assessed the possibility of using different electrophiles including acrylamides, chloroacetamides, or aryl fluorosulfates to target His residues covalently. Targeting His224 of hMcl-1 with model peptides, we demonstrate that both aryl fluorosulfates and chloroacetamides can be used to target His residues efficiently. Our studies also report strategies and biophysical approaches useful for the design and characterization of such His-covalent agents.
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Apr 2026
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I04-1-Macromolecular Crystallography (fixed wavelength)
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Diamond Proposal Number(s):
[25402]
Open Access
Abstract: Human protein kinase CK2 is a constitutively active serine/threonine kinase implicated in numerous cancers. Although ATP-competitive inhibitors such as CX-4945 show therapeutic potential, they are limited by off-target effects and incomplete or transient CK2 suppression. PROTACs offer an alternative strategy by inducing proteasome-mediated degradation, with potential advantages in potency, selectivity, and duration of action. Herein, a series of CK2-targeting PROTACs has been designed and synthesised. By conjugating a CAM4066-derived warhead to CRBN or VHL ligands, four VHL-recruiting PROTACs, were prepared using PEG and alkyl linkers, alongside two CRBN-recruiting analogues featuring constrained linkers. A ligand–linker analogue in which a linker is projected from the solvent-exposed region of CK2α retained binding affinity comparable to CAM4066, confirming that linker installation is tolerated and preserves key interactions in the αD and ATP sites.
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Apr 2026
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