Molecure Sa · GPW:MOC
Molecure Sa — Investor Day (February 2024)
91 slides from Molecure Sa's investor day presentation, Feb 6, 2024
Slide 1: R&D Day
R&D Day Fate can be altered February 6, 2024
Slide 2: Disclaimer
Molecure SA (”MOC") and its Board and employees make no warranties or assurances regarding the accuracy or completeness of any representations or other materials contained herein. The information and data contained herein reflect the current conditions and assessments of the market situation as at t
Slide 3: Agenda
Agenda 13.00-13.05 (5 min) 13.45-14.00 (15 min) Welcome and Agenda Future of the mRNA platform Katarzyna Mucha, CCGroup dr Zbigniew Zasłona, CSO (Part 1) 14.00-14.20 (20 min) 13.05 – 13.15 (10 min) MoleCuring diseases by targeting RNA - methods for Introduction – Plans for 2024 identifying compounds
Slide 4: Molecure | Experienced company leadership
Molecure | Experienced company leadership Marcin Szumowski Zbigniew Zasłona Samson Fung Chairman of the Board & CEO Chief Scientific Officer Chief Medical Officer Sławomir Broniarek Barbara Dymek Chief Financial Officer R&D Operations Director 06/02/2024 4
Slide 5: Molecure | Scientific support of experts
Molecure | Scientific support of experts Paul Van der Horst, PhD Luke O’Neill, PhD President of the Supervisory Board Member of the Scientific Advisory Board A world expert on innate immunity and Dr Van der Horst has a strong track record of inflammation listed in the top 1% of biotech business deve
Slide 6: Plans and key upcoming catalysts
2024 Plans and key upcoming catalysts 06/02/2024 6
Slide 7: Molecure | Balanced pipeline
Molecure | Balanced pipeline Current status Clinical Candidate 2025 estimate Selection Molecular Target Lead In Vivo Preclinical Regulatory Target Indications Validation Optimization POC Development Process Phase I Phase II Hit-to-lead PROTEIN TARGETS OATD-01 Sarcoidosis, other OATD-01 OATD-01 CSR*
Slide 8: Molecure | Key near term milestones
Molecure | Key near term milestones OATD-01 OATD-02 o Reaching the therapeutic dose (Ph2RD) o First patient dosed in the KITE Phase (end of 2024 / early 2025) II PoC study (expected March) o Interim data analysis by an independent AI unblinded committee after completion o Following the implementatio
Slide 9: OATD-01 & OATD-02 Clinical update
OATD-01 & OATD-02 Clinical update 06/02/2024 9
Slide 10: OATD-01 | Phase 2 in sarcoidosis
OATD-01 | Phase 2 in sarcoidosis Double-blind, randomized, placebo-controlled multi-center study to assess the safety and efficacy of an oral inhibitor of CHIT1 (OATD-01) in patients with active pulmonary sarcoidosis. Central Bioethic Application FPFV (US/UK) / Intermediate FDA MHRA LPLV approval Co
Slide 11: OATD-01 | current status
OATD-01 | current status Approval from the U.S. FDA and Central Bioethics Commission Clinical trial in Europe: o Approval of Medicines and Healthcare products Regulatory Agency (MHRA) in UK o Resubmission for EU and Norway Site initations process in US and UK in final stage Grant application to PARP
Slide 12: OATD-02 | Phase I FIH clinical trial
OATD-02 | Phase I FIH clinical trial Design: Open-label single-arm Patient population (30-40 pts): dose-escalation monotherapy Relapsed/refractory advanced and/or study (Bayesian design, 2.5-30mg) metastatic solid tumors Location: 3 sites in Poland: OATD-02 | Addressing cancers with unmet Warsaw, Ot
Slide 13: OATD-02 | administered to the third cohort of patients
OATD-02 | administered to the third cohort of patients Sixth cohort Min. 3-6 patients Fifth cohort Min. 3-9 patients 30 mg Fourth cohort Min. 3-9 patients 25 mg Third cohort 101-115 weeks for 6 pts Min. 3 patients 2 weeks for SRC 20 mg Second cohort 78-99 weeks for 9 pts Min. 3 patients 2 weeks for
Slide 14: Deubiquitinase (DUBs): An important group of targets for anticancer therapies
Deubiquitinase (DUBs) An important group of targets for anticancer therapies 06/02/2024 14
Slide 15: USP7i | stabilizes p53 and upregulates p21 expression
USP7i | stabilizes p53 and upregulates p21 expression – we have utlilzed this mechanism in a context of T cell activation o The MDM2/MDMX-p53 circuitry plays a pivotal role in cell proliferation, cell cycle progression, apoptosis, and senescence o USP7 directly interacts with MDM2 and MDMX, regardle
Slide 16: USP7i | enhances anti-tumor CD4 T cell response
USP7i | enhances anti-tumor CD4 T cell response In isolated CD4 T cells from naive mice treated with USP7i and stimulated with CD3/CD28 for 3 days, the pre-harvest treatment led to an increase of IFNg a marker of Th1 response, and a decrease of both IL-4 and IL-13, markers of Th2 response. 06/02/202
Slide 17: USP7i | increases cytotoxicity of CD8+ T cells
USP7i | increases cytotoxicity of CD8+ T cells in tumor and spleen by increasing the expression of Granzyme B in the tumors and INFg in stimulated splenocytes Ex vivo analysis Tumor Stimulated splenocytes from OAT-4828 treated mice 06/02/2024 17
Slide 18: Increased killing of CT26 cells by effector cells from USP7i treated mice
Increased killing of CT26 cells by effector cells from USP7i treated mice Live CT26 OAT-4828 Green dye + Live CT26 Mice-treated splenocytes 36h co-culture Live/Dead staining And FACS analysis Effector cells isolated from the spleen of mice treated with USP7i for 20 days bear the potential to induce
Slide 19: USP7i | efficacy in in vivo experiments
USP7i | efficacy in in vivo experiments B16F10 melanoma E0771 breast A20 lymphoma CT26 colon Vehicle 1000 2500 Vehicle 2000 Vehicle Vehicle (mm3, median with IQR) (mm3, median with IQR) OAT-4828 (mm3, median with IQR) OAT-4828 (25 mg/kg) (mm3, median with IQR) 1000 800 OAT-4828 (50 mg/kg) OAT-4828 (
Slide 20: USP7i | strongly enhances IFN-γ production in vivo
USP7i | strongly enhances IFN-γ production in vivo B16F10 melanoma CTR CD4+IFNy+ T cells CTR CD8+IFNy+ T cells [%; median] of CD4+ [%; median] of CD8+ 20 50 ✱✱✱ ✱ 40 15 30 10 20 5 10 IFN-γ IFN-γ 0 0 veh 100 veh 100 OAT-4828 OAT-4828 [mg/kg] OAT-4828 OAT-4828 [mg/kg] CD4 CD8 06/02/2024 20
Slide 21: Significant survival increase in CT26 model after USP7i with aPD-1
Significant survival increase in CT26 model after USP7i treatment in combination with aPD-1 OAT-4828 αPD-1 αPD-1 Vehicle αPD-1 100 mg/kg + OAT-4828 (25 mg/kg) + OAT-4828 (100 mg/kg) Median survival 21 days 24 days Median survival 23 days 25 days 29 days Statistical significance ** Statistical signif
Slide 22: USP7i | boosts human T cells activation and proliferation
USP7i | boosts human T cells activation and proliferation Not activated CTR OAT-4828 (0.125 μM) human CD3+ T cells Read out after 72 hrs IL-2 CD69hi CD25hi + 1h OAT-4828 15000 ✱ 60 hIL2 [pg/ml] 10000 20 washed 40 % of CD8 % of CD8 15 5000 20 αCD3 + αCD28 0 10 CTR OAT-4828 0 5 0 uM 0.125 uM 0.25 uM 0
Slide 23: Human primary co-cultures validate anti-tumor properties of USP7i
Human primary co-cultures validates anti-tumor and immunomodulatory properties of USP7i In human primary co-culture of Fibroblasts Venular endothelial cells fibroblasts (stromal model) or venular + PBMCs + HT-29 (CRC) + PBMCs + HT-29 (CRC) endothelial cells (immune model) with PBMC and colorectal ca
Slide 24: Competitive landscape
Competitive landscape No USP7 has entered clinical development stage yet The main, active competitors, based on patent searches and business intelligence include: • RAPT Therapeutics - terminated • Hybrigenics • Ubiquigent • Almac Discovery • Forma Therapeutics • Shouyao Holdings • Servier • Schrödi
Slide 25: USP21 | attractive therapeutic target
USP21 | attractive therapeutic target USP21 controls STAT3 activation cancer cell metabolism. Genes of mitochondrial respiration chain complexes regulated by USP21 are marked in red. USP21 drives intracellular ATP production used by cancer cells for proliferation and migration 06/02/2024 25
Slide 26: USP21 | First lead-stage USP21i compounds inhibit STAT3 phosphorylation
USP21 | First lead-stage USP21i compounds inhibit STAT3 phosphorylation HAP-1 WT HAP-1 USP21 KO HAP-1 WT HAP-1 WT + DMSO + OAT cmpd 0 1 5 10 20 0 1 5 10 20 IL-6 (min) 86 pSTAT3 86 pSTAT3 56 USP21 45 β-actin 45 β-actin Pharmacological inhibition of usp21 results in blockage of STAT3 phosphorylation -
Slide 27: Competitive landscape
Competitive landscape The main, active competitors based on patent searches and business intelligence include: • Mission Therapeutics • Bayer 06/02/2024 27
Slide 28: Our small molecule mRNA discovery platform
Our small molecule mRNA discovery platform with potential to disrupt direct mRNA targeting approaches 06/02/2024 28
Slide 29: mRNA platform | discovery workflow
mRNA platform | discovery workflow PoC achieved PoC achieved Hit to Lead and Lead Druggable 3D Active Optimization Optimized In silico In silico Docking Small Validation Sequence Secondary structure Small Lead molecule compound SHAPE- structures CryoEM of RNA (virtual (wet lab) molecule MaP X-ray sc
Slide 30: mRNA platform | druggable regions selection
mRNA platform | druggable regions selection Druggable Secondary structures Robust structures: SHAPE-MaP, prediction Functionaly: • literature • omics data OR • verification experiments Conservation RNA isoforms analysis 06/02/2024 30
Slide 31: mRNA platform | two ways to the target
mRNA platform | two ways to the target PoC achieved Hit to Lead and With Druggable 3D Docking Active Lead Optimization Optimized In silico In silico Validation virtual Sequence Secondary structure Small molecule Small Lead compound screening SHAPE- MaP structures CryoEM X-ray of RNA (virtual screeni
Slide 32: mRNA platform | two ways to the target
mRNA platform | two ways to the target PoC achieved Hit to Lead and Lead Druggable 3D Active Optimization Optimized With HTS Sequence In silico Secondary In silico structure Docking Small Validation Small Lead DEL SHAPE- structures CryoEM of RNA (virtual molecule (wet lab) molecule compound MaP X-ra
Slide 33: mRNA platform | verification of binding in wet lab
mRNA platform | verification of binding in wet lab In silico Druggable In silico 3D Docking Active Hit to Lead and Small Validation Lead Optimized Sequence Secondary structure Small SHAPE- CryoEM (virtual molecule Optimization Lead structures of RNA (wet lab) molecule compound MaP X-ray screening) D
Slide 34: PoC First milestone achieved
PoC First milestone achieved 06/02/2024 34
Slide 35: mRNA platform | first step - druggable region selection
mRNA platform | first step - druggable region selection SHAPE-MaP Predicted robust structure Shannon entropy In vitro In vitro – in cellulo Conservation Region with a stable structure Publications CLIP-seq Protein coding RNA isoforms from patient’s tissues and cell lines contain the target region Fu
Slide 36: mRNA platform | wet lab screening cascade – biophysical tests
mRNA platform | wet lab screening cascade – biophysical tests 5.5M compounds AI-based preselection VS SPR ~ 30 HITs with KD < 100 µM 500k 376 VS HITs 5 Different chemotypes RNA-Targeted Library 6 HITs with KD ≤ 10 µM 06/02/2024 36
Slide 37: mRNA platform | preliminary MedChem analysis of HITs
mRNA platform | preliminary MedChem analysis of the properties of the obtained HITs o No PAINs o No aggregation o No redox o Molecular Weight < 500 g/mol o LogP < 5 o PSA < 100 Å2 o No solubility issues in assay o Reasonable IP space o No safety concerns (no hERG, no CYP inhibition, no genotoxicity)
Slide 38: mRNA platform | cellular tests (cytotoxicity)
mRNA platform | wet lab screening cascade – cellular tests (cytotoxicity) Compound no. 1 Compound no. 2 Compound no. 3 Compound no. 4 Resynthetized HITs SPR KD = 11 µM SPR KD = 31 µM SPR KD = 5 µM SPR KD = 1.3 µM Compound no. 1 Compound no. 2 Compound no. 3 Compound no. 4 24h EC50 >100 µM EC50 >100
Slide 39: mRNA platform | cellular tests (target protein level)
mRNA platform | wet lab screening cascade – cellular tests (target protein level) Compound no. 1 Compound no. 2 Compound no. 3 Compound no. 4 target 36h housekeeping protein Compound no. 1 OAT-5932 Compound no. 2 OAT-5944 Compound no. 3 OAT-5953 Compound no. 4 OAT-5954 target (normalized to β-actin)
Slide 40: mRNA platform | our path to success
mRNA platform | our path to success in discovering RNA targeting small molecules >20 confirmed hit compounds Screening results of > 100,000s small molecules Q4 2023 3 targets selected for structural analysis 30 targets selected Q3 2022 for computational analysis Q1 2022 Q2 2021 In vitro proof of con
Slide 41: The RNA space is advancing rapidly
The RNA space is advancing rapidly Targeting RNA with small molecules has led to multiple significant biotech / big pharma partnerships More than $920M up-front payments 54M/ 42M/ 540M 30M/ 1BLN More than $14B total deals in value 45M/ >2BLN 75M/ 1BLN 17M/ >1BLN In under 6 years 10M/ no data 120M/ 1
Slide 42: mRNA platform | multiple collaborations with RNA experts
mRNA platform | multiple collaborations with RNA experts Field of cooperation RNA bioinformatics Prof. Janusz Bujnicki, head of the Laboratory of Bioinformatics and Protein Engineering (LBIB) at the International Institute of Molecular and Cell Biology (IIMCB) in Warsaw. RNA secondary structure pred
Slide 43: MoleCuring diseases by targeting RNA - methods for identifying compounds
MoleCuring diseases by targeting RNA - methods for identifying compounds Joanna Sztuba-Solińska, PhD 06/02/2024 43
Slide 44: Why target RNA?
Why target RNA? o Most drugs on the market target enzymes (1.5% of human genome ~ 20,000 proteins) o Within 20,000 proteins encoded roughly 12% is disease related = 3,000 proteins o Currently approved drugs interact with ~ 700 proteins (0.05 of the genome) o Mutations, drug resistance o We have no o
Slide 45: RNA regulates myriad of biological processes!
RNA regulates myriad of biological processes! o Transcription of other RNAs and self Interactions with chromatin o Degradation of other RNAs and self Interactions with proteins o Translation = protein expression o Interaction with chromatin o Localization of other RNAs, proteins Interactions with pr
Slide 46: Why is RNA lagging behind?
Why is RNA lagging behind? o Viewed as a simple messenger (1947, Boivin and Vendrely) o 14 years later, mRNA was isolated for the first time (Brenner, Jacob, Meselson) o The discovery of regulatory RNAs began to change that view (T. Cech and S. Althman, 1980) o 1980 - first FDA-approved antibacteria
Slide 47: RNA is a challenging target
RNA is a challenging target Challenge 1: “simplicity” of RNA molecule Challenge 2: most RNAs exist not as static folds but as ensembles of structurally and temporally dynamic molecule Challenge 3: the polyanionic backbone of RNA makes it difficult to form a deep hydrophobic pocket that would provide
Slide 48: What makes RNA druggable?
What makes RNA druggable? 06/02/2024 48
Slide 49: Structures in human mRNAs regulate key biological processes
Structures in human mRNAs regulate key biological processes Synucleozid - binds an A bulge in the IRE of Branaplam – leads to the SNCA mRNA, inhibiting inclusion of an extra exon in the translation (Parkinson) HTT. This extra exon makes the mRNA molecule unstable, and it ends up getting rapidly dest
Slide 50: RNA binding pockets have unique properties
RNA binding pockets have unique properties o similar propertied to protein binding pockets, i.e., volume, buriedness (lack of solvent exposure), o recognition by small molecules is driven by: ✓ stacking interactions of nucleobases (34.8%) ✓ hydrogen bonding (34.4%) – RNA binding pockets have wide va
Slide 51: Good RNA targets must have high information content
Good RNA targets must have high information content 1. REVERSE TRANSCRIPTION cDNA with SHAPE induced mutations 2. PCR AMPLIFICATION Illumina adapters are added in LNPs 3. NEXT-GENERATION SEQUENCING in in vitro cellulo ↑ information content = ↑ structural complexity that specifies targets uniqueness
Slide 52: What about ligands aka small molecules?
What about ligands aka small molecules? o RNA-binding small molecules have unique properties: ✓ ↓ octanol–water partition coefficients than protein-binding compounds (measure of lipophilicity) ✓ ↑ topological polar surface areas (drug absorption) ✓ ↑ hydrogen bond donors and acceptors ✓ ↑ heteroatom
Slide 53: Strategies for RNA targeting: Framework 1
Strategies for RNA targeting Framework 1: RNA motifs used as modules for ligand binding Focused on modules of RNA structure that can be used as units in designing selective small molecules 1. Hergenrother lab - discovered wedge-shaped compounds binding RNA bulges and other compounds binding to octal
Slide 54: Strategies for RNA targeting: Framework 2
Strategies for RNA targeting Framework 2: RNA-biased chemical space RNA molecule is considered as a whole unique target that forms defined binding pockets that can be target by specifically designed small molecule 1. Scaffold-based synthesis - molecular scaffold known to interact with an RNA molecul
Slide 55: Strategies for RNA targeting: Framework 2
Strategies for RNA targeting Framework 2: RNA-biased chemical space 2. RNA-biased libraries – o focused on studying structural properties of RNA- binding molecules and using these properties to design RNA-biased libraries o These guiding principles can facilitate the future design of selective ligan
Slide 56: Strategies for RNA targeting: Framework 3
Strategies for RNA targeting Framework 3: RNA-targeted small molecules may look like typical drugs o Traditional medicinal chemistry approaches are applied to RNA targets, i.e., Lipinski’s and Veber’s rules o Determinants for RNA binding are sufficiently similar to those for protein targeting to war
Slide 57: Screening and validating assays
Screening and validating assays Fluorescence-based assays ✓Displacement of a fluorescent dye or compound by a small molecule, e.g., TO-PRO-1 ✓2-aminopurine – 2-AP is incorporated into the target RNA; its fluorescence depends on the microenvironment, which changes upon small-molecule binding. The pos
Slide 58: Screening and validating assays
Screening and validating assays Small molecule microarray o A library of 20,000 small molecules ( primary, secondary alcohols, amines) are printed onto a functionalized glass surface. o Then, the array is incubated with a Cy5-labeled target RNA molecule as well as another Cy5-labeled control RNA, ir
Slide 59: Screening and validating assays
Screening and validating assays DNA-encoded compound libraries ✓ small molecules are synthesized on beads, and each compound is encoded with a DNA tag. ✓ Compound-beads are screened for binding to a fluorescently labeled target, often in the presence of an off-target that is differentially labeled (
Slide 60: Screening and validating assays
Screening and validating assays Biophysical assays o surface plasmon resonance [SPR] - directly measures changes in resonance angle or wavelength of a glass surface containing the immobilized RNA of interest upon addition of a small molecule o microscale thermophoresis [MST] - measures the directed
Slide 61: Screening and validating assays
Screening and validating assays Molecular assays Difference in total transcript o EMSA- electrophoretic mobility shift assays expression of SMN- C3 (500 nM) versus o RNA-seq - Differential expression analysis from DMSO-treated SMA type I patient RNA-Seq can also be used to quantitatively fibroblasts
Slide 62: Screening and validating assays
Screening and validating assays Phenotypic screening o Identification of compounds that affect pathways associated with a specific phenotype. o Designed around a biological process, e.g., alternative splicing, translation o No knowledge of the RNA structure or small molecule’s mode of action is need
Slide 63: Thank you for your attention!
Thank you for your attention! The global RNA targeting small molecule drug discovery market size was valued at USD 1.1 billion in 2022 and is expected to expand at a compound annual growth rate (CAGR) of 20.9% from 2023 to 2030 06/02/2024 63
Slide 64: Studying RNA conformations with DyRNA Thermometry and cryo-EM
Studying RNA conformations with DyRNA Thermometry and cryo-EM Jakub Nowak, PhD 06/02/2024 64
Slide 65: What do we know about structured RNA domains
What’s do we know about structured RNA domains ~ 5.400 known 3D ~ 24.000.000 RNA ~ 150.000 RNA structures of RNA and sequences sequences with RNA-protein complexes experimentally > 4.000 non-coding determined 127 non-coding RNA RNA families (Rfam) secondary structure families 3 RNA Cryo-EM structure
Slide 66: Is there a single structure of a given RNA sequence
Is there a single structure of a given RNA sequence Ganser et al., Nat Rev Mol Biol 2019 06/02/2024 66
Slide 67: Fluorimetry based detection of conformational changes
Fluorimetry based detection of conformational changes External vs Internal fluorescence probes Introduction/Results/Summary inDyRNA Thermometry ExDyRNA Thermometry Temperature 06/02/2024 67
Slide 68: DyRNA Thermometry introduction
DyRNA Thermometry introduction Standard experimental set-up Introduction/Results/Summary 06/02/2024 68
Slide 69: DyRNA Thermometry principle
DyRNA Thermometry principle Definition of TRIC effect Introduction/Results/Summary 𝜕𝐹 Physical components: 𝑐 𝜕𝑇 Quantum Yield Peak wavelength (ie. Stoke shift) TRIC – temperature effect strongly dependent on fluorophore environment 06/02/2024 69
Slide 70: DyRNA Thermometry principle
DyRNA Thermometry principle Combining temperature effect on fluorescence and stability of the RNA Introduction/Results/Summary TRIC response coincide with the RNA unfolding 06/02/2024 70
Slide 71: DyRNA Thermometry principle
DyRNA Thermometry principle Combining temperature effect on fluorescence read-out and stability of the RNA Introduction/Results/Summary DyRNA Thermometry senses the stability of the RNA hairpin 06/02/2024 71
Slide 72: inDyRNA Thermometry sensitivity
inDyRNA Thermometry sensitivity Global stability of unlabeled and an internally labeled hairpins Introduction/Results/Summary External fluorescent probe Incorporation of the probe does not affect global stability readout 06/02/2024 72
Slide 73: inDyRNA Thermometry sensitivity
inDyRNA Thermometry sensitivity Label position effect Introduction/Results/Summary Position of the dye is linked to the stability readout 06/02/2024 73
Slide 74: inDyRNA Thermometry sensitivity
inDyRNA Thermometry sensitivity Stem extension effect Introduction/Results/Summary Extension of the probe-loop distance increases the stability 06/02/2024 74
Slide 75: inDyRNA Thermometry sensitivity
inDyRNA Thermometry sensitivity Loop size effect Introduction/Results/Summary Larger loop size is correlated with a decrease of the stability 06/02/2024 75
Slide 76: inDyRNA Thermometry sensitivity
inDyRNA Thermometry sensitivity Sensing direction of hairpin opening Introduction/Results/Summary Extension of the distance between the loop and the probe shows increase in stability 06/02/2024 76
Slide 77: inDyRNA Thermometry sensitivity
inDyRNA Thermometry sensitivity Global vs local stability sensing at the domain level Introduction/Results/Summary inDyRNA Thermometry allows for the specific local-domain probing of conformational changes 06/02/2024 77
Slide 78: inDyRNA Thermometry validation with biological models
inDyRNA Thermometry validation with biological models Function of riboswitches in cell biology Introduction/Results/Summary Hallber et al, Annu. Rev. Biochem. 2017 Kavita et al, Trends in Biochemical Sciences 2023 06/02/2024 78
Slide 79: inDyRNA Thermometry validation with biological models
inDyRNA Thermometry validation with biological models SAM-II Riboswitch conformational switch effect Introduction/Results/Summary inDyRNA Thermometry could be used to monitor higher order conformation rearrangements – hairpin->pseudoknot 06/02/2024 79
Slide 80: inDyRNA Thermometry validation with biological models
inDyRNA Thermometry validation with biological models SAM-II Riboswitch conformational switch effect Introduction/Results/Summary Core stability of the hairpin driven by Mg ions Pseudoknot stability cooperation of Mg and SAM 06/02/2024 80
Slide 81: inDyRNA Thermometry validation with biological models
inDyRNA Thermometry validation with biological models SAM-II Riboswitch conformational switch ligand specificity Introduction/Results/Summary inDyRNA Thermometry could be used to screen for ligands introducing conformational switch 06/02/2024 81
Slide 82: exDyRNA Thermometry analysis of tRNA modifications
exDyRNA Thermometry analysis of tRNA modifications Position specific stabilization of tGln with pseudouridylation Introduction/Results/Summary exDyRNA Thermometry identifies key Pus driven pseudouridylation spots Anna Biela 06/02/2024 82
Slide 83: Cryo-EM structures of unmodified tRNAs
Cryo-EM structures of unmodified tRNAs Introduction/Results/Summary tGln cryo-EM density display characteristic for tRNA features Anna Biela 06/02/2024 83
Slide 84: Cryo-EM structures of modified tRNAs
Cryo-EM structures of modified tRNAs Introduction/Results/Summary Introduction of Ψ leads to changes in the elbow region and distinct densities for the phosphates groups Anna Biela 06/02/2024 84
Slide 85: Local structural stabilization of tRNAs by Ψ
Local structural stabilization of tRNAs by Ψ Introduction/Results/Summary Ψ39 rigidifies the ASL Ψ55 induces the compaction of the elbow region Anna Biela 06/02/2024 85
Slide 86: inDyRNA Thermometry analysis of the Ψ local effects in tRNA
inDyRNA Thermometry analysis of the Ψ local effects in tRNA Introduction/Results/Summary Ψ13 stabilizes the proximal nucleotides around the core of tRNA 06/02/2024 86
Slide 87: inDyRNA Thermometry analysis of the Ψ local effects in tRNA
inDyRNA Thermometry analysis of the Ψ local effects in tRNA Introduction/Results/Summary Complete Ψ pattern leads to high level of destabilization of tRNA conformation 06/02/2024 87
Slide 88: Summary
Summary Introduction/Results/Summary Conformational dynamics of RNA could be probed with a site-specific fluorescent labeling and the temperature gradient • exDyRNA Thermometry global effect assessment • inDyRNA Thermometry domain effect assessment Pseuodouridylation placed at specific position in t
Slide 89: RNA domains as drug targets
RNA domains as drug targets Childs-Disney et al., Nat Rev Drug Discovery 2022 06/02/2024 89
Slide 90: Acknowledgements
Acknowledgements Glatt Lab Janusz Bujnicki (IIMCB Warsaw) Norhane Abbassi Anna Biela Magdalena Bożek Zhaoming Su (Sichuan University) Agnieszka Broniec Andrzej Chramiec-Głąbik Priyanka Dahate Andrea Flores Ibarra Monika Gaik Tomasz Gawda Przemyslaw Grudnik Sebastian Guzman Perez Alexander Hammermeis
Slide 91: Q&A
Q&A 06/02/2024 91