The neuroscience landscape is being reshaped, and psychedelics are at the centre of this transformation. Once seen as controversial, compounds like psilocybin, MDMA, and ketamine are now serious contenders in psychiatry and neurology. Early clinical successes, together with advances in imaging and molecular profiling, are opening new frontiers for conditions where current therapies fall short.
In the biology team of o2h Discovery we are committed to quickly translating bold new ideas into executable science on the bench. In this edition of our Therapeutic Spotlight, we highlight how our biology team supports neuroscience and psychedelic research from receptor pharmacology to neurite outgrowth and high-content imaging platforms.
We have a range of collaborative models and low friction start points you can start working with us including Kickstarter, Biology Match Funding and more collaborative models for capital efficient ‘proof of concept’ such as Inflexion Tx. We are happy to use our experience with cascade and panel design and can get started on new projects on the bench in a matter of days/weeks.
At o2h Discovery, we seed new ideas in life science. We have an insight driven integrated multi-modality drug discovery capability based out of Cambridge, UK and Ahmedabad, India. Please contact me on gayathri.sadasivam@o2h.com and we would be happy to move at speed on the design and study execution.
Kind regards,
Gayathri Sadasivam,
Vice President - Biology, o2h Discovery
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Why Psychedelics Matter in CNS R&D?
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Despite decades of effort, CNS disorders remain some of the most underserved in medicine. Nearly 970 million people worldwide suffer from depression, anxiety, PTSD, schizophrenia, or neurodegenerative diseases, yet most treatments provide only symptomatic relief.
Psychedelics are gaining traction because of their rapid, durable effects in treatment-resistant conditions. Unlocking their full therapeutic potential, however, requires a deeper understanding of receptor selectivity, neural plasticity, and translational safety pharmacology.
Researchers are particularly focused on:
- Mechanism-driven psychiatric therapies (serotonin, glutamate, sigma receptors)
- Neuroinflammation and microglial activation in neurodegeneration
- Synaptic remodelling and neurogenesis in trauma recovery
- Biomarker-driven patient stratification
- Glial pathway modulation in Alzheimer’s, MS, and related disorders
This complexity demands robust cell-based models, functional assays, and quantitative biology - all of which are being systematically built and refined at o2h Discovery.
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o2h Discovery Capabilities in Psychedelics
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At o2h Discovery, stable 5HT2A, 5HT2B, and 5HT2C receptor cell lines have been created using a Flp-In TREx HEK293 system, enabling highly reproducible and quantitative assays targeting key psychedelic receptors. The 5-HT2A receptor - a GPCR pivotal in mood, cognition, and perception - triggers both Gq- and β-arrestin-dependent signaling cascades. Dissecting the complexity of this biased signaling is essential to separate the therapeutic benefits from the hallucinogenic effects of compounds such as LSD and psilocybin.
With the integration of advanced platforms like FLIPR Penta® for real-time calcium flux analysis and PathHunter® assays for β-arrestin recruitment, o2h Discovery is mapping ligand-specific receptor signaling in detail. Systematic profiling of small molecules with distinct preferences for Gq or β-arrestin pathways is revealing how differential activation underpins diverse biological outcomes.
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These approaches are advancing the mechanistic understanding of serotonin receptor pharmacology and establishing a foundation for the discovery of next-generation neuropsychiatric therapeutics.
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Figure 1: (A) Chemiluminescent detection of β-arrestin recruitment in U2OS PathHunter cells following stimulation with increasing concentrations of 5-HT (serotonin). Rising serotonin levels produce a dose-dependent increase in luminescence, indicating progressive recruitment of β-arrestin to the 5-HT2A receptor. Data were analyzed using non-linear regression (four-parameter logistic model).
(B) U2OS PathHunter cells were exposed to varying concentrations of test compounds and assessed for β-arrestin recruitment.
Distinct compounds produced differential, dose-dependent luminescence responses, reflecting varied capacities to promote β-arrestin recruitment to the 5-HT2A receptor.
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Figure 2: Tetracycline-regulated stable cell lines expressing 5-HT₂ receptors
(a–c) Stable cell lines expressing 5-HT₂A, 5-HT₂B, or 5-HT₂C receptors were stimulated with increasing concentrations of serotonin, and intracellular Ca²⁺ responses were measured using the FLIPR Penta platform. The resulting dose–response curves show robust serotonin-induced calcium signalling and reveal subtype-specific differences in both potency and efficacy for standard reference compounds.
(d) The 5HT2A cell line also exhibited dose-dependent IP1 accumulation, downstream of Ca²⁺ signaling, following serotonin stimulation. IP1 levels were quantified using an HTRF assay.
(e) Test compounds were evaluated for IP signalling activity and classified based on their agonist properties, demonstrating a range of potencies and efficacies in the IP assay.
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Figure 3:(A) Cells treated with increasing concentrations of the antagonist ketanserin show a dose-dependent reduction in IP1 production following stimulation with an EC80 concentration of 5-HT (serotonin).
(B, C) Screening of test compounds for antagonistic activity was performed by measuring intracellular levels of IP1 (B) and Ca²⁺ (C), respectively.
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Figure 4: (A) Schematic illustration of the NanoBiT methodology. The 5HT2A receptor is C-terminally tagged with LgBiT, while Barr2 is N-terminally tagged with SmBiT. Ligand binding by serotonin (5HT) induces interaction between these tagged proteins, leading to luciferase complementation and luminescence signal generation.
(B) Kinetic luminescence recordings over two hours from cells transfected with 5HT2A receptor-LgBiT and SmBiT-Barr2. Solid blue lines indicate samples with 1 μM 5HT added at time zero; dashed lines represent buffer controls. The transfected DNA amounts (for 5HT2A R and Barr2) were serially diluted two-fold, visualized as a heatmap reflecting decreasing DNA quantities.(C) The relative area under the curve (AUC) for the recordings shown in (B). Despite decreasing DNA amounts, the fold change in luminescence upon 5HT addition remains consistent. The negative control consisted of cells transfected with 5HT2A R-LgBiT and HaloTag-SmBiT, which showed no interaction upon 5HT stimulation.
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Beyond Psychedelics: Neurite Outgrowth & HCI
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Neuroregeneration represents one of the most challenging frontiers in neuroscience, with the limited capacity of terminally differentiated neurons to self-repair or divide posing a major hurdle. Identifying compounds that can promote neuroplasticity - the ability of neurons to form new connections and recover from trauma such as spinal cord injury requires robust, reproducible assays capable of modelling neuronal regeneration.
At o2h Discovery, we are addressing this need by developing a scalable neurite outgrowth assay using differentiated SH-SY5Y neurons, immunostained with MAP2 and HuC/D markers to closely mimic mature human neurons. Our high-content imaging (HCI) pipeline enables quantitative assessment of neurite extension, branching, and axonal regeneration, providing precise insights into compound-induced neuroplasticity.
Through these innovations, o2h Discovery’s biology team is contributing to the development of next-generation therapeutics that target neuronal growth and regeneration pathways.
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We are developing high content imaging (HCI)-based assays for quantification of neurite outgrowths in order to screen for compounds modulating neurite outgrowths and axonal regeneration. Using an HCI-based pipeline we capture images of neurite phenotypes in basic and blebbistatin-differentiated SH-SY5Y cells. Blebbistatin, a myosin II inhibitor, promotes neurite outgrowth by reducing actinomyosin II contractility. Control and treated SH-SY5Y cells were probed with Hoechst dye (for nucleus), MAP2 antibody (a microtubule-associated protein which act as a marker of neurons), HuC/D (an RNA-binding protein which induce neuronal differentiation & expressed in neuronal cell body) to create a mark for the respective markers. Image analysis algorithms then quantify morphological responses in the images.
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Figure 5: Analysis of SH-SY5Y cells grown under 3% serum, 20 uM retinoic acid, and cultured on ECM matrix treated with blebbistatin for 24 hours.
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Figure 6: Preliminary analysis of primary rat cortical neurons using our HCI platform; cells were cultured in neurobasal media + B-27, grown on poly-D-lysine and treated with blebbistatin for 20 hours.
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Following our early data with the SH-SY5Y model, we have extended our analysis pipeline to test similar neurite outgrowth conditions in primary rat cortical neurons. These cells responded with equally robust results, suggesting our assay is sensitive to morphological outcomes in both transformed and primary cell models.
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With future utility in models such as iPSC and primary cell cultures, our neurite outgrowth analytical pipeline is leveraging information-rich image data to report dose-responsive shifts in morphological changes relevant to neuronal processes. Building on our experience with recent cell assay innovations, this platform is likely to be enhanced through integration with SemaCyte® microcarrier technology, which enables miniaturized, barcoded, and assay-ready cell monolayers.
We look forward to making these advances into powerful, scalable tools for drug discovery in neurodegeneration and neural repair.
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From receptor pharmacology in psychedelics to neurite outgrowth and advanced imaging, o2h Discovery delivers translational biology to support CNS research.
Get in touch to learn how our customized assays and screening platforms can support your discovery pipeline.
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Exploring Psychedelics in CNS research?
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At o2h Discovery, we have developed and tested a series of small molecules that target the 5-HT2A receptor, each with different strengths when it comes to activating Gq and β-arrestin signaling pathways.
Read our blog to see how o2h Discovery is supporting this groundbreaking research.
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