This case study demonstrates Cellomatics’ capability to integrate:
- High-throughput screening (HTS)
- Complex primary human cell models
- Multiparametric functional readouts
- Cross-validation of hits in disease-relevant systems
Scientific objective
To systematically evaluate the functional impact of 100 siRNA-mediated gene knockdowns in primary human keratinocytes derived from both healthy and psoriatic donors, across multiple disease-relevant biological pathways and functional endpoints including
Cellomatics' approach
1.High-throughput screening design
This design ensured:
- Strong statistical robustness through the incorporation of technical replicates, internal assay controls, and highly standardised experimental conditions, enabling confident interpretation of screening outcomes and minimising assay variability.
- High biological reproducibility across multiple experimental runs and independent donor-derived samples, ensuring reliable comparison of functional responses and consistent identification of biologically relevant effects.
- Comprehensive assessment of donor-to-donor variability through the inclusion of multiple psoriatic patient samples, enabling the identification of patient-specific response patterns and improving the translational relevance of the screening platform.
2.Human translational disease model
- Primary human keratinocytes isolated from both healthy individuals and psoriatic patients were utilised to establish physiologically relevant in vitro disease models that more accurately reflect patient biology and clinical disease heterogeneity.
- Disease-relevant cellular stimulation was achieved using well-characterised pro-inflammatory cytokine combinations to induce psoriasis-associated phenotypes and downstream signalling pathways.
- IL-1α, TNFα, and Oncostatin M were used to establish a robust inflammatory model, driving the induction of key inflammatory mediators, cytokine release, and immune-associated transcriptional responses commonly observed in psoriatic skin lesions.
- IL-17A stimulation was employed to model keratinocyte hyperproliferation and altered epidermal biology, enabling assessment of pathways associated with abnormal tissue growth and disease progression.
- These disease-relevant stimulation models closely recapitulate the inflammatory and proliferative pathways observed in vivo in psoriatic skin, thereby providing a highly translational platform for early target discovery, mechanistic investigation, and functional validation of therapeutic candidates.
3.Multi-parametric functional readouts
Key results and insights
Robust and Reproducible HTS Execution
- Consistent and highly efficient siRNA transfection was achieved and maintained throughout the 72-hour assay duration, ensuring reliable target knockdown and uniform modulation of gene expression across all experimental and screening conditions.
- Optimised transfection protocols and assay workflows enabled stable assay performance with minimal technical variability, supporting high-confidence interpretation of screening data across large target sets.
- Assay performance remained robust and reproducible across independent donor-derived keratinocyte samples and technical replicates, demonstrating the suitability of the platform for scalable, high-throughput functional screening applications.
- The integration of rigorous quality control measures, standardised assay conditions, and replicate-based analysis ensured reproducible data generation and strengthened confidence in hit identification and downstream validation.
Clear Disease-Relevant Functional Responses
- Cytokine stimulation generated strong and reproducible inflammatory responses across multiple cytokine secretion and gene-expression markers, confirming successful induction of disease-relevant cellular phenotypes associated with psoriatic pathology.
- Functional profiling revealed activation of key inflammatory and proliferative pathways relevant to psoriasis biology, enabling comprehensive assessment of target-dependent cellular responses.
- Distinct response patterns were observed between healthy and psoriatic keratinocytes, demonstrating the platform’s ability to discriminate disease-associated biological phenotypes and capture clinically relevant functional differences.
- Additional donor-specific variations in inflammatory and proliferative responses were identified among individual patient samples, highlighting the importance of incorporating patient heterogeneity into translational drug discovery workflows.
- These findings demonstrate the capability of the platform to generate biologically meaningful, patient-relevant datasets that support target prioritisation, mechanistic understanding, and translational decision-making in early drug discovery.
Identification of Functional Hits
- The screening campaign successfully identified gene targets involved in important disease-related processes, including inflammatory cytokine production, keratinocyte proliferation, and epidermal differentiation pathways, all of which are key features of psoriasis progression.
- Functional analysis of these targets helped identify modulators capable of changing disease-relevant cellular responses, supporting the prioritisation of targets with potential therapeutic value.
- Several targets showed donor-specific and gene-specific effects, revealing clear differences in biological responses between individual patient-derived samples and highlighting patient variability within the disease.
- Differences in target responses between donors provided valuable insight into patient-specific biology and pathway dependence, supporting the development of more personalised therapeutic approaches.
- The ability to detect distinct target-specific responses across diverse patient samples further demonstrates the strength of the platform for translational target discovery, functional validation, and biomarker-driven drug development.
Built-in Pharmacological Validation
- Inclusion of clinically relevant reference compounds, including known pathway inhibitors and therapeutic modulators, enabled benchmarking of assay performance against established biological responses.
- Reference compounds such as Tofacitinib and Cryptotanshinone produced the expected pathway-specific effects in stimulated keratinocyte models, confirming successful activation of disease-relevant signalling pathways.
- Pharmacological benchmarking helped assess assay sensitivity, reproducibility, and consistency across different experimental conditions and donor samples.
- The responses observed with known modulators confirmed the biological relevance and reliability of the assay platform, increasing confidence in the identified screening hits and target prioritisation.
- Inclusion of validated pharmacological controls also improved the predictive value of the screening system by helping distinguish true biological effects from non-specific responses.
- This integrated validation approach demonstrates the ability of the platform to support functional screening together with pathway-level biological confirmation for translational drug discovery.
Cellomatics' capabilities demonstrated
High-Throughput Screening (HTS)
- The platform supports scalable screening of large target libraries while maintaining high assay quality and operational efficiency.
- Multiplexed assay formats enable simultaneous assessment of multiple biological endpoints within the same experimental workflow.
- High-content data generation combined with stringent quality control metrics ensures reliable and information-rich datasets for downstream analysis.
Human Translational Biology
- The use of primary patient-derived cells and disease-relevant stimulation systems provides a biologically meaningful and translationally relevant framework for early-stage drug discovery.
- Multi-donor validation further strengthens confidence in hit reproducibility and clinical relevance.
Integrated Functional Validation
Simultaneous evaluation of inflammation, proliferation, and tissue-function endpoints enables rapid differentiation of biologically meaningful targets from false positives or non-specific effects.
Cross-Therapeutic Applicability
While demonstrated in psoriasis, this platform is directly translatable to:
- Immunology & inflammation
- Oncology (tumour–immune interactions)
- Fibrosis
- Respiratory diseases
- Autoimmune disorders
Impact for drug discovery
This study highlights Cellomatics’ ability to:
- Accelerate early target discovery by combining high-throughput functional genomics with disease-relevant human biology.
- Reduce translational risk through early validation in primary human cellular systems rather than relying solely on simplified in vitro models.
- Integrate screening and mechanistic validation within a single workflow, improving efficiency and reducing development timelines.
- Generate actionable biological insights that support confident hit prioritisation and downstream decision-making.
Conclusion
Cellomatics Biosciences provides a comprehensive and integrated platform for early drug discovery that combines scalable high-throughput screening, complex human translational disease models, and multidimensional functional readouts. This approach enables clients to move beyond conventional screening strategies toward mechanistically informed, clinically relevant target discovery and validation.
