Pharmacodynamics (PD) Services for Glioma

Advanced Cell-Based Assays for Target Engagement

• Dose-Response Viability Curves: IC50 determination using CellTiter-Glo or RealTime-Glo MT assays, validated against clinical benchmarks (e.g., TMZ IC50≤50 µM in MGMT-unmethylated lines).
• Apoptosis and Cell Cycle Analysis: Annexin V/PI staining and EdU incorporation assays to quantify pro-apoptotic effects or G2/M arrest, critical for DNA-damaging agents.
• Target Phosphorylation Profiling: Multiplex Luminex or Wes Simple Western assays to measure EGFR, AKT, or ERK pathway inhibition.

3D Spheroid and Organoid Models Mimicking Tumor Microenvironment

• Invasive Growth Metrics: Quantified via confocal imaging of GFP-labeled spheroids infiltrating Matrigel or brain-mimetic hydrogels.
• Drug Penetration Studies: LC-MS/MS measures intracore drug distribution, identifying compounds with poor diffusion (e.g., >50% concentration gradient from periphery to core in bevacizumab-treated spheroids).
• High-Content Imaging and Multi-Omics Integration.
• Live-Cell Analysis: Longitudinal tracking of glioma cell migration and proliferation under treatment.
• Spatial Proteomics: maps PD-L1 or MGMT expression in hypoxic vs. normoxic spheroid regions.
• Metabolic Flux Analysis: quantify oxidative phosphorylation suppression by IDH1 inhibitors.

Orthotopic and Patient-Derived Xenograft (PDX) Model

• Orthotopic Models: U87MG or GL261 cells implanted into immunocompetent C57BL/6 mice, enabling evaluation of immune-modulating therapies.
• PDX Models: Tumors derived from primary or recurrent glioblastoma patients engrafted in NSG mice, retaining original mutations (e.g., EGFRvIII, PTEN loss) and drug response profiles.

Advanced Imaging for Real-Time PD Monitoring

Alfa Cytology integrates non-invasive imaging modalities to quantify tumor progression and therapeutic response:

• MRI: High-resolution T2-weighted and diffusion tensor imaging (DTI) for measuring tumor volume, edema, and white matter infiltration.
• Bioluminescence Imaging (BLI): Luciferase-expressing glioma cells enable longitudinal tracking of tumor burden in orthotopic models.
• PET/CT: [¹⁸F]FDG or [¹⁸F]FET tracers assess metabolic activity and amino acid uptake, correlating with treatment-induced necrosis.

Biomarker Analysis and PK/PD Integration

Alfa Cytology’s in vivo PD platform combines biofluid sampling and tissue analysis to elucidate mechanism-of-action (MoA):

• Liquid Biopsies: Longitudinal plasma ctDNA sequencing tracks clonal evolution and resistance mutations (e.g., EGFR T790M).
• Cerebrospinal Fluid (CSF) Sampling: LC-MS/MS quantifies BBB-penetrant drug concentrations.
• Immunohistochemistry (IHC): Spatial analysis of PD-L1, CD8+ T-cell infiltration, and Ki-67 indices in resected tumors.