At Oncocit, our ongoing research under the Seed Capital Project focuses on enhancing tumor treatment efficacy through advanced biotechnological methods. This involves a comparative evaluation of tumor proteins sourced from tissue biopsies and specific cancer cell lines, aiming to improve the induction of immune responses in dendritic cells (DCs) and pave the way for personalized immunotherapy and targeted therapy applications.
Methodology
Comparative Evaluation
Our study investigates tumor biopsies and corresponding cell lines from three types of cancer:
- Osteosarcoma: Comparison between fresh biopsies and standard SaOS-2 cell line.
- Colon Cancer: Patient tumor biopsies versus HT-29 and HCT116 cell lines.
- Breast Cancer: Comparisons with the MCF-7 cell line. Advanced antigen recovery protocols were applied to expose key epitopes like neoantigens linked to specific mutations, crucial for the targeted therapeutic approach.
Protein Quantification and Quality
Using standard quantification methods (Bradford, Lowry) and molecular characterization (SDS-PAGE and mass spectrometry), we assessed protein integrity with degradation indices showing significantly lower rates in cell lines (<10%) compared to biopsies (>30%). Quality monitoring ensured over 90% purity and functionality of key proteins such as BAX, BCL-2, P53, and mutated KRAS.
Enhancing Dendritic Cell Maturation
Cellular Maturation Induction
Tumor proteins from cell lines were co-cultured with immature dendritic cells from PBMCs using a Ficoll gradient. Maturation was evaluated through phenotypic markers HLADR, CD80, CD86, and the secretion of immunomodulatory cytokines (IL-12, TNF-α) was measured by ELISA. Flow cytometry was utilized to analyze the efficiency of dendritic cell activation.
Results
Improved Protein Yield and Quality
Cell lines demonstrated a 60-70% higher protein yield and greater molecular purity with less degradation, enhancing their functionality in immunotherapy. These proteins were more effective in activating dendritic cells, as evidenced by the high expression of maturation markers (CD80/CD86) and increased cytokine release (IL-12, TNF-α > 50 pg/mL).
Advancing Targeted Therapy
The study highlights the potential of mutated proteins from cell lines in developing accessible and effective targeted therapies. These proteins, offering higher immunogenic functionality compared to traditional recombinant proteins, reduce the reliance on excipients and preservatives that can impede immune response.
Conclusions
Superior Molecular Efficiency
Tumor proteins derived from cell lines surpass those obtained from biopsies in performance, quality, and functionality, proving critical in advanced immunotherapy applications. The optimization of dendritic cell activation confirms the potential of these proteins in clinical settings.
High-Impact Biotechnological Alternative
Mutated proteins from cell lines represent a cost-effective and efficient solution for directed therapies, enhancing production capabilities and reducing costs compared to traditional methods.
Future Implications
This project sets a new standard in the development of personalized immunotherapies based on specific tumor proteins from cell lines. Potential applications include targeted therapies for high mutational burden cancers and the design of more cost-effective and efficient protocols for clinical and translational research laboratories. Dr. Ramon Gutiérrez.