Investigation of Subcutaneous Retinoblastoma Tumor in a Mouse Model Following htsFLT01 Chimeric VEGF Receptor Therapy
Helaleh Vaezi Heris1 *, Zahra-Soheila Soheili2 , Mozhgan Rezaei Kanavi3 , Hamid Latifi-Navid2 , Shahram Samiei4
- Department of Molecular Medicine, National Institute of Genetic Engineering and Biotechnology,
Tehran 1497716316, Iran.
- Department of Molecular Medicine, National Institute of Genetic Engineering and Biotechnology, Tehran 1497716316, Iran.
- Ocular Tissue Engineering Research Center, Research Institute for Ophthalmology and Vision Science, Shahid Beheshti University of Medical Sciences, Tehran, Iran.
- Blood Transfusion Research Center, High Institute for Research and Education in Transfusion Medicine, Tehran, Iran.
Abstract: Retinoblastoma, a rapidly progressing pediatric retinal cancer caused by RB1 gene mutations, is characterized by angiogenesis, which drives tumor growth and metastasis. Anti-angiogenic therapies targeting vascular endothelial growth factor (VEGF) and placental growth factor (PLGF) hold promise for treatment. This study evaluates the efficacy of the htsFLT01 chimeric construct, delivered via lentiviral (LV) particles, in suppressing angiogenesis in a retinoblastoma mouse model.
Methods: The htsFLT01 construct was cloned into a pLenti vector through PCR amplification with Nhe I restriction sites, followed by transfection into HEK293T cells to generate high-titer LV particles. Y79 retinoblastoma cells will be injected subcutaneously into mice to establish xenografted tumors, which will be subsequently transduced with LV-htsFLT01 particles. Tumor angiogenesis and therapeutic efficacy will be assessed via real-time PCR, western blotting, immunohistochemistry, and histological analysis.
Results: Plasmid quality and PCR amplification were confirmed via gel electrophoresis. Sequencing will validate the identity of the construct. Successful production of high-titer LV particles enables effective tumor transduction, which is expected to demonstrate reduced angiogenesis and tumor growth.
Conclusion: This study establishes a robust xenograft model and demonstrates the feasibility of using htsFLT01-based gene therapy to inhibit tumor angiogenesis, providing a foundation for novel anti-angiogenic strategies in retinoblastoma treatment.
