Introduction:
Neuroblastoma is among the most common malignant tumors in childhood. Long-term survival for children with advanced disease older than 18 months of age is poor despite aggressive multimodal therapy. IL-12 and IL-18 (IFN-ginducing factor) are pleiotropic cytokines which function in immune modulation and thereby potentiate anti-tumor immunity. A technique of locally produced specific delivery of immunostimulatory interleukins within a tumor bed remains elusive and difficult.

Project Aims:
1) Construct recombinant retrovirus vectors expressing IL-12, IL-18, and B7.
2) Create mouse Neuroblastoma cell lines (Neuro-2A) with stable-expression IL-12, IL-18 and B7 termed Neuro-2A (IL-12), Neuro-2A (IL-18) and Neuro-2A (B7).
3) Create mouse dendritic cell lines with stable-expression B7 termed DC (B7).
4) Fuse Neuroblastoma cells with DC (B7).
5) Determine the capacity of these genetically-modified chimeric fusion cells as autologous tumor cell vaccines in eliminating established tumor burden in a mouse model.

Results/Conclusion:
We constructed retrovirus vectors expressing IL-12, IL-18 and B7, respectively. Neuro-2A cells with stable expression of IL-12, IL-18 and surface molecule B7 via the Invitrogen Neon™ electroporation device system were established:
• 29 clones Neuro-2A expressing IL-18
• 12 clones expressing IL-12
• 4 clones expressing B7
• 9 clones of dendritic cells expressing IL-18
• 6 clones of dendritic cells expressing B7.
We constructed a dual-expression vector for IL-18 and IL-12; screening for stable dual-expressing Neuro-2A is underway.
Neuro-2A drove tumor formation in immune-deficient mice (NSG), but did not drive tumor formation in immune-competent strains C57BL/6 and C3H/HeJ mice; however, it did drive tumor formation in immune-competent mouse strain (AJ). Both Neuro-2A (IL-12) and Neuro-2A (IL-18) did not drive tumor formation in immune-competent strain mice (AJ); however, Neuro-2A (B7) drove significant tumor formation.
Future experiments are to electrically fuse Neuro-2A (IL-12), Neuro-2A (IL-18) and DC (B7) together to create a more effective vaccine against neuroblastoma.
The era of computer simulation and cognitive computing may allow easier and safer translation of animal studies to human trials and use. It may be possible to use current knowledge of cellular function and cellular recognition in combination with a large and growing database of cellular signaling to create cellular interaction simulations and predictability models of tumoricidal activity.

 

PRECISION MEDICINE & DRUG DISCOVERY

Author: Kain Sosa II

Coauthor(s): Shengwen Calvin Li, Ph.D. Mustafa H. Kabeer, MD CHOC Children’s Research Institute, Orange University of California – Irvine School of Medicine, Irvine

Status: Project Concept