Transcriptome Signature of Immune Cells Post Reovirus Treatment in KRAS Mutated Colorectal Cancer

Abstract

Purpose

Reovirus propagates with high efficiency in KRAS mutated colorectal cancer (CRC). About 45–50% of CRC patients possess a KRAS mutation. Oncolytic reovirus treatment in combination with chemotherapy was tested in patients possessing KRAS mutated metastatic CRC. This study evaluates the biological responses to reovirus treatment by determining the gene expression patterns in RAS-related signaling pathways.

Methods

Reovirus was administered as a 60-min intravenous infusion for 5 consecutive days every 28 days, at a tissue culture infective dose (TCID₅₀) of 3×10¹⁰. Peripheral blood mononuclear cells (PBMCs) were isolated from whole-blood pre- and post-reovirus administration at 48 hr, day-8, and day-15. Clariom_D_Human_Assay was used to determine the expression of vital genes compared to pre-reovirus treatment by RNA sequencing. Using exported sample signals, ΔΔCt method was used to analyze the fold changes of genes within seven gene pathways. Significance was calculated by students-two-tail-t-test. Hierarchical clustering dendrogram was constructed by calculating Pearson’s correlation coefficients.

Results

As compared to the control, SOS1[48 hr; 2.49X], RRAS [48 hr; 2.24X], PIK3CB [D8, D15; 2.27X, 3.16X], MIR 16–2 [D15; 1.70X], CHORDC1 [48 hr, D15; 1.89X, 4.54X], RTN4 [48 hr; 4.66X], FAM96A [48 hr; 4.54X], NFKB [D8, D15; 19.0X, 1.42X], CASP8 [D8, D15; 2.11X, 1.77X], and CASP9 [D8; 1.45X] are upregulated post-reovirus. NOS3 [D15; 0.61X], SYNE1 [D8, D15; 0.78X, 0.71X], ANGPT1 [D8; 0.62X], VEGFB [48 hr, D8, D15; 0.44X, 0.28X, 0.28X], JUN [D15; 0.69X], and IGF2 [D8; 0.73X] are downregulated post-reovirus. Fold change values were significant [p<0.05].

Conclusion

This study highlights reovirus as a novel treatment option for KRAS mutated CRC and showcases its effect on the expression of crucial genes.

Keywords:

• transcriptome
• reovirus
• CRC
• KRAS
• CASP8
• CHORDC1
• RTN4
• VEGFB

Acknowledgments

We gratefully acknowledge the contribution of the patients and their family members in participating in the clinical trial, which made this study possible. We are thankful to genomic facility of Albert Einstein College of Medicine for performing transcriptome assay. We would like to thank Dr Matt Coffey (Oncolytics Biotech, Calgary, Canada) and Dr Selma Botman, the Yeshiva University Provost and Vice President for Academic Affairs, for graciously providing funding to RM for this research.

Author Contributions

All authors contributed to data analysis, drafting or revising the article, have agreed on the journal to which the article will be submitted, gave final approval for the version to be published, and agree to be accountable for all aspects of the work. Conception and design: SG, RM. Analysis and interpretation of data: EF, AS, RM. Writing, review, and/or revision of the manuscript: EF, AS, SG, RM. Administrative, technical, or material support: SG, RM. Study supervision: SG, RM.

Disclosure

The authors declare that there is no conflict of interest.