Experimental Adaptation of Rotaviruses to Tumor Cell Lines

Abstract

A number of viruses show a naturally extended tropism for tumor cells whereas other viruses have been genetically modified or adapted to infect tumor cells. Oncolytic viruses have become a promising tool for treating some cancers by inducing cell lysis or immune response to tumor cells. In the present work, rotavirus strains TRF-41 (G5) (porcine), RRV (G3) (simian), UK (G6-P5) (bovine), Ym (G11-P9) (porcine), ECwt (murine), Wa (G1-P8), Wi61 (G9) and M69 (G8) (human), and five wild-type human rotavirus isolates were passaged multiple times in different human tumor cell lines and then combined in five different ways before additional multiple passages in tumor cell lines. Cell death caused by the tumor cell-adapted isolates was characterized using Hoechst, propidium iodide, 7-AAD, Annexin V, TUNEL, and anti-poly-(ADP ribose) polymerase (PARP) and -phospho-histone H2A.X antibodies. Multiple passages of the combined rotaviruses in tumor cell lines led to a successful infection of these cells, suggesting a gain-of-function by the acquisition of greater infectious capacity as compared with that of the parental rotaviruses. The electropherotype profiles suggest that unique tumor cell-adapted isolates were derived from reassortment of parental rotaviruses. Infection produced by such rotavirus isolates induced chromatin modifications compatible with apoptotic cell death.

Assessing the oncolytic potential of rotavirus on mouse myeloma cell line Sp2/0-Ag14

“…Rotavirus isolates Wt1-5, WWM, TRUYO, ECwt-O, and WTEW, selected as previously reported 36 , were trypsin-activated (10 mg/ml) and separately inoculated (MOI of 2) in Sp2/0-Ag14 cells (1 x 10 7 ) in DMEM (10 ml) without FBS. The cells were cultured at 37 °C until their lysis (about 24 h).…”

“…ELISA analyses were conducted as previously described 36 . Briefly, Sp2/0-Ag14 cells were separately infected with the rotavirus isolates described above.…”

“…Several studies have shown rotavirus tropism in cells expressing Hsc70 33 , PDI 34 , and integrins such as αVβ3 35 . Recently, it was described how some reassorted rotavirus isolates can successfully replicate in some tumor cell lines 36 .…”

Repurposing rotavirus vaccines for intratumoral immunotherapy can overcome resistance to immune checkpoint blockade

“…Recently, the team of Guerrero et al (32) has reported similar results on the oncolytic properties of rotavirus against several cancer lines although with reduced potency compared to our results. We examined cells after 24 hours or more instead of 12 hours and also used strains selected for commercial vaccines, which were not the same as the Guerrero et al (32) study.…”

Virus oncolíticos: un arma contra el cáncer

“…GM-CSF: Factor estimulador de colonias monocito-macrófago; NSCLC: cáncer de pulmón de células no pequeñas; RGD: Arg-Gly-Asp motif; TK: timidina quinasa; US11: glicoproteína corta única 11. En el Laboratorio de Biología Molecular de Virus de la Facultad de Medicina de la Universidad Nacional de Colombia se han realizado estudios en los cuales se encontró que cepas rotavirales aisladas en el laboratorio infectan y lisan las líneas celulares Sp2/0-Ag14 cells (mieloma de ratón), U-937 (linfoma histiocítico), REH (leucemia linfocítica aguda), Kato III (carcinoma gástrico), MCF-7 (adenocarcinoma de mama), PC-3 (adenocarcinoma grado IV de próstata) y A549 (carcinoma de pulmón humano), lo que sugiere que las cepas estudiadas son potenciales virus oncolíticos (18).…”