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Differential microRNA Expression in Newcastle Disease Virus-Infected HeLa Cells and Its Role in Regulating Virus

Yu Chen¹, Shanshan Zhu¹, Yuru Pei¹

¹Animal Infectious Disease Laboratory, College of Veterinary Medicine, Yangzhou University, Yangzhou, China.

Frontiers in Oncology

|June 21, 2021

Abstract:

As an oncolytic virus, Newcastle disease virus (NDV) can specifically kill tumor cells and has been tested as an attractive oncolytic agent for cancer virotherapy. Virus infection can trigger the changes of the cellular microRNA (miRNA) expression profile, which can greatly influence viral replication and pathogenesis. However, the interplay between NDV replication and cellular miRNA expression in tumor cells is still largely unknown. In the present study, we compared the profiles of cellular miRNAs in uninfected and NDV-infected HeLa cells by small RNA deep sequencing. Here we report that NDV infection in HeLa cells significantly changed the levels of 40 miRNAs at 6 h post-infection (hpi) and 62 miRNAs at 12 hpi. Among 23 highly differentially expressed miRNAs, NDV infection greatly promoted the levels of 3 miRNAs and suppressed the levels of 20 miRNAs at both time points. These 23 miRNAs are predicted to target various genes involved in virus replication and antiviral immunity such as ErbB, Jak-STAT, NF-kB and RIG-I-like receptor. Verification of deep sequencing results by quantitative RT-PCR showed that 9 out of 10 randomly selected miRNAs chosen from this 23-miRNA pool were consistent with deep sequencing data, including 6 down-regulated and 3 up-regulated. Further functional research revealed that hsa-miR-4521, a constituent in this 23-miRNA pool, inhibited NDV replication in HeLa cells. Moreover, dual-luciferase and gene expression array uncovered that the member A of family with sequence similarity 129 (FAM129A) was directly targeted by hsa-miR-4521 and positively regulated NDV replication in HeLa cells, indicating that hsa-miR-4521 may regulate NDV replication via interaction with FAM129A. To our knowledge, this is the first report of the dynamic cellular miRNA expression profile in tumor cells after NDV infection and may provide a valuable basis for further investigation on the roles of miRNAs in NDV-mediated oncolysis.

Insights

Newcastle disease virus (NDV) infection alters cellular microRNA (miRNA) expression in tumor cells. One specific miRNA, hsa-miR-4521, was found to inhibit NDV replication by targeting FAM129A.

Area of Science:

Oncology
Virology
Molecular Biology

Background:

Newcastle disease virus (NDV) is an oncolytic virus with potential for cancer virotherapy.
Cellular microRNA (miRNA) expression changes during viral infections, impacting viral replication and pathogenesis.
The interaction between NDV and cellular miRNAs in tumor cells remains largely unexplored.

Purpose of the Study:

To investigate the dynamic changes in cellular miRNA expression profiles following NDV infection in tumor cells.
To identify specific miRNAs involved in the interplay between NDV replication and host cell response.
To elucidate the mechanism by which miRNAs regulate NDV replication.

Main Methods:

Small RNA deep sequencing was employed to compare miRNA profiles in uninfected and NDV-infected HeLa cells.
Quantitative RT-PCR was used to validate sequencing results for differentially expressed miRNAs.
Dual-luciferase assays and gene expression arrays were utilized to identify miRNA targets and their functional roles.

Main Results:

NDV infection significantly altered the expression of 40 miRNAs at 6 hours post-infection and 62 miRNAs at 12 hours post-infection.
Among 23 highly differentially expressed miRNAs, 3 were upregulated and 20 were downregulated by NDV infection.
hsa-miR-4521 was identified as an inhibitor of NDV replication, targeting FAM129A, which positively regulates viral replication.

Conclusions:

NDV infection induces dynamic changes in the cellular miRNA landscape of tumor cells.
hsa-miR-4521 plays a role in regulating NDV replication, potentially through its interaction with FAM129A.
These findings provide a foundation for understanding miRNA involvement in NDV-mediated oncolysis and cancer therapy.

Read full analysis

Keywords:

FAM129A HeLa cells NDV hsa-miR-4521 miRNA expression profile oncologic virus

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