Abstract Title
Overcoming host restriction to enable continuous passaging of human noroviruses in human intestinal enteroids
Presenter
Gurpreet Kaur, Baylor college of medicine
Co-Author(s)
Gurpreet Kaur- Department of Molecular Virology and Microbiology, Baylor College of Medicine, Houston, TX
Sue Crawford- Department of Molecular Virology and Microbiology, Baylor College of Medicine, Houston, TX.
Sasirekha Ramani- Department of Molecular Virology and Microbiology, Baylor College of Medicine, Houston, TX.
Hoa Nguyen Phuc- Department of Molecular and Cellular Biology, Baylor College of Medicine, Houston, TX.
Cristian Coarfa-Department of Molecular and Cellular Biology, Baylor College of Medicine, Houston, TX.
Mary Estes- Department of Molecular Virology and Microbiology, Baylor College of Medicine, Houston, TX, Department of Medicine, Baylor College of Medicine, Houston, TX.
Abstract Category
Entry and Replication
Abstract
Human noroviruses (HuNoVs) are the leading cause of viral gastroenteritis. We established human intestinal enteroids (HIEs) as a robust model for HuNoV infection and replication (Ettayebi et al., Science 2016; Ettayebi et al., mBio 2024), overcoming a longstanding cultivation barrier. However, indefinite passaging of HuNoVs in HIEs remains a challenge, limiting the ability to generate virus stocks and necessitating the use of patient stool samples as viral inocula. Therefore, sought to overcome the barrier that limits continuous passaging of HuNoVs. We used comparative transcriptomics (RNA-seq) to define responses of HIEs to HuNoV infection and to identify host restriction factors limiting viral passaging.
The top upregulated pathway was the host innate immune response, primarily driven by interferons and CXC based chemokines. CXCL10, CXCL11, and CCL5 were among the most highly upregulated interferon-stimulated genes, suggesting their potential role as host restriction factors. To counteract this, we used TAK-779, a CXCR3/CCR5 antagonist, which enhanced GII.3 HuNoV replication (1.5 log10 genome equivalents) and viral spread in a dose- and time-dependent manner. For the first time, we have passaged GII.3 HuNoV in the presence of TAK-779 for over 10 passages in two different HIE lines. This breakthrough enables the generation of sufficient virus quantities for structural, biochemical, and therapeutic studies. Unexpectedly, TAK-779 treatment did not affect pandemic GII.4 HuNoV replication, suggesting a distinct host response or evolved immune evasion mechanisms. Ongoing studies are exploring the mechanism of TAK-779 through transcriptomics and secretome analysis and the strategies used by GII.4 to circumvent the innate immune response.
The top upregulated pathway was the host innate immune response, primarily driven by interferons and CXC based chemokines. CXCL10, CXCL11, and CCL5 were among the most highly upregulated interferon-stimulated genes, suggesting their potential role as host restriction factors. To counteract this, we used TAK-779, a CXCR3/CCR5 antagonist, which enhanced GII.3 HuNoV replication (1.5 log10 genome equivalents) and viral spread in a dose- and time-dependent manner. For the first time, we have passaged GII.3 HuNoV in the presence of TAK-779 for over 10 passages in two different HIE lines. This breakthrough enables the generation of sufficient virus quantities for structural, biochemical, and therapeutic studies. Unexpectedly, TAK-779 treatment did not affect pandemic GII.4 HuNoV replication, suggesting a distinct host response or evolved immune evasion mechanisms. Ongoing studies are exploring the mechanism of TAK-779 through transcriptomics and secretome analysis and the strategies used by GII.4 to circumvent the innate immune response.