Zika virus NS1 drives tunneling nanotube formation for mitochondrial transfer and stealth transmission in trophoblasts

• Published in: Nature communications Vol. 16; no. 1; pp. 1803 - 19
• Main Authors: Michita, Rafael T., Tran, Long B., Bark, Steven J., Kumar, Deepak, Toner, Shay A., Jose, Joyce, Mysorekar, Indira U., Narayanan, Anoop
• Format: Journal Article
• Language: English
• Published: London Nature Publishing Group UK 20.02.2025; Nature Publishing Group; Nature Portfolio
• Subjects: 101/1; 14/19; 38/44; 49; 49/79; 631/326/596/2555; 631/326/596/2558; 64; 96; 96/109; Amino acids; Animals; Antiviral drugs; Cell survival; Disease transmission; Female; Humanities and Social Sciences; Humans; Infections; Infectious Disease Transmission, Vertical; Interferon; Mass spectrometry; Mass spectroscopy; Mitochondria; Mitochondria - metabolism; Mitochondria - virology; multidisciplinary; Mutants; Nanotechnology; Nanotubes; Placenta; Placenta - virology; Pregnancy; Protein purification; Proteins; Science; Science (multidisciplinary); Trophoblasts; Trophoblasts - metabolism; Trophoblasts - virology; Vector-borne diseases; Vertical transmission (disease); Viral Nonstructural Proteins - genetics; Viral Nonstructural Proteins - metabolism; Viruses; Zika virus; Zika Virus - genetics; Zika Virus - metabolism; Zika Virus - physiology; Zika Virus Infection - metabolism; Zika Virus Infection - transmission; Zika Virus Infection - virology
• ISSN: 2041-1723; 2041-1723
• DOI: 10.1038/s41467-025-56927-2

Zika virus (ZIKV) is unique among orthoflaviviruses in its vertical transmission capacity in humans, yet the underlying mechanisms remain incompletely understood. Here, we show that ZIKV induces tunneling nanotubes (TNTs) in placental trophoblasts which facilitate transfer of viral particles, proteins, mitochondria, and RNA to neighboring uninfected cells. TNT formation is driven exclusively via ZIKV non-structural protein 1 (NS1). Specifically, the N-terminal 1-50 amino acids of membrane-bound ZIKV NS1 are necessary for triggering TNT formation in host cells. Trophoblasts infected with TNT-deficient ZIKV ΔTNT mutant virus elicited a robust antiviral IFN-λ 1/2/3 response relative to WT ZIKV, suggesting TNT-mediated trafficking allows ZIKV cell-to-cell transmission camouflaged from host defenses. Using affinity purification-mass spectrometry of cells expressing wild-type NS1 or non-TNT forming NS1, we found mitochondrial proteins are dominant NS1-interacting partners. We demonstrate that ZIKV infection or NS1 expression induces elevated mitochondria levels in trophoblasts and that mitochondria are siphoned via TNTs from healthy to ZIKV-infected cells. Together our findings identify a stealth mechanism that ZIKV employs for intercellular spread among placental trophoblasts, evasion of antiviral interferon response, and the hijacking of mitochondria to augment its propagation and survival and offers a basis for novel therapeutic developments targeting these interactions to limit ZIKV dissemination. Michita et al. show that Zika virus (ZIKV) NS1 induces tunneling nanotubes (TNTs) in placental cells, which facilitate viral spread and transport of mitochondria in placental cells. Infection with a NS1 mutant ZIKV not inducing TNTs results in a higher interferon response than wild-type ZIKV infection.