Niclosamide rescues microcephaly in a humanized in vivo model of Zika infection using human induced neural stem cells

Abstract

Zika virus (ZIKV) is a mosquito-transmitted flavivirus with a causative link to microcephaly, a condition resulting in reduced cranial size and brain abnormalities. Despite recent progress, there is a current lack of in vivo models that permit the study of systemic virus on human neurons in a developing organism that replicates the pathophysiology of human disease. Furthermore, no treatment to date has been reported to reduce ZIKV-induced microcephaly. We tested the effects of ZIKV on human induced neural stem cells (hiNSCs) in vitro and found that infected hiNSCs secrete inflammatory cytokines, display altered differentiation, and become apoptotic. We also utilized this in vitro system to assess the therapeutic effects of niclosamide, an FDA-approved anthelminthic, and found that it decreases ZIKV production, partially restores differentiation, and prevents apoptosis in hiNSCs. We intracranially injected hiNSCs into developing chicks, subjected them to systemic ZIKV infection via the chorioallantoic membrane (CAM), a tissue similar in structure and function to the mammalian placenta, and found that humanized ZIKV-infected embryos developed severe microcephaly including smaller crania, decreased forebrain volume and enlarged ventricles. Lastly, we utilized this humanized model to show that CAM-delivery of niclosamide can partially rescue ZIKV-induced microcephaly and attenuate infection of hiNSCs in vivo..

Discovery and development of safe-in-man broad-spectrum antiviral agents

“…For example, novel anti-ZIKV activities of enoxacin, amodiaquine and niclosamide were discovered recently using human neural progenitor cells, human pluripotent stem cell-derived cortical neural progenitor cells, and human induced neural stem cells, respectively (Cairns et al, 2018;Xu et al, 2019;Zhou et al, 2017).…”

“…Amodiaquine is an anti-malaria drug, which also possesses antiviral activities against DENV, HCV, RRV, SINV, WNV, EFV, EBOV, LASV, RABV, VZV, and HSV-1 in immortalized cell cultures (Boonyasuppayakorn et al, 2014;Hulseberg et al, 2019;Mazzon et al, 2019). Niclosamide is an orally bioavailable anthelmintic drug, which inhibits the broadest range of viruses in vitro and, in some cases, in vivo (Cairns et al, 2018;Fang et al, 2013;Huang et al, 2017;Hulseberg et al, 2019;Jurgeit et al, 2012;Kao et al, 2018;Mazzon et al, 2019;Stachulski et al, 2011;Wang et al, 2016;Wu et al, 2004). These safe-in-man BSAAs represent promising drug candidates.…”

Host-Directed Antivirals: a Realistic Alternative to Fight Zika Virus

“…Niclosamide, a category B antihelmintic drug approved by FDA, was capable to inhibit ZIKV infection and although its antiflaviviral effect has been associated to its ability to neutralize endolysosomal pH and interfere with pH-dependent membrane fusion, in the case of ZIKV it seems that it was affecting other postentry steps [80]. In addition, recently, it has been reported that niclosamide decreases ZIKV production, partially restores differentiation, and prevents apoptosis in human induced NSCs; even more, it can partially rescue ZIKV-induced microcephaly and attenuate infection in a developed humanized ZIKV-infected embryo model in vivo [81]. Likewise, tenovin-1, which represses cell growth and induces apoptosis in cells expressing p53 by inhibiting the protein-deacetylating activities of SirT1 and SirT2 and, thus, affects endosome functions, potently inhibits ZIKV infection in primary placental fibroblast cells [63].…”

Novel Antiviral Activities of Obatoclax, Emetine, Niclosamide, Brequinar, and Homoharringtonine

“…Niclosamide is an orally bioavailable anthelmintic drug and potential antineoplastic agent. In addition, it inhibits the broadest range of viruses, including HSV-2, in vitro and, in some cases, in vivo [28][29][30][31][32][33][34][35][36][37]. It was shown that niclosamide induces endosomal neutralization and prevents virus entry into host cells.…”