DNA Damage Response Signaling Is Crucial for Effective Chikungunya Virus Replication

Chatterjee, Sanchari; Kumar, Sameer; Mamidi, Prabhudutta; Datey, Ankita; Sengupta, Soumya; Mahish, Chandan; Laha, Eshna; De, Saikat; Keshry, Supriya Suman; Nayak, Tapas Kumar; Ghosh, Soumyajit; Singh, Sharad; Subudhi, Bharat Bhusan; Chattopadhyay, Subhasis; Chattopadhyay, Soma

doi:10.1128/jvi.01334-22

Cited by 10 publications

(15 citation statements)

References 38 publications

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“…Based on our previous reports, where it was established that maximum CHIKV infection occurs at 8 hours post-infection (hpi) time point in the RAW264.7 macrophages, 8 hpi was selected for cell harvesting to carry out all the experiments of viral infection ( 19 , 22 ). E2, an envelope protein of CHIKV, was taken as a marker to assess CHIKV infection in different host systems ( 19 , 40 , 41 , 45 , 46 ).…”

Section: Resultsmentioning

confidence: 99%

“…The RAW264.7 cell line, TLR4KO RAW cell line, BALB/c and C57BL/6 mice-derived peritoneal monocyte-macrophages were infected with CHIKV-IS as reported earlier with minute modifications ( 19 , 22 , 40 , 41 , 45 , 46 ). Briefly, 4.5x10 6 cells were seeded in 90 mm dishes and allowed to grow for 16-18 h. Next, the cells were washed with 1X PBS 2 times and pre-incubated with either TAK-242 or DMSO for 3 h. For TAK-242 treated conditions, the cells were incubated with 0.5 and/or 1 µM concentrations of TAK-242 for 3 h before infection, during infection and post-infection.…”

Section: Methodsmentioning

confidence: 99%

“…Human blood was drawn from healthy donors following the guidelines of the Institutional Ethics Committee, NISER, Bhubaneswar (NISER/IEC/2022-04). The procedure for generating myeloid adherent cells from human peripheral blood mononuclear cells (hPBMC) was followed as described elsewhere with little modifications (38)(39)(40)(41). Briefly, circulating monocytes were enriched by 2 h adherence after Hi-Sep LSM (catalog number: HiSep LSM ™ 1077-LS001; HiMedia Laboratories Pvt Ltd, India) based density gradient-centrifugation according to the manufacturer's instructions.…”

Section: Hpbmc Isolationmentioning

confidence: 99%

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TLR4 is one of the receptors for Chikungunya virus envelope protein E2 and regulates virus induced pro-inflammatory responses in host macrophages

Mahish

Chatterjee

et al. 2023

Front. Immunol.

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Toll like receptor 4 (TLR4), a pathogen-associated molecular pattern (PAMP) receptor, is known to exert inflammation in various cases of microbial infection, cancer and autoimmune disorders. However, any such involvement of TLR4 in Chikungunya virus (CHIKV) infection is yet to be explored. Accordingly, the role of TLR4 was investigated towards CHIKV infection and modulation of host immune responses in the current study using mice macrophage cell line RAW264.7, primary macrophage cells of different origins and in vivo mice model. The findings suggest that TLR4 inhibition using TAK-242 (a specific pharmacological inhibitor) reduces viral copy number as well as reduces the CHIKV-E2 protein level significantly using p38 and JNK-MAPK pathways. Moreover, this led to reduced expression of macrophage activation markers like CD14, CD86, MHC-II and pro-inflammatory cytokines (TNF, IL-6, MCP-1) significantly in both the mouse primary macrophages and RAW264.7 cell line, in vitro. Additionally, TAK-242-directed TLR4 inhibition demonstrated a significant reduction of percent E2-positive cells, viral titre and TNF expression in hPBMC-derived macrophages, in vitro. These observations were further validated in TLR4-knockout (KO) RAW cells. Furthermore, the interaction between CHIKV-E2 and TLR4 was demonstrated by immuno-precipitation studies, in vitro and supported by molecular docking analysis, in silico. TLR4-dependent viral entry was further validated by an anti-TLR4 antibody-mediated blocking experiment. It was noticed that TLR4 is necessary for the early events of viral infection, especially during the attachment and entry stages. Interestingly, it was also observed that TLR4 is not involved in the post-entry stages of CHIKV infection in host macrophages. The administration of TAK-242 decreased CHIKV infection significantly by reducing disease manifestations, improving survivability (around 75%) and reducing inflammation in mice model. Collectively, for the first time, this study reports TLR4 as one of the novel receptors to facilitate the attachment and entry of CHIKV in host macrophages, the TLR4-CHIKV-E2 interactions are essential for efficient viral entry and modulation of infection-induced pro-inflammatory responses in host macrophages, which might have translational implication for designing future therapeutics to regulate the CHIKV infection.

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Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Hpbmc Isolationmentioning

confidence: 99%

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TLR4 is one of the receptors for Chikungunya virus envelope protein E2 and regulates virus induced pro-inflammatory responses in host macrophages

Mahish

Chatterjee

et al. 2023

Front. Immunol.

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“…Additionally, combining these compounds with specific direct antiviral compounds may lead to synergistic effects with the potential to develop more comprehensive treatment plans that target multiple pathways. Chatterjee et al (2022) recently identified checkpoint kinase proteins Chk1 and Chk2 (also known as CHEK1 and CHEK2), which are commonly known as DNA damage response proteins, as host factors that are involved in CHIKV replication. Similarly, De Caluwé et al (2021) characterised the role of the CD147 protein complex in CHIKV entry into host cells.…”

Section: Antiviral Strategies Targeting the Virus Replication Cyclementioning

confidence: 99%

Evolution and immunopathology of chikungunya virus informs therapeutic development

Sousa

Komarudin

Findlay-Greene

et al. 2023

Disease Models &Amp; Mechanisms

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Chikungunya virus (CHIKV), a mosquito-borne alphavirus, is an emerging global threat identified in more than 60 countries across continents. The risk of CHIKV transmission is rising due to increased global interactions, year-round presence of mosquito vectors, and the ability of CHIKV to produce high host viral loads and undergo mutation. Although CHIKV disease is rarely fatal, it can progress to a chronic stage, during which patients experience severe debilitating arthritis that can last from several weeks to months or years. At present, there are no licensed vaccines or antiviral drugs for CHIKV disease, and treatment is primarily symptomatic. This Review provides an overview of CHIKV pathogenesis and explores the available therapeutic options and the most recent advances in novel therapeutic strategies against CHIKV infections.

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“…A wide array of host factors, in addition to the non-structural proteins, leads to effective CHIKV infection ( 1 , 2 ). However, there is still much to explore on how cellular proteins contribute to effective CHIKV infection.…”

Section: Lettermentioning

confidence: 99%

A hidden gem Catenin-α-1 is essential for Chikungunya virus infection

Chatterjee,

Subudhi,

Chattopadhyay

2023

Microbiol Spectr

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DNA Damage Response Signaling Is Crucial for Effective Chikungunya Virus Replication

Abstract: Being intracellular parasites, viruses require several host cell machineries for effectively replicating their genome, along with virus-encoded enzymes. One of the strategies involves hijacking of the DDR pathways.

Cited by 10 publications

References 38 publications

TLR4 is one of the receptors for Chikungunya virus envelope protein E2 and regulates virus induced pro-inflammatory responses in host macrophages

TLR4 is one of the receptors for Chikungunya virus envelope protein E2 and regulates virus induced pro-inflammatory responses in host macrophages

Evolution and immunopathology of chikungunya virus informs therapeutic development

A hidden gem Catenin-α-1 is essential for Chikungunya virus infection

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