1. Home
  2. PUBLICATIONS
  3. 2024

Publications by Year: 2024

2024

Linn, Aung Khine, Suwimon Manopwisedjaroen, Phongthon Kanjanasirirat, Suparerk Borwornpinyo, Suradej Hongeng, Phetcharat Phanthong, and Arunee Thitithanyanont. (2024) 2024. “Unveiling the Antiviral Properties of Panduratin A through SARS-CoV-2 Infection Modeling in Cardiomyocytes.”. International Journal of Molecular Sciences 25 (3). https://doi.org/10.3390/ijms25031427.
Publisher's Version

Establishing a drug-screening platform is critical for the discovery of potential antiviral agents against SARS-CoV-2. In this study, we developed a platform based on human induced pluripotent stem cell-derived cardiomyocytes (iPSC-CMs) to investigate SARS-CoV-2 infectivity, with the aim of evaluating potential antiviral agents for anti-SARS-CoV-2 activity and cardiotoxicity. Cultured myocytes of iPSC-CMs and immortalized human cardiomyocyte cell line (AC-16) were primarily characterized for the expression of cardiac markers and host receptors of SARS-CoV-2. An infectivity model for the wild-type SARS-CoV-2 strain was then established. Infection modeling involved inoculating cells with SARS-CoV-2 at varying multiplicities of infection (MOIs) and then quantifying infection using immunofluorescence and plaque assays. Only iPSC-CMs, not AC16 cells, expressed angiotensin-converting enzyme 2 (ACE-2), and quantitative assays confirmed the dose-dependent infection of iPSC-CMs by SARS-CoV-2, unlike the uninfectable AC16 cells lacking the expression of ACE2. Cytotoxicity was evaluated using MTT assays across a concentration range. An assessment of the plant-derived compound panduratin A (panA) showed cytotoxicity at higher doses (50% cytotoxic concentration (CC50) 10.09 μM) but promising antiviral activity against SARS-CoV-2 (50% inhibition concentration (IC50) 0.8-1.6 μM), suppressing infection at concentrations 10 times lower than its CC50. Plaque assays also showed decreased viral production following panA treatment. Overall, by modeling cardiac-specific infectivity, this iPSC-cardiomyocyte platform enables the reliable quantitative screening of compound cytotoxicity alongside antiviral efficacy. By combining disease pathogenesis and pharmacology, this system can facilitate the evaluation of potential novel therapeutics, such as panA, for drug discovery applications.

Purwono, Priyo Budi, Vimvara Vacharathit, Suwimon Manopwisedjaroen, Natali Ludowyke, Ampa Suksatu, and Arunee Thitithanyanont. 2024. “Infection Kinetics, Syncytia Formation, and Inflammatory Biomarkers As Predictive Indicators for the Pathogenicity of SARS-CoV-2 Variants of Concern in Calu-3 Cells”. PLOS ONE 19 (4): 1-24. https://doi.org/10.1371/journal.pone.0301330.
Publisher's Version
The ongoing COVID-19 pandemic has led to the emergence of new SARS-CoV-2 variants as a result of continued host-virus interaction and viral genome mutations. These variants have been associated with varying levels of transmissibility and disease severity. We investigated the phenotypic profiles of six SARS-CoV-2 variants (WT, D614G, Alpha, Beta, Delta, and Omicron) in Calu-3 cells, a human lung epithelial cell line. In our model demonstrated that all variants, except for Omicron, had higher efficiency in virus entry compared to the wild-type. The Delta variant had the greatest phenotypic advantage in terms of early infection kinetics and marked syncytia formation, which could facilitate cell-to-cell spreading, while the Omicron variant displayed slower replication and fewer syncytia formation. We also identified the Delta variant as the strongest inducer of inflammatory biomarkers, including pro-inflammatory cytokines/chemokines (IP-10/CXCL10, TNF-α, and IL-6), anti-inflammatory cytokine (IL-1RA), and growth factors (FGF-2 and VEGF-A), while these inflammatory mediators were not significantly elevated with Omicron infection. These findings are consistent with the observations that there was a generally more pronounced inflammatory response and angiogenesis activity within the lungs of COVID-19 patients as well as more severe symptoms and higher mortality rate during the Delta wave, as compared to less severe symptoms and lower mortality observed during the current Omicron wave in Thailand. Our findings suggest that early infectivity kinetics, enhanced syncytia formation, and specific inflammatory mediator production may serve as predictive indicators for the virulence potential of future SARS-CoV-2 variants.
Huang, Yu-Ching, Sheng-Fan Wang, Bo-Cheng Chen, Zih-Syuan Yang, Meng-Chi Li, Xun-Ying Wu, Meng-Jey Youh, et al. 2024. “Towards Cost-Effective and Lightweight Surface Plasmon Resonance Biosensing for H5N1 Avian Influenza Virus Detection: Integration of Novel Near-Infrared Organic Photodetectors”. Sensors and Actuators B: Chemical 400: 134898. https://doi.org/https://doi.org/10.1016/j.snb.2023.134898.
Publisher's Version
H5N1 avian influenza virus (AIV) persists in causing highly fatal human infections, demanding rapid and accurate diagnostic assessment. In this study, we introduce an intensity-based SPR sensor utilizing NIR wavelength excitation in tandem with a specifically engineered NIR-OPD. The active layer of the OPD consists of a PTB7-Th and COTIC-4 F blend, offering an optimized response for a 980 nm excitation wavelength. Key performance metrics of this OPD include a low Jd of 0.185 nA/cm2, a high responsivity of 0.35 A/W, an EQE of 44.74%, and an exceptional detectivity of 4.59 × 1013 Jones at 980 nm wavelength under zero bias. It also exhibits a wide LDR of 113 dB. The integration of such OPDs into our SPR sensor provides advantages in compactness and cost-effectiveness. Employing this sensor, we detected the H5N1 AIV using a custom high-affinity polyclonal antibody against HA envelope of the H5N1 virus, completing analyses of culture medium samples within 12 min. The detection limit of this biosensor for the H5N1 AIV in PBS-diluted culture medium is approximately 4.3 × 104 copies/mL. When compared to a commercial H5-Ag lateral flow test kit, our biosensor showed a sensitivity 37 times higher. Key attributes of our biosensor include 3D printing technology for easy alignment of optical components and a rapid, simplified detection procedure. Collectively, our findings open up the potential of our SPR biosensor as an efficient tool for detecting H5N1 AIV, promising advancements in on-site detection methodologies.
Pisuttinusart, Nuttapat, Balamurugan Shanmugaraj, Chanya Srisaowakarn, Chutitorn Ketloy, Eakachai Prompetchara, Arunee Thitithanyanont, and Waranyoo Phoolcharoen. (2024) 2024. “Immunogenicity of a Recombinant Plant-Produced Respiratory Syncytial Virus F Subunit Vaccine in Mice.”. Biotechnology Reports (Amsterdam, Netherlands) 41: e00826. https://doi.org/10.1016/j.btre.2023.e00826.
Publisher's Version

Respiratory syncytial virus (RSV) is a highly infectious respiratory virus that causes serious illness, particularly in young children, elderly people, and those with immunocompromised individuals. RSV infection is the leading cause of infant hospitalization and can lead to serious complications such as pneumonia and bronchiolitis. Currently, there is an RSV vaccine approved exclusively for the elderly population, but no approved vaccine specifically designed for infants or any other age groups. Therefore, it is crucial to continue the development of an RSV vaccine specifically tailored for these populations. In this study, the immunogenicity of the two plant-produced RSV-F Fc fusion proteins (Native construct and structural stabilized construct) were examined to assess them as potential vaccine candidates for RSV. The RSV-F Fc fusion proteins were transiently expressed in Nicotiana benthamiana and purified using protein A affinity column chromatography. The recombinant RSV-F Fc fusion protein was recognized by the monoclonal antibody Motavizumab specific against RSV-F protein. Moreover, the immunogenicity of the two purified RSV-F Fc proteins were evaluated in mice by formulating with different adjuvants. According to our results, the plant-produced RSV-F Fc fusion protein is immunogenic in mice. These preliminary findings, demonstrate the immunogenicity of plant-based RSV-F Fc fusion protein, however, further preclinical studies such as antigen dose and adjuvant optimization, safety, toxicity, and challenge studies in animal models are necessary in order to prove the vaccine efficacy.

Puthanakit, Thanyawee, Eakachai Prompetchara, Sivaporn Gatechompol, Chutitorn Ketloy, Arunee Thitithanyanont, Anan Jongkaewwattana, Supranee Buranapraditkun, et al. (2024) 2024. “Phase II Prefusion Non-Stabilised Covid-19 MRNA Vaccine Randomised Study.”. Scientific Reports 14 (1): 2373. https://doi.org/10.1038/s41598-023-49653-6.
Publisher's Version

ChulaCov19 mRNA vaccine demonstrated promising phase 1 results. Healthy adults aged 18-59 years were double-blind randomised 4:1 to receive two intramuscular doses of ChulaCov19 50 µg or placebo. Primary endpoints were safety and microneutralization antibody against-wild-type (Micro-VNT50) at day 50. One hundred fifty adults with median (IQR) age 37 (30-46) years were randomised. ChulaCov19 was well tolerated, and most adverse events were mild to moderate and temporary. Geometric mean titres (GMT) of neutralizing titre against wild-type for ChulaCov19 on day 50 were 1367 IU/mL. T-cell IFN-γ-ELISpot showed the highest responses at one week (Day29) after dose 2 then gradually declined. ChulaCov19 50 µg is well tolerated and elicited high neutralizing antibodies and strong T-cell responses in healthy adults.Trial registration number: ClinicalTrials.gov Identifier NCT04566276, 28/09/2020.