41 results on '"Alfajaro, Mia Madel"'
Search Results
2. Pharmacological disruption of mSWI/SNF complex activity restricts SARS-CoV-2 infection
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Wei, Jin, Patil, Ajinkya, Collings, Clayton K., Alfajaro, Mia Madel, Liang, Yu, Cai, Wesley L., Strine, Madison S., Filler, Renata B., DeWeirdt, Peter C., Hanna, Ruth E., Menasche, Bridget L., Ökten, Arya, Peña-Hernández, Mario A., Klein, Jon, McNamara, Andrew, Rosales, Romel, McGovern, Briana L., Luis Rodriguez, M., García-Sastre, Adolfo, White, Kris M., Qin, Yiren, Doench, John G., Yan, Qin, Iwasaki, Akiko, Zwaka, Thomas P., Qi, Jun, Kadoch, Cigall, and Wilen, Craig B.
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- 2023
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3. Genome-wide bidirectional CRISPR screens identify mucins as host factors modulating SARS-CoV-2 infection
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Biering, Scott B., Sarnik, Sylvia A., Wang, Eleanor, Zengel, James R., Leist, Sarah R., Schäfer, Alexandra, Sathyan, Varun, Hawkins, Padraig, Okuda, Kenichi, Tau, Cyrus, Jangid, Aditya R., Duffy, Connor V., Wei, Jin, Gilmore, Rodney C., Alfajaro, Mia Madel, Strine, Madison S., Nguyenla, Xammy, Van Dis, Erik, Catamura, Carmelle, Yamashiro, Livia H., Belk, Julia A., Begeman, Adam, Stark, Jessica C., Shon, D. Judy, Fox, Douglas M., Ezzatpour, Shahrzad, Huang, Emily, Olegario, Nico, Rustagi, Arjun, Volmer, Allison S., Livraghi-Butrico, Alessandra, Wehri, Eddie, Behringer, Richard R., Cheon, Dong-Joo, Schaletzky, Julia, Aguilar, Hector C., Puschnik, Andreas S., Button, Brian, Pinsky, Benjamin A., Blish, Catherine A., Baric, Ralph S., O’Neal, Wanda K., Bertozzi, Carolyn R., Wilen, Craig B., Boucher, Richard C., Carette, Jan E., Stanley, Sarah A., Harris, Eva, Konermann, Silvana, and Hsu, Patrick D.
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- 2022
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4. Tuft-cell-intrinsic and -extrinsic mediators of norovirus tropism regulate viral immunity
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Strine, Madison S., Alfajaro, Mia Madel, Graziano, Vincent R., Song, Jaewon, Hsieh, Leon L., Hill, Ryan, Guo, Jun, VanDussen, Kelli L., Orchard, Robert C., Baldridge, Megan T., Lee, Sanghyun, and Wilen, Craig B.
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- 2022
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5. Perfusion change in benign prostatic hyperplasia before and after castration in a canine model: Contrast enhanced ultrasonography and CT perfusion study
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Yoon, Sooa, Alfajaro, Mia Madel, Cho, Kyoung-Oh, Choi, Ul-Soo, Je, Hyejin, Jung, Jinwoo, Jang, Youjung, and Choi, Jihye
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- 2020
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6. Intestinal tuft cell immune privilege enables norovirus persistence.
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Strine, Madison S., Fagerberg, Eric, Darcy, Patrick W., Barrón, Gabriel M., Filler, Renata B., Alfajaro, Mia Madel, D'Angelo-Gavrish, Nicole, Wang, Fang, Graziano, Vincent R., Menasché, Bridget L., Damo, Martina, Wang, Ya-Ting, Howitt, Michael R., Lee, Sanghyun, Joshi, Nikhil S., Mucida, Daniel, and Wilen, Craig B.
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NOROVIRUSES ,T cell receptors ,T cells ,INTESTINES ,NOROVIRUS diseases ,SURGICAL site infections - Abstract
The persistent murine norovirus strain MNV
CR6 is a model for human norovirus and enteric viral persistence. MNVCR6 causes chronic infection by directly infecting intestinal tuft cells, rare chemosensory epithelial cells. Although MNVCR6 induces functional MNV-specific CD8+ T cells, these lymphocytes fail to clear infection. To examine how tuft cells promote immune escape, we interrogated tuft cell interactions with CD8+ T cells by adoptively transferring JEDI (just EGFP death inducing) CD8+ T cells into Gfi1b-GFP tuft cell reporter mice. Unexpectedly, some intestinal tuft cells partially resisted JEDI CD8+ T cell–mediated killing—unlike Lgr5+ intestinal stem cells and extraintestinal tuft cells—despite seemingly normal antigen presentation. When targeting intestinal tuft cells, JEDI CD8+ T cells predominantly adopted a T resident memory phenotype with decreased effector and cytotoxic capacity, enabling tuft cell survival. JEDI CD8+ T cells neither cleared nor prevented MNVCR6 infection in the colon, the site of viral persistence, despite targeting a virus-independent antigen. Ultimately, we show that intestinal tuft cells are relatively resistant to CD8+ T cells independent of norovirus infection, representing an immune-privileged niche that can be leveraged by enteric microbes. Editor's summary: Norovirus is a major cause of acute foodborne illness that can also lead to persistent infections. Specialized chemosensory epithelial cells called tuft cells are the target cell for murine norovirus, but how chronic infections are established remains unclear. Using mice with CD8+ T cells bearing a fluorescent protein-specific T cell receptor, Strine et al. found that intestinal tuft cells are intrinsically resistant to CD8+ T cell–mediated killing compared with other gut epithelial cells. Tuft cell–mediated immune evasion from CD8+ T cells enabled norovirus persistence in the colon. These findings indicate that intestinal tuft cells represent an immune-privileged niche that is intrinsically resistant to T cell responses. —Claire Olingy [ABSTRACT FROM AUTHOR]- Published
- 2024
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7. Human iPSC-Based Model of COPD to Investigate Disease Mechanisms, Predict SARS-COV-2 Outcome, and Test Preventive Immunotherapy.
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Dagher, Rania, Moldobaeva, Aigul, Gubbins, Elise, Clark, Sydney, Alfajaro, Mia Madel, Wilen, Craig B, Hawkins, Finn, Qu, Xiaotao, Chiang, Chia Chien, Li, Yang, Clarke, Lori, Ikeda, Yasuhiro, Brown, Charles, Kolbeck, Roland, Ma, Qin, Rojas, Mauricio, Koff, Jonathan L, and Ghaedi, Mahboobe
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CHRONIC obstructive pulmonary disease ,SARS-CoV-2 ,IMMUNOGLOBULINS ,LUNGS ,TISSUE remodeling ,IMMUNOTHERAPY - Abstract
Chronic inflammation and dysregulated repair mechanisms after epithelial damage have been implicated in chronic obstructive pulmonary disease (COPD). However, the lack of ex vivo-models that accurately reflect multicellular lung tissue hinders our understanding of epithelial-mesenchymal interactions in COPD. Through a combination of transcriptomic and proteomic approaches applied to a sophisticated in vitro iPSC-alveolosphere with fibroblasts model, epithelial-mesenchymal crosstalk was explored in COPD and following SARS-CoV-2 infection. These experiments profiled dynamic changes at single-cell level of the SARS-CoV-2-infected alveolar niche that unveiled the complexity of aberrant inflammatory responses, mitochondrial dysfunction, and cell death in COPD, which provides deeper insights into the accentuated tissue damage/inflammation/remodeling observed in patients with SARS-CoV-2 infection. Importantly, this 3D system allowed for the evaluation of ACE2-neutralizing antibodies and confirmed the potency of this therapy to prevent SARS-CoV-2 infection in the alveolar niche. Thus, iPSC-alveolosphere cultured with fibroblasts provides a promising model to investigate disease-specific mechanisms and to develop novel therapeutics. [ABSTRACT FROM AUTHOR]
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- 2024
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8. Development of a live attenuated trivalent porcine rotavirus A vaccine against disease caused by recent strains most prevalent in South Korea
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Park, Jun-Gyu, Alfajaro, Mia Madel, Cho, Eun-Hyo, Kim, Ji-Yun, Soliman, Mahmoud, Baek, Yeong-Bin, Park, Chul-Ho, Lee, Ju-Hwan, Son, Kyu-Yeol, Cho, Kyoung-Oh, and Kang, Mun-Il
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- 2019
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9. Genetic diversity of the VP7, VP4 and VP6 genes of Korean porcine group C rotaviruses
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Jeong, Young-Ju, Matthijnssens, Jelle, Kim, Deok-Song, Kim, Ji-Yun, Alfajaro, Mia Madel, Park, Jun-Gyu, Hosmillo, Myra, Son, Kyu-Yeol, Soliman, Mahmoud, Baek, Yeong-Bin, Kwon, Joseph, Choi, Jong-Soon, Kang, Mun-Il, and Cho, Kyoung-Oh
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- 2015
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10. Comparison of pathogenicities and nucleotide changes between porcine and bovine reassortant rotavirus strains possessing the same genotype constellation in piglets and calves
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Park, Jun-Gyu, Kim, Deok-Song, Matthijnssens, Jelle, Kwon, Hyoung-Jun, Zeller, Mark, Alfajaro, Mia Madel, Son, Kyu-Yeol, Hosmillo, Myra, Ryu, Eun-Hye, Kim, Ji-Yun, Lee, Ju-Hwan, Park, Su-Jin, Kang, Mun-Il, Kwon, Joseph, Choi, Jong-Soon, and Cho, Kyoung-Oh
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- 2014
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11. Anti-rotavirus effects by combination therapy of stevioside and Sophora flavescens extract
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Alfajaro, Mia Madel, Rho, Mun-Chual, Kim, Hyun-Jeong, Park, Jun-Gyu, Kim, Deok-Song, Hosmillo, Myra, Son, Kyu-Yeol, Lee, Ju-Hwan, Park, Sang-Ik, Kang, Mun-Il, Ryu, Young Bae, Park, Ki Hun, Oh, Hyun-Mee, Lee, Seung Woong, Park, Su-Jin, Lee, Woo Song, and Cho, Kyoung-Oh
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- 2014
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12. The KDM6A-KMT2D-p300 axis regulates susceptibility to diverse coronaviruses by mediating viral receptor expression.
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Wei, Jin, Alfajaro, Mia Madel, Cai, Wesley L., Graziano, Vincent R., Strine, Madison S., Filler, Renata B., Biering, Scott B., Sarnik, Sylvia A., Patel, Sonam, Menasche, Bridget L., Compton, Susan R., Konermann, Silvana, Hsu, Patrick D., Orchard, Robert C., Yan, Qin, and Wilen, Craig B.
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CORONAVIRUS diseases , *CORONAVIRUSES , *VIRUS diseases , *COVID-19 , *SARS-CoV-2 - Abstract
Identification of host determinants of coronavirus infection informs mechanisms of pathogenesis and may provide novel therapeutic targets. Here, we demonstrate that the histone demethylase KDM6A promotes infection of diverse coronaviruses, including SARS-CoV, SARS-CoV-2, MERS-CoV and mouse hepatitis virus (MHV) in a demethylase activity-independent manner. Mechanistic studies reveal that KDM6A promotes viral entry by regulating expression of multiple coronavirus receptors, including ACE2, DPP4 and Ceacam1. Importantly, the TPR domain of KDM6A is required for recruitment of the histone methyltransferase KMT2D and histone deacetylase p300. Together this KDM6A-KMT2D-p300 complex localizes to the proximal and distal enhancers of ACE2 and regulates receptor expression. Notably, small molecule inhibition of p300 catalytic activity abrogates ACE2 and DPP4 expression and confers resistance to all major SARS-CoV-2 variants and MERS-CoV in primary human airway and intestinal epithelial cells. These data highlight the role for KDM6A-KMT2D-p300 complex activities in conferring diverse coronaviruses susceptibility and reveal a potential pan-coronavirus therapeutic target to combat current and emerging coronaviruses. One Sentence Summary: The KDM6A/KMT2D/EP300 axis promotes expression of multiple viral receptors and represents a potential drug target for diverse coronaviruses. Author summary: Three highly pathogenic coronaviruses SARS-CoV, MERS-CoV, and SARS-CoV-2 have spilled over into the human population in the last two decades. It is important to determine the mechanisms underlying how these viruses cause infection in order to increase our understanding of how viruses cause disease and to aid in the development of new therapeutics against current and future coronaviruses. Here we identify that the host proteins KDM6A, KMT2D, and p300 are critical for coronavirus infection. KDM6A, KMT2D, and p300 are important proteins implicated in cancer and developmental syndromes. They modify histone proteins, which regulate expression of specific host genes. We show that the KDM6A-KMT2D-p300 axis turns on expression of the host genes ACE2 and DPP4, which encode the receptors for SARS-like coronaviruses and MERS-CoV, respectively. KDM6A, KMT2D, and p300 act on the enhancer regions of the coronavirus receptor genes to turn on receptor expression. Small molecule inhibitors of p300 activity block receptor expression in both cell lines and primary human airway cells. This inhibition is well tolerated and makes the cells resistant to infection. This highlights the potential for targeting the KDM6A-KMT2D-p300 axis for diverse coronavirus infections. [ABSTRACT FROM AUTHOR]
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- 2023
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13. Murine Norovirus: Additional Protocols for Basic and Antiviral Studies.
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Wobus, Christiane E., Peiper, Amy M., McSweeney, Alice M., Young, Vivienne L., Chaika, Maryna, Lane, Miranda Sophie, Lingemann, Marit, Deerain, Joshua M., Strine, Madison S., Alfajaro, Mia Madel, Helm, Emily W., Karst, Stephanie M., Mackenzie, Jason M., Taube, Stefan, Ward, Vernon K., and Wilen, Craig B.
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- 2023
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14. DYRK1A promotes viral entry of highly pathogenic human coronaviruses in a kinase-independent manner.
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Strine, Madison S., Cai, Wesley L., Wei, Jin, Alfajaro, Mia Madel, Filler, Renata B., Biering, Scott B., Sarnik, Sylvia, Chow, Ryan D., Patil, Ajinkya, Cervantes, Kasey S., Collings, Clayton K., DeWeirdt, Peter C., Hanna, Ruth E., Schofield, Kevin, Hulme, Christopher, Konermann, Silvana, Doench, John G., Hsu, Patrick D., Kadoch, Cigall, and Yan, Qin
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SARS-CoV-2 ,MERS coronavirus ,COVID-19 ,GENE enhancers ,CORONAVIRUSES ,INSULIN receptors ,GENE expression ,EPHRIN receptors - Abstract
Identifying host genes essential for Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) has the potential to reveal novel drug targets and further our understanding of Coronavirus Disease 2019 (COVID-19). We previously performed a genome-wide CRISPR/Cas9 screen to identify proviral host factors for highly pathogenic human coronaviruses. Few host factors were required by diverse coronaviruses across multiple cell types, but DYRK1A was one such exception. Although its role in coronavirus infection was previously undescribed, DYRK1A encodes Dual Specificity Tyrosine Phosphorylation Regulated Kinase 1A and is known to regulate cell proliferation and neuronal development. Here, we demonstrate that DYRK1A regulates ACE2 and DPP4 transcription independent of its catalytic kinase function to support SARS-CoV, SARS-CoV-2, and Middle East Respiratory Syndrome Coronavirus (MERS-CoV) entry. We show that DYRK1A promotes DNA accessibility at the ACE2 promoter and a putative distal enhancer, facilitating transcription and gene expression. Finally, we validate that the proviral activity of DYRK1A is conserved across species using cells of nonhuman primate and human origin. In summary, we report that DYRK1A is a novel regulator of ACE2 and DPP4 expression that may dictate susceptibility to multiple highly pathogenic human coronaviruses. This study reveals that Dual Specificity Tyrosine Phosphorylation Regulated Kinase 1A (DYRK1A) can facilitate coronavirus entry by driving chromatin accessibility and active transcription at coronavirus receptor gene loci, independent of its catalytic function. [ABSTRACT FROM AUTHOR]
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- 2023
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15. Rotavirus-Induced Early Activation of the RhoA/ROCK/MLC Signaling Pathway Mediates the Disruption of Tight Junctions in Polarized MDCK Cells
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Soliman, Mahmoud, Cho, Eun-Hyo, Park, Jun-Gyu, Kim, Ji-Yun, Alfajaro, Mia Madel, Baek, Yeong-Bin, Kim, Deok-Song, Kang, Mun-Il, Park, Sang-Ik, and Cho, Kyoung-Oh
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- 2018
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16. Porcine sapovirus Cowden strain enters LLC-PK cells via clathrin- and cholesterol-dependent endocytosis with the requirement of dynamin II
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Soliman, Mahmoud, Kim, Deok-Song, Kim, Chonsaeng, Seo, Ja-Young, Kim, Ji-Yun, Park, Jun-Gyu, Alfajaro, Mia Madel, Baek, Yeong-Bin, Cho, Eun-Hyo, Park, Sang-Ik, Kang, Mun-Il, Chang, Kyeong-Ok, Goodfellow, Ian, and Cho, Kyoung-Oh
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- 2018
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17. Pathogenicity characterization of a bovine triple reassortant rotavirus in calves and piglets
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Kim, Hyun-Jeong, Park, Jun-Gyu, Alfajaro, Mia Madel, Kim, Deok-Song, Hosmillo, Myra, Son, Kyu-Yeol, Lee, Ju-Hwan, Bae, You-Chan, Park, Sang-Ik, Kang, Mun-Il, and Cho, Kyoung-Oh
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- 2012
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18. Azuki bean (Vigna angularis) extract inhibits the development of experimentally induced atopic dermatitis-like skin lesions in NC/Nga mice
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Collantes, Therese Marie, Rho, Mun-Chual, Kwon, Hyoung-Jun, Jung, Bock-Gie, Alfajaro, Mia Madel, Kim, Deok-Song, Kim, Hyun-Jeong, Hosmillo, Myra, Park, Jun-Gyu, Son, Kyu-Yeol, Park, Sang-Ik, Kang, Mun-Il, Park, Su-Jin, Lee, Seung Woong, Lee, Woo-Song, and Cho, Kyoung-Oh
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- 2012
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19. Intestinal and extra-intestinal pathogenicity of a bovine reassortant rotavirus in calves and piglets
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Kim, Hyun-Jeong, Park, Jun-Gyu, Matthijnssens, Jelle, Lee, Ju-Hwan, Bae, You-Chan, Alfajaro, Mia Madel, Park, Sang-Ik, Kang, Mun-Il, and Cho, Kyoung-Oh
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- 2011
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20. Reassortment among bovine, porcine and human rotavirus strains results in G8P[7] and G6P[7] strains isolated from cattle in South Korea
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Park, Sang-Ik, Matthijnssens, Jelle, Saif, Linda J., Kim, Hyun-Jeong, Park, Jun-Gyu, Alfajaro, Mia Madel, Kim, Deok-Song, Son, Kyu-Yeol, Yang, Dong-Kun, Hyun, Bang-Hun, Kang, Mun-Il, and Cho, Kyoung-Oh
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- 2011
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21. Evidences and consequences of extra-intestinal spread of rotaviruses in humans and animals
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Alfajaro, Mia Madel and Cho, Kyoung-Oh
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- 2014
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22. Development of universal SYBR Green real-time RT-PCR for the rapid detection and quantitation of bovine and porcine toroviruses
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Hosmillo, Myra D.T., Jeong, Young-Ju, Kim, Hyun-Jeong, Collantes, Therese Marie, Alfajaro, Mia Madel, Park, Jun-Gyu, Kim, Ha-Hyun, Kwon, Hyung-Jun, Park, Su-Jin, Kang, Mun-Il, Park, Sang-Ik, and Cho, Kyoung-Oh
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- 2010
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23. High-affinity, neutralizing antibodies to SARS-CoV-2 can be made without T follicular helper cells.
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Chen, Jennifer S., Chow, Ryan D., Song, Eric, Mao, Tianyang, Israelow, Benjamin, Kamath, Kathy, Bozekowski, Joel, Haynes, Winston A., Filler, Renata B., Menasche, Bridget L., Wei, Jin, Alfajaro, Mia Madel, Song, Wenzhi, Peng, Lei, Carter, Lauren, Weinstein, Jason S., Gowthaman, Uthaman, Chen, Sidi, Craft, Joe, and Shon, John C.
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T helper cells ,PARAINFLUENZA viruses ,RABIES virus ,MERS coronavirus ,SARS-CoV-2 ,IMMUNOGLOBULINS ,FOLLICULAR dendritic cells - Abstract
However, we discovered that antibodies from I Bcl6 i SP fl/fl sp I Cd4 i SP Cre sp mice still demonstrated substantial affinity toward S and RBD (2.7- and 1.9-fold reduction against S and RBD, respectively, relative to I Bcl6 i SP fl/fl sp mice), suggesting that non-T SB FH sb CD4 SP + sp T cells could also promote high-affinity antibody production. These results suggest that T SB FH sb cells help focus the antibody response to particular SARS-CoV-2 epitopes, whereas non-T SB FH sb cells promote antibodies to a wider array of targets. Antibodies mediate protection against severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), the causative agent of the coronavirus disease 2019 (COVID-19) pandemic ([1]). TFH cells focus the antibody repertoire but are dispensable for broad coverage of SARS-CoV-2... We next characterized the antibody epitope repertoire of T SB FH sb -dependent versus T SB FH sb -independent responses to SARS-CoV-2. [Extracted from the article]
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- 2022
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24. Anti-rotaviral effects of Glycyrrhiza uralensis extract in piglets with rotavirus diarrhea
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Alfajaro Mia Madel, Kim Hyun-Jeong, Park Jun-Gyu, Ryu Eun-Hye, Kim Ji-Yun, Jeong Young-Ju, Kim Deok-Song, Hosmillo Myra, Son Kyu-Yeol, Lee Ju-Hwan, Kwon Hyung-Jun, Ryu Young Bae, Park Su-Jin, Park Sang-Ik, Lee Woo Song, and Cho Kyoung-Oh
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Rotavirus ,Enteritis ,Glycyrrhiza uralensis extract ,Anti-rotaviral drug ,Infectious and parasitic diseases ,RC109-216 - Abstract
Abstract Background Since rotavirus is one of the leading pathogens that cause severe gastroenteritis and represents a serious threat to human and animal health, researchers have been searching for cheap, safe, and effective anti-rotaviral drugs. There is a widespread of interest in using natural products as antiviral agents, and among them, licorice derived from Glycyrrhiza spp. has exerted antiviral properties against several viruses. In this study, anti-rotaviral efficacy of Glycyrrhiza uralensis extract (GUE) as an effective and cheaper remedy without side-effects was evaluated in colostrums-deprived piglets after induction of rotavirus diarrhea. Methods Colostrums-deprived piglets were inoculated with porcine rotavirus K85 (G5P[7]) strain. On the onset of diarrhea, piglets were treated with different concentration of GUE. To evaluate the antiviral efficacy of GUE, fecal consistency score, fecal virus shedding and histological changes of the small intestine, mRNA expression levels of inflammation-related cytokines (IL8, IL10, IFN-β, IFN-γ and TNF-α), signaling molecules (p38 and JNK), and transcription factor (NFκB) in the small intestine and spleen were determined. Results Among the dosages (100-400 mg/ml) administrated to animals, 400 mg/ml of GUE cured diarrhea, and markedly improved small intestinal lesion score and fecal virus shedding. mRNA expression levels of inflammation-related cytokines (IL8, IL10, IFN-β, IFN-γ and TNF-α), signaling molecules (p38 and JNK), and transcription factor (NFκB) in the small intestine and spleen were markedly increased in animals with RVA-induced diarrhea, but dose- dependently decreased in GUE treated animals after RVA-induced diarrhea. Conclusions GUE cures rotaviral enteritis by coordinating antiviral and anti-inflammatory effects. Therapy of this herbal medicine can be a viable medication for curing rotaviral enteritis in animals and humans.
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- 2012
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25. Nonsteroidal Anti-inflammatory Drugs Dampen the Cytokine and Antibody Response to SARS-CoV-2 Infection.
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Chen, Jennifer S., Alfajaro, Mia Madel, Chow, Ryan D., Jin Wei, Filler, Renata B., Eisenbarth, Stephanie C., and Wilen, Craig B.
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COVID-19 , *SARS-CoV-2 , *ANTI-inflammatory agents , *ANTIBODY formation , *HUMAN cell culture , *PROSTAGLANDIN receptors - Abstract
Identifying drugs that regulate severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection and its symptoms has been a pressing area of investigation during the coronavirus disease 2019 (COVID-19) pandemic. Nonsteroidal anti-inflammatory drugs (NSAIDs), which are frequently used for the relief of pain and inflammation, could modulate both SARS-CoV-2 infection and the host response to the virus. NSAIDs inhibit the enzymes cyclooxygenase-1 (COX-1) and cyclooxygenase-2 (COX-2), which mediate the production of prostaglandins (PGs). Since PGs play diverse biological roles in homeostasis and inflammatory responses, inhibiting PG production with NSAIDs could affect COVID-19 pathogenesis in multiple ways, including (i) altering susceptibility to infection by modifying expression of angiotensin-converting enzyme 2 (ACE2), the cell entry receptor for SARS-CoV-2; (ii) regulating replication of SARS-CoV-2 in host cells; and (iii) modulating the immune response to SARS-CoV-2. Here, we investigate these potential roles. We demonstrate that SARS-CoV-2 infection upregulates COX-2 in diverse human cell culture and mouse systems. However, suppression of COX-2 by two commonly used NSAIDs, ibuprofen and meloxicam, had no effect on ACE2 expression, viral entry, or viral replication. In contrast, in a mouse model of SARS-CoV-2 infection, NSAID treatment reduced production of proinflammatory cytokines and impaired the humoral immune response to SARS-CoV-2, as demonstrated by reduced neutralizing antibody titers. Our findings indicate that NSAID treatment may influence COVID-19 outcomes by dampening the inflammatory response and production of protective antibodies rather than modifying susceptibility to infection or viral replication. [ABSTRACT FROM AUTHOR]
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- 2021
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26. Single-cell longitudinal analysis of SARS-CoV-2 infection in human airway epithelium identifies target cells, alterations in gene expression, and cell state changes.
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Ravindra, Neal G., Alfajaro, Mia Madel, Gasque, Victor, Huston, Nicholas C., Wan, Han, Szigeti-Buck, Klara, Yasumoto, Yuki, Greaney, Allison M., Habet, Victoria, Chow, Ryan D., Chen, Jennifer S., Wei, Jin, Filler, Renata B., Wang, Bao, Wang, Guilin, Niklason, Laura E., Montgomery, Ruth R., Eisenbarth, Stephanie C., Chen, Sidi, and Williams, Adam
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VIRAL tropism , *COVID-19 , *SARS-CoV-2 , *CORONAVIRUS disease treatment , *GENE expression , *TYPE I interferons , *EPITHELIAL cells , *VIRAL antibodies - Abstract
There are currently limited Food and Drug Administration (FDA)-approved drugs and vaccines for the treatment or prevention of Coronavirus Disease 2019 (COVID-19). Enhanced understanding of Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) infection and pathogenesis is critical for the development of therapeutics. To provide insight into viral replication, cell tropism, and host–viral interactions of SARS-CoV-2, we performed single-cell (sc) RNA sequencing (RNA-seq) of experimentally infected human bronchial epithelial cells (HBECs) in air–liquid interface (ALI) cultures over a time course. This revealed novel polyadenylated viral transcripts and highlighted ciliated cells as a major target at the onset of infection, which we confirmed by electron and immunofluorescence microscopy. Over the course of infection, the cell tropism of SARS-CoV-2 expands to other epithelial cell types including basal and club cells. Infection induces cell-intrinsic expression of type I and type III interferons (IFNs) and interleukin (IL)-6 but not IL-1. This results in expression of interferon-stimulated genes (ISGs) in both infected and bystander cells. This provides a detailed characterization of genes, cell types, and cell state changes associated with SARS-CoV-2 infection in the human airway. Single-cell analysis of human airway epithelial cells infected with SARS-CoV-2 provides novel insights into viral replication, cell tropism, and host-viral interactions. This study reveals novel polyadenylated viral transcripts, preferential tropism for ciliated cells that later extends to other epithelial cell types, and cell-intrinsic expression of type I and type III IFNs and IL6 induced by infection, resulting in expression of interferon-stimulated genes in both infected and bystander cells. [ABSTRACT FROM AUTHOR]
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- 2021
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27. CD300lf Conditional Knockout Mouse Reveals Strain-Specific Cellular Tropism of Murine Norovirus.
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Graziano, Vincent R., Alfajaro, Mia Madel, Schmitz, Cameron O., Filler, Renata B., Strine, Madison S., Jin Wei, Hsieh, Leon L., Baldridge, Megan T., Nice, Timothy J., Lee, Sanghyun, Orchard, Robert C., and Wilena, Craig B.
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KNOCKOUT mice , *VIRAL tropism , *NOROVIRUS diseases , *TROPISMS , *EPITHELIAL cells , *LYMPHOID tissue , *CELL receptors , *B cells - Abstract
Noroviruses are a leading cause of gastrointestinal infection in humans and mice. Understanding human norovirus (HuNoV) cell tropism has important implications for our understanding of viral pathogenesis. Murine norovirus (MNoV) is extensively used as a surrogate model for HuNoV. We previously identified CD300lf as the receptor for MNoV. Here, we generated a Cd300lf conditional knockout (CD300lfF/F) mouse to elucidate the cell tropism of persistent and nonpersistent strains of murine norovirus. Using this mouse model, we demonstrated that CD300lf expression on intestinal epithelial cells (IECs), and on tuft cells in particular, is essential for transmission of the persistent MNoV strain CR6 (MNoVCR6) in vivo. In contrast, the nonpersistent MNoV strain CW3 (MNoVCW3) does not require CD300lf expression on IECs for infection. However, deletion of CD300lf in myelomonocytic cells (LysM Cre1) partially reduces CW3 viral load in lymphoid and intestinal tissues. Disruption of CD300lf expression on B cells (CD19 Cre), neutrophils (Mrp8 Cre), and dendritic cells (CD11c Cre) did not affect MNoVCW3 viral RNA levels. Finally, we show that the transcription factor STAT1, which is critical for the innate immune response, partially restricts the cell tropism of MNoVCW3 to LysM1 cells. Taken together, these data demonstrate that CD300lf expression on tuft cells is essential for MNoVCR6; that myelomonocytic cells are a major, but not exclusive, target cell of MNoVCW3; and that STAT1 signaling restricts the cellular tropism of MNoVCW3. This study provides the first genetic system for studying the cell type-specific role of CD300lf in norovirus pathogenesis. IMPORTANCE Human noroviruses (HuNoVs) are a leading cause of gastroenteritis resulting in up to 200,000 deaths each year. The receptor and cell tropism of HuNoV in immunocompetent humans are unclear. We use murine norovirus (MNoV) as a model for HuNoV. We recently identified CD300lf as the sole physiologic receptor for MNoV. Here, we leverage this finding to generate a Cd300lf conditional knockout mouse to decipher the contributions of specific cell types to MNoV infection. We demonstrate that persistent MNoVCR6 requires CD300lf expression on tuft cells. In contrast, multiple CD300lf1 cell types, dominated by myelomonocytic cells, are sufficient for nonpersistent MNoVCW3 infection. CD300lf expression on epithelial cells, B cells, neutrophils, and dendritic cells is not critical for MNoVCW3 infection. Mortality associated with the MNoVCW3 strain in Stat12/2 mice does not require CD300lf expression on LysM1 cells, highlighting that both CD300lf receptor expression and innate immunity regulate MNoV cell tropism in vivo. [ABSTRACT FROM AUTHOR]
- Published
- 2021
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28. Dual Recognition of Sialic Acid and αGal Epitopes by the VP8 Domains of the Bovine Rotavirus G6P[5] WC3 and of Its Mono-reassortant G4P[5] RotaTeq Vaccine Strains.
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Alfajaro, Mia Madel, Ji-Yun Kim, Barbé, Laure, Eun-Hyo Cho, Jun-Gyu Park, Soliman, Mahmoud, Yeong-Bin Baek, Mun-Il Kang, Soo Hyun Kim, Geun-Joong Kim, Sang-Ik Park, Le Pendu, Jacques, and Kyoung-Oh Cho
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SIALIC acids , *NEURAMINIDASE , *ROTAVIRUSES , *EPITOPES , *TIGHT junctions , *INTEGRINS , *VACCINES , *PROTEIN binding - Abstract
Group A rotaviruses, an important cause of severe diarrhea in children and young animals, initiate infection via interactions of the VP8* domain of the VP4 spike protein with cell surface sialic acids (SAs) or histo-blood group antigens (HBGAs). Although the bovine G6P[5] WC3 strain is an important animal pathogen and is also used in the bovine-human reassortant RotaTeq vaccine, the receptor(s) for the VP8* domain of WC3 and its reassortant strains have not yet been identified. In the present study, HBGA- and saliva-binding assays showed that both G6P[5] WC3 and mono-reassortant G4P[5] strains recognized the αGal HBGA. The infectivity of both P[5]-bearing strains was significantly reduced in αGal-free MA-104 cells by pretreatment with a broadly specific neuraminidase or by coincubation with the α2,6- linked SA-specific Sambucus nigra lectin, but not by the α2,3-linked specific sialidase or by Maackia amurensis lectin. Free NeuAc and the αGal trisaccharide also prevented the infectivity of both strains. This indicated that both P[5]-bearing strains utilize α2,6-linked SA as a ligand on MA104 cells. However, the two strains replicated in differentiated bovine small intestinal enteroids and in their human counterparts that lack α2,6-linked SA or αGal HBGA, suggesting that additional or alternative receptors such as integrins, hsp70, and tight-junction proteins bound directly to the VP5* domain can be used by the P[5]-bearing strains to initiate the infection of human cells. In addition, these data also suggested that P[5]-bearing strains have potential for cross-species transmission. [ABSTRACT FROM AUTHOR]
- Published
- 2019
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29. Use of computed tomography and minimum intensity projection in the detection of lobar pneumonia mimicking lung lobe torsion in a dog.
- Author
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Lee, Sang‐Kwon, Cho, Kyoung‐Oh, Alfajaro, Mia Madel, Lee, Juhwan, Yu, Dohyeon, and Choi, Jihye
- Abstract
A 10‐year‐old female spayed Dachshund was referred with progressive coughing for 1 month. The dog was tentatively diagnosed with right middle lung torsion based on pleural effusion, vesicular emphysema, abruptly ending bronchus in consolidated right middle lung, and no contrast enhancement of the affected lobe on radiography and computed tomography (CT). There was no evidence of torsion upon thoracotomy, and histological examination confirmed lobar pneumonia. The CT images were reevaluated using minimum intensity projection and revealed normal bronchial courses. The minimum intensity projection technique can be to assist in evaluation of the bronchial tree for dogs with suspected lung lobe torsion and other pulmonary diseases. [ABSTRACT FROM AUTHOR]
- Published
- 2019
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30. Early Porcine Sapovirus Infection Disrupts Tight Junctions and Uses Occludin as a Coreceptor.
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Alfajaro, Mia Madel, Eun-Hyo Cho, Deok-Song Kim, Ji-Yun Kim, Jun-Gyu Park, Soliman, Mahmoud, Yeong-Bin Baek, Chul-Ho Park, Mun-Il Kang, Sang-Ik Park, and Kyoung-Oh Cho
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CALICIVIRUSES , *VIRUS diseases , *OCCLUDINS , *TIGHT junctions , *SIALIC acids - Abstract
The genus Sapovirus belongs to the family Caliciviridae, and its members are common causative agents of severe acute gastroenteritis in both humans and animals. Some caliciviruses are known to use either terminal sialic acids or histoblood group antigens as attachment factors and/or cell surface proteins, such as CD300lf, CD300ld, and junctional adhesion molecule 1 of tight junctions (TJs), as receptors. However, the roles of TJs and their proteins in sapovirus entry have not been examined. In this study, we found that porcine sapovirus (PSaV) significantly decreased transepithelial electrical resistance and increased paracellular permeability early in infection of LLC-PK cells, suggesting that PSaV dissociates TJs of cells. This led to the interaction between PSaV particles and occludin, which traveled in a complex into late endosomes via Rab5- and Rab7-dependent trafficking. Inhibition of occludin using small interfering RNA (siRNA), a specific antibody, or a dominantnegative mutant significantly blocked the entry of PSaV. Transient expression of occludin in nonpermissive Chinese hamster ovary (CHO) cells conferred susceptibility to PSaV, but only for a limited time. Although claudin-1, another TJ protein, neither directly interacted nor was internalized with PSaV particles, it facilitated PSaV entry and replication in the LLC-PK cells. We conclude that PSaV particles enter LLC-PK cells by binding to occludin as a coreceptor in PSaV-dissociated TJs. PSaV and occludin then form a complex that moves to late endosomes via Rab5- and Rab7- dependent trafficking. In addition, claudin-1 in the TJs opened by PSaV infection facilitates PSaV entry and infection as an entry factor. [ABSTRACT FROM AUTHOR]
- Published
- 2019
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31. Feline calicivirus- and murine norovirus-induced COX-2/PGE2 signaling pathway has proviral effects.
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Alfajaro, Mia Madel, Cho, Eun-Hyo, Park, Jun-Gyu, Kim, Ji-Yun, Soliman, Mahmoud, Baek, Yeong-Bin, Kang, Mun-Il, Park, Sang-Ik, and Cho, Kyoung-Oh
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CYCLOOXYGENASE 2 , *NOROVIRUS diseases , *FELINE calicivirus , *ANTIVIRAL agents , *CELLULAR signal transduction , *THERAPEUTICS - Abstract
Cyclooxygenases (COXs)/prostaglandin E2 (PGE2) signaling pathways are known to modulate a variety of homeostatic processes and are involved in various pathophysiological conditions. COXs/PGE2 signaling pathways have also been demonstrated to have proviral or antiviral effects, which appeared different even in the same virus family. A porcine sapovirus Cowden strain, a member of genus Sapovirus within the Caliciviridae family, induces strong COX-2/PGE2 but transient COX-1/PGE2 signaling to enhance virus replication. However, whether infections of other viruses in the different genera activate COXs/PGE2 signaling, and thus affect the replication of viruses, remains unknown. In the present study, infections of cells with the feline calicivirus (FCV) F9 strain in the genus Vesivirus and murine norovirus (MNV) CW-1 strain in the genus Norovirus only activated the COX-2/PGE2 signaling in a time-dependent manner. Treatment with pharmacological inhibitors or transfection of small interfering RNAs (siRNAs) against COX-2 enzyme significantly reduced the production of PGE2 as well as FCV and MNV replications. The inhibitory effects of these pharmacological inhibitors against COX-2 enzyme on the replication of both viruses were restored by the addition of PGE2. Silencing of COX-1 via siRNAs and inhibition of COX-1 via an inhibitor also decrease the production of PGE2 and replication of both viruses, which can be attributed to the inhibition COX-1/PGE2 signaling pathway. These data indicate that the COX-2/PGE2 signaling pathway has proviral effects for the replication of FCV and MNV, and pharmacological inhibitors against these enzymes serve as potential therapeutic candidates for treating FCV and MNV infections. [ABSTRACT FROM AUTHOR]
- Published
- 2018
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32. Activation of PI3K, Akt, and ERK during early rotavirus infection leads to V-ATPase-dependent endosomal acidification required for uncoating.
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Soliman, Mahmoud, Seo, Ja-Young, Kim, Deok-Song, Kim, Ji-Yun, Park, Jun-Gyu, Alfajaro, Mia Madel, Baek, Yeong-Bin, Cho, Eun-Hyo, Kwon, Joseph, Choi, Jong-Soon, Kang, Mun-Il, Park, Sang-Ik, and Cho, Kyoung-Oh
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ROTAVIRUSES ,VIRUSES ,ACIDIFICATION ,IMMUNOPRECIPITATION ,IMMUNOGLOBULINS - Abstract
The cellular PI3K/Akt and/or MEK/ERK signaling pathways mediate the entry process or endosomal acidification during infection of many viruses. However, their roles in the early infection events of group A rotaviruses (RVAs) have remained elusive. Here, we show that late-penetration (L-P) human DS-1 and bovine NCDV RVA strains stimulate these signaling pathways very early in the infection. Inhibition of both signaling pathways significantly reduced production of viral progeny due to blockage of virus particles in the late endosome, indicating that neither of the two signaling pathways is involved in virus trafficking. However, immunoprecipitation assays using antibodies specific for pPI3K, pAkt, pERK and the subunit E of the V-ATPase co-immunoprecipitated the V-ATPase in complex with pPI3K, pAkt, and pERK. Moreover, Duolink proximity ligation assay revealed direct association of the subunit E of the V-ATPase with the molecules pPI3K, pAkt, and pERK, indicating that both signaling pathways are involved in V-ATPase-dependent endosomal acidification. Acidic replenishment of the medium restored uncoating of the RVA strains in cells pretreated with inhibitors specific for both signaling pathways, confirming the above results. Isolated components of the outer capsid proteins, expressed as VP4-VP8* and VP4-VP5* domains, and VP7, activated the PI3K/Akt and MEK/ERK pathways. Furthermore, psoralen-UV-inactivated RVA and CsCl-purified RVA triple-layered particles triggered activation of the PI3K/Akt and MEK/ERK pathways, confirming the above results. Our data demonstrate that multistep binding of outer capsid proteins of L-P RVA strains with cell surface receptors phosphorylates PI3K, Akt, and ERK, which in turn directly interact with the subunit E of the V-ATPase to acidify the late endosome for uncoating of RVAs. This study provides a better understanding of the RVA-host interaction during viral uncoating, which is of importance for the development of strategies aiming at controlling or preventing RVA infections. [ABSTRACT FROM AUTHOR]
- Published
- 2018
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33. Activation of COX-2/PGE2 Promotes Sapovirus Replication via the Inhibition of Nitric Oxide Production.
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Alfajaro, Mia Madel, Jong-Soon Choi, Deok-Song Kim, Ja-Young Seo, Ji-Yun Kim, Jun-Gyu Park, Soliman, Mahmoud, Yeong-Bin Baek, Eun-Hyo Cho, Kwon, Joseph, Hyung-Jun Kwon, Su-Jin Park, Woo Song Lee, Mun-Il Kang, Hosmillo, Myra, Goodfellow, Ian, and Kyoung-Oh Cho
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CYCLOOXYGENASE 2 , *ENZYME activation , *CALICIVIRUSES , *VIRAL replication , *NITRIC oxide - Abstract
Enteric caliciviruses in the genera Norovirus and Sapovirus are important pathogens that cause severe acute gastroenteritis in both humans and animals. Cyclooxygenases (COXs) and their final product, prostaglandin E2 (PGE2), are known to play important roles in the modulation of both the host response to infection and the replicative cycles of several viruses. However, the precise mechanism(s) by which the COX/PGE2 pathway regulates sapovirus replication remains largely unknown. In this study, infection with porcine sapovirus (PSaV) strain Cowden, the only cultivable virus within the genus Sapovirus, markedly increased COX-2 mRNA and protein levels at 24 and 36 h postinfection (hpi), with only a transient increase in COX-1 levels seen at 24 hpi. The treatment of cells with pharmacological inhibitors, such as nonsteroidal anti-inflammatory drugs or small interfering RNAs (siRNAs) against COX-1 and COX-2, significantly reduced PGE2 production, as well as PSaV replication. Expression of the viral proteins VPg and ProPol was associated with activation of the COX/PGE2 pathway. We observed that pharmacological inhibition of COX-2 dramatically increased NO production, causing a reduction in PSaV replication that could be restored by inhibition of nitric oxide synthase via the inhibitor N-nitro-L-methylarginine ester. This study identified a pivotal role for the COX/PGE2 pathway in the regulation of NO production during the sapovirus life cycle, providing new insights into the life cycle of this poorly characterized family of viruses. Our findings also reveal potential new targets for treatment of sapovirus infection. [ABSTRACT FROM AUTHOR]
- Published
- 2017
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34. Both α2,3- and α2,6-Linked Sialic Acids on O-Linked Glycoproteins Act as Functional Receptors for Porcine Sapovirus.
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Kim, Deok-Song, Hosmillo, Myra, Alfajaro, Mia Madel, Kim, Ji-Yun, Park, Jun-Gyu, Son, Kyu-Yeol, Ryu, Eun-Hye, Sorgeloos, Frederic, Kwon, Hyung-Jun, Park, Su-Jin, Lee, Woo Song, Cho, Duck, Kwon, Joseph, Choi, Jong-Soon, Kang, Mun-Il, Goodfellow, Ian, and Cho, Kyoung-Oh
- Subjects
SIALIC acids ,GLYCOPROTEINS ,GASTROENTERITIS ,VIBRIO cholerae ,PROTEOLYTIC enzymes ,GLYCOSYLATION - Abstract
Sapovirus, a member of the Caliciviridae family, is an important cause of acute gastroenteritis in humans and pigs. Currently, the porcine sapovirus (PSaV) Cowden strain remains the only cultivable member of the Sapovirus genus. While some caliciviruses are known to utilize carbohydrate receptors for entry and infection, a functional receptor for sapovirus is unknown. To characterize the functional receptor of the Cowden strain of PSaV, we undertook a comprehensive series of protein-ligand biochemical assays in mock and PSaV-infected cell culture and/or piglet intestinal tissue sections. PSaV revealed neither hemagglutination activity with red blood cells from any species nor binding activity to synthetic histo-blood group antigens, indicating that PSaV does not use histo-blood group antigens as receptors. Attachment and infection of PSaV were markedly blocked by sialic acid and Vibrio cholerae neuraminidase (NA), suggesting a role for α2,3-linked, α2,6-linked or α2,8-linked sialic acid in virus attachment. However, viral attachment and infection were only partially inhibited by treatment of cells with sialidase S (SS) or Maackia amurensis lectin (MAL), both specific for α2,3-linked sialic acid, or Sambucus nigra lectin (SNL), specific for α2,6-linked sialic acid. These results indicated that PSaV recognizes both α2,3- and α2,6-linked sialic acids for viral attachment and infection. Treatment of cells with proteases or with benzyl 4-O-β-D-galactopyranosyl-β-D-glucopyranoside (benzylGalNAc), which inhibits O-linked glycosylation, also reduced virus binding and infection, whereas inhibition of glycolipd synthesis or N-linked glycosylation had no such effect on virus binding or infection. These data suggest PSaV binds to cellular receptors that consist of α2,3- and α2,6-linked sialic acids on glycoproteins attached via O-linked glycosylation. [ABSTRACT FROM AUTHOR]
- Published
- 2014
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35. Genome-wide CRISPR Screens Reveal Host Factors Critical for SARS-CoV-2 Infection.
- Author
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Wei, Jin, Alfajaro, Mia Madel, DeWeirdt, Peter C., Hanna, Ruth E., Lu-Culligan, William J., Cai, Wesley L., Strine, Madison S., Zhang, Shang-Min, Graziano, Vincent R., Schmitz, Cameron O., Chen, Jennifer S., Mankowski, Madeleine C., Filler, Renata B., Ravindra, Neal G., Gasque, Victor, de Miguel, Fernando J., Patil, Ajinkya, Chen, Huacui, Oguntuyo, Kasopefoluwa Y., and Abriola, Laura
- Subjects
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COVID-19 , *SARS-CoV-2 , *GENE libraries , *CRISPRS , *VESICULAR stomatitis , *MERS coronavirus - Abstract
Identification of host genes essential for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection may reveal novel therapeutic targets and inform our understanding of coronavirus disease 2019 (COVID-19) pathogenesis. Here we performed genome-wide CRISPR screens in Vero-E6 cells with SARS-CoV-2, Middle East respiratory syndrome CoV (MERS-CoV), bat CoV HKU5 expressing the SARS-CoV-1 spike, and vesicular stomatitis virus (VSV) expressing the SARS-CoV-2 spike. We identified known SARS-CoV-2 host factors, including the receptor ACE2 and protease Cathepsin L. We additionally discovered pro-viral genes and pathways, including HMGB1 and the SWI/SNF chromatin remodeling complex, that are SARS lineage and pan-coronavirus specific, respectively. We show that HMGB1 regulates ACE2 expression and is critical for entry of SARS-CoV-2, SARS-CoV-1, and NL63. We also show that small-molecule antagonists of identified gene products inhibited SARS-CoV-2 infection in monkey and human cells, demonstrating the conserved role of these genetic hits across species. This identifies potential therapeutic targets for SARS-CoV-2 and reveals SARS lineage-specific and pan-CoV host factors that regulate susceptibility to highly pathogenic CoVs. • Developed monkey CRISPR library to screen pathogenic coronaviruses in Vero-E6 cells • Screens identified genes that are SARS-CoV-2, MERS-CoV, and pan-coronavirus specific • Therapeutic targets, including SMARCA4, identified for SARS-CoV-2 infection • HMGB1 is novel regulator of ACE2 expression and critical for viral entry To identify potential therapeutic targets for SARS-CoV-2 and related pathogenic coronaviruses, Wei et al. conduct genome-wide CRISPR screens in Vero-E6 cells using SARS-CoV-2, MERS-CoV, and pseudoviruses presenting SARS-CoV-1 or SARS-CoV-2 spike proteins. They identify pro-viral genes and pathways, including HMGB1 and the SWI/SNF chromatin remodeling complex, that are SARS lineage and pan-coronavirus specific, respectively, and demonstrate that HMGB1 is critical for SARS lineage viral entry because it has a critical role in ACE2 expression. [ABSTRACT FROM AUTHOR]
- Published
- 2021
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- View/download PDF
36. Bovine Nebovirus Interacts with a Wide Spectrum of Histo-Blood Group Antigens.
- Author
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Eun-Hyo Cho, Soliman, Mahmoud, Alfajaro, Mia Madel, Ji-Yun Kim, Ja-Young Seo, Jun-Gyu Park, Deok-Song Kim, Yeong-Bin Baek, Mun-Il Kang, Sang-Ik Park, Pendu, Jacques Le, and Kyoung-Oh Choa
- Subjects
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CALICIVIRUSES , *CARBOHYDRATES , *BLOOD group antigens , *FUCOSE , *BOVINE anatomy - Abstract
Some viruses within the Caliciviridae family initiate their replication cycle by attachment to cell surface carbohydrate moieties, histo-blood group antigens (HBGAs), and/or terminal sialic acids (SAs). Although bovine nebovirus (BNeV), one of the enteric caliciviruses, is an important causative agent of acute gastroenteritis in cattle, its attachment factors and possibly other cellular receptors remain unknown. Using a comprehensive series of protein-ligand biochemical assays, we sought to determine whether BNeV recognizes cell surface HBGAs and/or SAs as attachment factors. It was found that BNeV virus-like particles (VLPs) bound to A type/H type 2/Ley HBGAs expressed in the bovine digestive tract and are related to HBGAs expressed in humans and other host species, suggesting a wide spectrum of HBGA recognition by BNeV. BNeV VLPs also bound to a large variety of different bovine and human saliva samples of all ABH and Lewis types, supporting previously obtained results and suggesting a zoonotic potential of BNeV transmission. Removal of α1,2-linked fucose and α1,3/4-linked fucose epitopes of target HBGAs by confirmation-specific enzymes reduced the binding of BNeV VLPs to synthetic HBGAs, bovine and human saliva, cultured cell lines and bovine small intestine mucosa, further supporting a wide HBGA binding spectrum of BNeV through recognition of α1,2- linked fucose and α1,3/4-linked fucose epitopes of targeted HBGAs. However, removal of terminal α2,3- and α2,6-linked SAs by their specific enzyme had no inhibitory effects on binding of BNeV VLPs, indicating that BNeV does not use terminal SAs as attachment factors. Further details of the binding specificity of BNeV remain to be explored. IMPORTANCE Enteric caliciviruses such as noroviruses, sapoviruses, and recoviruses are the most important etiological agents of severe acute gastroenteritis in humans and many other mammalian host species. They initiate infection by attachment to cell surface carbohydrate moieties, HBGAs, and/or terminal SAs. However, the attachment factor(s) for BNeV, a recently classified enteric calicivirus genus/type species, remains unexplored. Here, we demonstrate that BNeV VLPs have a wide spectrum of binding to synthetic HBGAs, bovine and human saliva samples and bovine duodenal sections. We further discovered that α1,2-linked fucose and α1,3/4-linked fucose epitopes are essential for binding of BNeV VLPs. However, BNeV VLPs do not bind to terminal SAs on cell carbohydrates. Continued investigation regarding the proteinaceous receptor(s) will be necessary for better understanding of the tropism, pathogenesis, and host range of this important viral genus. [ABSTRACT FROM AUTHOR]
- Published
- 2018
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37. Glycan-specificity of four neuraminidase-sensitive animal rotavirus strains.
- Author
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Kim, Ji-Yun, Kim, Deok-Song, Seo, Ja-Young, Park, Jun-Gyu, Alfajaro, Mia Madel, Soliman, Mahmoud, Baek, Yeong-Bin, Cho, Eun-Hyo, Kwon, Hyung-Jun, Park, Su-Jin, Kang, Mun-Il, and Cho, Kyoung-Oh
- Subjects
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GLYCANS , *NEURAMINIDASE , *ROTAVIRUS diseases , *SIALIC acids , *GLYCOPROTEINS - Abstract
Group A rotaviruses (RVAs) are divided into neuraminidase (NA)-sensitive and NA-insensitive strains depending upon their binding affinity to the VP8* domain in the terminal sialic acids (SAs) of cell surface carbohydrates. Although NA-sensitive strains are known to use terminal SAs as an attachment factor, the exact nature of this attachment factor is largely unknown. Here we show that the specific linkage of SA-containing glycan to glycoprotein or glycolipid is an attachment factor used by NA-sensitive porcine G9P[7] PRG9121 and G9P[23] PRG942, bovine G6P[1] NCDV, and canine G3P[3] strains. Infectivity of porcine G9P[7] and G9P[23] strains was markedly blocked by α2,3-linkage and α2,6-linkage inhibitors, indicating that these strains bind to both α2,3- and α2,6-linked SAs. However, the infectivity of bovine G6P[1] and canine G3P[3] strains was significantly reduced by α2,6-linkage inhibitor but not by α2,3-linkage blockers, demonstrating a predilection of these strains for α2,6-linked SAs. The infectivity of four NA-sensitive strains was equally reduced by inhibitors of lipid membrane and N -linked glycoprotein but not by an inhibitor of O -linked glycoprotein, indicating that these strains utilize both glycolipid and N -linked glycoprotein. Our study demonstrates that four NA-sensitive animal strains could have a strain-dependent binding preference toward α2,6-linked SAs (P[1] NCDV and P[3] CU-1 strains) or both α2,3- and α2,6-linked SAs (P[7] PRG9121 and P[23] PRG942 strains) to the glycolipid and N -linked glycoprotein. [ABSTRACT FROM AUTHOR]
- Published
- 2017
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38. Comparison of elastography, contrast-enhanced ultrasonography, and computed tomography for assessment of lesion margin after radiofrequency ablation in livers of healthy dogs.
- Author
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Sohyeon Moon, Seungjo Park, Sang-kwon Lee, Byunggyu Cheon, Sunghwa Hong, Hyun Cho, Jun-Gyu Park, Alfajaro, Mia Madel, Kyoung-Oh Cho, Dong Woo Chang, and Jihye Choi
- Subjects
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ELASTOGRAPHY , *CONTRAST-enhanced ultrasound , *CATHETER ablation , *ANIMAL health , *DOGS , *NECROSIS , *GRANULATION tissue - Abstract
OBJECTIVE To assess by use of various diagnostic imaging modalities acute changes in livers of healthy dogs after radiofrequency ablation (RFA) and determine the capability of each imaging modality to monitor ablation lesion changes. ANIMALS 6 healthy Beagles. PROCEDURES 12 ablation lesions were created in the liver of the dogs (2 lesions/dog). Ablation lesions were evaluated by use of conventional ultrasonography, strain elastography, and contrastenhanced ultrasonography immediately after (time 0), 30 to 60 minutes after, and 3 days after RFA, and by use of CT 30 minutes and 3 days after RFA. Three dogs were euthanized shortly after RFA, and the other 3 dogs were euthanized on day 3. Lesion size measured by each imaging modality was compared with necropsy findingsRESULTS Immediately after RFA, clear margins were more visible with elastography and contrast-enhanced ultrasonography than with conventional ultrasonography, which had acoustic shadowing. On triphasic contrast CT. the ablation zone, which indicated necrosis and hemorrhage, was not enhanced and rould be measured. Marked enhancement of the periablation rim was observed during the venous phase and was identified as granulation tissue. Size of the ablation area measured on enhanced CT images was Strongly correlated with actual lesion size. CONCLUSIONS AND CLINICAL RELEVANCE For dogs of this study. CT was the most reliable method for lesion size determination. Although ultrasonographic imaging measurements underestimated lesion size, all modalities could be used to provide additional real time guidance for RFA procedures of the liver as well as for other RFA procedures. [ABSTRACT FROM AUTHOR]
- Published
- 2017
- Full Text
- View/download PDF
39. Porcine Sapelovirus Uses α2,3-Linked Sialic Acid on GD1a Ganglioside as a Receptor.
- Author
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Deok-Song Kim, Kyu-Yeol Son, Kyung-Min Koo, Ji-Yun Kim, Alfajaro, Mia Madel, Jun-Gyu Park, Hosmillo, Myra, Soliman, Mahmoud, Yeong-Bin Baek, Eun-Hyo Cho, Ju-Hwan Lee, Mun-Il Kang, Ian Goodfellow, and Kyoung-Oh Cho
- Subjects
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SIALIC acids , *GANGLIOSIDES , *PICORNAVIRUSES , *DIARRHEA , *PNEUMONIA , *NEURAMINIDASE , *OLIGOSACCHARIDES , *THERAPEUTICS - Abstract
The receptor(s) for porcine sapelovirus (PSV), which causes diarrhea, pneumonia, polioencephalomyelitis, and reproductive disorders in pigs, remains largely unknown. Given the precedent for other picornaviruses which use terminal sialic acids (SAs) as receptors, we examined the role of SAs in PSV binding and infection. Using a variety of approaches, including treating cells with a carbohydrate-destroying chemical (NaIO4), mono- or oligosaccharides (N-acetylneuraminic acid, galactose, and 6'-sialyllactose), linkage-specific sialidases (neuraminidase and sialidase S), lectins (Maakia amurensis lectin and Sambucus nigra lectin), proteases (trypsin and chymotrypsin), and glucosylceramide synthase inhibitors (DL-threo-1-phenyl-2-decanoylamino-3-morpholino-1-propanol and phospholipase C), we demonstrated that PSV could recognize α2,3-linked SA on glycolipids as a receptor. On the other hand, PSVs had no binding affinity for synthetic histo-blood group antigens (HBGAs), suggesting that PSVs could not use HBGAs as receptors. Depletion of cell surface glycolipids followed by reconstitution studies indicated that GD1a ganglioside, but not other gangliosides, could restore PSV binding and infection, further confirming α2,3-linked SA on GD1a as a PSV receptor. Our results could provide significant information on the understanding of the life cycle of sapelovirus and other picornaviruses. For the broader community in the area of pathogens and pathogenesis, these findings and insights could contribute to the development of affordable, useful, and efficient drugs for anti-sapelovirus therapy. [ABSTRACT FROM AUTHOR]
- Published
- 2016
- Full Text
- View/download PDF
40. Comparison of pathogenicities and nucleotide changes between porcine and bovine reassortant rotavirus strains possessing the same genotype constellation in piglets and calves.
- Author
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Jun-Gyu Park, Deok-Song Kim, Matthijnssens, Jelle, Hyoung-Jun Kwon, Zeller, Mark, Alfajaro, Mia Madel, Kyu-Yeol Son, Hosmillo, Myra, Eun-Hye Ryu, Ji-Yun Kim, Ju-Hwan Lee, Su-Jin Park, Mun-Il Kang, Joseph Kwon, Jong-Soon Choi, and Kyoung-Oh Cho
- Subjects
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PATHOGENIC microorganisms , *COMPARATIVE studies , *NUCLEOTIDES , *ROTAVIRUSES , *BOS , *PIGLETS , *MICROBIAL virulence , *PHYSIOLOGY - Abstract
Although reassortment is one of the most important characteristics of group A rotavirus (RVA) evolution, the host range restriction and/or virulence of reassortant RVAs remain largely unknown. The porcine 174-1 strain isolated from a diarrheic piglet was identified as a reassortant strain, harboring the same genotype constellation as the previously characterized bovine strain KJ56-1. Owing to its same genotype constellation, the pathogenicity of the porcine strain 174-1 in piglets and calves was examined for comparison with that of the bovine reassortant KJ56-1 strain, whose pathogenicity has already been demonstrated in piglets and calves. The porcine 174-1 strain induced diarrhea and histopathological changes in the small intestine of piglets and calves, whereas KJ56-1 had been reported to be virulent only in piglets, but not in calves. Therefore, full genomic sequences of 174-1 and KJ56-1 strains were analyzed to determine whether specific mutations might be associated with clinical and pathological phenotypes. Sequence alignment between the 174-1 and KJ56-1 strains detected one nucleotide substitution at the 3′ untranslated region of the NSP3 gene and 16 amino acid substitutions at the VP7, VP4, VP1, VP3, NSP1 and NSP4 genes. These mutations may be critical molecular determinants for different virulence and/or pathogenicity of each strain. This study presents new insights into the host range restriction and/or virulence of RVAs. [ABSTRACT FROM AUTHOR]
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- 2014
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41. Discovery and functional interrogation of SARS-CoV-2 RNA-host protein interactions.
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Flynn, Ryan A., Belk, Julia A., Qi, Yanyan, Yasumoto, Yuki, Wei, Jin, Alfajaro, Mia Madel, Shi, Quanming, Mumbach, Maxwell R., Limaye, Aditi, DeWeirdt, Peter C., Schmitz, Cameron O., Parker, Kevin R., Woo, Elizabeth, Chang, Howard Y., Horvath, Tamas L., Carette, Jan E., Bertozzi, Carolyn R., Wilen, Craig B., and Satpathy, Ansuman T.
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SARS-CoV-2 , *PROTEIN-protein interactions , *RNA-binding proteins , *RNA viruses , *VIRAL proteins - Abstract
SARS-CoV-2 is the cause of a pandemic with growing global mortality. Using comprehensive identification of RNA-binding proteins by mass spectrometry (ChIRP-MS), we identified 309 host proteins that bind the SARS-CoV-2 RNA during active infection. Integration of this data with ChIRP-MS data from three other RNA viruses defined viral specificity of RNA-host protein interactions. Targeted CRISPR screens revealed that the majority of functional RNA-binding proteins protect the host from virus-induced cell death, and comparative CRISPR screens across seven RNA viruses revealed shared and SARS-specific antiviral factors. Finally, by combining the RNA-centric approach and functional CRISPR screens, we demonstrated a physical and functional connection between SARS-CoV-2 and mitochondria, highlighting this organelle as a general platform for antiviral activity. Altogether, these data provide a comprehensive catalog of functional SARS-CoV-2 RNA-host protein interactions, which may inform studies to understand the host-virus interface and nominate host pathways that could be targeted for therapeutic benefit. [Display omitted] • ChIRP-MS of SARS-CoV-2 RNA identifies viral RNA-host protein interaction networks • Comparative analysis identifies SARS-specific and multi-viral RNA-protein complexes • SARS-CoV-2 interactome-focused CRISPR screens reveal a broad antiviral response • Host mitochondria serve as a general organelle platform for anti-SARS-CoV-2 immunity Interrogation of SARS-CoV-2 RNA-host protein interaction networks by ChIRP-MS and CRISPR screens, in comparison with other human viruses such as flaviviruses, picornavirus, and rhinovirus, identifies complexes specific to SARS-CoV-2 infection and highlights the role of mitochondria in mediating antiviral immunity. [ABSTRACT FROM AUTHOR]
- Published
- 2021
- Full Text
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