https://scholar.dgist.ac.kr/handle/20.500.11750/8970 2026-04-04T13:36:43Z https://scholar.dgist.ac.kr/handle/20.500.11750/59228 Title: Reconciling a Kinetic Model for Dimerization of the EGFR Using Single-Molecule Tracking in Living Cells Author(s): Kim, Kiwook; Jang, Juhee; Cho, Juhyeong; Ahn, Yongdeok; Jeong, Seunghyeon; Shin, Jiwon; Yea, Kyungmoo; Lee, Wonhee John; Seo, Daeha Abstract: Epidermal growth factor receptor (EGFR) dimerization plays a pivotal role in cellular signaling, influencing proliferation and disease progression, particularly in cancer. Despite extensive studies, the quantitative relationship between EGFR expression levels and dimerization efficiency remains incompletely understood. In this study, we investigated EGFR dimerization kinetics using ensemble-level biochemical assays and single-molecule tracking (SMT) in living cells. Our findings revealed noncanonical negative cooperative dimerization, where the monomer-to-dimer transition rate decreased as EGFR expression increased, challenging the assumptions of a simplistic reaction model. Furthermore, we identified a dimer-specific degradation pathway highlighting the open-system nature of the plasma membrane environment. These findings establish a quantitative framework for understanding EGFR dimerization dynamics, offering insights into the complex regulatory principles governing membrane protein interactions. This model not only improves our understanding of EGFR-mediated signaling but also suggests broader applicability for the therapeutic targeting of membrane protein systems. 2025-08-31T15:00:00Z https://scholar.dgist.ac.kr/handle/20.500.11750/59090 Title: Surface-Engineered Natural Killer Cell-Derived Small Extracellular Vesicles Induce Potent Anti-Tumour Effects in Lung Cancer Cells Author(s): Kang, Sung-Min; Jung, Dokyung; Noh, Soojeong; Shin, Sanghee; Kim, Minju; Cho, Hanchae; Lee, Byungheon; Yea, Kyungmoo; Baek, Moonchang Abstract: Small extracellular vesicles (sEVs) derived from natural killer (NK) cells possess inherent anti-tumour activity and offer the advantages of cell-free therapy. In this study, we genetically engineered NK-sEVs to express interleukin 15 (IL15), an anti-tumour cytokine, and the monoclonal antibody cetuximab on their surface, creating a potent anti-tumour immunotherapy with enhanced tumour-targeting capabilities. These IL15- and cetuximab-tethered NK-sEVs (eEVs) were generated using lentivirus-based modification. eEVs selectively bound to EGFR+ cancer cells in vitro, confirming cetuximab-mediated targeting. Compared to control NK-sEVs, eEVs exhibited significantly enhanced cytotoxicity by directly inducing cancer cell death and promoting NK cell-mediated killing. In a lung cancer mouse model, eEVs selectively accumulated in tumours and exhibited significant anti-tumour efficacy. Notably, their administration, alone or in combination with anti-PD-1 antibody therapy, effectively suppressed tumour growth. Overall, our results indicate that genetically engineered NK-sEVs, equipped with IL15 and cetuximab, exhibit potent anti-tumour activity and tumour-targeting capabilities. These findings suggest that eEVs hold significant potential as a novel immunotherapeutic strategy for cancer treatment. © 2025 Elsevier B.V., All rights reserved. 2025-07-31T15:00:00Z https://scholar.dgist.ac.kr/handle/20.500.11750/59054 Title: Inflammatory cytokine-primed MSC-derived extracellular vesicles ameliorate acute lung injury via enhanced immunomodulation and alveolar repair Author(s): Jeong, Jongwon; Park, Jun-Kook; Shin, Jiwon; Jung, Inseong; Kim, Hyun-Woo; Park, Anyeseu; Cho, Hanchae; Kang, Sung-Min; Shin, Sanghee; Park, Eunju; Kim, Jisuk; Noh, Soojeong; Ahn, Yongdeok; Kim, Do-Kyun; Lee, Jeong Yoon; Seo, Daeha; Baek, Moon-Chang; Yea, Kyungmoo Abstract: Background: Acute lung injury (ALI) is characterized by excessive inflammation and alveolar damage, arising from pathogens or systemic insults such as sepsis, and can progress to severe acute respiratory distress syndrome (ARDS). Despite its severity, effective pharmacological treatments remain unavailable, and current clinical interventions are limited to supportive care such as mechanical ventilation. Mesenchymal stem cell-derived extracellular vesicles (MSC-EVs) have emerged as promising candidates for lung repair, but insufficient immunosuppressive capacity often limits their efficacy. Methods: Human adipose-derived mesenchymal stem cells (hADMSCs) were primed with IFN-γ and TNF-α to enhance the immunomodulatory properties of their secreted EVs. We characterized unprimed control MSC-EVs (C-MEVs) and primed MSC-EVs (P-MEVs) by transmission electron microscopy, nanoparticle tracking analysis, and western blotting for EV markers. Functional assays in THP-1 and A549 cells examined anti-inflammatory potency and barrier regeneration against lipopolysaccharide (LPS)-induced damage. A preclinical mouse model of LPS-induced ALI was used to evaluate inflammatory cytokine expression, immune cell infiltration, pulmonary edema, and vascular leakage. Finally, severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2)-infected Vero E6 cells were tested whether P-MEVs could mitigate the inflammatory damage characteristic of virus-triggered acute lung injury. Results: Primed hADMSCs exhibited elevated expression of immunosuppressive molecules (e.g., COX-2, IDO, TSG-6), without changing EV morphology or yield. P-MEVs mitigated LPS-induced inflammation more effectively than C-MEVs in THP-1 and A549 cells. In vivo, P-MEVs more robustly attenuated inflammatory cytokines, immune cell recruitment, and lung injury markers in mice challenged with LPS. In SARS-CoV-2-infected Vero E6 cells, P-MEVs suppressed cytopathic effects and inflammatory responses more potently than C-MEVs. Mechanistic analyses revealed that these enhancements were associated with elevated miRNA levels, including miR-221-3p, involved in inhibiting inflammatory pathways. Conclusion: Inflammatory cytokine priming substantially augments the immunomodulatory and tissue-regenerative efficacy of hADMSC-derived EVs, offering superior therapeutic effects in ALI models and promising activity against SARS-CoV-2-induced lung damage. These findings underscore the therapeutic potential of P-MEVs as an innovative, cell-free platform for treating severe pulmonary disorders, including ARDS. © 2025 Elsevier B.V., All rights reserved. 2025-07-31T15:00:00Z https://scholar.dgist.ac.kr/handle/20.500.11750/58652 Title: T-Cell-Derived Extracellular Vesicles with an Antitransferrin Receptor Antibody for Multicancer Targeting Author(s): Cho, Hanchae; Ju, Hyunji; Shin, Sanghee; Ahn, Yongdeok; Park, Eunju; Jung, Inseong; Kang, Sung-Min; Noh, Soojeong; Shin, Jiwon; Park, Jun-Kook; Jeong, Jongwon; Seo, Daeha; Song, Byoung-Joon; Yea, Kyungmoo; Baek, Moon-Chang Abstract: Recent research has explored the anticancer properties of immune-cell-derived small extracellular vesicles (sEVs), but many challenges, like the need for improved targeting, remain. To address these challenges, we engineered T-cell-derived sEVs with antitransferrin receptor 1 (TfR1) antibodies (T-EVs). This modification enhanced the delivery of sEV to six types of cancer cells, as confirmed by flow cytometry, immunocytochemistry, live cell imaging, and blocking experiments in vitro. The T-EVs also reduced PD-L1 and Rab27a levels, decreased sEV production from breast cancer cells, and increased susceptibility to CD8+ T-cell-mediated cytotoxicity. Systemically administered T-EVs efficiently targeted breast, lung, and skin tumors in mouse models. Notably, T-EVs significantly inhibited tumor growth without systemic toxicity. Additionally, T-EVs reduced PD-L1 and Rab27a levels in cancer cells while enhancing the CD8+ T-cell cytotoxicity and proliferation. Overall, this study highlights the anticancer effects of T-EVs against multiple cancer types, underscoring their potential in developing targeted cancer therapies. © 2025 American Chemical Society 2025-06-30T15:00:00Z