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Lymphocytes-Associated Extracellular Vesicles Activate Natural Killer Cells in HNSCC

CANCER SCIENCE 2025年第3期116卷 633-642页

作者： li, chuwen liu, Chun Ma, Hailong Zhang, Zhiyuan Zhang, Jianjun Shanghai Jiao Tong Univ Sch Med Shanghai Peoples Hosp 9 Dept Oral & Maxillofacial Head & Neck Oncol Shanghai Peoples R China Shanghai Jiao Tong Univ Coll Stomatol Shanghai Peoples R China Natl Ctr Stomatol Shanghai Peoples R China Shanghai Res Inst Stomatol Natl Clin Res Ctr Oral Dis Shanghai Ctr Head & Neck Oncol Clin & Translat Sci Shanghai Key Lab Stomatol Shanghai Peoples R China Chinese Acad Med Sci Res Unit Oral & Maxillofacial Regenerat Med Shanghai Peoples R China

Small extracellular vesicles (sEVs) facilitate intercellular communication and play a pivotal role in tumor progression. Accumulated evidence has indicated the diversity of sEVs but with limited results revealing the landscape of heterogeneity of sEVs. The heterogeneity of cargo RNA in sEVs presents the different cell origins and indicates different functions. Here, we analyzed the heterogeneity of sEVs at droplet levels from single-cell RNA sequencing results of head and neck squamous cell carcinoma (HNSCC) with the previously reported algorithm SEVtras. With the sEVs secretion activity calculated by SEVtras, we also found that the T cells held the major role of sEVs secretion. In addition, we found these sEVs secreted by T cells increased the cytotoxic ability of natural killer cells (NK cells), which illustrated an indirect manner for the anti-tumor function of T cells. These results revealed the heterogeneity of cargo RNA of sEVs in HNSCC and underlined a sEVs-dependent manner in which T cells act on NK cells and anti-tumor immunity.

关键词： head and neck squamous cell carcinoma natural killer cells single-cell RNA-sequencing small extracellular vesicles T cells

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SPP1+ macrophages promote head and neck squamous cell carcinoma progression by secreting TNF-α and IL-1β

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JOURNAL OF EXPERIMENTAL & CliNICAL CANCER RESEARCH 2024年第1期43卷 332页

作者： liu, Chun Wu, Kun li, chuwen Zhang, Zhen Zhai, Peisong Guo, Haiyan Zhang, Jianjun Shanghai Jiao Tong Univ Sch Med Shanghai Peoples Hosp 9 Dept Oral & Maxillofacial Head & Neck Oncol Shanghai Peoples R China Shanghai Jiao Tong Univ Coll Stomatol Shanghai Peoples R China Shanghai Jiao Tong Univ Natl Ctr Stomatol Shanghai Peoples R China Shanghai Jiao Tong Univ Natl Clin Res Ctr Oral Dis Shanghai Peoples R China Shanghai Key Lab Stomatol Shanghai Peoples R China Shanghai Jiao Tong Univ Shanghai Res Inst Stomatol Shanghai Peoples R China Shanghai Jiao Tong Univ Shanghai Ctr Head & Neck Oncol Clin & Translat Sci Shanghai Peoples R China Cent South Univ Xiangya Hosp 2 Dept Oral & Maxillofacial Surg Changsha Peoples R China Shanghai Jiao Tong Univ Shanghai Peoples Hosp 9 Sch Med Dept Clin Lab Shanghai Peoples R China

Background Head and neck squamous cell carcinoma (HNSCC) is a very aggressive disease characterized by a heterogeneous tumor immune microenvironment (TIME). Tumor-associated macrophages (TAMs) constitute the major innate immune population in the TIME where they facilitate crucial regulatory processes that participate in malignant tumor progression. SPP1 + macrophages (SPP1 + Macs) are found in many cancers, but their effects on HNSCC remain unknown. This study aimed to identify and validate the role and function of SPP1 + Macs in the malignant progression of HNSCC. Methods In this study, we applied single-cell RNA sequencing (scRNA-seq) analyses of paired tumor and normal tissues from 5 HNSCC patients to identify tumor-specific SPP1 + Macs. RT-qPCR and multiplex immunohistochemical and multiplex immunofluorescence staining were used to verify the presence of SPP1 + Macs in the clinical samples. Gene set variation analysis suggested that SPP1 + Macs were actively involved in cytokine production. Thus, we constructed SPP1-OE macrophages and SPP1-KD macrophages (both differentiated from THP-1 cells), performed a Luminex liquid suspension chip detection assay to detect differential cytokines, and further assessed their biological functions and mechanisms in several HNSCC cell lines and adjacent macrophages. An in vivo experiment was used to verify the function of SPP1 + Macs in HNSCC progression. Results The scRNA-seq results revealed that myeloid cells were heterogeneous and strongly correlated with tumor cells in the TIME in HNSCC and identified tumor-specific SPP1 + Macs, which were positively correlated with poor prognosis of HNSCC patients. Gene set variation analysis (GSVA) suggested that SPP1 + Macs were actively involved in cytokine production. Luminex liquid suspension chip detection assay indicated that SPP1 + Mac-derived TNF-alpha and IL-1 beta played important roles. Both in vitro and in vivo experiments and the use of VGX-1027, an inhibitor of macrophage-de

关键词： Macrophages Single-cell RNA sequencing SPP1+macrophages TNF-alpha IL-1 beta

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Fabrication and characterization of W1/O/W2 double emulsions stabilized with Pleurotus geesteranus protein particles via one-step emulsification

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FOOD HYDROCOLLOIDS 2024年 151卷

作者： Jin, Manhong li, chuwen Ma, Menghuan Zheng, Qianwang Guo, liqiong lin, Junfang Ye, Zhiwei Zou, Yuan South China Agr Univ Coll Food Sci Dept Bioengn Guangzhou 510642 Peoples R China Guangdong Microecol Engn Res Ctr Guangzhou 510642 Peoples R China

In this study, Pleurotus geesteranus protein isolate (PPI) with low solubility in water was fabricated into nanoparticle (PPIP) that can be uniformly dispersed in pH 7.0 aqueous solution using pH cycle method. PPIP with a zeta potential of -19.21 mV had a mean size of 250.53 nm and two main peaks in particle size distribution profile. This was consistent with the TEM results, which existed both as individual particles and as particle aggregates. The main force responsible for maintaining the structure of protein particles was hydrogen bonding. Interestingly, these protein particles were successfully utilized to fabricate W1/O/W2 double emulsions using one-step homogenous emulsification, and the formation and properties of PPIP-stabilized W1/O/W2 double emulsions were significantly influenced by the protein particle concentration, oil content, and salt concentration in W2 continuous phase. Specifically, higher particle concentration and salt concentration caused a reduction in the number of W1/O droplets and the size of oil droplets. Conversely, an increase in oil content increased the number of W1/O droplets primarily due to the augmentation in the size of oil droplets. The gel strength of double emulsions was enhanced with particle concentration and oil content, as this led to the formation of stronger particle networks and droplet networks. While an opposite phenomenon was found in increasing salt concentration in W2 continuous phase, possibly resulting from the disruption of the ordered gel network. These findings not only provide new food-grade emulsifiers for W1/O/W2 double emulsions, but also broaden the application of edible mushroom proteins in food industry.

关键词： Pleurotus geesteranus protein particle W 1 /O/W 2 double emulsion One -step emulsification method Emulsion properties

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Pharmacological activity and clinical application analysis of traditional Chinese medicine ginger from the perspective of one source and multiple substances

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CHINESE MEDICINE 2024年第1期19卷 97页

作者： Zhang, Cheng Rao, Anyang Chen, Cui li, Yuqing Tan, Xiuchi Long, Jiaxin Wang, Xinyue Cai, Junjie Huang, Jiquan Luo, Hua li, chuwen Dang, Yuanye Chengdu Univ Tradit Chinese Med Dept Pharm Chengdu 611137 Peoples R China Guangzhou Med Univ Affiliated Hosp 5 Sch Pharmaceut Sci Guangdong Prov Key Lab Mol Target & Clin Pharmacol Guangzhou 511436 Peoples R China Guangzhou Med Univ Sch Pharmaceut Sci NMPA Guangzhou 511436 Peoples R China Guangzhou Med Univ Sch Pharmaceut Sci State Key Lab Guangzhou Peoples R China Hongxing Town Govt Ruoergai Cty Aba Tibetan & Qiang Autonomous Prefecture Chengdu 624504 Sichuan Peoples R China Univ Macau Inst Chinese Med Sci State Key Lab Qual Res Chinese Med Macau 999078 Peoples R China

All types of ginger have common fundamental components, although they possess distinct strengths and inclinations when it comes to effectiveness and medicinal applications. Fresh ginger possesses the ability to effectively stimulate movement within the body, alleviate the act of vomiting, induce sweating, and provide relief for external syndromes. Dried ginger possesses both defensive and stimulant characteristics, which effectively raise the internal temperature and enhance the Yang energy. Fresh ginger is more hydrating than dried ginger, highly skilled at heating the Middle-jiao, alleviating pain, halting bleeding, and managing diarrhea. Dried ginger possesses the ability to alleviate coldness when consumed in a heated form, as well as to alleviate diarrhea when consumed in a heated form. It thrives in warm conditions and has a tendency to revert back to its warm nature. The moisture content of baked ginger is inferior to that of dried ginger, but it is highly effective in alleviating pain, bleeding, and diarrhea by warming the Middle-jiao. Ginger charcoal and stir-fried charcoal, produced through carbonization, have excellent heat retention properties and are effective in warming meridians and stopping bleeding. The potency and ability to spread of roasted ginger is less intense compared to fresh ginger, and its moisture content is not as low as that of dried ginger. The medicinal characteristics of this substance are gentle, making it beneficial for alleviating vomiting in patients who are physically frail. Its primary mode of action is on the Middle-jiao. Nevertheless, the main chemical compositions of various traditional Chinese medicines are nearly identical due to their shared base element. Ginger, in particular, possesses a range of pharmacological activities including antioxidant, anti-inflammatory, anti-tumor, anti-bacterial, and anticoagulant properties. However, modern pharmacological research has not fully acknowledged the clinical medicinal value of

关键词： Ginger One source and multiple substances Performance and efficacy Pharmacological activity Clinical application

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The efficacy and safety of dalpiciclib, a cyclin-dependent kinase 4/6 inhibitor, in patients with advanced head and neck mucosal melanoma harboring CDK4 amplification

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BMC MEDICINE 2024年第1期22卷 1页

作者： Shi, Chaoji Ju, Houyu Zhou, Rong Xu, Shengming Wu, Yunteng Gu, Ziyue Wang, Ying Chen, Wanling Huang, Xinyi Han, Yong Sun, Shuyang li, chuwen Wang, Min Zhou, Guoyu Zhang, Zhiyuan li, Jiang Ren, Guoxin Shanghai Jiao Tong Univ Shanghai Peoples Hosp 9 Sch Med Dept Oral & Maxillofacial Head Neck OncolColl Sto 639 Zhi Zao Ju Rd Shanghai 200011 Peoples R China Natl Ctr Stomatol Natl Clin Res Ctr Oral Dis Shanghai 200011 Peoples R China Shanghai Key Lab Stomatol Shanghai 200011 Peoples R China Shanghai Res Inst Stomatol Shanghai 200011 Peoples R China Chinese Acad Med Sci Res Unit Oral & Maxillofacial Regenerat Med Shanghai 200011 Peoples R China Jiangsu Hengrui Pharmaceut Co Ltd Clin Res & Dev Shanghai 200011 Peoples R China Shanghai Jiao Tong Univ Peoples Hosp 9 Dept Oral Pathol Sch Med 639 Zhi Zao Ju Rd Shanghai 200011 Peoples R China

Background Mucosal melanoma (MM) is a rare but devastating subtype of melanoma. Our previous studies have demonstrated robust anti-tumor effects of cyclin-dependent kinase 4/6 (CDK 4/6) inhibitors in head and neck MM (HNMM) patient-derived xenograft models with CDK4 amplification. Herein, we aimed to investigate the efficacy and safety of dalpiciclib (SHR6390), a CDK4/6 inhibitor, in HNMM patients harboring CDK4 *** The anti-tumor efficacy of dalpiciclib was assessed by HNMM patient-derived xenograft (PDX) models and patient-derived tumor cells (PDC) in vivo and in vitro. Immunohistochemical analyses and western blot were then performed to assess the markers of cell proliferation and CDK4/6 signaling pathway. For the clinical trial, advanced recurrent and/or metastatic HNMM patients with CDK4 amplification were treated with dalpiciclib 125 mg once daily for 21 consecutive days in 28-day cycles. The primary endpoint was disease control rate (DCR). Secondary endpoints included safety, objective response rate (ORR), progression-free survival (PFS), and overall survival (OS).Results Dalpiciclib profoundly suppressed growth of HNMM-PDX and PDC with CDK4 amplification, whereas it showed relatively weak suppression in those with CDK4 wild type compared with vehicle. And dalpiciclib resulted in a remarkable reduction in the expression levels of Ki-67 and phosphorylated Rb compared with control group. In the clinical trial, a total of 17 patients were enrolled, and 16 patients were evaluable. The ORR was 6.3%, and the DCR was 81.3%. The estimated median PFS was 9.9 months (95% CI, 4.8-NA), and the median OS was not reached. The rate of OS at 12 months and 24 months was 68.8% (95% CI, 0.494-0.957) and 51.6% (95% CI, 0.307-0.866), respectively. The most frequent adverse events were neutrophil count decrease, white blood cell count decrease, and *** Dalpiciclib was well-tolerated and displayed a durable benefit for HNMM patients with CDK4 ampli

关键词： CDK4/6 inhibitor Dalpiciclib Head and neck mucosal melanoma Clinical trial CDK4 amplification

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H3K9 lactylation in malignant cells facilitates CD8⁺ T cell dysfunction and poor immunotherapy response

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CELL REPORTS 2024年第9期43卷 114957页

作者： Wang, Ruijie li, chuwen Cheng, Zhongyi li, Mingyu Shi, Jianbo Zhang, Zhiyuan Jin, Shufang Ma, Hailong Shanghai Jiao Tong Univ Shanghai Peoples Hosp 9 Coll Stomatol Dept Oral Maxillofacial Head & Neck OncolSch Med Shanghai 200011 Peoples R China Natl Ctr Stomatol Shanghai 200011 Peoples R China Natl Clin Res Ctr Oral Dis Shanghai 200011 Peoples R China Shanghai Key Lab Stomatol Shanghai 200011 Peoples R China Shanghai Res Inst Stomatol Shanghai 200011 Peoples R China Jingjie PTM Biolab Hangzhou Hangzhou 310018 Zhejiang Peoples R China Shanghai Jiao Tong Univ Dept Dent Ctr 2 Shanghai Peoples Hosp 9 Sch Med Shanghai 200011 Peoples R China Shanghai Jiao Tong Univ Coll Stomatol Shanghai 200011 Peoples R China

Histone lysine lactylation (Kla) is a post-translational modification, and its role in tumor immune escape remains unclear. Here, we find that increased histone lactylation is associated with poor response to immunotherapy in head and neck squamous cell carcinoma (HNSCC). H3K9la is identified as a specific modification site in HNSCC. Using cleavage under targets and tagmentation analyses, interleukin-11 (IL-11) is identified as a downstream regulatory gene of H3K9la. IL-11 transcriptionally activates immune checkpoint genes through JAK2/STAT3 signaling in CD8(+) T cells. Additionally, IL-11 overexpression promotes tumor progression and CD8(+) T cell dysfunction in vivo. Moreover, IL11 knockdown reverses lactate-induced CD8(+) T cell exhaustion, and cholesterol-modified siIL11 restores CD8(+) T cell killing activity and enhances immunotherapy efficacy. Clinically, H3K9la positively correlates with IL-11 expression and unfavorable immunotherapy responses in patients. This study reveals the crucial role of histone lactylation in immune escape, providing insights into immunotherapy strategies for HNSCC.

关键词： head and neck squamous cell carcinoma histone lactylation interleukin-11 tumor microenvironment T cell dysfunction

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Phthalide metabolites produced by mangrove endophytic fungus Pestalotiopsis sp. SAS4

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MAGNETIC RESONANCE IN CHEMISTRY 2022年第5期60卷 525-529页

作者： Peng, Jiayin Chen, Peiwen li, chuwen liu, Juntao Sun, Yijun Yang, Mingze Ding, Bo Huang, Hongbo Tao, Yiwen Guangzhou Med Univ Key Lab Mol Target & Clin Pharmacol Guangzhou 511436 Peoples R China Guangzhou Med Univ State Key Lab Resp Dis Sch Pharmaceut Sci Guangzhou 511436 Peoples R China Guangzhou Med Univ Affiliated Hosp 5 Guangzhou 511436 Peoples R China

Two new phthalide derivatives (1-2) and four known phthalide compounds (3-6) were purified from the culture of a mangrove endophytic fungus Pestalotiopsis sp. SAS4. Their chemical structures were established by analyses of 1D and 2D nuclear magnetic resonance (NMR) and high resolution mass spectrometry (HR-MS) spectroscopic data. All of these compounds were evaluated in vitro for antibacterial, cytotoxicity, and resistance to hypoxic-ischemic injury activities.

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Acetyltanshinone IIA reduces the synthesis of cell cycle-related proteins by degrading p70S6K and subsequently inhibits drug-resistant lung cancer cell growth

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PHARMACOLOGICAL RESEARCH 2022年第0期179卷 106209页

作者： Huang, Bin Hao, Meng li, chuwen Luo, Kathy Qian Univ Macau Fac Hlth Sci Taipa Macao Peoples R China Univ Macau Inst Chinese Med Sci State Key Lab Qual Res Chinese Med Taipa Macao Peoples R China

Targeted therapies using tyrosine kinase inhibitors (TKIs) against epidermal growth factor receptor (EGFR) have improved the outcomes of patients with non-small cell lung cancer (NSCLC). However, due to genetic mutations of EGFR or activation of other oncogenic pathways, cancer cells can develop resistance to TKIs, resulting in usually temporary and reversible therapeutic effects. Therefore, new anticancer agents are urgently needed to treat drug-resistant NSCLC. In this study, we found that acetyltanshinone IIA (ATA) displayed much stronger potency than erlotinib in inhibiting the growth of drug-resistant NSCLC cells and their-derived xenograft tumors. Our analyses revealed that ATA achieved this effect by the following mechanisms. First, ATA could bind p70S6K at its ATP-binding pocket to prevent phosphorylation, and second by increasing the ubiquitination of p70S6K to cause its degradation. Since phosphorylation of S6 ribosome protein (S6RP) by p70S6K can induce protein synthesis at the ribosome, the dramatic reduction of p70S6K after ATA treatment led to great reductions of new protein synthesis on several cell cycle-related proteins including cyclin D3, aurora kinase A, polo-like kinase, cyclin B1, survivin;and reduced the levels of EGFR and MET. In addition, ATA treatment increased the levels of p53 and p21 proteins, which blocked cell cycle progression in the G1/S phase. Taken together, as ATA can effectively block multiple signaling pathways essential for protein synthesis and cell proliferation, ATA can potentially be developed into a multi-target anti-cancer agent to treat TKI-resistant NSCLC.

关键词： Drug-resistant lung cancer Acetyltanshinone IIA EGFR P70S6K Aurora kinase A

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Candidate therapeutic agents in a newly established triple wild-typemucosalmelanoma cell line

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Cancer Communications 2022年第7期42卷 627-647页

作者： Chaoji Shi Ziyue Gu Shengming Xu Houyu Ju Yunteng Wu Yong Han Jiayi li chuwen li Jing Wu lizhen Wang Jiang li Guoyu Zhou Weimin Ye Guoxin Ren Zhiyuan Zhang Rong Zhou Department of Oral and Maxillofacial-Head Neck Oncology Shanghai Ninth People’s HospitalCollege of StomatologyShanghai Jiao Tong University School of MedicineShanghai 200011P.R.China National Clinical Research Center for Oral Diseases Shanghai 200011P.R.China Shanghai Key Laboratory of Stomatology&Shanghai Research Institute of Stomatology Shanghai 200011P.R.China Research Unit of Oral and Maxillofacial Regenerative Medicine Chinese Academy of Medical SciencesShanghai 200011P.R.China Department of Oral Pathology Ninth People’s HospitalShanghai Jiao Tong UniversitySchool of MedicineShanghai 200011P.R.China

Background:Mucosalmelanoma has characteristically distinct genetic features and typically poor *** lack of representativemucosal melanoma models,especially cell lines,has hindered translational research on this melanoma *** this study,we aimed to establish and provide the biological properties,genomic features and the pharmacological profiles of a mucosal melanoma cell line that would contribute to the understanding and treatment optimization of molecularly-defined mucosal melanoma ***:The sample was collected from a 67-year-old mucosal melanoma patient and processed into pieces for the establishment of cell line and patientderived xenograft(PDX)*** proliferation and tumorigenic property of cancer cells from different passageswere evaluated,andwhole-genome sequencing(WGS)was performed on the original tumor,PDX,established cell line,and the matched blood to confirm the establishment and define the genomic features of this cell *** was conducted to depict the architecture of amplified regions detected by ***-throughput drug screening(HTDS)assay including a total of 103 therapeutic agents was implemented on the established cell line,and selected candidate agents were validated in the corresponding PDX ***:A mucosal melanoma cell line,MM9H-1,was established which exhibited robust proliferation and tumorigenicity after more than 100 serial *** analysis of MM9H-1,corresponding PDX,and the original tumor showed genetic fidelity across genomes,and MM9H-1 was defined as a triple wild-type(TWT)melanoma subtype lacking well-characterized“driver mutations”.Instead,the amplification of several oncogenes,telomerase reverse transcriptase(TERT),v-Rafmurine sarcoma viral oncogene homolog B1(BRAF),melanocyte Inducing transcription factor(MITF)and INO80 complex ATPase subunit(INO80),via large-scale genomic rearrangement potentially contributed to oncogenesis of ***,HTDS identified proteasome

关键词： bortezomib cell line extrachromosomal DNA high-throughput drug screening mucosal melanoma patient derived tumor cells patient-derived xenograft whole-genome sequencing

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Nuclear miR-451a activates KDM7A and leads to cetuximab resistance in head and neck squamous cell carcinoma

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CELLULAR AND MOLECULAR liFE SCIENCES 2024年第1期81卷 282-282页

作者： Zhai, Peisong Tong, Tong Wang, Xiaoning li, chuwen liu, Chun Qin, Xing li, Shu Xie, Fei Mao, Jiayi Zhang, Jianjun Guo, Haiyan Shanghai Jiao Tong Univ Natl Clin Res Ctr Oral DisSch MedColl Stomatol Shanghai Peoples Hosp 9Shanghai Key Lab Stomat Dept Oral & Maxillofacial Head & Neck OncolNatl C 639 Zhizaoju Rd Shanghai 200011 Peoples R China Fudan Univ Shanghai Stomatol Hosp Dept Oral & Maxillofacial Surg Shanghai 200011 Peoples R China Fudan Univ Shanghai Key Lab Craniomaxillofacial Dev & Dis Shanghai 200002 Peoples R China Shanghai Jiao Tong Univ Shanghai Peoples Hosp 9 Dept Clin Lab Sch Med Shanghai 200011 Peoples R China Shanghai Jiao Tong Univ Shanghai Peoples Hosp 9 Dept Plast & Reconstruct Surg Sch Med Shanghai 200011 Peoples R China

Cetuximab resistance has been a major challenge for head and neck squamous cell carcinoma (HNSCC) patients receiving targeted therapy. However, the mechanism that causes cetuximab resistance, especially microRNA (miRNA) regulation, remains unclear. Growing evidence suggests that miRNAs may act as "nuclear activating miRNAs" for targeting promoter regions or enhancers related to target genes. This study elucidates a novel mechanism underlying cetuximab resistance in HNSCC involving the nuclear activation of KDM7A transcription via miR-451a. Herein, small RNA sequencing, quantitative real-time polymerase chain reaction (qRT-PCR) and fluorescence in situ hybridization (FISH) results provided compelling evidence of miR-451a nuclear enrichment in response to cetuximab treatment. Chromatin isolation via RNA purification, microarray analysis, and bioinformatic analysis revealed that miR-451a interacts with an enhancer region in KDM7A, activating its expression and further facilitating cetuximab resistance. It has also been demonstrated that the activation of KDM7A by nuclear miR-451a is induced by cetuximab treatment and is AGO2 dependent. Logistic regression analyses of 87 HNSCC samples indicated the significance of miR-451a and KDM7A in the development of cetuximab resistance. These discoveries support the potential of miR-451a and KDM7A as valuable biomarkers for cetuximab resistance and emphasize the function of nuclear-activating miRNAs.

关键词： Nuclear miRNAs Noncoding RNA Cetuximab Head and neck squamous cell carcinoma

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