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An integrated microfluidics platform with high-throughput single-cell cloning array and concentration gradient generator for efficient cancer drug effect screening

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Military Medical Research 2023年第3期10卷 325-341页

作者： Biao Wang Bang-Shun He Xiao-Lan Ruan Jiang Zhu Rui Hu Jie Wang Ying Li Yun-Huang Yang Mai-Li Liu Key Laboratory of Magnetic Resonance in Biological Systems State Key Laboratory of Magnetic Resonance and Atomic and Molecular PhysicsNational Center for Magnetic Resonance in WuhanWuhan Institute of Physics and MathematicsInnovation Academy for Precision Measurement Science and Technology-Wuhan National Laboratory for OptoelectronicsChinese Academy of SciencesWuhan 430071China Department of Laboratory Medicine Nanjing First HospitalNanjing Medical UniversityNanjing 210006China Department of Hematology Renmin HospitalWuhan University Wuhan 430060China University of Chinese Academy of Sciences Beijing 10049China Canary Center at Stanford for Cancer Early Detection Department of RadiologySchool of MedicineStanford UniversityPalo AltoCA 94304USA

Background:Tumor cell heterogeneity mediated drug resistance has been recognized as the stumbling block of cancer *** the cytotoxicity of anticancer drugs at single-cell level in a high-throughput way is thus of great value for developing precision ***,current techniques suffer from limitations in dynamically characterizing the responses of thousands of single cells or cell clones presented to multiple drug ***:We developed a new microfluidics-based“SMART”platform that is Simple to operate,able to generate a Massive single-cell array and Multiplex drug concentrations,capable of keeping cells Alive,Retainable and Trackable in the *** features are achieved by integrating a Microfluidic chamber Array(4320 units)and a sixConcentration gradient generator(MAC),which enables highly efficient analysis of leukemia drug effects on single cells and cell clones in a high-throughput ***:A simple procedure produces 6 on-chip drug gradients to treat more than 3000 single cells or single-cell derived clones and thus allows an efficient and precise analysis of cell *** statistic results reveal that Imatinib(Ima)and Resveratrol(Res)combination treatment on single cells or clones is much more efficient than Ima or Res single drug treatment,indicated by the markedly reduced half maximal inhibitory concentration(IC50).Additionally,single-cell derived clones demonstrate a higher IC_(50) in each drug treatment compared to single ***,primary cells isolated from two leukemia patients are also found with apparent heterogeneity upon drug treatment on ***:This microfluidics-based“SMART”platform allows high-throughput single-cell capture and culture,dynamic drug-gradient treatment and cell response monitoring,which represents a new approach to efficiently investigate anticancer drug effects and should benefit drug discovery for leukemia and other cancers.

关键词： Microfluidics Single-cell analysis Leukemia high-throughput drug screening Single-cell cloning

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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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