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Membrane lipid binding molecules for the isolation of bona fide extracellular vesicle types and associated biomarkers in liquid biopsy

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Journal of Cancer Metastasis and Treatment 2019年第9期4卷 1-9页

作者： Ruenn Chai Lai Kok Hian Tan sai kiang lim A*STAR Institute of Medical Biology 8A Biomedical Grove#05-05 ImmunosSingapore 138648Singapore Department of Maternal Fetal Medicine KK Women’s&Children’s HospitalSingapore 229899Singapore Department of Surgery YLL School of MedicineNUSSingapore 119074Singapore

Cancer exacts a heavy socioeconomic cost. Earlier detection and treatment are likely to mitigate this cost. Unfortunately, conventional tissue biopsy, the gold standard in cancer diagnosis cannot fulfill the goal of earlier detection. While liquid biopsy is a promising alternative to tissue biopsy, it has its challenges and limitations. A major challenge is the isolation of bona fide lipid membrane vesicles from biological fluids. In this review, we presented a new perspective of isolating different types of extracellular vesicles (EVs) by their affinity for membrane lipid binding ligands for liquid biopsy. EVs are lipid membrane particles naturally released by almost all cells and are found in almost all biological fluids suitable for liquid biopsy. They carry materials from the secreting cells that could affect the biology of the recipient cells and could thus inform on the state and progress of the disease. However, isolating bona fide EVs is a technical challenge as biological fluids have a complex composition and contain particles or aggregates that are physically similar to EVs. Here we review the use of membrane lipid-binding ligands to isolate different bona fide EV subtypes, and to circumvent the problem of co-isolating physically similar non-EV complexes in current EV isolation protocols. We will discuss the advantages of this technique and its potential for accelerated biomarker discovery and validation through examples of pre-clinical studies. We propose that isolating EV subtypes is a technically viable and robust strategy to overcome the current bottleneck of isolating EVs for liquid biopsy.

关键词： Membrane lipid binding molecules extracellular vesicle cholera toxin B shiga toxin B annexin V biomarker

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Generation and improvement of insulin-producing cells from differentiating embryonic stem cells for diabetes therapy

Generation and improvement of insulin-producing cells from d...

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中国生理学会第23届全国会员代表大会暨生理学学术大会

作者： Arun Sharma sai kiang lim Joslin Diabetes Center Harvard Medical School BostonUSA Institute of Medical Biology A*STAR Singapore

Most type 1 diabetic patients rely on daily insulin injections,yet still developing cardiovascular complications. Transplantation of human pancreas or isolated islets may cure the ***,shortage of cadaveric pancreata and requirement of life-long immune suppression are 2 major obstacles in transplantation therapy of insulin-dependent *** of functional insulin-producing cells from stem cells and immunoisolation by encapsulation may circumvent the *** had generated a dozen of unlimited and glucose-responsive insulin-producing cell lines(*** and ERoSHK) from mouse embryonic stem and progenitor *** of these cells in immune deficient diabetic mice could correct *** allo-transplantation studies were carried out in immune-competent,streptozotocin-induced diabetic ***-producing(MEPI-1) cells derived from mouse embryonic progenitor cells were immunoisolated by encapsulation with alginate and peritoneally transplanted into diabetic *** glucose and insulin levels,and glucose tolerance tests were *** were washed out to determine cell functions and *** was reversed within a few days after transplantation,followed by a period of stable normoglycemic or moderate hypoglycemic levels for 2.5 months before *** reappeared immediately upon washing out of capsules.A second transplantation in relapsed mice corrected hyperglycemia as in the first *** from washed-out capsules remained the capability of glucose-stimulated insulin *** spite of fibrotic overgrowth on the capsules,the permeability of capsules seemed not *** transplantation,the number of viable cells in capsules was increased initially and then decreased to less than the transplanted cell number after 75 *** examinations showed the presence of proliferating cells and a necrotic core in the encapsulated cell clusters beyond 40 *** chang

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Mesenchymal stromal cell-derived small extracellular vesicles modulate macrophage polarization and enhance angio-osteogenesis to promote bone healing

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Genes & Diseases 2022年第4期9卷 841-844页

作者： Shang Jiunn Chuah Chee Weng Yong Kristeen Ye Wen Teo Jacob Ren Jie Chew Yi Ann Cheow Shipin Zhang Raymond Chung Wen Wong Asher Ah Tong lim sai kiang lim Wei Seong Toh Faculty of Dentistry National University of Singapore119085Singapore Institute of Molecular and Cell Biology Agency for ScienceTechnology and Research138648Singapore Department of Orthopaedic Surgery Yong Loo Lin School of MedicineNational University of Singapore119288Singapore Department of Biomedical Engineering Faculty of EngineeringNational University of Singapore117583Singapore Tissue Engineering Program Life Sciences InstituteNational University of Singapore117456Singapore Integrative Sciences and Engineering Program NUS Graduate SchoolNational University of Singapore119077Singapore

Mesenchymal stem/stromal cells(MSCs)have demonstrated therapeutic efficacy for bone regeneration in animal and clinical *** MSCs were initially thought to differentiate to various cell types to replace the injured/diseased tissue,it is now accepted that these cells secrete factors to promote tissue repair.1 Among these factors,small extracellular vesicles(sEVs)of size 50–200 nm,which include the exosomes,have been identified as the principal agent mediating the wide-ranging therapeutic efficacy of MSCs.2 Several studies have also reported the therapeutic effects of MSC-sEVs to enhance bone repair in animal models,as recently reviewed.3 However,the cellular processes and mechanisms mediated by MSC-sEVs in bone regeneration remain to be fully elucidated.

关键词： healing clinical macrophage

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Bleeding complications after percutaneous kidney biopsies–nationwide experience from Brunei Darussalam

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World Journal of Nephrology 2023年第5期12卷 147-158页

作者： Chiao Yuen lim sai Laung Khay Department of Renal Services RIPAS HospitalBandar Seri Begawan BA1712Brunei Darussalam

BACKGROUND Kidney biopsy serves as a valuable method for both diagnosing and monitoring kidney *** studies have identified several risk factors associated with bleeding complications following the procedure,but these findings have shown inconsistency and *** To investigate the risk of bleeding complications following percutaneous kidney biopsy in Brunei *** sought to explore the relevant clinical and pathological risk factors associated with these complications while also considering the findings within the broader international literature *** We conducted a retrospective study of all adult patients who underwent kidney biopsy in Brunei Darussalam from October 2013 to September *** outcomes of interest were post-biopsy bleeding and the need for blood ***,clinical,laboratory and procedural-related data were *** regression analysis was used to identify predictors of *** A total of 255 kidney biopsies were included,with 11%being performed on transplanted *** majority of biopsies were done under ultrasound guidance(83.1%),with the rest under computer tomography guidance(16.9%).The most common indications for biopsy were chronic kidney disease of undefined cause(36.1%),nephrotic syndrome(24.3%)and acute kidney injury(11%).Rate of bleeding complication was 6.3%–2%frank hematuria and 4.3%perinephric *** transfusion was required in 2.8%of *** patient lost a kidney or died because of the *** logistic regression identified baseline hemoglobin[odds ratio(OR):4.11;95%confidence interval(95%CI):1.12-15.1;P=0.03 for hemoglobin≤11 g/dL vs.>11 g/dL)and the presence of microscopic hematuria(OR:5.24;95%CI:1.43-19.1;P=0.01)as independent risk factors for post-biopsy ***,low baseline platelet count was identified as the dominant risk factor for requiring postbiopsy ***,each 10109/L decrease in baseline pl

关键词： Kidney biopsy Bleeding complications Logistic regression Retrospective cohort study Risk

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DPHL:A DIA Pan-human Protein Mass Spectrometry Library for Robust Biomarker Discovery

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Genomics, Proteomics & Bioinformatics 2020年第2期18卷 104-119页

作者： Tiansheng Zhu Yi Zhu Yue Xuan Huanhuan Gao Xue Cai Sander R.Piersma Thang V.Pham Tim Schelfhorst Richard R.G.D.Haas Irene V.Bijnsdorp Rui Sun Liang Yue Guan Ruan Qiushi Zhang Mo Hu Yue Zhou Winan J.Van Houdt Tessa Y.S.Le Large Jacqueline Cloos Anna Wojtuszkiewicz Danijela Koppers-Lalic Franziska Bottger Chantal Scheepbouwer Ruud H.Brakenhoff Geert J.L.H.van Leenders Jan N.M.Ijzermans John W.M.Martens Renske D.M.Steenbergen Nicole C.Grieken Sathiyamoorthy Selvarajan Sangeeta Mantoo Sze S.Lee Serene J.Y.Yeow Syed M.F.Alkaff Nan Xiang Yaoting Sun Xiao Yi Shaozheng Dai Wei Liu Tian Lu Zhicheng Wu Xiao Liang Man Wang Yingkuan Shao Xi Zheng Kailun Xu Qin Yang Yifan Meng Cong Lu Jiang Zhu Jin'e Zheng Bo Wang sai Lou Yibei Dai Chao Xu Chenhuan Yu Huazhong Ying Tony K.lim Jianmin Wu Xiaofei Gao Zhongzhi Luan Xiaodong Teng Peng Wu Shi'ang Huang Zhihua Tao Narayanan G.Iyer Shuigeng Zhou Wenguang Shao Henry Lam Ding Ma Jiafu Ji Oi L.Kon Shu Zheng Ruedi Aebersold Connie R.Jimenez Tiannan Guo Zhejiang Provincial Laboratory of Life Sciences and Biomedicine Westlake UniversityHangzhou 310024China Key Laboratory of Structural Biology of Zhejiang Province School of Life SciencesWestlake UniversityHangzhou 310024China Institute of Basic Medical Sciences Westlake Institute for Advanced StudyHangzhou 310024China School of Computer Science Shanghai Key Laboratory of Intelligent Information ProcessingFudan UniversityShanghai 200433China Thermo Fisher Scientific(BREMEN)GmbH Bremen 28195Germany OncoProteomics Laboratory Department of Medical OncologyVU University Medical CenterVU UniversityAmsterdam 1011The Netherlands Amsterdam UMC Vrije Universiteit AmsterdamUrologyCancer Center AmsterdamAmsterdam 1011The Netherlands Thermo Fisher Scientific Shanghai 201206China The Netherlands Cancer Institute Surgical OncologyAmsterdam 1011The Netherlands Amsterdam UMC Vrije Universiteit AmsterdamSurgeryCancer Center AmsterdamAmsterdam 1011The Netherlands Amsterdam UMC Vrije Universiteit AmsterdamPediatric Oncology/HematologyCancer Center AmsterdamAmsterdam 1011The Netherlands Amsterdam UMC Vrije Universiteit AmsterdamHematologyCancer Center AmsterdamAmsterdam 1011The Netherlands Amsterdam UMC Vrije Universiteit AmsterdamMedical OncologyCancer Center AmsterdamAmsterdam 1011The Netherlands Amsterdam UMC Vrije Universiteit AmsterdamNeurosurgeryCancer Center AmsterdamAmsterdam 1011The Netherlands Amsterdam UMC Vrije Universiteit AmsterdamPathologyCancer Center AmsterdamAmsterdam 1011The Netherlands Amsterdam UMC Vrije Universiteit AmsterdamOtolaryngology/Head and Neck SurgeryCancer Center AmsterdamAmsterdam 1011The Netherlands Erasmus MC University Medical Center PathologyRotterdam 1016LVThe Netherlands Erasmus MC University Medical Center SurgeryRotterdam 1016LVThe Netherlands Erasmus MC University Medical Center Medical OncologyRotterdam 1016LVThe Netherlands Department of Anatomical Pathology Singapore General HospitalSingapore 169608Singapore Division of Medical Scie

To address the increasing need for detecting and validating protein biomarkers in clinical specimens,mass spectrometry(MS)-based targeted proteomic techniques,including the selected reaction monitoring(SRM),parallel reaction monitoring(PRM),and massively parallel dataindependent acquisition(DIA),have been *** optimal performance,they require the fragment ion spectra of targeted peptides as prior *** this report,we describe a MS pipeline and spectral resource to support targeted proteomics studies for human tissue *** build the spectral resource,we integrated common open-source MS computational tools to assemble a freely accessible computational workflow based on *** then applied the workflow to generate DPHL,a comprehensive DIA pan-human library,from 1096 data-dependent acquisition(DDA)MS raw files for 16 types of cancer *** extensive spectral resource was then applied to a proteomic study of 17 prostate cancer(PCa)***,PRM validation was applied to a larger study of 57 PCa patients and the differential expression of three proteins in prostate tumor was *** a second application,the DPHL spectral resource was applied to a study consisting of plasma samples from 19 diffuse large B cell lymphoma(DLBCL)patients and 18 healthy control *** expressed proteins between DLBCL patients and healthy control subjects were detected by DIA-MS and confirmed by *** data demonstrate that the DPHL supports DIA and PRM MS pipelines for robust protein biomarker *** is freely accessible at https://***/page/***?id=IPX0001400000.

关键词： Data-independent acquisition Parallel reaction monitoring Spectral library Prostate cancer Diffuse large B cell lymphoma

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