Single-Cell Multiplexed Proteomics on the IsoLight Resolves Cellular Functional Heterogeneity to Reveal Clinical Responses of Cancer Patients to Immunotherapies

Liu, Dong; Paczkowski, Patrick; Mackay, Sean; Ng, Colin

doi:10.1007/978-1-4939-9773-2_19

Cited by 33 publications

(25 citation statements)

References 22 publications

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“…Consequently, the hypothesis that one single trophozoite expresses one single VSP could be modified in the sense that one single trophozoite may express VSPs that may be encoded by one or more ORFs. This hypothesis could be verified only by single-cell proteomics (Dou et al ., 2019; Liu et al ., 2020), a method which has not been established for G. lamblia , so far.…”

Section: Discussionmentioning

confidence: 99%

Comparative proteomics of threeGiardia lambliastrains: investigation of antigenic variation in the post-genomic era

et al. 2020

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Giardia lamblia is a causative agent of persistent diarrhoea widespread in regions with low hygienic standards. Laboratory research is based on cloned lines issuing from various patient isolates typed in the late 1980s and 90s using restriction analysis and serology. In the present study, we compared the well-characterized strain WBC6 with another clone of the parent WB isolate termed WBA1 and with a clone from another isolate, GS/M-83-H7, using shotgun mass spectrometry proteomics. We identified 398 proteins differentially expressed between the GS and both WB isolates and 97 proteins differentially expressed between the two WB isolates. We investigated the expression levels of the predominant variant-specific surface proteins (VSPs) in each clone and matched the previously described major VSPs of each strain to the corresponding open reading frame sequences identified by whole-genome sequencing efforts. Furthermore, since the original WB isolate comes from a patient treated with metronidazole, we compared the susceptibilities of the strains to nitro compounds, as well the expression levels of enzymes involved in nitro reduction and on the corresponding enzyme activities and found distinct differences between the three strains.

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Section: Discussionmentioning

confidence: 99%

Comparative proteomics of threeGiardia lambliastrains: investigation of antigenic variation in the post-genomic era

et al. 2020

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“…Though enabling multi‐analyte profiling, the method is not easily applicable to single cells. However, another technique, called IsoLight, combines sandwich ELISA and fluorescence signal detection (Liu et al, 2020) and is based on a single‐cell, highly multiplexed chip consisting of an antibody barcode array. It enables the quantification of 40+ key secreted proteins from up to ~10,000 live single cells.…”

Section: Hallmarks Of Cellular Senescencementioning

confidence: 99%

Hallmarks and detection techniques of cellular senescence and cellular ageing in immune cells

2021

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The ageing of the global population brings about unprecedented challenges. Chronic age‐related diseases in an increasing number of people represent an enormous burden for health and social care. The immune system deteriorates during ageing and contributes to many of these age‐associated diseases due to its pivotal role in pathogen clearance, tissue homeostasis and maintenance. Moreover, in order to develop treatments for COVID‐19, we urgently need to acquire more knowledge about the aged immune system, as older adults are disproportionally and more severely affected. Changes with age lead to impaired responses to infections, malignancies and vaccination, and are accompanied by chronic, low‐degree inflammation, which together is termed immunosenescence. However, the molecular and cellular mechanisms that underlie immunosenescence, termed immune cell senescence, are mostly unknown. Cellular senescence, characterised by an irreversible cell cycle arrest, is thought to be the cause of tissue and organismal ageing. Thus, better understanding of cellular senescence in immune populations at single‐cell level may provide us with insight into how immune cell senescence develops over the life time of an individual. In this review, we will briefly introduce the phenotypic characterisation of aged innate and adaptive immune cells, which also contributes to overall immunosenescence, including subsets and function. Next, we will focus on the different hallmarks of cellular senescence and cellular ageing, and the detection techniques most suitable for immune cells. Applying these techniques will deepen our understanding of immune cell senescence and to discover potential druggable pathways, which can be modulated to reverse immune ageing.

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“…Thus, heterogeneity is the rule rather than the exception. Table 1 summarizes some of the known types of heterogeneity in solid tumors [1][2][3][4][5][6][7][8][9][10][11][12][13]. Table 1.…”

Section: Heterogeneity In the Tumor Microenvironment (Tme)mentioning

confidence: 99%

Resolving Metabolic Heterogeneity in Experimental Models of the Tumor Microenvironment from a Stable Isotope Resolved Metabolomics Perspective

et al. 2020

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The tumor microenvironment (TME) comprises complex interactions of multiple cell types that determines cell behavior and metabolism such as nutrient competition and immune suppression. We discuss the various types of heterogeneity that exist in solid tumors, and the complications this invokes for studies of TME. As human subjects and in vivo model systems are complex and difficult to manipulate, simpler 3D model systems that are compatible with flexible experimental control are necessary for studying metabolic regulation in TME. Stable Isotope Resolved Metabolomics (SIRM) is a valuable tool for tracing metabolic networks in complex systems, but at present does not directly address heterogeneous metabolism at the individual cell level. We compare the advantages and disadvantages of different model systems for SIRM experiments, with a focus on lung cancer cells, their interactions with macrophages and T cells, and their response to modulators in the immune microenvironment. We describe the experimental set up, illustrate results from 3D cultures and co-cultures of lung cancer cells with human macrophages, and outline strategies to address the heterogeneous TME.

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Single-Cell Multiplexed Proteomics on the IsoLight Resolves Cellular Functional Heterogeneity to Reveal Clinical Responses of Cancer Patients to Immunotherapies

Cited by 33 publications

References 22 publications

Comparative proteomics of threeGiardia lambliastrains: investigation of antigenic variation in the post-genomic era

Comparative proteomics of threeGiardia lambliastrains: investigation of antigenic variation in the post-genomic era

Hallmarks and detection techniques of cellular senescence and cellular ageing in immune cells

Resolving Metabolic Heterogeneity in Experimental Models of the Tumor Microenvironment from a Stable Isotope Resolved Metabolomics Perspective

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