Tracking the expression of therapeutic protein targets in rare cells by antibody-mediated nanoparticle labelling and magnetic sorting

Labib, Mahmoud; Wang, Zongjie; Ahmed, Sharif Uddin; Mohamadi, Reza M.; Duong, Bill; Green, Brenda; Sargent, Edward H.; Kelley, Shana O.

doi:10.1038/s41551-020-0590-1

Cited by 49 publications

(50 citation statements)

References 55 publications

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“…Furthermore, scRNA-Seq can be combined with scDNA barcoding, using CRISPR for lineage tracing ( Raj et al, 2018 ; Spanjaard et al, 2018 ), or with sc-epigenetics ( Ai et al, 2019 ; Chen et al, 2019 ; Li et al, 2019 ; Rooijers et al, 2019 ; Zhu et al, 2019 ). Single-cell proteomics is also being extensively developed ( Goltsev et al, 2018 ; Gut et al, 2018 ; Saka et al, 2019 ; Labib et al, 2020 ). In situ single-cell sequencing methods can identify spatial connections between cell types of interest that may be particularly useful for further inferences concerning the structural development of the pancreas or niche and pancreas crosstalk.…”

Section: Discussion: Concluding Remarks and Future Perspectivesmentioning

confidence: 99%

Endocrine Pancreas Development and Dysfunction Through the Lens of Single-Cell RNA-Sequencing

Szlachcic

Ziojla

Kizewska

et al. 2021

Front. Cell Dev. Biol.

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A chronic inability to maintain blood glucose homeostasis leads to diabetes, which can damage multiple organs. The pancreatic islets regulate blood glucose levels through the coordinated action of islet cell-secreted hormones, with the insulin released by β-cells playing a crucial role in this process. Diabetes is caused by insufficient insulin secretion due to β-cell loss, or a pancreatic dysfunction. The restoration of a functional β-cell mass might, therefore, offer a cure. To this end, major efforts are underway to generate human β-cells de novo, in vitro, or in vivo. The efficient generation of functional β-cells requires a comprehensive knowledge of pancreas development, including the mechanisms driving cell fate decisions or endocrine cell maturation. Rapid progress in single-cell RNA sequencing (scRNA-Seq) technologies has brought a new dimension to pancreas development research. These methods can capture the transcriptomes of thousands of individual cells, including rare cell types, subtypes, and transient states. With such massive datasets, it is possible to infer the developmental trajectories of cell transitions and gene regulatory pathways. Here, we summarize recent advances in our understanding of endocrine pancreas development and function from scRNA-Seq studies on developing and adult pancreas and human endocrine differentiation models. We also discuss recent scRNA-Seq findings for the pathological pancreas in diabetes, and their implications for better treatment.

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Section: Discussion: Concluding Remarks and Future Perspectivesmentioning

confidence: 99%

Endocrine Pancreas Development and Dysfunction Through the Lens of Single-Cell RNA-Sequencing

Szlachcic

Ziojla

Kizewska

et al. 2021

Front. Cell Dev. Biol.

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“…CTC characterization is a "liquid biopsy" rather than an Unlike tissue biopsy approach, allowing minimally invasive and real-time biomarker discovery and expression monitoring 31 . However, most of the current clinical applications of CTC detection and monitoring are based on simple enumeration of CTCs 20 .…”

Section: Discussionmentioning

confidence: 99%

Integrated microfluidic single-cell immunoblotting chip enables high-throughput isolation, enrichment and direct protein analysis of circulating tumor cells

Abdulla

Zhang

et al. 2022

Microsyst Nanoeng

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Effective capture and analysis of a single circulating tumor cell (CTC) is instrumental for early diagnosis and personalized therapy of tumors. However, due to their extremely low abundance and susceptibility to interference from other cells, high-throughput isolation, enrichment, and single-cell-level functional protein analysis of CTCs within one integrated system remains a major challenge. Herein, we present an integrated multifunctional microfluidic system for highly efficient and label-free CTC isolation, CTC enrichment, and single-cell immunoblotting (ieSCI). The ieSCI-chip is a multilayer microfluidic system that combines an inertia force-based cell sorter with a membrane filter for label-free CTC separation and enrichment and a thin layer of a photoactive polyacrylamide gel with microwell arrays at the bottom of the chamber for single-cell immunoblotting. The ieSCI-chip successfully identified a subgroup of apoptosis-negative (Bax-negative) cells, which traditional bulk analysis did not detect, from cisplatin-treated cells. Furthermore, we demonstrated the clinical application of the ieSCI-chip with blood samples from breast cancer patients for personalized CTC epithelial-to-mesenchymal transition (EMT) analysis. The expression level of a tumor cell marker (EpCAM) can be directly determined in isolated CTCs at the single-cell level, and the therapeutic response to anticancer drugs can be simultaneously monitored. Therefore, the ieSCI-chip provides a promising clinical translational tool for clinical drug response monitoring and personalized regimen development.

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“…In 2018, the FDA approved the PARP inhibitor, olaparib, for treatment of patients with advanced breast and ovarian cancers harboring BRCA mutations [158]. More recently, the MagRC platform was featured as a technology capable of analyzing intracellular proteins in the nucleus, mitochondria and cytoplasm of rare tumor cells, without interference from hematopoietic cells [54]. The approach relied on the use of magnetic amplification reagents to facilitate sensitive detection of low-abundance proteins (Fig.…”

Section: Altered Proteins and Resistancementioning

confidence: 99%

“…Immunomagnetic separation involves prelabeling the cells with magnetic beads-conjugated antibodies, followed by capturing CTCs within a microfluidic device exposed to a magnetic field [41]. This workflow has been adopted in the magnetic ranking cytometry (MagRC) technology, which can rank the cells according to the expression of cell-surface proteins [53], intracellular proteins [54], or mRNAs [55]. The MagRC devices contain capture zones to rank the cells according to their magnetic loading (Fig.…”

Section: Positive Enrichmentmentioning

confidence: 99%

Circulating tumor cell profiling for precision oncology

Labib

2021

Molecular Oncology

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Analysis of circulating tumor cells (CTCs) collected from patient's blood offers a broad range of opportunities in the field of precision oncology. With new advances in profiling technology, it is now possible to demonstrate an association between the molecular profiles of CTCs and tumor response to therapy. In this Review, we discuss mechanisms of tumor resistance to therapy and their link to phenotypic and genotypic properties of CTCs. We summarize key technologies used to isolate and analyze CTCs and discuss recent clinical studies that examined CTCs for genomic and proteomic predictors of responsiveness to therapy. We also point out current limitations that still hamper the implementation of CTCs into clinical practice. We finally reflect on how these shortcomings can be addressed with the likely contribution of multiparametric approaches and advanced data analytics.

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Tracking the expression of therapeutic protein targets in rare cells by antibody-mediated nanoparticle labelling and magnetic sorting

Cited by 49 publications

References 55 publications

Endocrine Pancreas Development and Dysfunction Through the Lens of Single-Cell RNA-Sequencing

Endocrine Pancreas Development and Dysfunction Through the Lens of Single-Cell RNA-Sequencing

Integrated microfluidic single-cell immunoblotting chip enables high-throughput isolation, enrichment and direct protein analysis of circulating tumor cells

Circulating tumor cell profiling for precision oncology

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