scholarly journals Application of single-cell sequencing technologies in pancreatic cancer

2021 ◽  
Author(s):  
Mastan Mannarapu ◽  
Begum Dariya ◽  
Obul Reddy Bandapalli
Keyword(s):  
Pancreatic Cancer ◽  
Single Cell ◽  
Tumor Cells ◽  
Circulating Tumor Cells ◽  
Single Cells ◽  
Evolutionary Relationship ◽  
Intratumor Heterogeneity ◽  
Sequencing Technology ◽  
Single Cell Sequencing ◽  
Sequencing Technologies

AbstractPancreatic cancer (PC) is the third lethal disease for cancer-related mortalities globally. This is mainly because of the aggressive nature and heterogeneity of the disease that is diagnosed only in their advanced stages. Thus, it is challenging for researchers and clinicians to study the molecular mechanism involved in the development of this aggressive disease. The single-cell sequencing technology enables researchers to study each and every individual cell in a single tumor. It can be used to detect genome, transcriptome, and multi-omics of single cells. The current single-cell sequencing technology is now becoming an important tool for the biological analysis of cells, to find evolutionary relationship between multiple cells and unmask the heterogeneity present in the tumor cells. Moreover, its sensitivity nature is found progressive enabling to detect rare cancer cells, circulating tumor cells, metastatic cells, and analyze the intratumor heterogeneity. Furthermore, these single-cell sequencing technologies also promoted personalized treatment strategies and next-generation sequencing to predict the disease. In this review, we have focused on the applications of single-cell sequencing technology in identifying cancer-associated cells like cancer-associated fibroblast via detecting circulating tumor cells. We also included advanced technologies involved in single-cell sequencing and their advantages. The future research indeed brings the single-cell sequencing into the clinical arena and thus could be beneficial for diagnosis and therapy of PC patients.

scholarly journals Using single-cell sequencing technology to detect circulating tumor cells in solid tumors

2021 ◽  
Vol 20 (1) ◽  
Author(s):  
Jiasheng Xu ◽  
Kaili Liao ◽  
Xi Yang ◽  
Chengfeng Wu ◽  
Wei Wu ◽  
...  
Keyword(s):  
Single Cell ◽  
Solid Tumors ◽  
Tumor Cells ◽  
Circulating Tumor Cells ◽  
Peripheral Blood ◽  
High Throughput Sequencing ◽  
Metastatic Potential ◽  
Sequencing Analysis ◽  
Sequencing Technology ◽  
Single Cell Sequencing

AbstractCirculating tumor cells are tumor cells with high vitality and high metastatic potential that invade and shed into the peripheral blood from primary solid tumors or metastatic foci. Due to the heterogeneity of tumors, it is difficult for high-throughput sequencing analysis of tumor tissues to find the genomic characteristics of low-abundance tumor stem cells. Single-cell sequencing of circulating tumor cells avoids interference from tumor heterogeneity by comparing the differences between single-cell genomes, transcriptomes, and epigenetic groups among circulating tumor cells, primary and metastatic tumors, and metastatic lymph nodes in patients' peripheral blood, providing a new perspective for understanding the biological process of tumors. This article describes the identification, biological characteristics, and single-cell genome-wide variation in circulating tumor cells and summarizes the application of single-cell sequencing technology to tumor typing, metastasis analysis, progression detection, and adjuvant therapy.

Capillary Force Driven Single-Cell Spiking Apparatus for Studying Circulating Tumor Cells

2018 ◽  
Author(s):  
Jacob Amontree ◽  
Kangfu Chen ◽  
Jose Varillas ◽  
Z. Hugh Fan
Keyword(s):  
Single Cell ◽  
Tumor Cells ◽  
Circulating Tumor Cells ◽  
Single Cells ◽  
Lymphoblastic Leukemia ◽  
Surface Membrane ◽  
Critical Element ◽  
Cell Capture ◽  
Biomedical Sciences ◽  
Low Concentrations

The characterization of single cells within heterogeneous populations has great impact on both biomedical sciences and cancer research. By investigating cellular compositions on a broad scale, pertinent outliers may be lost in the sample set. Alternatively, an investigation focused on the behavior of specific cells, such as circulating tumor cells (CTCs), will reveal genetic biomarkers or phenotypic characteristics associated with cancer and metastasis. On average, CTC concentration in peripheral blood is extremely low, as few as one to two per billion of healthy blood cells. Consequently, the critical element lacking in many methods of CTC detection is accurate cell capture efficiency at low concentrations. To simulate CTC isolation, researchers usually spike small amounts of tumor cells to healthy blood for separation. However, spiking tumor cells at extremely low concentrations is challenging in a standard laboratory setting. We report our study on an innovative apparatus and method designed for low-cost, precise, and replicable single-cell spiking (SCS). Our SCS method operates solely from capillary aspiration without the reliance on external laboratory equipment. To ensure that our method does not affect the viability of each cell, we investigated the effects of surface membrane tensions induced by aspiration. Finally, we performed affinity-based CTC isolation using human acute lymphoblastic leukemia cells (CCRF-CEM) spiked into healthy whole blood with the SCS technique. The results of the isolation experiments demonstrate the reliability of our method in generating low-concentration cell samples.

scholarly journals Research and application of single-cell sequencing in tumor heterogeneity and drug resistance of circulating tumor cells

2020 ◽  
Vol 8 (1) ◽  
Author(s):  
Zhe Dai ◽  
Xu-yu Gu ◽  
Shou-yan Xiang ◽  
Dan-dan Gong ◽  
Chang-feng Man ◽  
...  
Keyword(s):  
Drug Resistance ◽  
Single Cell ◽  
Tumor Cells ◽  
Circulating Tumor Cells ◽  
Tumor Heterogeneity ◽  
Malignant Tumors ◽  
Single Cell Sequencing ◽  
Tumor Drug Resistance ◽  
New Perspective ◽  
Tumor Drugs

Abstract Malignant tumor is a largely harmful disease worldwide. The cure rate of malignant tumors increases with the continuous discovery of anti-tumor drugs and the optimisation of chemotherapy options. However, drug resistance of tumor cells remains a massive obstacle in the treatment of anti-tumor drugs. The heterogeneity of malignant tumors makes studying it further difficult for us. In recent years, using single-cell sequencing technology to study and analyse circulating tumor cells can avoid the interference of tumor heterogeneity and provide a new perspective for us to understand tumor drug resistance.

scholarly journals A Combinatorial Approach for Single-cell Variant Detection via Phylogenetic Inference

2019 ◽  
Author(s):  
Mohammadamin Edrisi ◽  
Hamim Zafar ◽  
Luay Nakhleh
Keyword(s):  
Single Cell ◽  
Human Cancer ◽  
Evolutionary Relationship ◽  
Error Rates ◽  
Intratumor Heterogeneity ◽  
Combinatorial Approach ◽  
Cancer Dataset ◽  
Inference Problem ◽  
Single Cell Sequencing ◽  
Joint Inference

AbstractSingle-cell sequencing provides a powerful approach for elucidating intratumor heterogeneity by resolving cell-to-cell variability. However, it also poses additional challenges including elevated error rates, allelic dropout and non-uniform coverage. A recently introduced single-cell-specific mutation detection algorithm leverages the evolutionary relationship between cells for denoising the data. However, due to its probabilistic nature, this method does not scale well with the number of cells. Here, we develop a novel combinatorial approach for utilizing the genealogical relationship of cells in detecting mutations from noisy single-cell sequencing data. Our method, called scVILP, jointly detects mutations in individual cells and reconstructs a perfect phylogeny among these cells. We employ a novel Integer Linear Program algorithm for deterministically and efficiently solving the joint inference problem. We show that scVILP achieves similar or better accuracy but significantly better runtime over existing methods on simulated data. We also applied scVILP to an empirical human cancer dataset from a high grade serous ovarian cancer patient.

scholarly journals Effect of Blood Collection Tube Type and Time to Processing on the Enumeration and High-Content Characterization of Circulating Tumor Cells Using the High-Definition Single-Cell Assay

2017 ◽  
Vol 142 (2) ◽  
pp. 198-207 ◽  
Author(s):  
Mariam Rodríguez-Lee ◽  
Anand Kolatkar ◽  
Madelyn McCormick ◽  
Angel D. Dago ◽  
Jude Kendall ◽  
...  
Keyword(s):  
Single Cell ◽  
Tumor Cells ◽  
Circulating Tumor Cells ◽  
Single Cells ◽  
Blood Collection ◽  
High Definition ◽  
Cell Assay ◽  
Collection Tube ◽  
Blood Collection Tube

Context.— As circulating tumor cell (CTC) assays gain clinical relevance, it is essential to address preanalytic variability and to develop standard operating procedures for sample handling in order to successfully implement genomically informed, precision health care. Objective.— To evaluate the effects of blood collection tube (BCT) type and time-to-assay (TTA) on the enumeration and high-content characterization of CTCs by using the high-definition single-cell assay (HD-SCA). Design.— Blood samples of patients with early- and advanced-stage breast cancer were collected into cell-free DNA (CfDNA), EDTA, acid-citrate-dextrose solution, and heparin BCTs. Time-to-assay was evaluated at 24 and 72 hours, representing the fastest possible and more routine domestic shipping intervals, respectively. Results.— We detected the highest CTC levels and the lowest levels of negative events in CfDNA BCT at 24 hours. At 72 hours in this BCT, all CTC subpopulations were decreased with the larger effect observed in high-definition CTCs and cytokeratin-positive cells smaller than white blood cells. Overall cell retention was also optimal in CfDNA BCT at 24 hours. Whole-genome copy number variation profiles were generated from single cells isolated from all BCT types and TTAs. Cells from CfDNA BCT at 24-hour TTA exhibited the least noise. Conclusions.— Circulating tumor cells can be identified and characterized under a variety of collection, handling, and processing conditions, but the highest quality can be achieved with optimized conditions. We quantified performance differences of the HD-SCA for specific preanalytic variables that may be used as a guide to develop best practices for implementation into patient care and/or research biorepository processes.

scholarly journals The Single-Cell Sequencing: A Dazzling Light Shining on the Dark Corner of Cancer

2021 ◽  
Vol 11 ◽  
Author(s):  
Jing Li ◽  
Nan Yu ◽  
Xin Li ◽  
Mengna Cui ◽  
Qie Guo
Keyword(s):  
Single Cell ◽  
Tumor Cells ◽  
Growth Regulation ◽  
Immune Escape ◽  
Tumor Development ◽  
Normal Growth ◽  
Current Evidence ◽  
Intratumor Heterogeneity ◽  
Single Cell Sequencing ◽  
Diagnosis And Prognosis

Tumorigenesis refers to the process of clonal dysplasia that occurs due to the collapse of normal growth regulation in cells caused by the action of various carcinogenic factors. These “successful” tumor cells pass on the genetic templates to their generations in evolutionary terms, but they also constantly adapt to ever-changing host environments. A unique peculiarity known as intratumor heterogeneity (ITH) is extensively involved in tumor development, metastasis, chemoresistance, and immune escape. An understanding of ITH is urgently required to identify the diversity and complexity of the tumor microenvironment (TME), but achieving this understanding has been a challenge. Single-cell sequencing (SCS) is a powerful tool that can gauge the distribution of genomic sequences in a single cell and the genetic variability among tumor cells, which can improve the understanding of ITH. SCS provides fundamental ideas about existing diversity in specific TMEs, thus improving cancer diagnosis and prognosis prediction, as well as improving the monitoring of therapeutic response. Herein, we will discuss advances in SCS and review SCS application in tumors based on current evidence.

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