FRINK Linked Data Fragments server

Matches in SemOpenAlex for { <https://semopenalex.org/work/W2954624125> ?p ?o ?g. }

• W2954624125 endingPage "326" @default.
• W2954624125 startingPage "LB" @default.
• W2954624125 abstract "Abstract Background Understanding the genetic heterogeneity of circulating immune and tumors cells at single-cell resolution will enable one to decipher the interplay between cancer and immune cells, and ultimately lead to better design of immunotherapy assays. Marker-free methods for isolation of cells are attractive because they provide an opportunity to analyze a larger set of these cells that may otherwise be missed due to variable or no expression of protein (marker) markers. Integration of the Biolidics ClearCell® FX System and the Fluidigm® Polaris™ system enabled us not only to develop a marker-free workflow to isolate immune and circulating tumor cells (CTCs) but also to seamlessly prepare amplified cDNA from immune and CTCs for full-length mRNA-seq analysis. Method and Results The ClearCell FX System processes blood samples and isolates CTCs in a marker-free manner. During the CTC isolation process the immune cells are separated from CTCs on the microfluidic device. To differentiate larger blood cells from CTCs, we stained the enriched cells with Alexa Fluor® 647-conjugated CD45 and CD31 to identify leukocytes and endothelial cells, respectively. Calcein AM (live cell marker) and CellTracker™ Orange (universal cell marker) were added to identify live cells. Different enrichment strategies on Polaris allowed us to either specifically select for CTCs (live+, universal cell marker+, CD45-, CD31-) or immune cells (live+, universal cell marker+, CD45+). The enriched cells were then lysed, mRNA were reverse-transcribed, and cDNA were preamplified on a Polaris integrated fluidic circuit (IFC). Sequencing libraries were generated (off-IFC) using the Nextera® XT DNA library prep protocol and sequenced on Illumina® MiSeq™ systems. We successfully profiled full-length mRNA of 81 CTCs from stage III and IV subjects. Based on hormone receptor status, these six subjects were categorized as three types: (1) ER-/PR-/HER2- (triple-negative breast cancer; n=1), (2) ER+/PR+/HER2- (n=3), and (3) ER+/PR+/HER2- (n=2). For immune cell comparative analysis, we included CD45+ (FACS-sorted) from healthy donor. Unsupervised hierarchal clustering of gene expression data showed clustering by subject with a unique set of genes expressed by triple-negative breast cancer only. Heterogeneity was noted in immune cells from subjects at baseline (pretreatment), under treatment with drugs and a healthy subject without cancer. Using TraCeR computational method, we successfully reconstructed full-length, paired T cell receptor (TCR) sequences from CD45+ single-cell RNA sequence data from breast cancer subjects. Conclusion We present the feasibility of integrating two microfluidics platforms to isolate circulating immune and tumor cells for transcriptome and functional study. Our data suggests that the heterogeneity of immune and cancer cells can be elucidated from single-cell mRNA sequencing data and full-length, paired T cell receptor sequences can be reconstructed from immune cells of breast cancer subjects. Citation Format: Naveen Ramalingam, Yifang Lee, Lukasz Szpankowski, Anne Leyrat, Brian Fowler, Ninez Delos Angeles, Chad Sanada, Andrew Wu, Yoon Sim Yap, Jay West, Ali Asgar Bhagat, Kyle Hukari, Mark Lynch, David King. Full-Length mRNA transcriptome analysis of matched circulating tumor and immune cells from breast cancer subjects [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr LB-326." @default.
• W2954624125 created "2019-07-12" @default.
• W2954624125 creator A5011621241 @default.
• W2954624125 creator A5021015559 @default.
• W2954624125 creator A5022430976 @default.
• W2954624125 creator A5024057870 @default.
• W2954624125 creator A5036726954 @default.
• W2954624125 creator A5041015803 @default.
• W2954624125 creator A5054147614 @default.
• W2954624125 creator A5072578890 @default.
• W2954624125 creator A5077316309 @default.
• W2954624125 creator A5077950247 @default.
• W2954624125 creator A5079091432 @default.
• W2954624125 creator A5079386914 @default.
• W2954624125 creator A5086617305 @default.
• W2954624125 creator A5090030583 @default.
• W2954624125 date "2019-07-01" @default.
• W2954624125 modified "2023-09-27" @default.
• W2954624125 title "Abstract LB-326: Full-Length mRNA transcriptome analysis of matched circulating tumor and immune cells from breast cancer subjects" @default.
• W2954624125 doi "https://doi.org/10.1158/1538-7445.am2019-lb-326" @default.
• W2954624125 hasPublicationYear "2019" @default.
• W2954624125 type Work @default.
• W2954624125 sameAs 2954624125 @default.
• W2954624125 citedByCount "1" @default.
• W2954624125 countsByYear W29546241252022 @default.
• W2954624125 crossrefType "journal-article" @default.
• W2954624125 hasAuthorship W2954624125A5011621241 @default.
• W2954624125 hasAuthorship W2954624125A5021015559 @default.
• W2954624125 hasAuthorship W2954624125A5022430976 @default.
• W2954624125 hasAuthorship W2954624125A5024057870 @default.
• W2954624125 hasAuthorship W2954624125A5036726954 @default.
• W2954624125 hasAuthorship W2954624125A5041015803 @default.
• W2954624125 hasAuthorship W2954624125A5054147614 @default.
• W2954624125 hasAuthorship W2954624125A5072578890 @default.
• W2954624125 hasAuthorship W2954624125A5077316309 @default.
• W2954624125 hasAuthorship W2954624125A5077950247 @default.
• W2954624125 hasAuthorship W2954624125A5079091432 @default.
• W2954624125 hasAuthorship W2954624125A5079386914 @default.
• W2954624125 hasAuthorship W2954624125A5086617305 @default.
• W2954624125 hasAuthorship W2954624125A5090030583 @default.
• W2954624125 hasBestOaLocation W29546241251 @default.
• W2954624125 hasConcept C121608353 @default.
• W2954624125 hasConcept C153911025 @default.
• W2954624125 hasConcept C203014093 @default.
• W2954624125 hasConcept C2779013556 @default.
• W2954624125 hasConcept C502942594 @default.
• W2954624125 hasConcept C54355233 @default.
• W2954624125 hasConcept C61238886 @default.
• W2954624125 hasConcept C86803240 @default.
• W2954624125 hasConcept C8891405 @default.
• W2954624125 hasConceptScore W2954624125C121608353 @default.
• W2954624125 hasConceptScore W2954624125C153911025 @default.
• W2954624125 hasConceptScore W2954624125C203014093 @default.
• W2954624125 hasConceptScore W2954624125C2779013556 @default.
• W2954624125 hasConceptScore W2954624125C502942594 @default.
• W2954624125 hasConceptScore W2954624125C54355233 @default.
• W2954624125 hasConceptScore W2954624125C61238886 @default.
• W2954624125 hasConceptScore W2954624125C86803240 @default.
• W2954624125 hasConceptScore W2954624125C8891405 @default.
• W2954624125 hasIssue "13_Supplement" @default.
• W2954624125 hasLocation W29546241251 @default.
• W2954624125 hasOpenAccess W2954624125 @default.
• W2954624125 hasPrimaryLocation W29546241251 @default.
• W2954624125 hasRelatedWork W1988618052 @default.
• W2954624125 hasRelatedWork W2121022075 @default.
• W2954624125 hasRelatedWork W2394991672 @default.
• W2954624125 hasRelatedWork W2547401879 @default.
• W2954624125 hasRelatedWork W2582875230 @default.
• W2954624125 hasRelatedWork W4213166086 @default.
• W2954624125 hasRelatedWork W4283731173 @default.
• W2954624125 hasRelatedWork W4291367166 @default.
• W2954624125 hasRelatedWork W4308359949 @default.
• W2954624125 hasRelatedWork W4360983034 @default.
• W2954624125 hasVolume "79" @default.
• W2954624125 isParatext "false" @default.
• W2954624125 isRetracted "false" @default.
• W2954624125 magId "2954624125" @default.
• W2954624125 workType "article" @default.