Abstract 16226: Single-cell RNA Sequencing Coupled With Optical Imaging for Targeted Real-time Visualization of the Cardiac Conduction System

Circulation ◽  
2020 ◽  
Vol 142 (Suppl_3) ◽  
Author(s):  
William R Goodyer ◽  
Benjamin Beyersdorf ◽  
Nynke Van Den Berg ◽  
Nazan Puluca ◽  
Jan Buikema ◽  
...  
Keyword(s):  
Cell Surface ◽  
Optical Imaging ◽  
Real Time ◽  
Cardiac Conduction ◽  
Specific Cell ◽  
Surface Markers ◽  
Cell Surface Markers ◽  
Real Time Visualization ◽  
Specific Cell Surface

Introduction: Optical imaging has the potential to revolutionize cardiothoracic surgery by allowing for the real-time visualization of structures often inadvertently damaged due to inadequate visibility. The cardiac conduction system (CCS) consists of specialized cells embedded within the heart that are essential for cardiac function yet indistinguishable from heart muscle tissue. Intraoperative CCS injury is a major complication in cardiac surgery, representing a significant source of morbidity and mortality. To date, there exists no intraoperative method to visualize the CCS. Hypothesis: We hypothesized that unique, CCS-specific cell surface markers could be used for the in vivo labelling of the CCS. Objectives: Use single-cell RNA sequencing (scRNAseq) to discover cell surface markers that may serve as the basis for generating optical imaging agents for real-time CCS visualization. Methods/Results: Gene expression analysis of a comprehensive scRNAseq dataset of the entire murine CCS revealed significant enrichment of a host of CCS-specific cell surface genes. A subset of genes were subsequently validated in the CCS of mice and/or human tissue. In total, 7 novel cell surface markers were confirmed to have unique expression patterns throughout or within distinct components of the CCS. Next, optical imaging agents were created consisting of a near-infrared (NIR) dye conjugated to antibodies directed against two distinct CCS-specific cell surface markers. Each optical imaging agent demonstrated high sensitivity and specificity in labeling the entire CCS in vivo following a single intravenous injection in mice. Specificity was confirmed within intact, whole hearts using both closed-field NIR imaging and whole mount immunolabeling with volume imaging (iDISCO+). Dosage, timecourse and biodistribution analyses were performed as well as safety validation by surface ECG. Conclusions: In summary, we coupled scRNAseq with optical imaging to create novel tools for the real-time visualization of a complex tissue substructure. We provide a proof-of-principle for broadening the scope of optical imaging but also address a significant unmet clinical need, laying the foundation for translational opportunities in cardiac intervention and imaging.

Towards an indirect screening technique facilitating detection of cellular populations bearing specific cell surface markers

2010 ◽  
Vol 26 (6) ◽  
pp. 1544-1550
Author(s):  
Sandeep K. Arora ◽  
Rohit Sharma ◽  
Gagandeep Kaur ◽  
Preeti Bhoria ◽  
Maryada Sharma ◽  
...  
Keyword(s):  
Cell Surface ◽  
Specific Cell ◽  
Surface Markers ◽  
Cell Surface Markers ◽  
Screening Technique ◽  
Specific Cell Surface

scholarly journals Identification of Specific Cell-Surface Markers of Adipose-Derived Stem Cells from Subcutaneous and Visceral Fat Depots

2014 ◽  
Vol 2 (2) ◽  
pp. 171-179 ◽  
Author(s):  
Wee Kiat Ong ◽  
Chuen Seng Tan ◽  
Kai Li Chan ◽  
Grace Gandi Goesantoso ◽  
Xin Hui Derryn Chan ◽  
...  
Keyword(s):  
Stem Cells ◽  
Cell Surface ◽  
Visceral Fat ◽  
Specific Cell ◽  
Surface Markers ◽  
Adipose Derived Stem Cells ◽  
Cell Surface Markers ◽  
Fat Depots ◽  
Specific Cell Surface

P5369Alpha-V integrin regulates the contribution of PW1+ cells to cardiac fibrosis

2019 ◽  
Vol 40 (Supplement_1) ◽  
Author(s):  
M Bouvet ◽  
O Claude ◽  
M Roux ◽  
N Mougenot ◽  
V Duval ◽  
...  
Keyword(s):  
Cell Surface ◽  
Cardiac Fibrosis ◽  
Cardiac Fibroblasts ◽  
Surface Proteins ◽  
Specific Cell ◽  
Surface Markers ◽  
Cell Surface Markers ◽  
Proteomic Profiling ◽  
The Matrix ◽  
Almost All

Abstract Background Activated cardiac fibroblasts produce extracellular matrix proteins that accumulate during cardiac fibrosis. We have recently shown that PW1 is expressed in a subset of cardiac stromal cells and that cardiac PW1+ cells represent a cellular source of fibroblasts in the ischemic hearts. Purpose We aimed to further identify new cell surface markers expressed by cardiac PW1+ cells and to investigate their role in the fibrogenic behavior of these cells. Methods and results We first performed transcriptomic and proteomic profiling of FACS-isolated cardiac PW1+ from normal and ischemic hearts. RNA-sequencing output files were processed with bioinformatics algorithms to identify 378 specific cell-surface markers for cardiac PW1+ cells. By comparing these candidates with the proteomic profile, we then cross-identified 9 cell surface proteins primarily involved in cell motility, adhesion to the matrix, inflammatory response and response to wounding. One of these candidates (i.e., aV-integrin or CD51) was expressed in almost all cardiac PW1+ cells (93±1%), and was predominantly found in cells expressing PW1 in the myocardium. Cardiac PW1+ cells showed a predominant expression of aVβ1 complex which is known to mediate fibrosis through TGF-beta activation in a number of tissues. The transfer of isolated cardiac PW1+CD51+ cells into ischemic hearts was associated with fibrosis development. We further demonstrated that inhibition of aV-integrin in cardiac PW1+ cells reduces their profibrotic gene expression profile and their ability to differentiate into fibroblasts. Lastly, a pharmacological blockade of aV-integrin improved cardiac function and animal survival following myocardial infarction coupled with a reduced infarct size and fibrotic lesion. Conclusions These data identify a targetable pathway that regulates cardiac fibrosis in response to an ischemic injury and demonstrate that pharmacological inhibition of aV-integrin leads to reduced pathological outcomes following cardiac ischemia. Acknowledgement/Funding Fondation Leducq (grant 13CVD01, CardioStemNet project), Fédération Française de Cardiologie and Era-CVD (ANR-16-ECVD-0011-03, Clarify project)

scholarly journals Single-cell RNA-Sequencing uncovers transcriptional states and fate decisions in haematopoiesis

2017 ◽  
Author(s):  
Emmanouil I. Athanasiadis ◽  
Jan G. Botthof ◽  
Helena Andres ◽  
Lauren Ferreira ◽  
Pietro Lio ◽  
...  
Keyword(s):  
Cell Surface ◽  
Specific Cell ◽  
Surface Markers ◽  
Evolutionary Analysis ◽  
Cell Surface Markers ◽  
Lineage Differentiation ◽  
Lineage Tree ◽  
Stem And Progenitor Cells ◽  
A Cell ◽  
Fate Decision

ABSTRACTThe success of marker-based approaches for dissecting haematopoiesis in mouse and human is reliant on the presence of well-defined cell-surface markers specific for diverse progenitor populations. An inherent problem with this approach is that the presence of specific cell surface markers does not directly reflect the transcriptional state of a cell. Here we used a marker-free approach to computationally reconstruct the blood lineage tree in zebrafish and order cells along their differentiation trajectory, based on their global transcriptional differences. Within the population of transcriptionally similar stem and progenitor cells our analysis revealed considerable cell-to-cell differences in their probability to transition to another, committed state. Once fate decision was executed, the suppression of transcription of ribosomal genes and up-regulation of lineage specific factors coordinately controlled lineage differentiation. Evolutionary analysis further demonstrated that this haematopoietic program was highly conserved between zebrafish and higher vertebrates.

scholarly journals Systemic surfaceome profiling identifies target antigens for immune-based therapy in subtypes of advanced prostate cancer

2018 ◽  
Vol 115 (19) ◽  
pp. E4473-E4482 ◽  
Author(s):  
John K. Lee ◽  
Nathanael J. Bangayan ◽  
Timothy Chai ◽  
Bryan A. Smith ◽  
Tiffany E. Pariva ◽  
...  
Keyword(s):  
Prostate Cancer ◽  
Cell Surface ◽  
Cell Lines ◽  
Treatment Resistance ◽  
Specific Cell ◽  
Surface Markers ◽  
Mrna Levels ◽  
Cell Surface Markers ◽  
Cell Surface Antigens ◽  
Cancer Subtypes

Prostate cancer is a heterogeneous disease composed of divergent molecular and histologic subtypes, including prostate adenocarcinoma (PrAd) and neuroendocrine prostate cancer (NEPC). While PrAd is the major histology in prostate cancer, NEPC can evolve from PrAd as a mechanism of treatment resistance that involves a transition from an epithelial to a neurosecretory cancer phenotype. Cell surface markers are often associated with specific cell lineages and differentiation states in normal development and cancer. Here, we show that PrAd and NEPC can be broadly discriminated by cell-surface profiles based on the analysis of prostate cancer gene expression datasets. To overcome a dependence on predictions of human cell-surface genes and an assumed correlation between mRNA levels and protein expression, we integrated transcriptomic and cell-surface proteomic data generated from a panel of prostate cancer cell lines to nominate cell-surface markers associated with these cancer subtypes. FXYD3 and CEACAM5 were validated as cell-surface antigens enriched in PrAd and NEPC, respectively. Given the lack of effective treatments for NEPC, CEACAM5 appeared to be a promising target for cell-based immunotherapy. As a proof of concept, engineered chimeric antigen receptor T cells targeting CEACAM5 induced antigen-specific cytotoxicity in NEPC cell lines. Our findings demonstrate that the surfaceomes of PrAd and NEPC reflect unique cancer differentiation states and broadly represent vulnerabilities amenable to therapeutic targeting.

scholarly journals ASC-1, PAT2, and P2RX5 are cell surface markers for white, beige, and brown adipocytes

2014 ◽  
Vol 6 (247) ◽  
pp. 247ra103-247ra103 ◽  
Author(s):  
Siegfried Ussar ◽  
Kevin Y. Lee ◽  
Simon N. Dankel ◽  
Jeremie Boucher ◽  
Max-Felix Haering ◽  
...  
Keyword(s):  
Amino Acid ◽  
Cell Surface ◽  
Cell Types ◽  
Amino Acid Transporter ◽  
Specific Cell ◽  
Surface Markers ◽  
Cell Surface Markers ◽  
Brown Adipocytes ◽  
Beige Adipocytes ◽  
Acid Transporter

White, beige, and brown adipocytes are developmentally and functionally distinct but often occur mixed together within individual depots. To target white, beige, and brown adipocytes for diagnostic or therapeutic purposes, a better understanding of the cell surface properties of these cell types is essential. Using a combination of in silico, in vitro, and in vivo methods, we have identified three new cell surface markers of adipose cell types. The amino acid transporter ASC-1 is a white adipocyte–specific cell surface protein, with little or no expression in brown adipocytes, whereas the amino acid transporter PAT2 and the purinergic receptor P2RX5 are cell surface markers expressed in classical brown and beige adipocytes in mice. These markers also selectively mark brown/beige and white adipocytes in human tissue. Thus, ASC-1, PAT2, and P2RX5 are membrane surface proteins that may serve as tools to identify and target white and brown/beige adipocytes for therapeutic purposes.

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