Very rapid cloning, expression and identifying specificity of T-cell receptors for T-cell engineering

Shan Zong; Tiejuan Mi; Leo G. Flores; Amir Alpert; Simon Olivares; Krina Patel; Sourindra Maiti; George McNamara; Laurence J.N. Cooper; Hiroki Torikai

doi:10.1371/journal.pone.0228112

Very rapid cloning, expression and identifying specificity of T-cell receptors for T-cell engineering

Shan Zong, Tiejuan Mi, Leo G. Flores, Amir Alpert, Simon Olivares, Krina Patel, Sourindra Maiti, George McNamara, Laurence J.N. Cooper, Hiroki Torikai

Research output: Contribution to journal › Article › peer-review

7 Scopus citations

Abstract

Neoantigens can be predicted and in some cases identified using the data obtained from the whole exome sequencing and transcriptome sequencing of tumor cells. These sequencing data can be coupled with single-cell RNA sequencing for the direct interrogation of the transcriptome, surfaceome, and pairing of αβ T-cell receptors (TCRαβ) from hundreds of single T cells. Using these 2 large datasets, we established a platform for identifying antigens recognized by TCRαβs obtained from single T cells. Our approach is based on the rapid expression of cloned TCRαβ genes as Sleeping Beauty transposons and the determination of the introduced TCRαβs’ antigen specificity and avidity using a reporter cell line. The platform enables the very rapid identification of tumor-reactive TCRs for the bioengineering of T cells with redirected specificity.

Original language	English (US)
Article number	e0228112
Journal	PloS one
Volume	15
Issue number	2
DOIs	https://doi.org/10.1371/journal.pone.0228112
State	Published - Feb 1 2020

ASJC Scopus subject areas

General Biochemistry, Genetics and Molecular Biology
General Agricultural and Biological Sciences
General

MD Anderson CCSG core facilities

Flow Cytometry and Cellular Imaging Facility
Advanced Technology Genomics Core
Cytogenetics and Cell Authentication Core
Research Animal Support Facility
Tissue Biospecimen and Pathology Resource

Access to Document

10.1371/journal.pone.0228112

Cite this

@article{94b8104587ff4e1c8430cca3d6b08477,

title = "Very rapid cloning, expression and identifying specificity of T-cell receptors for T-cell engineering",

abstract = "Neoantigens can be predicted and in some cases identified using the data obtained from the whole exome sequencing and transcriptome sequencing of tumor cells. These sequencing data can be coupled with single-cell RNA sequencing for the direct interrogation of the transcriptome, surfaceome, and pairing of αβ T-cell receptors (TCRαβ) from hundreds of single T cells. Using these 2 large datasets, we established a platform for identifying antigens recognized by TCRαβs obtained from single T cells. Our approach is based on the rapid expression of cloned TCRαβ genes as Sleeping Beauty transposons and the determination of the introduced TCRαβs{\textquoteright} antigen specificity and avidity using a reporter cell line. The platform enables the very rapid identification of tumor-reactive TCRs for the bioengineering of T cells with redirected specificity.",

author = "Shan Zong and Tiejuan Mi and Flores, {Leo G.} and Amir Alpert and Simon Olivares and Krina Patel and Sourindra Maiti and George McNamara and Cooper, {Laurence J.N.} and Hiroki Torikai",

note = "Funding Information: Pediatric Research Funding ? MD Anderson Cancer Center. This is the department fund from the institution. This study used MD Anderson?s Flow Cytometry and Cellular Imaging Core, which is supported by the National Institutes of Health through MD Anderson?s Cancer Center Support Grant (P30 CA016672). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. We thank the personnel of MD Anderson Cancer Center? Flow Cytometry and Cellular Imaging Core for performing cell sorting. High-affinity NY-ESO-1/HLA-A2?specific TCR was provided by Dr. Steven Rosenberg (NCI, Bethesda, MD). Publisher Copyright: {\textcopyright} 2020 Zong et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.",

year = "2020",

month = feb,

day = "1",

doi = "10.1371/journal.pone.0228112",

language = "English (US)",

volume = "15",

journal = "PloS one",

issn = "1932-6203",

publisher = "Public Library of Science",

number = "2",

}

TY - JOUR

T1 - Very rapid cloning, expression and identifying specificity of T-cell receptors for T-cell engineering

AU - Zong, Shan

AU - Mi, Tiejuan

AU - Flores, Leo G.

AU - Alpert, Amir

AU - Olivares, Simon

AU - Patel, Krina

AU - Maiti, Sourindra

AU - McNamara, George

AU - Cooper, Laurence J.N.

AU - Torikai, Hiroki

N1 - Funding Information: Pediatric Research Funding ? MD Anderson Cancer Center. This is the department fund from the institution. This study used MD Anderson?s Flow Cytometry and Cellular Imaging Core, which is supported by the National Institutes of Health through MD Anderson?s Cancer Center Support Grant (P30 CA016672). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. We thank the personnel of MD Anderson Cancer Center? Flow Cytometry and Cellular Imaging Core for performing cell sorting. High-affinity NY-ESO-1/HLA-A2?specific TCR was provided by Dr. Steven Rosenberg (NCI, Bethesda, MD). Publisher Copyright: © 2020 Zong et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

PY - 2020/2/1

Y1 - 2020/2/1

N2 - Neoantigens can be predicted and in some cases identified using the data obtained from the whole exome sequencing and transcriptome sequencing of tumor cells. These sequencing data can be coupled with single-cell RNA sequencing for the direct interrogation of the transcriptome, surfaceome, and pairing of αβ T-cell receptors (TCRαβ) from hundreds of single T cells. Using these 2 large datasets, we established a platform for identifying antigens recognized by TCRαβs obtained from single T cells. Our approach is based on the rapid expression of cloned TCRαβ genes as Sleeping Beauty transposons and the determination of the introduced TCRαβs’ antigen specificity and avidity using a reporter cell line. The platform enables the very rapid identification of tumor-reactive TCRs for the bioengineering of T cells with redirected specificity.

AB - Neoantigens can be predicted and in some cases identified using the data obtained from the whole exome sequencing and transcriptome sequencing of tumor cells. These sequencing data can be coupled with single-cell RNA sequencing for the direct interrogation of the transcriptome, surfaceome, and pairing of αβ T-cell receptors (TCRαβ) from hundreds of single T cells. Using these 2 large datasets, we established a platform for identifying antigens recognized by TCRαβs obtained from single T cells. Our approach is based on the rapid expression of cloned TCRαβ genes as Sleeping Beauty transposons and the determination of the introduced TCRαβs’ antigen specificity and avidity using a reporter cell line. The platform enables the very rapid identification of tumor-reactive TCRs for the bioengineering of T cells with redirected specificity.

UR - http://www.scopus.com/inward/record.url?scp=85079216817&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85079216817&partnerID=8YFLogxK

U2 - 10.1371/journal.pone.0228112

DO - 10.1371/journal.pone.0228112

M3 - Article

C2 - 32040512

AN - SCOPUS:85079216817

SN - 1932-6203

VL - 15

JO - PloS one

JF - PloS one

IS - 2

M1 - e0228112

ER -

Very rapid cloning, expression and identifying specificity of T-cell receptors for T-cell engineering

Abstract

ASJC Scopus subject areas

MD Anderson CCSG core facilities

Access to Document

Other files and links

Fingerprint

Cite this