Functional TCR T cell screening using single-cell droplet microfluidics
Adoptive T cell transfer, in particular TCR T cell therapy, holds great promise for cancer immunotherapy with encouraging clinical results. However, finding the right TCR T cell clone is a tedious, time-consuming, and costly process. Thus, there is a critical need for single cell technologies to con...
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| Veröffentlicht in: | Lab on a chip 2018-12, Vol.18 (24), p.3733-3749 |
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| creator | Segaliny, Aude I Li, Guideng Kong, Lingshun Ren, Ci Chen, Xiaoming Wang, Jessica K Baltimore, David Wu, Guikai Zhao, Weian |
| description | Adoptive T cell transfer, in particular TCR T cell therapy, holds great promise for cancer immunotherapy with encouraging clinical results. However, finding the right TCR T cell clone is a tedious, time-consuming, and costly process. Thus, there is a critical need for single cell technologies to conduct fast and multiplexed functional analyses followed by recovery of the clone of interest. Here, we use droplet microfluidics for functional screening and real-time monitoring of single TCR T cell activation upon recognition of target tumor cells. Notably, our platform includes a tracking system for each clone as well as a sorting procedure with 100% specificity validated by downstream single cell reverse-transcription PCR and sequencing of TCR chains. Our TCR screening prototype will facilitate immunotherapeutic screening and development of T cell therapies.
Droplet-based single cell platform allows functional screening and sorting of desirable TCR T cells to accelerate development of adoptive T cell therapies. |
| doi_str_mv | 10.1039/c8lc00818c |
| format | Article |
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Droplet-based single cell platform allows functional screening and sorting of desirable TCR T cells to accelerate development of adoptive T cell therapies.</description><identifier>ISSN: 1473-0197</identifier><identifier>EISSN: 1473-0189</identifier><identifier>DOI: 10.1039/c8lc00818c</identifier><identifier>PMID: 30397689</identifier><language>eng</language><publisher>England: Royal Society of Chemistry</publisher><subject>Antigens, Neoplasm - chemistry ; Antigens, Neoplasm - metabolism ; Cell Line, Tumor ; Droplets ; Equipment Design ; Humans ; Immunotherapy, Adoptive ; Lymphocytes ; Microfluidic Analytical Techniques - instrumentation ; Microfluidics ; Neoplasms - therapy ; Receptors, Antigen, T-Cell - analysis ; Receptors, Antigen, T-Cell - chemistry ; Receptors, Antigen, T-Cell - metabolism ; Screening ; Single-Cell Analysis - instrumentation ; Single-Cell Analysis - methods ; T-Lymphocytes - chemistry ; T-Lymphocytes - cytology ; T-Lymphocytes - metabolism ; T-Lymphocytes - transplantation ; Target recognition ; Tracking systems</subject><ispartof>Lab on a chip, 2018-12, Vol.18 (24), p.3733-3749</ispartof><rights>Copyright Royal Society of Chemistry 2018</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c506t-4477aa51bd883a46afc567771685ffb4fa1e04085cca7b80dd5395af7a4595613</citedby><cites>FETCH-LOGICAL-c506t-4477aa51bd883a46afc567771685ffb4fa1e04085cca7b80dd5395af7a4595613</cites><orcidid>0000-0002-7794-9914 ; 0000-0003-1421-4969 ; 0000-0001-8723-8190 ; 0000-0001-9670-8350</orcidid></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>230,314,776,780,881,27903,27904</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/30397689$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Segaliny, Aude I</creatorcontrib><creatorcontrib>Li, Guideng</creatorcontrib><creatorcontrib>Kong, Lingshun</creatorcontrib><creatorcontrib>Ren, Ci</creatorcontrib><creatorcontrib>Chen, Xiaoming</creatorcontrib><creatorcontrib>Wang, Jessica K</creatorcontrib><creatorcontrib>Baltimore, David</creatorcontrib><creatorcontrib>Wu, Guikai</creatorcontrib><creatorcontrib>Zhao, Weian</creatorcontrib><title>Functional TCR T cell screening using single-cell droplet microfluidics</title><title>Lab on a chip</title><addtitle>Lab Chip</addtitle><description>Adoptive T cell transfer, in particular TCR T cell therapy, holds great promise for cancer immunotherapy with encouraging clinical results. However, finding the right TCR T cell clone is a tedious, time-consuming, and costly process. Thus, there is a critical need for single cell technologies to conduct fast and multiplexed functional analyses followed by recovery of the clone of interest. Here, we use droplet microfluidics for functional screening and real-time monitoring of single TCR T cell activation upon recognition of target tumor cells. Notably, our platform includes a tracking system for each clone as well as a sorting procedure with 100% specificity validated by downstream single cell reverse-transcription PCR and sequencing of TCR chains. Our TCR screening prototype will facilitate immunotherapeutic screening and development of T cell therapies.
Droplet-based single cell platform allows functional screening and sorting of desirable TCR T cells to accelerate development of adoptive T cell therapies.</description><subject>Antigens, Neoplasm - chemistry</subject><subject>Antigens, Neoplasm - metabolism</subject><subject>Cell Line, Tumor</subject><subject>Droplets</subject><subject>Equipment Design</subject><subject>Humans</subject><subject>Immunotherapy, Adoptive</subject><subject>Lymphocytes</subject><subject>Microfluidic Analytical Techniques - instrumentation</subject><subject>Microfluidics</subject><subject>Neoplasms - therapy</subject><subject>Receptors, Antigen, T-Cell - analysis</subject><subject>Receptors, Antigen, T-Cell - chemistry</subject><subject>Receptors, Antigen, T-Cell - metabolism</subject><subject>Screening</subject><subject>Single-Cell Analysis - instrumentation</subject><subject>Single-Cell Analysis - methods</subject><subject>T-Lymphocytes - chemistry</subject><subject>T-Lymphocytes - cytology</subject><subject>T-Lymphocytes - metabolism</subject><subject>T-Lymphocytes - transplantation</subject><subject>Target recognition</subject><subject>Tracking systems</subject><issn>1473-0197</issn><issn>1473-0189</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2018</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNpdkc9LwzAcxYMoTqcX70rBiwjVZE2a5CJIcVMYCDLPIU3TmZG2M2kF_3vTbdYfl28C75MvL-8BcIbgDYIJv1XMKggZYmoPHCFMkxgixveHO6cjcOz9CkJEcMoOwSgJz2jK-BGYTbtataappY0W2Uu0iJS2NvLKaV2behl1vp_9sDreaIVr1la3UWWUa0rbmcIofwIOSmm9Pt2dY_A6fVhkj_H8efaU3c9jRWDaxhhTKiVBecFYInEqS0VSSilKGSnLHJcSaYghI0pJmjNYFCThRJZUYsJJipIxuNvuXXd5pQul69ZJK9bOVNJ9ikYa8VepzZtYNh8inVBOOA0LrnYLXPPead-Kyvj-X7LWTefFBIVwYBJ8BvTyH7pqOheS6inMYfANSaCut1RIw3uny8EMgqLvR2Rsnm36yQJ88dv-gH4XEoDzLeC8GtSfgpMvWxyU1g</recordid><startdate>20181204</startdate><enddate>20181204</enddate><creator>Segaliny, Aude I</creator><creator>Li, Guideng</creator><creator>Kong, Lingshun</creator><creator>Ren, Ci</creator><creator>Chen, Xiaoming</creator><creator>Wang, Jessica K</creator><creator>Baltimore, David</creator><creator>Wu, Guikai</creator><creator>Zhao, Weian</creator><general>Royal Society of Chemistry</general><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7SP</scope><scope>7TB</scope><scope>7U5</scope><scope>8FD</scope><scope>FR3</scope><scope>L7M</scope><scope>7X8</scope><scope>5PM</scope><orcidid>https://orcid.org/0000-0002-7794-9914</orcidid><orcidid>https://orcid.org/0000-0003-1421-4969</orcidid><orcidid>https://orcid.org/0000-0001-8723-8190</orcidid><orcidid>https://orcid.org/0000-0001-9670-8350</orcidid></search><sort><creationdate>20181204</creationdate><title>Functional TCR T cell screening using single-cell droplet microfluidics</title><author>Segaliny, Aude I ; Li, Guideng ; Kong, Lingshun ; Ren, Ci ; Chen, Xiaoming ; Wang, Jessica K ; Baltimore, David ; Wu, Guikai ; Zhao, Weian</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c506t-4477aa51bd883a46afc567771685ffb4fa1e04085cca7b80dd5395af7a4595613</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2018</creationdate><topic>Antigens, Neoplasm - chemistry</topic><topic>Antigens, Neoplasm - metabolism</topic><topic>Cell Line, Tumor</topic><topic>Droplets</topic><topic>Equipment Design</topic><topic>Humans</topic><topic>Immunotherapy, Adoptive</topic><topic>Lymphocytes</topic><topic>Microfluidic Analytical Techniques - instrumentation</topic><topic>Microfluidics</topic><topic>Neoplasms - therapy</topic><topic>Receptors, Antigen, T-Cell - analysis</topic><topic>Receptors, Antigen, T-Cell - chemistry</topic><topic>Receptors, Antigen, T-Cell - metabolism</topic><topic>Screening</topic><topic>Single-Cell Analysis - instrumentation</topic><topic>Single-Cell Analysis - methods</topic><topic>T-Lymphocytes - chemistry</topic><topic>T-Lymphocytes - cytology</topic><topic>T-Lymphocytes - metabolism</topic><topic>T-Lymphocytes - transplantation</topic><topic>Target recognition</topic><topic>Tracking systems</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Segaliny, Aude I</creatorcontrib><creatorcontrib>Li, Guideng</creatorcontrib><creatorcontrib>Kong, Lingshun</creatorcontrib><creatorcontrib>Ren, Ci</creatorcontrib><creatorcontrib>Chen, Xiaoming</creatorcontrib><creatorcontrib>Wang, Jessica K</creatorcontrib><creatorcontrib>Baltimore, David</creatorcontrib><creatorcontrib>Wu, Guikai</creatorcontrib><creatorcontrib>Zhao, Weian</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Electronics & Communications Abstracts</collection><collection>Mechanical & Transportation Engineering Abstracts</collection><collection>Solid State and Superconductivity Abstracts</collection><collection>Technology Research Database</collection><collection>Engineering Research Database</collection><collection>Advanced Technologies Database with Aerospace</collection><collection>MEDLINE - Academic</collection><collection>PubMed Central (Full Participant titles)</collection><jtitle>Lab on a chip</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Segaliny, Aude I</au><au>Li, Guideng</au><au>Kong, Lingshun</au><au>Ren, Ci</au><au>Chen, Xiaoming</au><au>Wang, Jessica K</au><au>Baltimore, David</au><au>Wu, Guikai</au><au>Zhao, Weian</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Functional TCR T cell screening using single-cell droplet microfluidics</atitle><jtitle>Lab on a chip</jtitle><addtitle>Lab Chip</addtitle><date>2018-12-04</date><risdate>2018</risdate><volume>18</volume><issue>24</issue><spage>3733</spage><epage>3749</epage><pages>3733-3749</pages><issn>1473-0197</issn><eissn>1473-0189</eissn><abstract>Adoptive T cell transfer, in particular TCR T cell therapy, holds great promise for cancer immunotherapy with encouraging clinical results. However, finding the right TCR T cell clone is a tedious, time-consuming, and costly process. Thus, there is a critical need for single cell technologies to conduct fast and multiplexed functional analyses followed by recovery of the clone of interest. Here, we use droplet microfluidics for functional screening and real-time monitoring of single TCR T cell activation upon recognition of target tumor cells. Notably, our platform includes a tracking system for each clone as well as a sorting procedure with 100% specificity validated by downstream single cell reverse-transcription PCR and sequencing of TCR chains. Our TCR screening prototype will facilitate immunotherapeutic screening and development of T cell therapies.
Droplet-based single cell platform allows functional screening and sorting of desirable TCR T cells to accelerate development of adoptive T cell therapies.</abstract><cop>England</cop><pub>Royal Society of Chemistry</pub><pmid>30397689</pmid><doi>10.1039/c8lc00818c</doi><tpages>17</tpages><orcidid>https://orcid.org/0000-0002-7794-9914</orcidid><orcidid>https://orcid.org/0000-0003-1421-4969</orcidid><orcidid>https://orcid.org/0000-0001-8723-8190</orcidid><orcidid>https://orcid.org/0000-0001-9670-8350</orcidid><oa>free_for_read</oa></addata></record> |
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| source | MEDLINE; Royal Society Of Chemistry Journals 2008-; Alma/SFX Local Collection |
| subjects | Antigens, Neoplasm - chemistry Antigens, Neoplasm - metabolism Cell Line, Tumor Droplets Equipment Design Humans Immunotherapy, Adoptive Lymphocytes Microfluidic Analytical Techniques - instrumentation Microfluidics Neoplasms - therapy Receptors, Antigen, T-Cell - analysis Receptors, Antigen, T-Cell - chemistry Receptors, Antigen, T-Cell - metabolism Screening Single-Cell Analysis - instrumentation Single-Cell Analysis - methods T-Lymphocytes - chemistry T-Lymphocytes - cytology T-Lymphocytes - metabolism T-Lymphocytes - transplantation Target recognition Tracking systems |
| title | Functional TCR T cell screening using single-cell droplet microfluidics |
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