Profiling protein-protein interactions of single cancer cells with in situ lysis and co-immunoprecipitation

Ryu, Ji Young1,2,7; Kim, Jihye3; Shon, Min Ju1,2; Sun, Jiashu4; Jiang, Xingyu5; Lee, Wonhee3,6; Yoon, Tae-Young1,2

2019-06-07

10.1039/c9lc00139e

LAB ON A CHIP   影响因子和分区

1473-0197

1473-0189

Heterogeneity in a tumor allows a small portion of cancer cells to survive and regrow upon targeted cancer therapy, eventually leading to cancer relapse. Such drug-resistant cells often exhibit dynamic adaptation of their signaling pathways at the level of protein- protein interactions (PPIs). To probe the rewiring of signaling pathways and the heterogeneity across individual cancer cells, we developed a single-cell version of the co-immunoprecipitation (co-IP) analysis that examines the amount and PPIs of target proteins immunoprecipitated from individual cells. The method captures cancer cells at predefined locations using a microfluidic chip, pulls down target proteins on the surface using antibodies, and lyses the captured cells in situ. Then, subsequent addition of eGFP-labeled downstream proteins enables the determination of the corresponding PPIs for the minimal amount of target proteins sampled from a single cell. We applied the technique to probe epidermal growth factor receptors (EGFRs) in PC9 lung adenocarcinoma cells. The results reveal that the strength of EGFR PPIs can be largely uncorrelated with the expression level of EGFRs in single cells. In addition, the individual PC9 cells showed markedly different patterns of PPIs, indicating a high heterogeneity in EGFR signaling within a genetically homogeneous population.

SCI ; EI

英语

其他

Basic Research Program in Science and Engineering[NRF-2018R1A2B6003793]

Biochemistry & Molecular Biology ; Chemistry ; Science & Technology - Other Topics

Biochemical Research Methods ; Chemistry, Multidisciplinary ; Chemistry, Analytical ; Nanoscience & Nanotechnology

WOS:000472924500002

ROYAL SOC CHEMISTRY

20201708538460

Antibodies ; Diseases ; Signaling ; Cell signaling

Biomedical Engineering:461.1 ; Medicine and Pharmacology:461.6 ; Immunology:461.9.1

CHEMISTRY

Web of Science

1.Seoul Natl Univ, Sch Biol Sci, Seoul 08826, South Korea
2.Seoul Natl Univ, Inst Mol Biol & Genet, Seoul 08826, South Korea
3.Korea Adv Inst Sci & Technol, Grad Sch Nanosci & Technol, Daejeon 34141, South Korea
4.Natl Ctr Nanosci & Technol, CAS Ctr Excellence Nanosci, CAS Key Lab Standardizat & Measurement Nanotechno, Beijing 100190, Peoples R China
5.Southern Univ Sci & Technol, Dept Biomed Engn, 1088 Xueyuan Rd, Shenzhen, Guangdong, Peoples R China
6.Korea Adv Inst Sci & Technol, Dept Phys, Daejeon 34141, South Korea
7.Proteina Inc, R&D Ctr, Seoul 08826, South Korea

Ryu, Ji Young,Kim, Jihye,Shon, Min Ju,et al. Profiling protein-protein interactions of single cancer cells with in situ lysis and co-immunoprecipitation[J]. LAB ON A CHIP,2019,19(11):1922-1928.

Ryu, Ji Young.,Kim, Jihye.,Shon, Min Ju.,Sun, Jiashu.,Jiang, Xingyu.,...&Yoon, Tae-Young.(2019).Profiling protein-protein interactions of single cancer cells with in situ lysis and co-immunoprecipitation.LAB ON A CHIP,19(11),1922-1928.

Ryu, Ji Young,et al."Profiling protein-protein interactions of single cancer cells with in situ lysis and co-immunoprecipitation".LAB ON A CHIP 19.11(2019):1922-1928.