Coupling Lipid Labeling and Click Chemistry Enables Isolation of Extracellular Vesicles for Noninvasive Detection of Oncogenic Gene Alterations

Na Sun; Benjamin V Tran; Zishan Peng; Jing Wang; Ceng Zhang; Peng Yang; Tiffany X Zhang; Josephine Widjaja; Ryan Y Zhang; Wenxi Xia; Alexandra Keir; Jia-Wei She; Hsiao-Hua Yu; Jing-Jong Shyue; Hongguang Zhu; Vatche G Agopian; Renjun Pei; James S Tomlinson; Jeffrey A Toretsky; Steven J Jonas; Noah Federman; Shaohua Lu; Hsian-Rong Tseng; Yazhen Zhu

doi:10.1002/advs.202105853

Coupling Lipid Labeling and Click Chemistry Enables Isolation of Extracellular Vesicles for Noninvasive Detection of Oncogenic Gene Alterations

Adv Sci (Weinh). 2022 May;9(14):e2105853. doi: 10.1002/advs.202105853. Epub 2022 Mar 9.

Authors

Na Sun^{1

2}, Benjamin V Tran³, Zishan Peng⁴, Jing Wang⁵, Ceng Zhang¹, Peng Yang¹, Tiffany X Zhang¹, Josephine Widjaja¹, Ryan Y Zhang¹, Wenxi Xia¹, Alexandra Keir⁶, Jia-Wei She⁷, Hsiao-Hua Yu⁷, Jing-Jong Shyue⁸, Hongguang Zhu⁵, Vatche G Agopian³, Renjun Pei², James S Tomlinson³, Jeffrey A Toretsky⁹, Steven J Jonas^{6

10}, Noah Federman⁶, Shaohua Lu⁴, Hsian-Rong Tseng¹, Yazhen Zhu¹

Affiliations

¹ California NanoSystems Institute, Crump Institute for Molecular Imaging, Department of Molecular and Medical Pharmacology, University of California, Los Angeles, Los Angeles, CA, 90095, USA.
² Key Laboratory for Nano-Bio Interface, Suzhou Institute of Nano-Tech and Nano-Bionics, University of Chinese Academy of Sciences, Chinese Academy of Sciences, Suzhou, 215123, P. R. China.
³ Department of Surgery, University of California, Los Angeles, Los Angeles, CA, 90095, USA.
⁴ Department of Pathology, Zhongshan Hospital, Fudan University, Shanghai, 200032, P. R. China.
⁵ Department of Pathology, Shanghai Medical College, Fudan University, Shanghai, 200032, P. R. China.
⁶ Department of Pediatrics, David Geffen School of Medicine, Eli and Edythe Broad Center of Regenerative Medicine and Stem Cell Research, and Children's Discovery and Innovation Institute, University of California, Los Angeles, Los Angeles, CA, 90095, USA.
⁷ Smart Organic Materials Laboratory, Institute of Chemistry, Academia Sinica, Nankang, Taipei, 115, Taiwan.
⁸ Research Center for Applied Sciences, Academia Sinica, Nankang, Taipei, 115, Taiwan.
⁹ Departments of Oncology and Pediatrics, Georgetown University, Washington, DC, 20057, USA.
¹⁰ California NanoSystems Institute, Departments of Chemistry and Biochemistry and of Materials Science and Engineering, University of California, Los Angeles, Los Angeles, CA, 90095, USA.

Abstract

Well-preserved molecular cargo in circulating extracellular vesicles (EVs) offers an ideal material for detecting oncogenic gene alterations in cancer patients, providing a noninvasive diagnostic solution for detection of disease status and monitoring treatment response. Therefore, technologies that conveniently isolate EVs with sufficient efficiency are desperately needed. Here, a lipid labeling and click chemistry-based EV capture platform ("Click Beads"), which is ideal for EV message ribonucleic acid (mRNA) assays due to its efficient, convenient, and rapid purification of EVs, enabling downstream molecular quantification using reverse transcription digital polymerase chain reaction (RT-dPCR) is described and demonstrated. Ewing sarcoma protein (EWS) gene rearrangements and kirsten rat sarcoma viral oncogene homolog (KRAS) gene mutation status are detected and quantified using EVs isolated by Click Beads and matched with those identified in biopsy specimens from Ewing sarcoma or pancreatic cancer patients. Moreover, the quantification of gene alterations can be used for monitoring treatment responses and disease progression.

Keywords: click chemistry; extracellular vesicles; gene alteration quantification; lipid labeling.

Publication types

Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't

MeSH terms

Carcinogenesis / genetics
Click Chemistry
Extracellular Vesicles* / genetics
Extracellular Vesicles* / metabolism
Genes, ras
Humans
Lipids
RNA-Binding Protein EWS / genetics
RNA-Binding Protein EWS / metabolism
Sarcoma, Ewing* / genetics
Sarcoma, Ewing* / metabolism

Substances

Lipids
RNA-Binding Protein EWS

Abstract

Publication types

MeSH terms

Substances

Grants and funding