Engineered nanointerfaces for microfluidic isolation and molecular profiling of tumor-specific extracellular vesicles

Eduardo Reátegui; Kristan E van der Vos; Charles P Lai; Mahnaz Zeinali; Nadia A Atai; Berent Aldikacti; Frederick P Floyd Jr; Aimal H Khankhel; Vishal Thapar; Fred H Hochberg; Lecia V Sequist; Brian V Nahed; Bob S Carter; Mehmet Toner; Leonora Balaj; David T Ting; Xandra O Breakefield; Shannon L Stott

doi:10.1038/s41467-017-02261-1

Engineered nanointerfaces for microfluidic isolation and molecular profiling of tumor-specific extracellular vesicles

Nat Commun. 2018 Jan 12;9(1):175. doi: 10.1038/s41467-017-02261-1.

Authors

Eduardo Reátegui^{1

2

3

4

5

6}, Kristan E van der Vos^{7

8}, Charles P Lai^{7

9}, Mahnaz Zeinali^{1

2

3

4}, Nadia A Atai⁷, Berent Aldikacti^{1

2

4}, Frederick P Floyd Jr^{1

2

3}, Aimal H Khankhel^{1

2

3}, Vishal Thapar⁴, Fred H Hochberg¹⁰, Lecia V Sequist^{4

11}, Brian V Nahed^{4

12}, Bob S Carter¹⁰, Mehmet Toner^{1

2

3

5}, Leonora Balaj⁷, David T Ting⁴, Xandra O Breakefield^{7

13}, Shannon L Stott^{14

15

16

17

18

19}

Affiliations

¹ Center for Engineering in Medicine, Massachusetts General Hospital, Harvard Medical School, Charlestown, MA, 02129, USA.
² Harvard Medical School, Boston, MA, 02114, USA.
³ Department of Surgery, Massachusetts General Hospital, Harvard Medical School, Boston, MA, 02114, USA.
⁴ Massachusetts General Hospital Cancer Center, Harvard Medical School, Boston, MA, 02114, USA.
⁵ Shriners Hospital for Children, Harvard Medical School, Boston, MA, 02114, USA.
⁶ William G. Lowrie Department of Chemical and Biomolecular Engineering, Comprehensive Cancer Center, The Ohio State University, Columbus, OH, 43210, USA.
⁷ Neurodiscovery Center, Massachusetts General Hospital, Harvard Medical School, Boston, MA, 02124, USA.
⁸ Department of Molecular Carcinogenesis, Netherlands Cancer Institute, 1066, CX, Amsterdam, The Netherlands.
⁹ Institute of Atomic and Molecular Sciences, Academia Sinica, Taipei, 10617, Taiwan.
¹⁰ Department of Neurosurgery, University of California San Diego, La Jolla, CA, 92093, USA.
¹¹ Department of Medicine, Massachusetts General Hospital, Harvard Medical School, Boston, MA, 02114, USA.
¹² Department of Neurosurgery, Massachusetts General Hospital, Harvard Medical School, Boston, MA, 02124, USA.
¹³ Department of Neurology and Radiology, Massachusetts General Hospital, Harvard Medical School, Boston, MA, 02114, USA.
¹⁴ Center for Engineering in Medicine, Massachusetts General Hospital, Harvard Medical School, Charlestown, MA, 02129, USA. [email protected].
¹⁵ Harvard Medical School, Boston, MA, 02114, USA. [email protected].
¹⁶ Department of Surgery, Massachusetts General Hospital, Harvard Medical School, Boston, MA, 02114, USA. [email protected].
¹⁷ Massachusetts General Hospital Cancer Center, Harvard Medical School, Boston, MA, 02114, USA. [email protected].
¹⁸ Shriners Hospital for Children, Harvard Medical School, Boston, MA, 02114, USA. [email protected].
¹⁹ Department of Medicine, Massachusetts General Hospital, Harvard Medical School, Boston, MA, 02114, USA. [email protected].

Abstract

Extracellular vesicles (EVs) carry RNA, DNA, proteins, and lipids. Specifically, tumor-derived EVs have the potential to be utilized as disease-specific biomarkers. However, a lack of methods to isolate tumor-specific EVs has limited their use in clinical settings. Here we report a sensitive analytical microfluidic platform (^EVHB-Chip) that enables tumor-specific EV-RNA isolation within 3 h. Using the ^EVHB-Chip, we achieve 94% tumor-EV specificity, a limit of detection of 100 EVs per μL, and a 10-fold increase in tumor RNA enrichment in comparison to other methods. Our approach allows for the subsequent release of captured tumor EVs, enabling downstream characterization and functional studies. Processing serum and plasma samples from glioblastoma multiforme (GBM) patients, we can detect the mutant EGFRvIII mRNA. Moreover, using next-generation RNA sequencing, we identify genes specific to GBM as well as transcripts that are hallmarks for the four genetic subtypes of the disease.

Publication types

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

MeSH terms

Biological Transport
Brain Neoplasms / chemistry
Brain Neoplasms / genetics
Brain Neoplasms / metabolism*
Cell Line, Tumor
ErbB Receptors / genetics
ErbB Receptors / metabolism
Extracellular Vesicles / chemistry*
Extracellular Vesicles / metabolism
Glioblastoma / chemistry
Glioblastoma / genetics
Glioblastoma / metabolism*
Humans
Microfluidics / instrumentation
Microfluidics / methods*
RNA / genetics
RNA / metabolism

Substances

epidermal growth factor receptor VIII
RNA
ErbB Receptors

Abstract

Publication types

MeSH terms

Substances

Grants and funding