Principal Investigator

Hakho
Lee
Awardee Organization

Massachusetts General Hospital
United States

Fiscal Year
2022
Activity Code
U01
Early Stage Investigator Grants (ESI)
Not Applicable
Project End Date

Early Detection through Novel OCEAN Technology - Ovarian Cancer Exosomal Analysis with Nanoplasmonics

Extracellular vesicles (EVs) are a new class of circulating biomarker that promises non-invasive, real-time cancer monitoring. Many studies have shown that i) EVs may function as reliable surrogates of parental cancer cells, and ii) these vesicles can reflect global tumor burden, overcoming limitations of tumor heterogeneity and sampling bias. Translating EV analyses into a clinically relevant cancer test, however, is limited by the lack of standardized, practical methods. Conventional assay tools (e.g., ultracentrifugation, Western blotting, ELISA) require large amounts of EVs and extensive processing, making them impractical for clinical workflows. Moreover, variations in sample handling and testing protocols often lead to inconsistent and confounding results. The goal of this proposal is to i) address such technical challenges in EV analyses, and ii) rigorously evaluate EVs' clinical value as a novel biomarker for early detection of ovarian cancer. We formed a strategic academic-industry partnership to achieve this goal: Exosome Diagnostics, an industry leader in EV-based cancer diagnostics, offering ready capacity to develop and manufacture in-vitro diagnostic medical devices; and the Center for Systems Biology at the Massachusetts General Hospital, a pioneer in developing novel analytical technologies for EV analyses. These teams will bring their multidisciplinary expertise, innovative technologies and complementary resources to carry out the following translational projects. First, we will advance a standardized EV assay platform. We will adopt our recently developed ExoLution platform to streamline EV collection, and the nPLEX (nano-plasmonic exosome) technology for high-throughput EV protein screening. Our initial study showed that nPLEX achieved >1000-fold higher sensitivity than conventional methods and yet consumed scant sample volumes (0.1 μL). We now seek to advance nPLEX to the instrument level by i) improving its robustness and throughput, and ii) establishing standardized assay protocols. Leveraging the developmental and regulatory expertise of Exosome Diagnostics, the resulting platform will be ready for translation into clinical diagnostic laboratories. Second, we will perform a targeted clinical study, particularly testing whether EVs can be exploited as a biomarker for early detection of ovarian cancer and progression monitoring. Our preclinical study will use patient-derived ovarian cancer cells and novel genetically engineered mouse models to identify EV protein signatures for ovarian cancer. We will next profile circulating EVs from ovarian cancer patients and assess the correlation between tumor burden and EV protein signature. Our study will be designed to ensure assay reliability and reproducibility, thereby delivering clinically translatable EV diagnostics. We will impose stringent quality controls on device design and sample processing, accrue well-annotated patient and control samples, and perform multisite testing. The technical and scientific outcomes of this research could have a significant translational impact in cancer management, establishing a robust, highly specific liquid biopsy for early detection of ovarian cancer.

Publications

  • Yang KS, Lin HY, Curley C, Welch MW, Wolpin BM, Lee H, Weissleder R, Im H, Castro CM. Bead-Based Extracellular Vesicle Analysis Using Flow Cytometry. Advanced biosystems. 2020 Dec;4(12):e2000203. Epub 2020 Oct 25. PMID: 33103361
  • Batool SM, Yekula A, Khanna P, Hsia T, Gamblin AS, Ekanayake E, Escobedo AK, You DG, Castro CM, Im H, Kilic T, Garlin MA, Skog J, Dinulescu DM, Dudley J, Agrawal N, Cheng J, Abtin F, Aberle DR, Chia D, Elashoff D, Grognan T, Krysan K, Oh SS, Strom C, Tu M, Wei F, Xian RR, Skates SJ, Zhang DY, Trinh T, Watson M, Aft R, Rawal S, Agarwal A, Kesmodel SB, Yang C, Shen C, Hochberg FH, Wong DTW, Patel AA, Papadopoulos N, Bettegowda C, Cote RJ, Srivastava S, Lee H, Carter BS, Balaj L. The Liquid Biopsy Consortium: Challenges and opportunities for early cancer detection and monitoring. Cell reports. Medicine. 2023 Oct 17;4(10):101198. Epub 2023 Sep 15. PMID: 37716353
  • Yu H, Lee H, Cheong J, Woo SW, Oh J, Oh HK, Lee JH, Zheng H, Castro CM, Yoo YE, Kim MG, Cheon J, Weissleder R, Lee H. A rapid assay provides on-site quantification of tetrahydrocannabinol in oral fluid. Science translational medicine. 2021 Oct 20;13(616):eabe2352. Epub 2021 Oct 20. PMID: 34669441
  • Kilic T, Weissleder R, Lee H. Molecular and Immunological Diagnostic Tests of COVID-19: Current Status and Challenges. iScience. 2020 Aug 21;23(8):101406. Epub 2020 Jul 25. PMID: 32771976
  • Huang CH, Park YI, Lin HY, Pathania D, Park KS, Avila-Wallace M, Castro CM, Weissleder R, Lee H. Compact and Filter-Free Luminescence Biosensor for Mobile in Vitro Diagnoses. ACS nano. 2019 Oct 22;13(10):11698-11706. Epub 2019 Sep 6. PMID: 31461265
  • Gerton TJ, Green A, Campisi M, Chen M, Gjeci I, Mahadevan N, Lee CAA, Mishra R, Vo HV, Haratani K, Li ZH, Hasselblatt KT, Testino B, Connor T, Lian CG, Elias KM, Lizotte P, Ivanova EV, Barbie DA, Dinulescu DM. Development of a Patient-Derived 3D Immuno-Oncology Platform to Potentiate Immunotherapy Responses in Ascites-Derived Circulating Tumor Cells. Cancers. 2023 Aug 16;15. (16). PMID: 37627156
  • Chin LK, Son T, Hong JS, Liu AQ, Skog J, Castro CM, Weissleder R, Lee H, Im H. Plasmonic Sensors for Extracellular Vesicle Analysis: From Scientific Development to Translational Research. ACS nano. 2020 Nov 24;14(11):14528-14548. Epub 2020 Oct 29. PMID: 33119256
  • Park J, Woo S, Kim J, Lee H, Yoo YE, Hong S. Rapid and simple single-chamber nucleic acid detection system prepared through nature-inspired surface engineering. Theranostics. 2021 May 3;11(14):6735-6745. doi: 10.7150/thno.57153. eCollection 2021. PMID: 34093850
  • Batool SM, Hsia T, Beecroft A, Lewis B, Ekanayake E, Rosenfeld Y, Escobedo AK, Gamblin AS, Rawal S, Cote RJ, Watson M, Wong DTW, Patel AA, Skog J, Papadopoulos N, Bettegowda C, Castro CM, Lee H, Srivastava S, Carter BS, Balaj L. Extrinsic and intrinsic preanalytical variables affecting liquid biopsy in cancer. Cell reports. Medicine. 2023 Oct 17;4(10):101196. Epub 2023 Sep 18. PMID: 37725979
  • Peterson HM, Chin LK, Iwamoto Y, Oh J, Carlson JCT, Lee H, Im H, Weissleder R. Integrated Analytical System for Clinical Single-Cell Analysis. Advanced science (Weinheim, Baden-Wurttemberg, Germany). 2022 Jul;9(20):e2200415. Epub 2022 May 4. PMID: 35508767
  • Min J, Im H, Allen M, McFarland PJ, Degani I, Yu H, Normandin E, Pathania D, Patel JM, Castro CM, Weissleder R, Lee H. Computational Optics Enables Breast Cancer Profiling in Point-of-Care Settings. ACS nano. 2018 Sep 25;12(9):9081-9090. Epub 2018 Aug 20. PMID: 30113824
  • Lee CY, Degani I, Cheong J, Weissleder R, Lee JH, Cheon J, Lee H. Development of Integrated Systems for On-Site Infection Detection. Accounts of chemical research. 2021 Nov 2;54(21):3991-4000. Epub 2021 Oct 22. PMID: 34677927
  • Woo HK, Cho YK, Lee CY, Lee H, Castro CM, Lee H. Characterization and modulation of surface charges to enhance extracellular vesicle isolation in plasma. Theranostics. 2022 Jan 31;12(5):1988-1998. doi: 10.7150/thno.69094. eCollection 2022. PMID: 35265194
  • Zaborowski MP, Lee K, Na YJ, Sammarco A, Zhang X, Iwanicki M, Cheah PS, Lin HY, Zinter M, Chou CY, Fulci G, Tannous BA, Lai CP, Birrer MJ, Weissleder R, Lee H, Breakefield XO. Methods for Systematic Identification of Membrane Proteins for Specific Capture of Cancer-Derived Extracellular Vesicles. Cell reports. 2019 Apr 2;27(1):255-268.e6. PMID: 30943406
  • Jo A, Green A, Medina JE, Iyer S, Ohman AW, McCarthy ET, Reinhardt F, Gerton T, Demehin D, Mishra R, Kolin DL, Zheng H, Cheon J, Crum CP, Weinberg RA, Rueda BR, Castro CM, Dinulescu DM, Lee H. Inaugurating High-Throughput Profiling of Extracellular Vesicles for Earlier Ovarian Cancer Detection. Advanced science (Weinheim, Baden-Wurttemberg, Germany). 2023 Sep;10(27):e2301930. Epub 2023 Jul 23. PMID: 37485618
  • Zaborowski MP, Cheah PS, Zhang X, Bushko I, Lee K, Sammarco A, Zappulli V, Maas SLN, Allen RM, Rumde P, György B, Aufiero M, Schweiger MW, Lai CP, Weissleder R, Lee H, Vickers KC, Tannous BA, Breakefield XO. Membrane-bound Gaussia luciferase as a tool to track shedding of membrane proteins from the surface of extracellular vesicles. Scientific reports. 2019 Nov 22;9(1):17387. PMID: 31758005
  • Park JH, Park EK, Cho YK, Shin IS, Lee H. Normalizing the Optical Signal Enables Robust Assays with Lateral Flow Biosensors. ACS omega. 2022 May 19;7(21):17723-17731. doi: 10.1021/acsomega.2c00793. eCollection 2022 May 31. PMID: 35664567
  • Gessner I, Park JH, Lin HY, Lee H, Weissleder R. Magnetic Gold Nanoparticles with Idealized Coating for Enhanced Point-Of-Care Sensing. Advanced healthcare materials. 2022 Jan;11(2):e2102035. Epub 2021 Nov 16. PMID: 34747576
  • Min J, Allen M, Castro CM, Lee H, Weissleder R, Im H. Computational Optics for Point-of-Care Breast Cancer Profiling. Methods in molecular biology (Clifton, N.J.). 2022;2393:153-162. PMID: 34837178
  • Kwak TJ, Son T, Hong JS, Winter UA, Jeong MH, McLean C, Weissleder R, Lee H, Castro CM, Im H. Electrokinetically enhanced label-free plasmonic sensing for rapid detection of tumor-derived extracellular vesicles. Biosensors & bioelectronics. 2023 Oct 1;237:115422. Epub 2023 Jun 3. PMID: 37301179
  • Cho YK, Kim H, Bénard A, Woo HK, Czubayko F, David P, Hansen FJ, Lee JI, Park JH, Schneck E, Weber GF, Shin IS, Lee H. Electrochemiluminescence in paired signal electrode (ECLipse) enables modular and scalable biosensing. Science advances. 2022 Sep 23;8(38):eabq4022. Epub 2022 Sep 21. PMID: 36129990
  • Lee H, Castro CM. Thermophoretically enriched detection. Nature biomedical engineering. 2019 Mar;3(3):163-164. PMID: 30948812
  • Van Deun J, Jo A, Li H, Lin HY, Weissleder R, Im H, Lee H. Integrated Dual-Mode Chromatography to Enrich Extracellular Vesicles from Plasma. Advanced biosystems. 2020 Dec;4(12):e1900310. Epub 2020 Apr 30. PMID: 32351054
  • Stollmann A, Garcia-Guirado J, Hong JS, Im H, Lee H, Arroyo JO, Quidant R. Molecular fingerprinting of biological nanoparticles with a label-free optofluidic platform. Research square. 2023 Oct 2. PMID: 37886549
  • Min J, Son T, Hong JS, Cheah PS, Wegemann A, Murlidharan K, Weissleder R, Lee H, Im H. Plasmon-Enhanced Biosensing for Multiplexed Profiling of Extracellular Vesicles. Advanced biosystems. 2020 Dec;4(12):e2000003. Epub 2020 Aug 19. PMID: 32815321
  • Kilic T, Gessner I, Cho YK, Jeong N, Quintana J, Weissleder R, Lee H. Zwitterionic Polymer Electroplating Facilitates the Preparation of Electrode Surfaces for Biosensing. Advanced materials (Deerfield Beach, Fla.). 2022 Feb;34(8):e2107892. Epub 2022 Jan 17. PMID: 34890082
  • Lee CY, Degani I, Cheong J, Lee JH, Choi HJ, Cheon J, Lee H. Fluorescence polarization system for rapid COVID-19 diagnosis. Biosensors & bioelectronics. 2021 Apr 15;178:113049. Epub 2021 Jan 30. PMID: 33540323
  • Stollmann A, Garcia-Guirado J, Hong JS, Im H, Lee H, Arroyo JO, Quidant R. Molecular fingerprinting of biological nanoparticles with a label-free optofluidic platform. ArXiv. 2023 Aug 11. PMID: 37608933
  • Liebel M, Ortega Arroyo J, Beltrán VS, Osmond J, Jo A, Lee H, Quidant R, van Hulst NF. 3D tracking of extracellular vesicles by holographic fluorescence imaging. Science advances. 2020 Nov 4;6. (45). Print 2020 Nov. PMID: 33148645
  • Oh J, Carlson JCT, Landeros C, Lee H, Ferguson S, Faquin WC, Clark JR, Pittet MJ, Pai SI, Weissleder R. Rapid Serial Immunoprofiling of the Tumor Immune Microenvironment by Fine Needle Sampling. Clinical cancer research : an official journal of the American Association for Cancer Research. 2021 Sep 1;27(17):4781-4793. Epub 2021 Jul 7. PMID: 34233961
  • Pathania D, Landeros C, Rohrer L, D'Agostino V, Hong S, Degani I, Avila-Wallace M, Pivovarov M, Randall T, Weissleder R, Lee H, Im H, Castro CM. Point-of-care cervical cancer screening using deep learning-based microholography. Theranostics. 2019 Nov 26;9(26):8438-8447. doi: 10.7150/thno.37187. eCollection 2019. PMID: 31879529
  • Min J, Chin LK, Oh J, Landeros C, Vinegoni C, Lee J, Lee SJ, Park JY, Liu AQ, Castro CM, Lee H, Im H, Weissleder R. CytoPAN-Portable cellular analyses for rapid point-of-care cancer diagnosis. Science translational medicine. 2020 Aug 5;12. (555). PMID: 32759277
  • Ortiz-Orruño U, Jo A, Lee H, van Hulst NF, Liebel M. Precise Nanosizing with High Dynamic Range Holography. Nano letters. 2021 Jan 13;21(1):317-322. Epub 2020 Dec 21. PMID: 33346670
  • Kim SJ, Wang C, Zhao B, Im H, Min J, Choi HJ, Tadros J, Choi NR, Castro CM, Weissleder R, Lee H, Lee K. Deep transfer learning-based hologram classification for molecular diagnostics. Scientific reports. 2018 Nov 19;8(1):17003. PMID: 30451953
  • Huber S, Min C, Staat C, Oh J, Castro CM, Haase A, Weissleder R, Gleich B, Lee H. Multichannel digital heteronuclear magnetic resonance biosensor. Biosensors & bioelectronics. 2019 Feb 1;126:240-248. Epub 2018 Oct 27. PMID: 30445298
  • Cheong J, Yu H, Lee CY, Lee JU, Choi HJ, Lee JH, Lee H, Cheon J. Fast detection of SARS-CoV-2 RNA via the integration of plasmonic thermocycling and fluorescence detection in a portable device. Nature biomedical engineering. 2020 Dec;4(12):1159-1167. Epub 2020 Dec 3. PMID: 33273713
  • Park J, Park JS, Huang CH, Jo A, Cook K, Wang R, Lin HY, Van Deun J, Li H, Min J, Wang L, Yoon G, Carter BS, Balaj L, Choi GS, Castro CM, Weissleder R, Lee H. An integrated magneto-electrochemical device for the rapid profiling of tumour extracellular vesicles from blood plasma. Nature biomedical engineering. 2021 Jul;5(7):678-689. Epub 2021 Jun 28. PMID: 34183802