Program Official

Principal Investigator

Bob S
Carter
Awardee Organization

Massachusetts General Hospital
United States

Fiscal Year
2022
Activity Code
R01
Early Stage Investigator Grants (ESI)
Not Applicable
Project End Date

Standardized Molecular Analyses of Glioma EVs

Extracellular vesicles (EVs) have emerged as a promising surrogate for tissue biopsy, potentially enabling non-invasive, real-time cancer monitoring. Most cancer cells release large numbers of EVs into circulation that carry molecular constituents reflective of the heterogeneity of the parent tumor. This project is designed to optimize a liquid biopsy to diagnose malignant glioma tumors. Currently, such tumors are diagnosed through a brain tissue biopsy which involves considerable risk for patients and doesn’t allow for longitudinal follow up of clinical care. Current EV isolation and characterization methods yield inconsistent results and render data reproducibility challenging, often leading to unpredictable conclusions. The ​goals​ of this project are to i) address variability among the different EV isolation methods and platforms currently available, and to ii) pinpoint to the “best” method to validate candidate biomarkers for glioma diagnosis. Our exceptional investigative team brings together experts in malignant glioma treatment, the field of nano-engineering, vesicular research, assay development and droplet digital PCR technology to optimize the necessary elements for the development of a blood-based assay capable of moving towards clinical settings. Through a simple blood test, clinicians will be able to diagnose, stratify and monitor a tumor without the need for tissue biopsy. Our strategic partnership with ​Exosome Diagnostics​, an industry leader in EV-based cancer diagnostics, offers us venues allowing for the translation of our findings, coupled with access to clinical grade kits, platforms and study design. The D​ epartment of Neurosurgery​ and the ​Center for Systems Biology​ at the Massachusetts General Hospital comprise multidisciplinary clinical expertise, innovative technologies and complementary resources to carry out the following translational projects: ​First,​ based on our prior kit comparison work, we have picked two top EV isolation kits and enrichment platforms to test in a series of well controlled, reference standards to determine an optimal EV isolation method. ​Second,​ we will test whether EV gene signatures can be used as biomarkers for cancer detection as well as tracking recurrence. By following quality control on device design and sample processing, accruing well-annotated patient and control samples, and performing multi site testing, we will ​ensure assay reliability and reproducibility​ to deliver clinically translatable EV diagnostics. Fourteen genes were selected through literature data mining based on the putative evidence that they can distinguish gliomas from controls. ​Finally,​ a gene’s signature with the highest sensitivity and specificity will be validated in a large cohort of patient samples. The technical and scientific outcomes of this research could have a significant ​translational impact in gliomas​, establishing a robust, highly specific assay to guide treatment decision and assess tumor recurrence.

Publications

  • 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
  • Yekula A, Muralidharan K, Rosh ZS, Youngkin AE, Kang KM, Balaj L, Carter BS. Liquid Biopsy Strategies to Distinguish Progression from Pseudoprogression and Radiation Necrosis in Glioblastomas. Advanced biosystems. 2020 Dec;4(12):e2000029. Epub 2020 Jun 2. PMID: 32484293
  • 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
  • Batool SM, Muralidharan K, Hsia T, Falotico S, Gamblin AS, Rosenfeld YB, Khanna SK, Balaj L, Carter BS. Highly Sensitive EGFRvIII Detection in Circulating Extracellular Vesicle RNA of Glioma Patients. Clinical cancer research : an official journal of the American Association for Cancer Research. 2022 Sep 15;28(18):4070-4082. PMID: 35849415
  • Verweij FJ, Balaj L, Boulanger CM, Carter DRF, Compeer EB, D'Angelo G, El Andaloussi S, Goetz JG, Gross JC, Hyenne V, Krämer-Albers EM, Lai CP, Loyer X, Marki A, Momma S, Nolte-'t Hoen ENM, Pegtel DM, Peinado H, Raposo G, Rilla K, Tahara H, Théry C, van Royen ME, Vandenbroucke RE, Wehman AM, Witwer K, Wu Z, Wubbolts R, van Niel G. The power of imaging to understand extracellular vesicle biology in vivo. Nature methods. 2021 Sep;18(9):1013-1026. Epub 2021 Aug 26. PMID: 34446922
  • Hsia T, Yekula A, Batool SM, Rosenfeld YB, You DG, Weissleder R, Lee H, Carter BS, Balaj L. Glioblastoma-derived extracellular vesicle subpopulations following 5-aminolevulinic acid treatment bear diagnostic implications. Journal of extracellular vesicles. 2022 Nov;11(11):e12278. PMID: 36404434
  • Yeo AT, Rawal S, Delcuze B, Christofides A, Atayde A, Strauss L, Balaj L, Rogers VA, Uhlmann EJ, Varma H, Carter BS, Boussiotis VA, Charest A. Single-cell RNA sequencing reveals evolution of immune landscape during glioblastoma progression. Nature immunology. 2022 Jun;23(6):971-984. Epub 2022 May 27. PMID: 35624211
  • Kilic T, Cho YK, Jeong N, Shin IS, Carter BS, Balaj L, Weissleder R, Lee H. Multielectrode Spectroscopy Enables Rapid and Sensitive Molecular Profiling of Extracellular Vesicles. ACS central science. 2022 Jan 26;8(1):110-117. Epub 2022 Jan 7. PMID: 35111901
  • Kang KM, Muralidharan K, Yekula A, Small JL, Rosh ZS, Jones PS, Carter BS, Balaj L. Blood-Based Detection of BRAF V600E in Gliomas and Brain Tumor Metastasis. Cancers. 2021 Mar 11;13. (6). PMID: 33799709
  • 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
  • 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
  • Muralidharan K, Yekula A, Small JL, Rosh ZS, Kang KM, Wang L, Lau S, Zhang H, Lee H, Bettegowda C, Chicoine MR, Kalkanis SN, Shankar GM, Nahed BV, Curry WT, Jones PS, Cahill DP, Balaj L, Carter BS. TERT Promoter Mutation Analysis for Blood-Based Diagnosis and Monitoring of Gliomas. Clinical cancer research : an official journal of the American Association for Cancer Research. 2021 Jan 1;27(1):169-178. Epub 2020 Oct 13. PMID: 33051308
  • 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
  • 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
  • Batool SM, Hsia T, Khanna SK, Gamblin AS, Rosenfeld Y, You DG, Carter BS, Balaj L. Decoding vesicle-based precision oncology in gliomas. Neuro-oncology advances. 2022 Nov 11;4(Suppl 2):ii53-ii60. doi: 10.1093/noajnl/vdac035. eCollection 2022 Nov. PMID: 36380860