HCC EV ECG score: An extracellular vesicle-based protein assay for detection of early-stage hepatocellular carcinoma

Na Sun; Ceng Zhang; Yi Te Lee; Benjamin V. Tran; Jing Wang; Hyoyong Kim; Junseok Lee; Ryan Y. Zhang; Jasmine J. Wang; Junhui Hu; Zhicheng Zhang; Manaf S. Alsudaney; Kuan Chu Hou; Hubert Tang; Tiffany X. Zhang; Icy Y. Liang; Ziang Zhou; Mengxiang Chen; Angela Hsiao Jiun Yeh; Wenyuan Li; Xianghong Jasmine Zhou; Helena R. Chang; Steven Huy B. Han; Saeed Sadeghi; Richard S. Finn; Sammy Saab; Ronald W. Busuttil; Mazen Noureddin; Walid S. Ayoub; Alexander Kuo; Vinay Sundaram; Buraq Al-Ghaieb; Juvelyn Palomique; Kambiz Kosari; Irene K. Kim; Tsuyoshi Todo; Nicholas N. Nissen; Maria Lauda Tomasi; Sungyong You; Edwin M. Posadas; James X. Wu; Madhuri Wadehra; Myung Shin Sim; Yunfeng Li; Hanlin L. Wang; Samuel W. French; Shelly C. Lu; Lily Wu; Renjun Pei; Li Liang; Ju Dong Yang; Vatche G. Agopian; Hsian Rong Tseng; Yazhen Zhu

doi:10.1002/hep.32692

HCC EV ECG score: An extracellular vesicle-based protein assay for detection of early-stage hepatocellular carcinoma

Na Sun, Ceng Zhang, Yi Te Lee, Benjamin V. Tran, Jing Wang, Hyoyong Kim, Junseok Lee, Ryan Y. Zhang, Jasmine J. Wang, Junhui Hu, Zhicheng Zhang, Manaf S. Alsudaney, Kuan Chu Hou, Hubert Tang, Tiffany X. Zhang, Icy Y. Liang, Ziang Zhou, Mengxiang Chen, Angela Hsiao Jiun Yeh, Wenyuan LiXianghong Jasmine Zhou, Helena R. Chang, Steven Huy B. Han, Saeed Sadeghi, Richard S. Finn, Sammy Saab, Ronald W. Busuttil, Mazen Noureddin, Walid S. Ayoub, Alexander Kuo, Vinay Sundaram, Buraq Al-Ghaieb, Juvelyn Palomique, Kambiz Kosari, Irene K. Kim, Tsuyoshi Todo, Nicholas N. Nissen, Maria Lauda Tomasi, Sungyong You, Edwin M. Posadas, James X. Wu, Madhuri Wadehra, Myung Shin Sim, Yunfeng Li, Hanlin L. Wang, Samuel W. French, Shelly C. Lu, Lily Wu, Renjun Pei, Li Liang, Ju Dong Yang, Vatche G. Agopian, Hsian Rong Tseng, Yazhen Zhu

Research output: Contribution to journal › Article › peer-review

23 Scopus citations

Abstract

Background and Aims: The sensitivity of current surveillance methods for detecting early-stage hepatocellular carcinoma (HCC) is suboptimal. Extracellular vesicles (EVs) are promising circulating biomarkers for early cancer detection. In this study, we aim to develop an HCC EV-based surface protein assay for early detection of HCC. Approach and Results: Tissue microarray was used to evaluate four potential HCC-associated protein markers. An HCC EV surface protein assay, composed of covalent chemistry-mediated HCC EV purification and real-time immuno-polymerase chain reaction readouts, was developed and optimized for quantifying subpopulations of EVs. An HCC EV ECG score, calculated from the readouts of three HCC EV subpopulations (E pCAM+ CD63+, C D147+ CD63+, and G PC3+ CD63+ HCC EVs), was established for detecting early-stage HCC. A phase 2 biomarker study was conducted to evaluate the performance of ECG score in a training cohort (n = 106) and an independent validation cohort (n = 72). Overall, 99.7% of tissue microarray stained positive for at least one of the four HCC-associated protein markers (EpCAM, CD147, GPC3, and ASGPR1) that were subsequently validated in HCC EVs. In the training cohort, HCC EV ECG score demonstrated an area under the receiver operating curve (AUROC) of 0.95 (95% confidence interval [CI], 0.90-0.99) for distinguishing early-stage HCC from cirrhosis with a sensitivity of 91% and a specificity of 90%. The AUROCs of the HCC EV ECG score remained excellent in the validation cohort (0.93; 95% CI, 0.87-0.99) and in the subgroups by etiology (viral: 0.95; 95% CI, 0.90-1.00; nonviral: 0.94; 95% CI, 0.88-0.99). Conclusion: HCC EV ECG score demonstrated great potential for detecting early-stage HCC. It could augment current surveillance methods and improve patients' outcomes.

Original language	English (US)
Pages (from-to)	774-788
Number of pages	15
Journal	Hepatology
Volume	77
Issue number	3
DOIs	https://doi.org/10.1002/hep.32692
State	Published - Mar 2023

Keywords

Humans
Carcinoma, Hepatocellular/diagnosis
Liver Neoplasms/diagnosis
Biomarkers, Tumor/analysis
Extracellular Vesicles/chemistry
Membrane Proteins
Electrocardiography
Glypicans

ASJC Scopus subject areas

Hepatology

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10.1002/hep.32692

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Sun, N., Zhang, C., Lee, Y. T., Tran, B. V., Wang, J., Kim, H., Lee, J., Zhang, R. Y., Wang, J. J., Hu, J., Zhang, Z., Alsudaney, M. S., Hou, K. C., Tang, H., Zhang, T. X., Liang, I. Y., Zhou, Z., Chen, M., Yeh, A. H. J., ... Zhu, Y. (2023). HCC EV ECG score: An extracellular vesicle-based protein assay for detection of early-stage hepatocellular carcinoma. Hepatology, 77(3), 774-788. https://doi.org/10.1002/hep.32692

Sun, N, Zhang, C, Lee, YT, Tran, BV, Wang, J, Kim, H, Lee, J, Zhang, RY, Wang, JJ, Hu, J, Zhang, Z, Alsudaney, MS, Hou, KC, Tang, H, Zhang, TX, Liang, IY, Zhou, Z, Chen, M, Yeh, AHJ, Li, W, Zhou, XJ, Chang, HR, Han, SHB, Sadeghi, S, Finn, RS, Saab, S, Busuttil, RW , Noureddin, M, Ayoub, WS, Kuo, A, Sundaram, V, Al-Ghaieb, B, Palomique, J, Kosari, K, Kim, IK, Todo, T, Nissen, NN, Tomasi, ML, You, S, Posadas, EM, Wu, JX, Wadehra, M, Sim, MS, Li, Y, Wang, HL, French, SW, Lu, SC, Wu, L, Pei, R, Liang, L, Yang, JD, Agopian, VG, Tseng, HR & Zhu, Y 2023, 'HCC EV ECG score: An extracellular vesicle-based protein assay for detection of early-stage hepatocellular carcinoma', Hepatology, vol. 77, no. 3, pp. 774-788. https://doi.org/10.1002/hep.32692

@article{b7c82f3432384194b2c195cf05b35364,

title = "HCC EV ECG score: An extracellular vesicle-based protein assay for detection of early-stage hepatocellular carcinoma",

abstract = "Background and Aims: The sensitivity of current surveillance methods for detecting early-stage hepatocellular carcinoma (HCC) is suboptimal. Extracellular vesicles (EVs) are promising circulating biomarkers for early cancer detection. In this study, we aim to develop an HCC EV-based surface protein assay for early detection of HCC. Approach and Results: Tissue microarray was used to evaluate four potential HCC-associated protein markers. An HCC EV surface protein assay, composed of covalent chemistry-mediated HCC EV purification and real-time immuno-polymerase chain reaction readouts, was developed and optimized for quantifying subpopulations of EVs. An HCC EV ECG score, calculated from the readouts of three HCC EV subpopulations (E pCAM+ CD63+, C D147+ CD63+, and G PC3+ CD63+ HCC EVs), was established for detecting early-stage HCC. A phase 2 biomarker study was conducted to evaluate the performance of ECG score in a training cohort (n = 106) and an independent validation cohort (n = 72). Overall, 99.7% of tissue microarray stained positive for at least one of the four HCC-associated protein markers (EpCAM, CD147, GPC3, and ASGPR1) that were subsequently validated in HCC EVs. In the training cohort, HCC EV ECG score demonstrated an area under the receiver operating curve (AUROC) of 0.95 (95% confidence interval [CI], 0.90-0.99) for distinguishing early-stage HCC from cirrhosis with a sensitivity of 91% and a specificity of 90%. The AUROCs of the HCC EV ECG score remained excellent in the validation cohort (0.93; 95% CI, 0.87-0.99) and in the subgroups by etiology (viral: 0.95; 95% CI, 0.90-1.00; nonviral: 0.94; 95% CI, 0.88-0.99). Conclusion: HCC EV ECG score demonstrated great potential for detecting early-stage HCC. It could augment current surveillance methods and improve patients' outcomes.",

keywords = "Humans, Carcinoma, Hepatocellular/diagnosis, Liver Neoplasms/diagnosis, Biomarkers, Tumor/analysis, Extracellular Vesicles/chemistry, Membrane Proteins, Electrocardiography, Glypicans",

author = "Na Sun and Ceng Zhang and Lee, {Yi Te} and Tran, {Benjamin V.} and Jing Wang and Hyoyong Kim and Junseok Lee and Zhang, {Ryan Y.} and Wang, {Jasmine J.} and Junhui Hu and Zhicheng Zhang and Alsudaney, {Manaf S.} and Hou, {Kuan Chu} and Hubert Tang and Zhang, {Tiffany X.} and Liang, {Icy Y.} and Ziang Zhou and Mengxiang Chen and Yeh, {Angela Hsiao Jiun} and Wenyuan Li and Zhou, {Xianghong Jasmine} and Chang, {Helena R.} and Han, {Steven Huy B.} and Saeed Sadeghi and Finn, {Richard S.} and Sammy Saab and Busuttil, {Ronald W.} and Mazen Noureddin and Ayoub, {Walid S.} and Alexander Kuo and Vinay Sundaram and Buraq Al-Ghaieb and Juvelyn Palomique and Kambiz Kosari and Kim, {Irene K.} and Tsuyoshi Todo and Nissen, {Nicholas N.} and Tomasi, {Maria Lauda} and Sungyong You and Posadas, {Edwin M.} and Wu, {James X.} and Madhuri Wadehra and Sim, {Myung Shin} and Yunfeng Li and Wang, {Hanlin L.} and French, {Samuel W.} and Lu, {Shelly C.} and Lily Wu and Renjun Pei and Li Liang and Yang, {Ju Dong} and Agopian, {Vatche G.} and Tseng, {Hsian Rong} and Yazhen Zhu",

note = "Funding Information: This work is supported by American College of Gastroenterology Junior Faculty Development Award, Department of Defense Peer Reviewed Cancer Research Program Career Development Award (CA191051), and National Institutes of Health (R01 CA218356, U01 CA198900, P01 CA233452, R01 CA255727, R01 CA253651, R01 CA246304, U01 CA230705, U01 EB026421, U54 CA14391, R21 CA240887, and R21 CA235340). The funders had no role in the collection of data; the design and conduct of the study; management, analysis, and interpretation of the data; preparation, review, or approval of the manuscript; and decision to submit the manuscript for publication. Funding Information: Hsian‐Rong Tseng owns stock in CytoLumina and Pulsar. Vinay Sundaram consults for Saol. He is on the speakers{\textquoteright} bureau for Gilead, AbbVie, and Intercept. Mazen Noureddin owns stock in, consults for, and received grants from Viking. He consults for and received grants from Gilead, Pfizer, and Madrigal. He consults for 89 Bio, Altimmune, CoBar, Cytodyn, Intercept, Novo Nordisk, Blade, EchoSens, Fractyl, NorthSea, Perspecturm, Terns, Sami‐Sabina Group, Siemens, and Roche. He received grants from Allergan, BMS, Galmed, Galectin, Genfit, Conatus, Enanta, Novartis, Shire, and Zydus. Richard S. Finn consults for AstraZeneca, Bayer, BMS, Exelixis, CStone, Eisai, Eli Lilly, Pfizer, Merck, and Roche/Genentech. His institution receives grants from Bayer, Eisai, Bristol Myers Squibb, Eli Lilly, Roche/Genentech, and Pfizer. Saeed Sadeghi consults for and is in the speakers{\textquoteright} bureau for Eisai. Wenyuan Li is a co‐founder and shareholder in Early Diagnostics Inc. Publisher Copyright: {\textcopyright} 2023 American Association for the Study of Liver Diseases.",

year = "2023",

month = mar,

doi = "10.1002/hep.32692",

language = "English (US)",

volume = "77",

pages = "774--788",

journal = "Hepatology",

issn = "0270-9139",

publisher = "Wiley",

number = "3",

}

TY - JOUR

T1 - HCC EV ECG score

T2 - An extracellular vesicle-based protein assay for detection of early-stage hepatocellular carcinoma

AU - Sun, Na

AU - Zhang, Ceng

AU - Lee, Yi Te

AU - Tran, Benjamin V.

AU - Wang, Jing

AU - Kim, Hyoyong

AU - Lee, Junseok

AU - Zhang, Ryan Y.

AU - Wang, Jasmine J.

AU - Hu, Junhui

AU - Zhang, Zhicheng

AU - Alsudaney, Manaf S.

AU - Hou, Kuan Chu

AU - Tang, Hubert

AU - Zhang, Tiffany X.

AU - Liang, Icy Y.

AU - Zhou, Ziang

AU - Chen, Mengxiang

AU - Yeh, Angela Hsiao Jiun

AU - Li, Wenyuan

AU - Zhou, Xianghong Jasmine

AU - Chang, Helena R.

AU - Han, Steven Huy B.

AU - Sadeghi, Saeed

AU - Finn, Richard S.

AU - Saab, Sammy

AU - Busuttil, Ronald W.

AU - Noureddin, Mazen

AU - Ayoub, Walid S.

AU - Kuo, Alexander

AU - Sundaram, Vinay

AU - Al-Ghaieb, Buraq

AU - Palomique, Juvelyn

AU - Kosari, Kambiz

AU - Kim, Irene K.

AU - Todo, Tsuyoshi

AU - Nissen, Nicholas N.

AU - Tomasi, Maria Lauda

AU - You, Sungyong

AU - Posadas, Edwin M.

AU - Wu, James X.

AU - Wadehra, Madhuri

AU - Sim, Myung Shin

AU - Li, Yunfeng

AU - Wang, Hanlin L.

AU - French, Samuel W.

AU - Lu, Shelly C.

AU - Wu, Lily

AU - Pei, Renjun

AU - Liang, Li

AU - Yang, Ju Dong

AU - Agopian, Vatche G.

AU - Tseng, Hsian Rong

AU - Zhu, Yazhen

N1 - Funding Information: This work is supported by American College of Gastroenterology Junior Faculty Development Award, Department of Defense Peer Reviewed Cancer Research Program Career Development Award (CA191051), and National Institutes of Health (R01 CA218356, U01 CA198900, P01 CA233452, R01 CA255727, R01 CA253651, R01 CA246304, U01 CA230705, U01 EB026421, U54 CA14391, R21 CA240887, and R21 CA235340). The funders had no role in the collection of data; the design and conduct of the study; management, analysis, and interpretation of the data; preparation, review, or approval of the manuscript; and decision to submit the manuscript for publication. Funding Information: Hsian‐Rong Tseng owns stock in CytoLumina and Pulsar. Vinay Sundaram consults for Saol. He is on the speakers’ bureau for Gilead, AbbVie, and Intercept. Mazen Noureddin owns stock in, consults for, and received grants from Viking. He consults for and received grants from Gilead, Pfizer, and Madrigal. He consults for 89 Bio, Altimmune, CoBar, Cytodyn, Intercept, Novo Nordisk, Blade, EchoSens, Fractyl, NorthSea, Perspecturm, Terns, Sami‐Sabina Group, Siemens, and Roche. He received grants from Allergan, BMS, Galmed, Galectin, Genfit, Conatus, Enanta, Novartis, Shire, and Zydus. Richard S. Finn consults for AstraZeneca, Bayer, BMS, Exelixis, CStone, Eisai, Eli Lilly, Pfizer, Merck, and Roche/Genentech. His institution receives grants from Bayer, Eisai, Bristol Myers Squibb, Eli Lilly, Roche/Genentech, and Pfizer. Saeed Sadeghi consults for and is in the speakers’ bureau for Eisai. Wenyuan Li is a co‐founder and shareholder in Early Diagnostics Inc. Publisher Copyright: © 2023 American Association for the Study of Liver Diseases.

PY - 2023/3

Y1 - 2023/3

N2 - Background and Aims: The sensitivity of current surveillance methods for detecting early-stage hepatocellular carcinoma (HCC) is suboptimal. Extracellular vesicles (EVs) are promising circulating biomarkers for early cancer detection. In this study, we aim to develop an HCC EV-based surface protein assay for early detection of HCC. Approach and Results: Tissue microarray was used to evaluate four potential HCC-associated protein markers. An HCC EV surface protein assay, composed of covalent chemistry-mediated HCC EV purification and real-time immuno-polymerase chain reaction readouts, was developed and optimized for quantifying subpopulations of EVs. An HCC EV ECG score, calculated from the readouts of three HCC EV subpopulations (E pCAM+ CD63+, C D147+ CD63+, and G PC3+ CD63+ HCC EVs), was established for detecting early-stage HCC. A phase 2 biomarker study was conducted to evaluate the performance of ECG score in a training cohort (n = 106) and an independent validation cohort (n = 72). Overall, 99.7% of tissue microarray stained positive for at least one of the four HCC-associated protein markers (EpCAM, CD147, GPC3, and ASGPR1) that were subsequently validated in HCC EVs. In the training cohort, HCC EV ECG score demonstrated an area under the receiver operating curve (AUROC) of 0.95 (95% confidence interval [CI], 0.90-0.99) for distinguishing early-stage HCC from cirrhosis with a sensitivity of 91% and a specificity of 90%. The AUROCs of the HCC EV ECG score remained excellent in the validation cohort (0.93; 95% CI, 0.87-0.99) and in the subgroups by etiology (viral: 0.95; 95% CI, 0.90-1.00; nonviral: 0.94; 95% CI, 0.88-0.99). Conclusion: HCC EV ECG score demonstrated great potential for detecting early-stage HCC. It could augment current surveillance methods and improve patients' outcomes.

AB - Background and Aims: The sensitivity of current surveillance methods for detecting early-stage hepatocellular carcinoma (HCC) is suboptimal. Extracellular vesicles (EVs) are promising circulating biomarkers for early cancer detection. In this study, we aim to develop an HCC EV-based surface protein assay for early detection of HCC. Approach and Results: Tissue microarray was used to evaluate four potential HCC-associated protein markers. An HCC EV surface protein assay, composed of covalent chemistry-mediated HCC EV purification and real-time immuno-polymerase chain reaction readouts, was developed and optimized for quantifying subpopulations of EVs. An HCC EV ECG score, calculated from the readouts of three HCC EV subpopulations (E pCAM+ CD63+, C D147+ CD63+, and G PC3+ CD63+ HCC EVs), was established for detecting early-stage HCC. A phase 2 biomarker study was conducted to evaluate the performance of ECG score in a training cohort (n = 106) and an independent validation cohort (n = 72). Overall, 99.7% of tissue microarray stained positive for at least one of the four HCC-associated protein markers (EpCAM, CD147, GPC3, and ASGPR1) that were subsequently validated in HCC EVs. In the training cohort, HCC EV ECG score demonstrated an area under the receiver operating curve (AUROC) of 0.95 (95% confidence interval [CI], 0.90-0.99) for distinguishing early-stage HCC from cirrhosis with a sensitivity of 91% and a specificity of 90%. The AUROCs of the HCC EV ECG score remained excellent in the validation cohort (0.93; 95% CI, 0.87-0.99) and in the subgroups by etiology (viral: 0.95; 95% CI, 0.90-1.00; nonviral: 0.94; 95% CI, 0.88-0.99). Conclusion: HCC EV ECG score demonstrated great potential for detecting early-stage HCC. It could augment current surveillance methods and improve patients' outcomes.

KW - Humans

KW - Carcinoma, Hepatocellular/diagnosis

KW - Liver Neoplasms/diagnosis

KW - Biomarkers, Tumor/analysis

KW - Extracellular Vesicles/chemistry

KW - Membrane Proteins

KW - Electrocardiography

KW - Glypicans

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U2 - 10.1002/hep.32692

DO - 10.1002/hep.32692

M3 - Article

C2 - 35908246

AN - SCOPUS:85137367051

SN - 0270-9139

VL - 77

SP - 774

EP - 788

JO - Hepatology

JF - Hepatology

IS - 3

ER -

HCC EV ECG score: An extracellular vesicle-based protein assay for detection of early-stage hepatocellular carcinoma

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