621P - Effective identification of primary liver cancer from cirrhosis or chronic hepatitis virus infection using eight methylated plasma DNA markers: Marker discovery, phase I pilot, and phase II clinical validation

Background

Early detection of primary liver cancer (PLC) in patients with liver cirrhosis (LC) or chronic hepatitis virus infection (CHVI) improves survival. The urgent need is convenient and affordable tools with high sensitivity to facilitate timely diagnosis.

Methods

Tissue and plasma samples from 159 healthy individuals and 89 PLC, LC, or CHVI patients were sequenced by a targeted methylation panel (∼70,000 CpGs) to identify candidate methylated DNA markers (MDMs). In phase I, the performance of each selected MDMs was validated in 175 plasma samples (PLC, n=101; LC/CHVI, n=74) by CO-methylation aMplification rEal-Time PCR (COMET) assay. A logistic model was then trained and validated in phase II with 310 plasma samples (hepatocellular carcinoma [HCC], n=212; combined hepatocellular-cholangiocarcinoma [cHCC-CC], n=12; CHVI, n=106; training vs. validation, 2:1).

Results

The 11 selected MDMs with top performance consistently showed significant differences in tissue samples between PLC and LC/CHVI, as well as in plasma samples between PLC and LC/CHVI (P<0.05). In phase I, 8 of the above 11 MDMs with an area under the curve (AUC) over 0.80 to differentiate PLC and LC/CHVI were selected for further investigation. In phase II, the MDM-based model achieved sensitivity of 87.2% (95% confidence interval [CI], 80.8%–92.4%) and 88.0% (78.4%–94.4%), at respective specificity of 97.1% (90.1%–99.7%) and 100% (90.3%–100%) in the training and validation sets. In the validation set, sensitivity in patients with BCLC stage 0, diameter<3 cm, AFP-negative and PIVKA-II-negative was 90.0% (55.5%–99.7%), 88.9% (65.3%–98.6%), 80.6% (64.0%–91.8%), and 81.3% (54.4%–96.0%), respectively. Additionally, our model detected 19 of 24 (79.3%, 57.8%–92.9%) intrahepatic cholangiocarcinoma. Combining AFP and PIVKA-II, the model achieved higher sensitivity of 93.3% (85.1%–97.8%) and specificity of 100.0% (90.3%–100%).

Conclusions

COMET featuring low cost and high accuracy exhibits preferable potential for PLC detection in patients with LC or CHVI. Further validation in a prospective cohort is warranted.

Clinical trial identification

NCT05996666, release date 08/10/2023.

Editorial acknowledgement

Legal entity responsible for the study

Tian Yang.

Funding

National Natural Science Foundation of China (No: 81972726 and 82273074), Dawn Project Foundation of Shanghai (No: 21SG36), Shanghai Health Academic Leader Program (No. 2022XD001).

Disclosure

X. Zhu, L. Zhang, Q. You, J. Xu, Y. Xu, H. Lu, B. Li, G. Wang, S. Cai: Financial Interests, Personal, Full or part-time Employment: Burning Rock Biotech. All other authors have declared no conflicts of interest.