Multimodal Approach for Identification and Validation of Hepatocellular Carcinoma Targets for Radiotheranostics
Abstract:
Identifying tumor selective targets is critical for the development of precision diagnostic and therapeutic agents in oncology. Despite advances in precision oncology elsewhere, there are no FDA-approved hepatocellular carcinoma (HCC)-selective treatments. HCC is the most common type of liver cancer and accounts for significant morbidity and mortality worldwide. Here, we sought to integrate bulk (371 cases) and single cell RNA sequencing (scRNAseq, n =2 datasets, 34 cases, 102,956 cells) of patient samples to enrich for molecules that are overexpressed in HCC, which could serve as HCC-selective targets. To guide definitions of tumor and normal cell clusters with higher fidelity, we also imported a normal liver scRNAseq dataset. Using this integrated approach, we identified several HCC-selective plasma membrane molecules. To validate these targets, we performed immunohistochemical staining of HCC and normal tissue microarrays and confirmed HCC-selective staining of identified targets. Next, we verified the presence of these targets in several commercially available HCC cell lines by flow cytometry and western blot. Finally, we designed, engineered, and tested novel antibody-based positron emission tomography (immunoPET) agents to these targets in various murine models of liver cancer. Our findings confirm that we can leverage this multimodal approach to identify and validate of HCC-selective targets, which can be used to develop tumor-selective diagnostic and therapeutic radiopharmaceuticals, or radiotheranostics, and other precision oncology agents.
One Sentence Summary:
A multimodal pipeline defines and validates tumor-selective surface targets for radiotheranostic use in hepatocellular carcinoma.
Insights
Researchers identified new surface targets specific to hepatocellular carcinoma (HCC), a common liver cancer. These targets enable the development of precision radiopharmaceuticals for diagnosing and treating HCC.
Area of Science:
- Oncology
- Molecular Biology
- Biotechnology
Background:
- Hepatocellular carcinoma (HCC) lacks tumor-selective treatments despite advances in precision oncology.
- Identifying HCC-specific targets is crucial for developing targeted diagnostic and therapeutic agents.
- Current treatment options for HCC are limited in their selectivity.
Purpose of the Study:
- To identify and validate novel plasma membrane molecules overexpressed in HCC for targeted therapies.
- To develop HCC-selective diagnostic and therapeutic radiopharmaceuticals (radiotheranostics).
- To establish a multimodal approach for discovering and validating tumor-selective targets.
Main Methods:
- Integrated analysis of bulk and single-cell RNA sequencing data from HCC and normal liver samples.
- Immunohistochemical staining of tissue microarrays for target validation.
- Flow cytometry and western blot analysis to confirm target expression in HCC cell lines.
- Development and testing of novel antibody-based positron emission tomography (immunoPET) agents in murine models.
Main Results:
- Identification of several HCC-selective plasma membrane molecules.
- Validation of target selectivity through immunohistochemistry and cell line analysis.
- Successful development and preclinical testing of immunoPET agents targeting identified HCC markers.
- Demonstration of a multimodal pipeline for identifying and validating tumor-selective targets.
Conclusions:
- A multimodal approach effectively identifies and validates HCC-selective surface targets.
- Validated targets can be utilized for developing novel HCC-specific diagnostic and therapeutic radiopharmaceuticals.
- This strategy supports the advancement of precision oncology for hepatocellular carcinoma treatment.
- The identified targets hold potential for creating advanced radiotheranostic agents.
