Wenjing Sun, Xin You, Xinjia Zhao, Xiaoyu Zhang, Chunhui Yang, Fusheng Zhang, Jiaqi Yu, Kaiguang Yang, Jixia Wang, Fangfang Xu, Yongxin Chang, Boxin Qu, Xinmiao Zhao, Yuxuan He, Qi Wang,* Jinghua Chen,* Guangyan Qing*

Adv. Mater. 2024, DOI: 10.1002/adma.202402379

https://onlinelibrary.wiley.com/doi/10.1002/adma.202402379

  Recently, our group has made new progress in the early diagnosis of liver cancer based on circulating tumor cells (CTCs) in collaboration with Professor Wang Qi from the Second Affiliated Hospital of Dalian Medical University and Professor Chen Jinghua from the School of Life Science and Health Engineering of Jiangnan University. The cooperation team has prepared a new type of cell imprinted hydrogel material by highly selective binding of affinity ligands and target glycans, and by using the precise shape and size matching and good biocompatibility of the cell imprinted polymer, which has realized the accurate capture and non-destructive release of CTCs in peripheral blood, and is expected to promote the early diagnosis of liver cancer and single cell multi-component analysis.

  According to statistics, there are 906000 newly diagnosed liver cancer patients and 830000 deaths worldwide each year, with China accounting for 45% and 47% of cases, respectively. One important reason for the low survival rate of liver cancer is the expensive screening cost and delayed liver pain. Most patients are diagnosed in the middle and late stages, missing the optimal treatment period. Serum marker alpha fetoprotein (AFP) is an existing clinical testing method for liver cancer screening, diagnosis, recurrence risk assessment, and treatment efficacy observation. However, AFP sensitivity is weak, and there are more positive cases in patients with low AFP values.

  CTCs in peripheral blood are recognized as gold markers in liquid biopsy due to carrying genetic and epigenetic information of tumor tissue. The precise capture of CTCs is of great clinical significance for early diagnosis, disease monitoring, efficacy evaluation, and postoperative evaluation of cancer. However, there are only 0 to 3 CTCs per milliliter of blood, making the detection results susceptible to interference from the complex blood environment. In addition, CTCs are also susceptible to damage during capture and processing. Therefore, achieving precise capture and non-destructive release of CTCs is extremely challenging.

  The existing CTC capture materials are divided into negative, physical, and positive enrichment materials. Negative and physically enriched materials have inherent issues such as low purity, efficiency, and specificity of CTCs. Positive enrichment materials are based on the principle of antigen antibody biological affinity. Targeting the surface target antigens of CTCs, their specific binding antibodies are modified onto the material surface to achieve specific capture of CTCs. However, the strong binding ability of antibodies makes it difficult for CTCs to be released. In addition, the high cost and poor stability of antibodies also limit their widespread application.

  In this work, the research team developed a super strong affinity ligand His His for the sialylated glycan on the surface of CTC, and fused it with the cell imprinted polymer based on PEGDMA to prepare PP-co-AHH+hydrogel with low swelling rate, good compression performance, good reversibility, good blood and cell compatibility, strong anti fouling ability and clear cell imprinting. When 10 CTCs were evaluated, the capture efficiency of the hydrogel reached 93%. More importantly, the captured CTCs can be released using trypsin solution to maintain high cell activity, which is crucial for single-cell multi omics analysis.

  In addition, the team used the hydrogel material to conduct the capture CTC test in 100 human samples, and the AFP values of the test population's peripheral blood were all lower than 20 ng/mL, that is, the low sensitive area of AFP. The results indicate that CTCs counting has the ability to distinguish between liver cancer, cirrhosis, and healthy individuals, with an accuracy of 90%. The team further combined the results of AFP, CTCs, and natural killer cells to improve the accuracy of the test to 94% and the sensitivity to 90%, which can meet the needs of clinical testing. At the same time, the hydrogel material can be recycled for ten times, and the capture cost of each sample is only 50 yuan, far below the hospital price of 3500 yuan. In addition, this hydrogel material can also be used to obtain high-purity and highly active CTCs from the patient's blood, which is connected with the downstream single-cell multigroup analysis, helping to promote the precise treatment of liver cancer. The new cell imprinting hydrogel prepared in this work is expected to become a valuable diagnostic tool for early diagnosis of liver cancer.

  The related research, titled "Precise Capture and Dynamic Release of Circulating Life Cancer Cells with Dual histidine based Cells Imprinted Hydrogels," was recently published in Advanced Materials. The first author of this work is Sun Wenjing, a joint doctoral student from our 1824 group. The above work has been supported by projects such as the National Natural Science Foundation of China, the National Key Research and Development Program, and the Innovation Fund of our institute. (Text/Image Sun Wenjing)