BIT Makes Progress in Gene Editing Treatment of Tumors

News Source: Advanced Research Institute of Multidisciplinary Science

Editor: News Agency of BIT

Translator: Li Xinxin, News Agency of BIT


Figure 1 Schematic diagram of iLP181/psgPLK1 research and in vitro and in vivo effects

Beijing Institute of Technology, July 27th, 2021: Recently, Huang Yuanyu’s research group from Advanced Research Institute of Multidisciplinary Science, Beijing Institute of Technology (BIT), has made important research progress in the delivery of CRISPR / Cas9 gene editing element mediated by ionizable lipid and its efficacy in tumor therapy. The research results were published in the top journal Bioactive Materials (IF 2020="14.593) in the field of biomedicine as “Ionizable lipid-assisted efficient hepatic delivery of gene editing elements for oncotherapy”. The first author of the paper includes Li Chunhui and Yang Tongren, doctor students of the School of Life Science / Advanced Research Institute of Multidisciplinary Science, BIT; Weng Yuhua, associated vice researcher  of Institute of Engineering Medicine, BIT; and the corresponding author is researcher Huang Yuanyu.

The development of CRISPR / Cas gene editing technology has entered a promising era, which provides a new treatment for incurable diseases at present. To a certain extent, the emergence of CRISPR / Cas reflects the major advance in human gene editing technology, and provides an effective and powerful platform for further understanding the gene function, potential biological or pathological mechanism of various diseases, and ultimately the development of new therapeutic schemes. However, CRISPR / Cas gene editing system needs to enter the cell to be effective. How to enter the cell efficiently and further escape from the endosome to the cytoplasm is a very important and challenging scientific problem. In terms of the delivery of biomolecules, ionizable lipid nanoparticles (iLNP) are the most promising nucleic acid delivery vehicles. The first siRNA drug Onpattro® approved for market in the world, and the new coronary pneumonia mRNA (mRNA-1273, BNT162b2) vaccine developed by Moderna and BioNTechis are both liposome preparations. At the same time, iLNP is also the most effective delivery system of CRISPR / Cas, and has been used to deliver gene editing elements to cancer cells, liver cells, lung and brain tissues.

In this study, researchers have developed a new type of ionizable lipid nanoparticles (iLNP) called iLP181, and constructed a sgRNA plasmid named Cas9-sgPLK1 or psgPLK1, which simultaneously expresses Cas9 protein and targets PLK1 (polo-like kinase 1) gene. Besides, they have researched the efficiency and gene editing of Cas9-sgPLK1 delivered by iLP181 in vitro and in vivo. Finally, the therapeutic effect and safety of iLP181 / psgPLK1 preparation in tumor-bearing mice are evaluated (Figure 1).

It has been proved that when the pKa value of the delivery carrier system is between 6.2 and 6.5, it can ionize rapidly in response to the change of the pH value of the endosomes, thus interact with the anionic lipids (such as phosphatidylserine) in the endosomal membrane components, destroying the stability of the endosomes, promoting nucleic acid molecules to escape from endosomes to the cytoplasm, and releasing the nucleic acid molecules at the same time. This study proves that the pKa value of the iLP181 system is 6.43, which meets the requirements of ionizable liposomes (Figure 1B). The co-localization analysis of the fluorescent signal in the cell proves that the iLP181/psgPLK1 shows efficient endosomal escape after entering the cell 3 hours (Figure 1C). Furthermore, the researchers evaluate the long-acting gene editing of iLP181/psgPLK1 in vitro, and the 32% gene editing efficiency can still be achieved on the 7th day after transfection of HepG2-Luc cells into iLP181/psgPLK1 (Figure 1D). In order to determine whether iLP181/psgPLK1 has longer-term stability, the researchers also observe the long-term stability of iLP181/psgPLK1 in different buffer solutions. The data proves that iLP181/psgPLK1 shows good stability in either PBS buffer solution or buffer solution containing 10% human serum for up to 14 days (Figure 1E and F).

Subsequently, the researchers inject the iLP181/psgPLK1 lipid nanocomposite into mice via the tail vein to evaluate the effect of its distribution in the body. The data shows that iLP181/psgPLK1 mainly accumulates in the tumor site in 2-5 days after administration (Figure 1G), indicating that iLP181 has good quality of tumor targeting. At the same time, this paper also studies the in vivo safety of the liposome nucleic acid system. Pathological slices, serum biochemistry and other indicators shows that the system has good in vivo safety.

In conclusion, this study proves that iLP181 can efficiently deliver CRISPR / Cas system to cells and tumor tissues, achieve efficient gene editing, and has good safety. This study not only provides an efficient and safe lipid nanocarrier for CRISPR /Cas system (and even other biologically active molecules), but also provides a potential treatment plan for DNA editing of tumor genes. The research results above have been submitted for patent application recently.

Attached brief introduction to the author:

Huang Yuanyu, researcher, project leader and doctoral supervisor of BIT. Research interests mainly focus on nucleic acid technology and drugs/ vaccines. He has published more than 40 papers as the corresponding/first author in Signal Transduct Target Ther, Nano Lett, Nano Today, Adv Funct Mater and so on, and 20 papers with IF>10; published more than 70 papers in total, wrote 9 monograph chapters, and applied for 12 Patents, including 2 authorized PCT patents and 1 pending PCT patent. He has successively presided more than 10 projects, including projects from the National Natural Science Foundation of China. He was selected as a member of Beijing Science and Technology Rising Star (2020) and won the "Future Star" Award (2019) of the Sino-American Society of Nanomedicine and Nanobiotechnology and other honors. He is the secretary-general of the Nanobiology Branch of the Chinese Biophysical Society, and director or committee member of the other 4 societies; he is the deputy editor-in-chief of Exploration (published by Wiley), (youth) editorial board member of Chinese Chemical Letters and other journals .

Paper details:

Chunhui Li#, Tongren Yang#, Yuhua Weng#, Mengjie Zhang, Deyao Zhao, Shuai Guo, Bo Hu, Wanxuan Shao, Xiaoxia Wang, Abid Hussain, Xing-Jie Liang, Yuanyu Huang∗. Ionizable lipid-assisted efficient hepatic delivery of gene editing elements for oncotherapy. Bioact Mater. 2021, x(x): xxxx-xxxx.

DOI: 10.1016/j.bioactmat.2021.05.051

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