Beijing Institute of Technology, Dec. 28th, 2020:Recently, Huang Yuanyu's research group from the Advanced Research Institute of Multidisciplinary Science of Beijing Institute of Technology (BIT) has made important research progress in nucleic acid drug delivery and tumor immunotherapy. The research results were published in the top international journal “Nano Today” (impact factor 16.907) in the field of nanotechnology and biomedicine as “Rolling microneedle electrode array (RoMEA) empowered nucleic acid delivery and cancer immunotherapy”. The first authors of this paper are PhD student Yang Tongren from School of Life Sciences/Advanced Research Institute of Multidisciplinary Science of BIT and Dr. Huang Dong from Peking University. The corresponding authors are Researcher Huang Yuanyu of BIT and Professor Li Zhihong of PKU.

Nucleic acid drugs based on small interfering RNA (siRNA) have been continuously approved for marketing in recent years and have become a representative technology leading the third wave of pharmaceuticals. The key issue in the development of siRNA drugs is how to achieve efficient enrichment and endocytosis, rapid access to the cytoplasm, and efficient and long-term silencing of target gene expression. This process requires research and development of efficient and safe drug delivery technologies. At present, a slew of delivery vehicles have the problems such as low administration efficiency, poor safety, high cost, and lack of universality.

Figure 1 Schematic diagram of RoMEA and the research ideas

In 2018, Huang Yuanyu’s group and Li Zhihong’s group cooperated, proposing and verifying a rolling microneedle-assisted flexible electroporation chip-mediated siRNA transdermal delivery strategy in vivo (Theranostics 2018; 8(9):2361-2376). This strategy combines the microneedle roller and the flexible finger electrode chip (FIEA) in the field of medical cosmetology. First, the microneedle roller is used to create microchannels in the mouse skin tissue to form liquid microelectrodes, and then a flexible electroporation chip is used to give a pulsed electric field, promoting transdermal delivery of nucleic acid drug molecules. Furthermore, the researchers conceived to "two-in-one" a rolling microneedle and an electroporation chip, and designed a rolling microneedle electrode array (RoMEA, Figure 1). The outstanding advantage of this device is that it applies a synchronous electric field while rolling, which can support large-area electroporation of living tissue on the surface of irregular living tissue. On the same time, the device boasts the advantages of low invasiveness, convenient operation, safety, low manufacturing cost, and easy mass production.

Figure 2 Research on RoMEA-mediated nucleic acid delivery in vivo

Researchers used RoMEA to deliver plasmids expressing red fluorescent protein (RFP) or Cy5-labeled siRNA, or siRNA that inhibits the expression of SCD1 gene, exploring the parameters of RoMEA's nucleic acid delivery in vivo (Figure 2). The results showed that when the voltage applied by RoMEA was in the range of 30V-90V, the protein of RFP was significantly expressed, and the protein expression intensity of RFP reached the peak under the voltage of 50V (Figure 2c). And when the voltage applied by RoMEA was 50V, it significantly increased the retention time of Cy5-siRNA in the legs and tumors of mice (Figure 2d, e). Similarly, the authors found that siRNA has a good gene suppression efficiency when the voltage applied by RoMEA is 50V (Figure 2f, g). Under these conditions, electroporation CpG2395 (immune adjuvant) can effectively stimulate the secretion of IL-6 and IFN-g in mice (Figure 2h). Therefore, RoMEA can achieve local and efficient nucleic acid transfection efficiency at a voltage of 50V.

Figure 3 RoMEA-mediated immunotherapy of B16-F10 melanoma

Blocking the PD-L1/PD-1 immune checkpoint pathway is the most representative tumor immunotherapy approach in clinical treatment. Combined with the optimal parameters of RoMEA, the researchers further selected siRNA (siPD-L1) that inhibits PD-L1 expression and monoclonal antibody (PD-1 mAb) that antagonizes PD-1 expression in the B16-F10 melanoma mouse model for research (Figure 3a, b). The treatment is started when the tumor is small (simulating early clinical tumor, Figure 3c-i) and when the tumor is large (simulating mid-clinical tumor, Figure 3j-l). The results show that siPD-L1 alone or in combination with PD-1 mAb can both effectively inhibit tumor growth and prolong the life cycle of animals. At the same time, the results of flow cytometry showed that the number of CD4+ T cells and CD8+ T cells in the tumor microenvironment increased significantly, confirming the mechanism and effect of the PD-L1/PD-1 pathway.

The rolling microneedle electrode array (RoMEA) designed by this work is simple to process and has strong versatility, which lays a foundation for the application and development of various therapeutic agents (not limited to nucleic acid drugs) in the future, showing a good practical prospect.

Attached introduction to the author:

Huang Yuanyu is a researcher, project leader and doctoral supervisor of BIT. Research interests mainly focus on nucleic acid technology and drugs, including the design and screening of nucleic acid drugs (such as siRNA, mRNA, etc.), drug delivery, and innovative diagnosis and treatment technologies for major diseases. Currently, he has published more than 50 SCI papers in journals such as Nano Today, Adv Funct Mater, Nano Lett, Biotechnol Adv, etc. Among them, there are more than 30 first or corresponding author papers, including 13 IF>10 papers and 3 ESI highly cited papers. There are 6 monographs, 1 Chinese textbook; 6 patent applications, including 2 authorized PCT patents. He has presided over more than 10 projects including the National Natural Science Foundation of China. Selected as Beijing Science and Technology Star (2020), Beijing Association for Science and Technology Advanced Worker (2020), and Beijing Association for Science and Technology Young Talents (2019), and won the "Future Star" Award from the Chinese American Society of Nanomedicine and Nano Biotechnology (2019), academic newcomers of the Ministry of Education and other honors. He is the secretary-general and director of the Nanobiology Branch of the Chinese Biophysical Society, and a director or member of the other 4 societies; the associate editor of Advances (New Cooperative Journal of Wiley), and the editorial board member of many journals such as Chinese Chemical Letters.

Paper details:

Tongren Yang1, Dong Huang1, Chunhui Li, Deyao Zhao, Junshi Li, Mengjie Zhang, Yufeng Chen, Qining Wang, Zicai Liang, Xing-Jie Liang, Zhihong Li∗, Yuanyu Huang∗. Rolling microneedle electrode array (RoMEA) empowered nucleic acid delivery and cancer immunotherapy. Nano Today 2021; 36: 101017. doi: 10.1016/j.nantod.2020.101017.

Teacher Homepage:http://arims.bit.edu.cn/xzdw/qnggjs/tbyjy/178349.htm

Paper link:https://doi.org/10.1016/j.nantod.2020.101017