Overview of Hydroporation-Based Gene Delivery

Gene delivery refers to the process of effectively transporting exogenous genes or other biomacromolecules into target cells and expressing or functioning in them. Gene delivery is the core technology in the fields of gene therapy, gene vaccines, and gene function research, and it is also an important means of biomedical research. However, the currently commonly used gene delivery methods, such as viral vectors, liposomes, and nanoparticles, all have certain limitations, such as problems in safety, efficiency, stability, and cost. Therefore, developing new gene delivery methods to improve the effect and application range of gene delivery is an urgent need and a research hotspot in the field of biomedicine.

Hydroporation is a new gene delivery method that uses high-pressure water flow to create micropores outside the cell, allowing foreign genes or other biomacromolecules to enter the cell. Hydroporation has some advantages, such as no carrier, no immune reaction, no toxic side effects, simple operation, and low cost. Hydroporation can be used for gene delivery in superficial tissues such as skin and muscle, as well as in solid organs such as the liver. Hydroporation has potential application value in gene therapy, gene vaccines, gene function research, and so on.

The Method, Principle, and Mechanism of Hydroporation

Hydroporation is a novel method of gene delivery. The principle is to use high-pressure water flow to generate shear force and tension on the cell membrane, causing the cell membrane to deform and rupture, thereby forming micropores. These micropores allow foreign genes or other biomacromolecules to enter the cell through the cell membrane. These micropores usually close automatically within seconds, restoring the integrity of the cell membrane.

The process of Hydroporation consists of five steps. First, prepare a solution containing exogenous genes or other biomacromolecules, and adjust the volume, concentration, pH value, temperature, and other parameters of the solution according to different goals and needs. Second, the solution is filled into a syringe or other device connected to a pump or source of compressed gas that creates a high-pressure stream of water. Third, aim the nozzle of the syringe or other device at the tissue surface where the target cells are located, maintaining a certain distance and angle. Fourth, start the pump or compressed gas source, spray the solution onto the tissue surface in the form of high-pressure water flow, and control the pressure, speed, time, and other parameters of the water flow. Finally, stop spraying and observe the distribution and expression of genes or other biomacromolecules on the tissue surface and in cells.

The effect of hydroporation is affected by many factors, mainly including water flow parameters, solution parameters, tissue parameters, and cell parameters. These factors determine the size and duration of the shear force and tension generated by the water flow on the cell membrane, the distribution and stability of exogenous genes or other biomacromolecules in the water flow, the diffusion and resistance of the water flow in the tissue, and the effect of the cell membrane on the water flow. Sensitivity and reactivity to shear and tension forces generated. There may be interactions and influences among different factors, so experimental conditions need to be optimized according to different goals and the need to achieve the best gene delivery effect.

The molecular mechanism and cellular effects of hydroporation are not completely clear, but some studies have shown that hydroporation can induce some signaling pathways on the cell membrane, such as calcium signaling, and MAPK signaling, thereby affecting cell survival, proliferation, differentiation, apoptosis, and other processes. Hydroporation can also alter gene expression levels and patterns within cells, thereby affecting cell function and properties.

Progress of Hydroporation in Gene Delivery

Hydroporation, as a new gene delivery method, has made some progress and achievements in gene therapy, gene vaccines, gene function research, and so on. For example, Hydroporation can be used to transfect anticancer genes, antiviral genes, and immune regulatory genes into the liver cells of mice or rats so as to achieve the treatment or prevention of diseases such as liver cancer, hepatitis B, and liver fibrosis. Hydroporation can also be used to transfect fluorescent genes, reporter genes, knockout genes, and so on. into the skin or muscle cells of mice or rats, so as to realize research on cell function, signaling pathways, epigenetics, etc. Hydroporation can also be used to transfect DNA or RNA vaccines into the skin or muscle cells of mice or rats, thereby achieving immune protection against infectious diseases such as influenza, AIDS, and tuberculosis.

Compared with other gene delivery methods, Hydroporation has some advantages, such as no carrier, no immune reaction, no toxic side effects, simple operation, and low cost. These advantages make Hydroporation more suitable for some special application scenarios, such as gene delivery of macromolecules or multiple molecules, large-scale or high-frequency gene delivery, and acute or temporary gene delivery. However, Hydroporation also has some disadvantages and limitations, such as unstable efficiency, greater cell damage, lower tissue selectivity, a smaller transfection range, and a shorter expression duration. These shortcomings make it difficult for Hyroporation to be applied to some gene delivery scenarios that require high efficiency, low damage, high selectivity, wide coverage, and long-term expression, such as the treatment of chronic or genetic diseases and transfection of internal organs or deep tissues.

Table 1. Comparison of Hydroporation with Other Gene Delivery Methods

In order to overcome the limitations of hydroporation in the field of gene delivery, some researchers have proposed some innovative solutions, such as optimizing experimental conditions, combining other methods, developing new devices and technologies, and conducting in-depth research on mechanisms and effects. These solutions aim to improve the efficiency, stability, selectivity, coverage, and persistence of hydroporation; reduce the damage, toxicity, side effects, and cost of hydroporation; expand the application range and potential of hydroporation; and reveal the impact of hydroporation on cell function, characteristics, and regulation. These solutions provide new ideas and methods for the development and progress of Hyroporation in the field of gene delivery.

Hydroporation for Disease Treatment

Hydroporation is a novel gene delivery method that uses a high-pressure water jet to create pores in the cell membrane and allow the entry of exogenous genes or other biomolecules into the cell. Hydroporation has been applied for the treatment of various types of diseases, such as genetic, infectious, and tumorous diseases.

Hydroporation has been used to deliver therapeutic genes or vaccines to different organs or tissues in animal models of various diseases. For example, hydroporation has been used to deliver anti-cancer genes to the liver of mice with hepatocellular carcinoma, anti-viral genes to the liver of mice with hepatitis B virus infection, immune-regulatory genes to the skin of mice with allergic contact dermatitis, DNA vaccines to the muscle of mice with influenza virus infection, and RNA vaccines to the skin of mice with tuberculosis infection. The data from these studies have shown that hydroporation can achieve high efficiency, high specificity, high expression, and high therapeutic efficacy of gene delivery in vivo. Hydroporation has some advantages over other gene delivery methods, such as no need for carriers, no immune response, no toxicity, simple operation, low cost, high efficiency, and high cell viability. However, hydroporation also has some limitations and risks, such as unstable efficiency, high cell damage, low tissue selectivity, small delivery range, and short-expression duration. Moreover, hydroporation may cause some adverse effects, such as bleeding, inflammation, infection, tissue damage, or organ failure. Therefore, hydroporation should be carefully optimized and evaluated for each specific application and target.

Hydroporation has not yet been tested in human clinical trials for disease treatment. However, some preclinical studies have suggested that hydroporation may be a promising technique for human gene therapy or vaccination. For example, a study has shown that hydroporation can deliver a therapeutic gene to the liver of non-human primates with minimal invasiveness and toxicity. Another study has shown that hydroporation can deliver a DNA vaccine to the skin of non-human primates with high immunogenicity and protection. These studies indicate that hydroporation may be feasible and safe for human use in the future.

Based on the above, hydroporation can achieve high efficiency, high specificity, high expression, and high therapeutic efficacy of gene delivery in vivo, without the need for carriers, immune response, or toxicity. Hydroporation can deliver different types of genes or biomolecules, such as anti-cancer genes, anti-viral genes, immune-regulatory genes, DNA vaccines, or RNA vaccines, to different organs or tissues, such as the liver, skin, or muscle. Hydroporation can overcome some physical barriers that limit other gene delivery methods, such as spatial barriers, structural barriers, or plasma membrane barriers. Hydroporation can be optimized and evaluated for each specific application and target by adjusting the water jet parameters and solution parameters. Hydroporation may be feasible and safe for human use in the future, as suggested by some preclinical studies in non-human primates. In conclusion, hydroporation is a promising technique for gene therapy or vaccination that may have significant impacts on the prevention and treatment of various diseases in the future.

References

1. Yudin MA, et al. Study of the Efficiency of the Hydroporation for Delivery of Plasmid DNA to the Cells on the Model of Toxic Neuropathy. Bull Exp Biol Med. 2018 Apr;164(6):798-802.
2. Zhang G, et al. Hydroporation as the mechanism of hydrodynamic delivery. Gene Ther. 2004 Apr;11(8):675-82.
3. Chakrabarty P, et al. Microfluidic mechanoporation for cellular delivery and analysis. Mater Today Bio. 2021 Dec 20;13:100193.
4. André FM, et al. Variability of naked DNA expression after direct local injection: the influence of the injection speed. Gene Ther. 2006 Dec;13(23):1619-27.
5. Kizer ME, et al. Hydroporator: a hydrodynamic cell membrane perforator for high-throughput vector-free nanomaterial intracellular delivery and DNA origami biostability evaluation. Lab Chip. 2019 May 14;19(10):1747-1754.
6. Magre M, et al. s-Block Metal Catalysts for the Hydroboration of Unsaturated Bonds. Chem Rev. 2022 May 11;122(9):8261-8312.
7. Suda T, et al. Hydrodynamic gene delivery: its principles and applications. Mol Ther. 2007 Dec;15(12):2063-9.
8. Liu F, et al. Hydrodynamics-based transfection in animals by systemic administration of plasmid DNA. Gene Ther. 1999 Aug;6(7):1258-66.
9. Zhang G, et al. Efficient expression of naked DNA delivered intraarterially to limb muscles of nonhuman primates. Hum Gene Ther. 2004 Feb;15(2):131-41.