Johns Hopkins Medicine scientists report that they have created tiny nanoparticles that can penetrate cells and make proteins and proteins in even the most difficult cells, based on a genetic correction tool called CRISPR.
If their genes – derived from biodegradable polymers – pass the next lab test, they could provide a way to transfer large complex cells to well-selected target cells.
Accounts of their work appeared in the December 6 issue of Science Advances.
“Many drugs spread unnecessarily throughout the body and do not target specific cells,” said biomedical engineer Jordan Green, Ph.D., lead researcher on the same team. “Some drugs, such as antibiotics, target cell receptors, but we don’t have an adequate mechanism for the immediate transfer of antibiotics to cells, where treatment has the best chance of working well and poorly. Negative impact. Small.”
Many scientists and scientists have found a better mode of transport for treatment, says Green, professor of biomedical engineering, ophthalmology, oncology, neurosurgery, engineering and mechanical engineering, chemistry, and biomolecular engineering at Johns Hopkins School of Medicine and a Bloomberg member ~ Kimmel Cancer Immunotherapy Institute at Johns Hopkins.
Many markets on the market use dispersed strains – known for their ability to “pass” cells immediately – to provide treatment, although these types of non-infectious transplants may be responses to immune system development. Other treatments designed for infected blood cells, for example, are more complex and require removing the patient’s blood and then electrically opening the pores and cell membranes to allow them to enter.
The nano-sized containers produced by Green and his team at Johns Hopkins borrow ideas from the properties of bacteria, many of which are nearly spherical in shape and have negative and positive charges. With complete remission, the virus can move closer to the cell. This is not the case with many bioactive drugs, which contain high levels of proteins and nucleic acids that are often needed to regenerate cells.
To solve this problem, graduate student Yuan Rui developed a new polymer composite material. Polymer is a general term for the composition of many molecules. To create a polymer, Rui is tied together – like a tree branch – into four molecular elements that break down and dissolve in water over time. Molecules have both positive and negative charges.
Using the balance of positive and negative charges, the molecule moves and attracts according to its charge on its hydrogen atom and surrounding bacterial treatment system. This results in nanostructures containing bioavailable therapeutic agents.
The positively charged nanoparticle cell attaches to the skin cell and the cell fills with cell turns called endosomes.
Once inside, the rupturing of the nanoparticle membrane opens the damaged endosomes and polymers, causing the drug to function inside the cell.
To test their product, Rui made small protein nanocontainers and delivered them to mouse kidney cells in a traditional dish. It attaches a green fluorescent marker to a small protein and detects it spread across multiple cells, indicating that the protein is well absorbed.
Rui then collected another protein: human immunoglobulin, a drug commonly used to strengthen the immune system as well as a model for immunosuppression. Currently, he found that 90 percent of the kidney cells he treated were treated with immunoglobulin, a green fluorescent marker.
“When nanoparticles enter cells, they are usually trapped in endosomes, which degrades their content, but our study shows that protein packets are distributed across multiple cells and do not block them with endosomes,” he said. Rui.
For an even bigger challenge, Rui created a nano package containing a CRISPR-based protein and a nucleic acid that can suppress green light signals or brighten red blood cells when CRISPR is activated from parts of the genome. The researchers found that genetic editing worked to remove genes from about 77 percent of laboratory cells and to add or repair genes in about 4 percent of cases of cells.
Rui said, “It’s great when you consider that, through other genetic editing techniques, you can achieve less genetic reduction results in less than 10 percent.” CRISPR-based therapies can improve the quality of treatment because of their ability to focus on specific genetic defects. Several CRISPR therapies are being tested in clinical trials.
In the latest experiment, Rui and his colleagues injected brain tumor cells into mice. It stimulated the nanoreceptors of the gene directly into the mouse brain and checked their cells for a red light indicating successful cell formation. He found red cancer cells a few millimeters from the injection site.
Rui said, “When I started this project five years ago, scientists did not think that other non-bacterial drugs would be used to inject these drugs into cells.” The development of new technologies can help us to better understand this disease, but also for the development of new drugs. “
Rui and Green wanted to stabilize the nanotubes so they could be injected into the bloodstream and focus on cells that have some genetic markers.
Scientists are applying for a license for this project.
Funding from the National Science Foundation, Johns Hopkins University School of Medicine, National Institutes of Health (R01CA228133, R01EB022148, and P30 EY001765), Bloomberg ~ Kimmel Institute for Cancer Immunotherapy at Johns Hopkins and Free Research in Prevention. The Catalyst Award blinded James and Carole.
Other scientists who contributed to the project include David Wilson, John Choi, Mahita Varanasi, Katie Sanders, Johan Karlsson, and Michael Lim of Johns Hopkins.