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Taming the insulin gene – with nanoelectroporation

A woman in lab coat and gloves is working on an experiment. Photo.
Frida Ekstrand et al recently published the study “Nanopore Electroporation: A New Delivery Method Within the Field of Epigenetic Editing”. Photo: Kennet Ruona

More than half a billion people are affected by type 2 diabetes – and the number is growing. In a recent publication in the journal Small, researchers explain how nanoelectroporation can be used to deliver molecules that can affect insulin expression through epigenetic means, potentially leading to new treatments.

Genetics cannot explain type 2 diabetes, and epigenetics (i.e. chemical modifications of the DNA not affecting the genetic code) is expected to play a key role. The group of Professor Charlotte Ling in Malmö has identified a series of epigenetic differences between people with type 2 diabetes and people without the disease. It is not fully clear whether these changes are the cause or the consequence of the disease. To assess this, they need better methods to implement specific epigenetic changes and study the corresponding effects on cells. This study is an important step toward achieving this. In a proof-of-concept study, the authors deliver DNA plasmids coding for CRISPRi and show downregulation of the insulin gene in beta cells.

How would you describe this study?

“This study explores the possibility of achieving a change in insulin gene expression by delivering a set of certain DNA molecules (CRISPRi plasmids) to beta cells (insulin-producing cells in the pancreas) using nanoelectroporation. By utilising gentle electric pulses and nanoporous membranes, we were able to deliver custom-designed DNA molecules to beta cells, where they yielded a desired reduction of insulin gene expression. This is a first step towards successful epigenetic editing in beta cells using nanoelectroporation,” says Frida Ekstrand, one of the NanoLundian PhD students who defended her doctoral thesis last year. 

Understanding more about the disease mechanisms is crucial for developing better and more targeted treatments.

“Epigenetics is thought to play a key role in type 2 diabetes. Enabling the changing of specific epigenetic signatures and studying the corresponding effect could expand our knowledge about the disease and potentially lead to new treatments. Type 2 diabetes is a group of diseases that affects the lives of over half a billion people every day, a number that is expected to increase further. Understanding more about the disease mechanisms is crucial for developing better and more targeted treatments. Our study suggests a new delivery method for studying these mechanisms in beta cells.”

Previously, delivering these large DNA molecules has only been achieved using empty virus shells. 

“Using viruses poses several safety concerns, and they cannot be used for therapeutic purposes. We show in this study that we can alter gene expression using a non-viral method, which could open the possibility for therapeutic applications in type 2 diabetes,” says Frida Ekstrand.

What is the most important thing you have learned from this work?

“That a specific change in beta cell gene expression can be achieved using the non-viral method nanoelectroporation.” 

Something surprising in the results was the significant effect that was observed: 

“These cells express very high levels of insulin, and we estimate that we managed to affect about 25% of the cells. That a lower gene expression in only 25% of the cells, with the rest expressing normal levels of insulin, was significant was a bit surprising.”  

The main conclusions of the study are that the authors showed that a significant downregulation of insulin gene expression through epigenetic modifications could be achieved by using nanoelectroporation.

“This is the first step towards assessing the causal effects of epigenetic marks in type 2 diabetes using nanotechnology. In the future, one could imagine targeting these effects for therapeutic purposes,” says Christelle Prinz, Professor of biophysics at the Division of solid state physics. 

“I look forward to further developing this nanotechnology-based method for editing of specific epigenetic marks together with Professor Prinz’s group,” says Professor Charlotte Ling.

Facts

Type 2 diabetes 

Happens when the body cannot secrete and/or use insulin correctly, and sugar builds up in the blood. This can damage the eyes, kidneys, nerves, and heart. There is currently no cure. Losing weight, eating well, and exercising can help manage blood sugar, but if this isn’t enough, diabetes therapies or insulin therapy may help.

Insulin

A hormone made in the beta cells in the pancreas that helps sugar enter the cells in the rest of the body.

CRISPRi

Clustered regularly interspaced short palindromic repeats interference, a genetic perturbation technique that makes it possible to repress sequence-specific gene expression in certain cells.

DNA plasmids

A small, extrachromosomal DNA molecule, separated from chromosomal DNA, that can replicate independently.