Revolutionizing Therapeutics with RNA delivery platform : FlashRNA®

FlashRNA® is a non-integrative viral RNA delivery system allowing transient, efficient and safe gene expression which marks a significant milestone in the field of gene and cell therapy. Its non-toxic, non-immunogenic nature, high cellular uptake, and ability to transduce various RNAs showcase its value. This versatile technology, compliant with Good Manufacturing Practices (cGMPs), creates new therapeutic possibilities with enhanced safety features.

Recent advances in gene and cell therapy highlight the complex challenge of delivering therapeutic agents tailored to specific diseases. While DNA-based therapies have gained notoriety (ILV & AAV administration), the focus is now shifting to RNA-based therapies as a versatile alternative, that can target undruggable pathways(1). RNA therapeutics are a rapidly expanding class of medicines that will change the standard of care for many diseases and bring personalized medicine up to date. It is a disruptive therapeutic technology. This shift is driven by the need for a universal delivery tool that can be used for a wide range of applications while minimising risks, particularly genotoxicity. RNA-based therapies thus offer the flexibility required to target specific therapeutic applications requiring transient expression, whether for gene modification, regenerative medicine or immuno-oncology. This field of application encompasses triggering cellular processes, modifying genetic sequences or directing cells along specific differentiation pathways. Once optimised, these genetically modified cells become effective medicines, illustrating an innovative approach in the landscape of next-generation 2.0 therapies(2).

To overcome the safety limitations of AAV and lentiviral vectors, a revolutionary RNA delivery tool called FlashRNA® (formerly LentiFlash®) has been developed, providing safe and effective solution. This innovative bacteriophage-lentivirus chimera combines the RNA packaging system of bacteriophages with the structure of lentiviral particles, delivering several species of RNA in a single FlashRNA® particle(3). This not only protects the RNAs from degradation but also enables them to be delivered efficiently into the cell cytoplasm, thanks to VSV-G pseudotyping that ensures broad tropism or alternatively use of particular pseudotyping to target specific cell types. RNA bioavailability and protein translation is fast, without any risk of integration into the host genome. As a result, transduction by FlashRNA® does not result in GMO generation. Furthermore, the packaged RNAs are devoid of any lentiviral sequence and are from biological origin: indeed, as they are synthesized in human producer cells, they undergo all cellular post-transcriptional modifications, thus preventing adverse immune responses upon delivery into target cells.

To highlight the three innovative therapeutic strategies – gene editing, regenerative medicine and immuno-oncology – we invite you to discover concrete examples that illustrate each of these approaches.

GENE EDITING 

FlashRNA® delivers the CRISPR-Cas9 machinery in a single step in many types of primary cells, from stem cells to immune cells, as well as human induced pluripotent stem cells (iPSCs). This approach guarantees efficient gene editing without compromising cell viability or the differentiation capacity of stem cells. FlashRNA® was successfully used to generate knock-out and gene conversion in iPSCs in vitro (4) as well as to perform base editing in vivo (5).

REGENERATIVE THERAPY 

Within the current Theralymph clinical project, FlashRNA® is being used to express two different mRNAs to restore lymphatic function in patients suffering from secondary lymphedema. Preclinical results obtained in mouse models demonstrated the potential to abolish lymphedema when 2 mRNAs are co-delivered, enabling lymphatic flow to be restored. This paves the way for a Phase I/II clinical trial, that will be conduced next year at the Toulouse University Hospital, France.

IMMUNO-ONCOLOGY 

Through direct in vivo delivery of tumour antigen-carrying FlashRNA® or FlashRNA® modified-Dendritic cells, the immune system can then efficiently block tumour development in wild-type mice. This method allows the expression of multiple antigens, enhancing specific immune responses and minimising the risk of tumour relapse.

Flash BioSolutions

About FlashRNA® :

FlashRNA® is a non-integrative viral RNA delivery system allowing transient, efficient and safe gene expression which marks a significant milestone in the field of gene and cell therapy. This valuable technology is demonstrated by its non-toxic, non-immunogenic nature, combined with a high uptake due to its viral characteristics and its possibility to deliver multiple distinct RNAs. Its versatility, coupled with a production platform compliant with current cGMPs, opens new avenues for therapeutic approaches, providing additional safety considerations compared to other methods.

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Alexandra ICHE

Gene Engineering Project Manager 

Flash Therapeutics expert since 2006

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Poster
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From research to clinical application; successful & reproducible process for lentiviral vector manufacturing

Discover the scientific poster presented at the Advanced Therapies 2023 congress in London, about our manufacturing process through a continuum from the R&D to the clinic.

Gene and cell therapies have emerged as a viable medical and commercial modality for treating not only genetic diseases but also cancers. As such, Flash Therapeutics, a French biotech company, is positioned as a global CDMO expert in the manufacturing of lentiviral vectors used for gene addition or gene editing therapeutic approaches. Our proprietary lentiviral platform includes both regular integrative lentiviral vectors (iLV) and our proprietary non-integrative LentiFlash® technology. Through a continuum from research to clinic, we offer high quality vector manufacturing and personalized support.

Our patented production and purification process guarantees the same specifications from R&D to GMP grade, thus ensuring the success of your clinical study. Our successful and reproducible continuum includes 4 bio-production steps from small scale research to large scale GMP. These stages are supported by regulatory documentation based on quality controls, equipment qualification and certificates of analysis. Each of these stages is punctuated by regular meetings with the scientific team, global account manager and project manager.

Since 2005, Flash Therapeutics has produced more than 9,000 custom batches including 50 large scale (30 to 200mL at about 1E9 IG/mL). A detailed analytical characterization of product titer, process impurities & quality attributes was established, showing the removal of 99% of proteins and DNA impurities. The methodology allows an efficient gene transfer even into delicate or hard-to-transduce cells, while preserving the viability and phenotypic profile of transduced cells.

Combining know-how and high-tech processes, Flash Therapeutics provides small, medium and large-scale production of high-quality lentiviral vectors with the same attribute specifications throughout the process from small scale R&D grade to large scale GMP grade.

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Lucille Lamouroux, PhD

Project Manager 

Flash Therapeutics expert since 2012

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Poster
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What are the differences between transfection and transduction ?

Two different letters and what else ?

Everyone uses the word Transfection to designate the deliberate introduction of genetic material (DNA, RNA) into eukaryotic cells. The term is often used whatever the delivery tool but actually it should not!

Gene transfer using a chemical carrier is called transfection

Transfection of DNA or RNA into animal cells has to be restricted to non-viral systems that transiently open pores into the cell membrane to permit the entry of nucleic acids.

It non only includes chemical-based products such as calcium phosphate, cationic polymers (PEI) and liposomes, but also non-chemical methods such as electroporation.

Transduction means gene transfer using viral vector carriers

If DNA or RNA is introduced into cells by using viral vector carriers, then the technique is called Transduction, and the resulting cells are said to be transduced!

It includes virus-based vectors such as lentiviral vectors, adeno-associated viruses (AAV) and adenoviruses.

Transfection and transduction methods are optimal for different types of experiments

Both transfection and transduction can lead to a transient or stable expression of DNA into cells, depending on the method or the viral tool.

If a stable expression is required to maintain the foreign nucleic acid sequence in the genome of the cells and its daughter cells, then there are two options:

  • Co-transfect cells with a marker gene, which gives the cell some selectable advantage, such as resistance or labeling
  • Use a lentiviral vector to transduce target cells which naturally integrates its DNA into the host cell genome in a random manner.

On the other hand, transfection or transduction of RNA is always transient.

Transfection is efficient on adherent immortalized cells but primary and stem cells require transduction.

A limitation of the transfection approach lies in the toxicity of transfection for delicate cells, and its suspected effect on the expression of other genes or proteins.

For more information on the comparison between those two techniques and the bases necessary to know lentiviral vectors better, you can visit our Lentiviral vectors page.