Scalable synthesis of lipid nanoparticles for nucleic acid drug delivery using an isometric channel-size enlarging strategy

Lipid nanoparticles(LNPs)have emerged as highly effective delivery systems for nucleic acid-based therapeutics.However,the broad clinical translation of LNP-based drugs is hampered by the lack of robust and scalable synthesis techniques that can consistently produce formulations from early development to clinical application.In this work,we proposed a method to achieve scalable synthesis of LNPs by scaling inertial microfluidic mixers isometrically in three dimensions.Moreover,a theoretical predictive method,which controls the mixing time to be equal across different chips,is developed to ensure consistent particle size and size distribution of the synthesized LNPs.LNPs loaded with small interfering RNA(siRNA)were synthesized at different flow rates,exhibiting consistent physical properties,including particle size,size distribution and encapsulation efficiency.This work provides a practical approach for scalable synthesis of LNPs consistently,offering the potential to accelerate the transition of nucleic acid drug development into clinical application.

Noncationic polymer-assisted carrier for nucleic acid drug delivery

Nucleic acid drugs are emerging as a novel biotherapeutic modality for disease treatment,targeting nucleic acids to regulate the protein translation process and thereby facilitating disease management.They hold significant promise in biomedical applications and treatment avenues.Given their negative charge,high molecular weight,and hydrophilic properties,nucleic acid drugs require carriers to traverse multiple biological barriers and facilitate intracellular delivery.Cationic material-based carriers present an unprecedented opportunity to address these challenges through electrostatic interactions with nucleic acids.However,concerns regarding the biosafety and cytotoxic responses of cationic materials have emerged in early clinical studies.As a result,the use of non-cationic polymer carriers,by controlling or circumventing the use of cationic materials,represents a promising approach for nucleic acid delivery.In this review,we highlight various designs of non-cationic polymer carriers that go beyond the principle of electrostatic interactions,including conjugation,chemical bonding,physical crosslinking,hydrophobic interactions,and coordination bonding with nucleic acids.Additionally,we discuss strategies for enhancing the efficiency of nucleic acid delivery and therapeutic effects of non-cationic polymer carriers,focusing on targeted delivery,cellular internalization,and endosomal escape.

Lipid-nanoparticle-enabled nucleic acid therapeutics for liver disorders

Inherited genetic disorders of the liver pose a significant public health burden.Liver transplantation is often limited by the availability of donor livers and the exorbitant costs of immunosuppressive therapy.To overcome these limitations,nucleic acid therapy provides a hopeful alternative that enables gene repair,gene supplementation,and gene silencing with suitable vectors.Though viral vectors are the most efficient and preferred for gene therapy,pre-existing immunity debilitating immune responses limit their use.As a potential alternative,lipid nanoparticle-mediated vectors are being explored to deliver multiple nucleic acid forms,including pDNA,mRNA,siRNA,and proteins.Herein,we discuss the broader applications of lipid nanoparticles,from protein replacement therapy to restoring the disease mechanism through nucleic acid delivery and gene editing,as well as multiple preclinical and clinical studies as a potential alternative to liver transplantation.

Reduction-responsive nucleic acid nanocarrier-mediated miR-22 inhibition of PI3K/AKT pathway for the treatment of patient-derived tumor xenograft osteosarcoma

miRNAs are important regulators of gene expression and play key roles in the development of cancer, including osteosarcoma. During the development of osteosarcoma, the expression of miR-22 is significantly downregulated, making miR-22 as a promising therapeutic target against osteosarcoma. To design and fabricate efficient delivery carriers of miR-22 into osteosarcoma cells, a hydroxyl-rich reduction-responsive cationic polymeric nanoparticle, TGIC-CA (TC), was developed in this work, which also enhanced the therapeutic effects of Volasertib on osteosarcoma. TC was prepared by the ring-opening reaction between amino and epoxy groups by one-pot method, which had the good complexing ability with nucleic acids, reduction-responsive degradability and gene transfection performance. TC/miR-22 combined with volasertib could inhibit proliferation, migration and promote apoptosis of osteosarcoma cells in vitro. The anti-tumor mechanisms were revealed as TC/ miR-22 and volasertib could inhibit the PI3K/Akt signaling pathway synergistically. Furthermore, this strategy showed outstanding tumor suppression performance in animal models of orthotopic osteosarcoma, especially in patient-derived chemo-resistant and chemo-intolerant patient-derived xenograft (PDX) models, which reduced the risk of tumor lung metastasis and overcame drug resistance. Therefore, it has great potential for efficient treatment of metastasis and drug resistance of osteosarcoma by the strategy of localized, sustained delivery of miR-22 using the cationic nanocarriers combined with non-traditional chemotherapy drugs.

In Situ Loading and Delivery of Short Single- and Double-Stranded DNA by Supramolecular Organic Frameworks

Short DNA represents an important class of bioma-cromolecules that arewidely applied in gene therapy,editing,and modulation.However,the development of simple and reliable methods for their intracellular delivery remains a challenge.Herein,we describe that seven water-soluble,homogeneous supramole-cular organic frameworks(SOFs)with a well-defined pore size and high stability in water that can accom-plish in situ inclusion of single-stranded(ss)and double-stranded(ds)DNA(21,23,and 58 nt)and effective intracellular delivery(including two non-cancerous and six cancerous cell lines).

Assembling the RNA therapeutics toolbox

From the approval of COVID-19 mRNA vaccines to the 2023 Nobel Prize awarded for nucleoside base modifications,RNA therapeutics have entered the spotlight and are transforming drug development.While the term“RNA therapeutics”has been used in various contexts,this review focuses on treatments that utilize RNA as a component or target RNA for therapeutic effects.We summarize the latest advances in RNA-targeting tools and RNA-based technologies,including but not limited to mRNA,antisense oligos,siRNAs,small molecules and RNA editors.We focus on the mechanisms of current FDA-approved therapeutics but also provide a discussion on the upcoming workforces.The clinical utility of RNA-based therapeutics is enabled not only by the advances in RNA technologies but in conjunction with the significant improvements in chemical modifications and delivery platforms,which are also briefly discussed in the review.We summarize the latest RNA therapeutics based on their mechanisms and therapeutic effects,which include expressing proteins for vaccination and protein replacement therapies,degrading deleterious RNA,modulating transcription and translation efficiency,targeting noncoding RNAs,binding and modulating protein activity and editing RNA sequences and modifications.This review emphasizes the concept of an RNA therapeutic toolbox,pinpointing the readers to all the tools available for their desired research and clinical goals.As the field advances,the catalog of RNA therapeutic tools continues to grow,further allowing researchers to combine appropriate RNA technologies with suitable chemical modifications and delivery platforms to develop therapeutics tailored to their specific clinical challenges.