Lipid-based GalNac-RNA Delivery Service
Overview
As the landscape of genomic medicine shifts toward extra-hepatic and cell-specific delivery, the liver remains the primary frontier for systemic RNA therapeutics. Traditional lipid nanoparticles (LNPs) rely heavily on the endogenous adsorption of Apolipoprotein E (ApoE) for uptake via the low-density lipoprotein receptor (LDLR). However, this pathway is limited in patients with homozygous familial hypercholesterolemia (HoFH) or other LDLR-deficient conditions. CD BioGlyco provides a specialized lipid-based GalNac-RNA delivery service that bypasses these limitations by utilizing N-acetylgalactosamine (GalNAc) ligands. By targeting the asialoglycoprotein receptor (ASGPR), our technology ensures robust, high-efficiency delivery of mRNA, siRNA, and base editors even in the absence of functional LDL receptors.
Core Technologies
Our platform integrates advanced molecular engineering with proprietary lipid chemistry to maximize therapeutic index and targeting precision:
- Trivalent GalNAc Ligand Engineering
We utilize a lysine-based trivalent ligand architecture that offers superior binding affinity to ASGPR compared to traditional TRIS-based designs.
- GL6 Lipid Anchor Systems
Our proprietary GL6 lipid anchor, featuring a 1,2-di-O-octadecyl-sn-glyceryl moiety and optimized PEG spacers, ensures stable ligand orientation on the LNP surface.
- ASGPR-Mediated Endocytosis
By decoupling delivery from the LDLR pathway, our particles achieve rapid internalization into hepatocytes through the highly expressed asialoglycoprotein receptors.
- Optimized Surface Density Control
Precision titration of GalNAc-lipid concentrations (down to 0.05 mol %) ensures maximal rescue of liver editing while maintaining particle stability and low immunogenicity.
Lipid-based GalNAc-RNA Delivery: Precision Liver Targeting and Gene Editing
Our service portfolio is designed to meet the diverse needs of biopharmaceutical researchers:
- Custom GalNAc-LNP synthesis: Tailored formulations for specific RNA modalities, including CRISPR/Cas9 and adenine base editors.
- Ligand optimization studies: Evaluation of different PEG spacer lengths and lipid anchors to enhance tissue-specific uptake.
- Targeting efficiency assays: Quantitative analysis of liver accumulation and hepatocyte-specific internalization.
- Model-specific delivery: Specialized services for delivering RNA to LDLR knockout (KO) non-human primate and rodent models.
Workflow
We employ a rigorous, multi-stage process to transform your RNA cargo into a clinic-ready delivery vehicle:
Cargo Assessment & Design
We begin by analyzing your specific RNA sequence, whether mRNA, gRNA, or siRNA, to evaluate its structural and chemical properties. Based on this assessment, we determine the optimal LNP size and lipid composition required to ensure maximum stability, efficient encapsulation, and functional integrity of the cargo.
Ligand Customization
Depending on the target species, such as rodent, NHP, or human cell lines, we select the most suitable GalNAc ligand structure and optimize its spacer length. This tailored approach maximizes binding affinity to the ASGPR, enabling precise and effective cellular targeting.
Automated Microfluidic Formulation
Using advanced, scalable microfluidic mixing technology, we incorporate the GalNAc-conjugated lipid into the LNP shell. This automated process ensures uniform distribution of the targeting moieties, resulting in consistent particle morphology and reproducible ligand presentation.
Physicochemical Characterization
Each production batch undergoes quality control testing. This includes measurement of Z-average size, polydispersity index (PDI), encapsulation efficiency, and pKa verification. Our rigorous characterization ensures that every formulation meets strict standards for stability, uniformity, and performance.
In Vitro / In Vivo Validation
We provide screening services using relevant models, such as LDLR-deficient systems, to evaluate the biological activity of the formulation. This stage generates quantitative data on target gene knockdown or base editing efficiency, with performance benchmarked against published data to confirm efficacy and translatability.
Scalable Process Development
Once a lead formulation is identified, we seamlessly transition the optimized process to larger batch production. This scale-up phase is designed to support advanced pre-clinical studies, including toxicology assessments, while maintaining the quality and consistency achieved at smaller scales.
Publication Data
Journal: Nature communications
DOI: 10.1038/s41467-023-37465-1
IF: 15.7
Published: 2023
Results: This research article presents the development and optimization of a novel GalNAc-conjugated LNP platform designed to enable efficient, LDLR-independent delivery of a CRISPR base editing therapy to the liver. The authors employed a structure-guided rational design approach to create and screen various GalNAc-lipid conjugates, identifying an optimal configuration (GL6) that facilitates hepatic uptake via the ASGPR. This strategy is particularly vital for treating patients with homozygous familial hypercholesterolemia (HoFH), who have deficient LDLR activity. The optimized GalNAc-LNPs, loaded with an adenine base editor (ABE8.8) mRNA and a guide RNA targeting the ANGPTL3 gene, were evaluated in both LDLR-deficient and wild-type mouse and non-human primate (NHP) models. The results demonstrated that the GalNAc-LNPs rescued high-efficiency liver editing (~61%) and produced durable reductions in ANGPTL3 protein (~89%) in LDLR-deficient NHPs, where standard LNPs failed. The study concludes that this GalNAc-LNP platform represents a potent and promising delivery system for liver-directed CRISPR therapies, overcoming the limitation of LDLR dependency.
Applications
Homozygous Familial Hypercholesterolemia (HoFH)
Our GalNAc-LNP service is essential for treating research for the lack of functional LDL receptors, providing a reliable pathway for liver-directed gene editing and cholesterol management.
Atherosclerotic Cardiovascular Disease (ASCVD)
We facilitate the delivery of RNA therapies targeting ANGPTL3 or PCSK9, helping to reduce systemic lipid levels and prevent the progression of cardiovascular plaques in high-risk populations.
Hypertriglyceridemia Research
By delivering base editors or siRNAs that knock down triglyceride-regulating proteins, our platform supports the development of next-generation therapies for severe metabolic disorders and pancreatitis prevention.
Alpha-1 Antitrypsin Deficiency (AATD)
Our targeted delivery systems can be used to deliver gene-correcting mRNA or siRNAs to hepatocytes, addressing both the lung and liver manifestations of this genetic condition.
Advantages
Scalable Manufacturing Excellence
We employ a pre-mixing strategy for GalNAc-lipids, ensuring a uniform distribution across the LNP surface. This process is fully scalable, maintaining particle integrity from benchtop to large-scale production.
Versatile Cargo Accommodation
Whether you are delivering small siRNAs or large mRNA transcripts encoding base editors, our lipid-based systems provide robust encapsulation and protection from nucleases in the systemic circulation.
High-Fidelity Hepatocyte Targeting
ASGPR is expressed almost exclusively on hepatocytes. Our service ensures that your RNA therapy is sequestered by the liver, reducing accumulation in the spleen or kidneys compared to non-targeted LNPs.
Rapid Internalization Kinetics
The high affinity of our trivalent GalNAc ligands for ASGPR leads to rapid receptor-mediated endocytosis, ensuring the RNA payload is delivered to the cytoplasm before particle degradation occurs.
Frequently Asked Questions
Customer Review
The GalNAc-LNP platform completely changed our approach to HoFH. We saw a 90% reduction in the target protein where standard LNPs previously failed. Their expertise in ligand density optimization was important.
— By Dr. A.L., Senior Scientist, Therapeutic Development
The potency of the GL6-anchored GalNAc ligands is remarkable. We were able to reduce our total RNA dose by 5-fold while maintaining therapeutic levels of base editing in the liver.
— By Dr. R.S., Principal Investigator, Gene Therapy
CD BioGlyco provided not just the LNPs, but deep scientific insight. Their ability to deliver large Cas9 mRNA payloads to the liver with high specificity is industry-leading.
— By Director, Pre-clinical Research
Associated Services
RNA-Cell Binding Interaction Analysis Services
Quantitative assessment of the affinity between your targeted RNA-LNP and cell-surface receptors.
Off-Target Activity Assessment Service
Comprehensive screening for unintended genomic modifications or non-specific tissue accumulation.
Conjugate Potency Assay Service
Specialized in vitro assays to determine the biological activity and potency of your GalNAc-RNA conjugates.
Absorption, distribution, metabolism, and excretion studies to support pharmacological characterization.
CD BioGlyco is committed to advancing the future of liver-targeted RNA medicine. Whether you are targeting rare metabolic disorders or common cardiovascular diseases, our lipid-based GalNac-RNA delivery service provides the precision and potency your research demands. Contact us for more information and to discuss your project.
Reference
- Kasiewicz, L.N.; et al. GalNAc-lipid nanoparticles enable non-LDLR dependent hepatic delivery of a CRISPR base editing therapy. Nature Communications. 2023, 14(1): 2776. (Open Access)