<i>In vitro</i>and<i>ex vivo</i>nano-enabled immunomodulation by the protein corona

Giulimondi, Francesca; Digiacomo, Luca; Vulpis, Elisabetta; Loconte, Luisa; Ferri, Gianmarco; Cardarelli, Francesco; Pozzi, Daniela; Zingoni, Alessandra; Caracciolo, Giulio

doi:10.1039/d2nr01878k

Cited by 7 publications

(4 citation statements)

References 57 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…To develop optimized gene delivery systems, current investigations have focused on exploiting protein coatings on lipid-based gene delivery systems (GDSs) as a promising strategy. − The design of precoated GDSs capitalizes on the targeting ability of plasma proteins obtained directly from the biological source. This protein coating acts as a functionalization acquired by LNPs upon exposure to the biological milieu .…”

Section: Resultsmentioning

confidence: 99%

Optimizing Transfection Efficiency in CAR-T Cell Manufacturing through Multiple Administrations of Lipid-Based Nanoparticles

Giulimondi,

Digiacomo,

Renzi

et al. 2024

ACS Appl. Bio Mater.

Self Cite

View full text Add to dashboard Cite

The existing manufacturing protocols for CAR-T cell therapies pose notable challenges, particularly in attaining a transient transfection that endures for a significant duration. To address this gap, this study aims to formulate a transfection protocol utilizing multiple lipid-based nanoparticles (LNPs) administrations to enhance transfection efficiency (TE) to clinically relevant levels. By systematically fine-tuning and optimizing our transfection protocol through a series of iterative refinements, we have accomplished a remarkable one-order-of-magnitude augmentation in TE within the immortalized T-lymphocyte Jurkat cell line. This enhancement has been consistently observed over 2 weeks, and importantly, it has been achieved without any detrimental impact on cell viability. In the subsequent phase of our study, we aimed to optimize the gene delivery system by evaluating three lipid-based formulations tailored for DNA encapsulation using our refined protocol. These formulations encompassed two LNPs constructed from ionizable lipids and featuring systematic variations in lipid composition (iLNPs) and a cationic lipoplex (cLNP). Our findings showcased a notable standout among the three formulations, with cLNP emerging as a frontrunner for further refinement and integration into the production pipeline of CAR-T therapies. Consequently, cLNP was scrutinized for its potential to deliver CAR-encoding plasmid DNA to the HEK-293 cell line. Confocal microscopy experiments demonstrated its efficiency, revealing substantial internalization compared to iLNPs. By employing a recently developed confocal image analysis method, we substantiated that cellular entry of cLNP predominantly occurs through macropinocytosis. This mechanism leads to heightened intracellular endosomal escape and mitigates lysosomal accumulation. The successful expression of anti-CD19-CD28-CD3z, a CAR engineered to target CD19, a protein often expressed on the surface of B cells, was confirmed using a fluorescence-based assay. Overall, our results indicated the effectiveness of cLNP in gene delivery and suggested the potential of multiple administration transfection as a practical approach for refining T-cell engineering protocols in CAR therapies. Future investigations may focus on refining outcomes by adjusting transfection parameters like nucleic acid concentration, lipid-to-DNA ratio, and incubation time to achieve improved TE and increased gene expression levels.

show abstract

Section: Resultsmentioning

confidence: 99%

Optimizing Transfection Efficiency in CAR-T Cell Manufacturing through Multiple Administrations of Lipid-Based Nanoparticles

Giulimondi,

Digiacomo,

Renzi

et al. 2024

ACS Appl. Bio Mater.

Self Cite

View full text Add to dashboard Cite

show abstract

“…This protein corona significantly affects not only the NPs’ fate (pharmacokinetics, biodistribution, and target recognition), but also their effectiveness as therapeutic biocarriers and/or immunomodulators [ 154 , 155 , 156 ]. Interestingly, recent works have now opened the possibility to engineer the “protein corona” to modulate NP interaction with the immune system [ 157 , 158 ]. If this strategy could be used to improve NPs’ potential as immunomodulators deserves further investigation.…”

Section: Nanomedicine-based Strategies For Immunomodulationmentioning

confidence: 99%

Nano-Immunomodulation: A New Strategy for Skeletal Muscle Diseases and Aging?

Millozzi

Papait

Bouché

et al. 2023

IJMS

View full text Add to dashboard Cite

The skeletal muscle has a very remarkable ability to regenerate upon injury under physiological conditions; however, this regenerative capacity is strongly diminished in physio-pathological conditions, such as those present in diseased or aged muscles. Many muscular dystrophies (MDs) are characterized by aberrant inflammation due to the deregulation of both the lymphoid and myeloid cell populations and the production of pro-inflammatory cytokines. Pathological inflammation is also observed in old muscles due to a systemic change in the immune system, known as “inflammaging”. Immunomodulation represents, therefore, a promising therapeutic opportunity for different skeletal muscle conditions. However, the use of immunomodulatory drugs in the clinics presents several caveats, including their low stability in vivo, the need for high doses to obtain therapeutically relevant effects, and the presence of strong side effects. Within this context, the emerging field of nanomedicine provides the powerful tools needed to control the immune response. Nano-scale materials are currently being explored as biocarriers to release immunomodulatory agents in the damaged tissues, allowing therapeutic doses with limited off-target effects. In addition, the intrinsic immunomodulatory properties of some nanomaterials offer further opportunities for intervention that still need to be systematically explored. Here we exhaustively review the state-of-the-art regarding the use of nano-sized materials to modulate the aberrant immune response that characterizes some physio-pathological muscle conditions, such as MDs or sarcopenia (the age-dependent loss of muscle mass). Based on our learnings from cancer and immune tolerance induction, we also discuss further opportunities, challenges, and limitations of the emerging field of nano-immunomodulation.

show abstract

“…75 The protein corona regulates the interaction with immune system cells in vitro and ex vivo. 76 These findings prompted us to create biomimetic lipoplexes with an artificial opsonin-deficient protein corona that minimizes the uptake by immune cells in vivo. 16 We have also demonstrated that the protein corona of lipoplexes locks onto receptors of cancer cells, promoting internalization by a receptor-mediated mechanism.…”

Section: ■ Future Research Directionsmentioning

confidence: 99%

“…In previous works, we have shown that the protein corona acquired by lipid vesicles has a composition that is simultaneously regulated by the electrical charge and by the affinity of lipids for individual proteins or specific classes of plasma proteins . The protein corona regulates the interaction with immune system cells in vitro and ex vivo …”

Section: Future Research Directionsmentioning

confidence: 99%

Looking Back, Moving Forward: Lipid Nanoparticles as a Promising Frontier in Gene Delivery

Pozzi,

Caracciolo

2023

ACS Pharmacol. Transl. Sci.

Self Cite

View full text Add to dashboard Cite

Lipid nanoparticles (LNPs) have shown remarkable success in delivering genetic materials like COVID-19 LNP vaccines, such as mRNA-1273/SpikeVax by Moderna and BNT162b2/Comirnaty by BioNTech/Pfizer, as well as siRNA for rare inherited diseases, such as Onpattro from Alnylam Pharmaceuticals. These LNPs are advantageous since they minimize side effects, target specific cells, and regulate payload delivery. There has been a surge of interest in these particles due to their success stories; however, we still do not know much about how they work. This perspective will recapitulate the evolution of lipid-based gene delivery, starting with Felgner’s pioneering 1987 PNAS paper, which introduced the initial DNA-transfection method utilizing a synthetic cationic lipid. Our journey takes us to the early 2020s, a time when advancements in bionano interactions enabled us to create biomimetic lipoplexes characterized by a remarkable ability to evade capture by immune cells in vivo. Through this overview, we propose leveraging previous achievements to assist us in formulating improved research goals when optimizing LNPs for medical conditions such as infectious diseases, cancer, and heritable disorders.

show abstract

In vitroandex vivonano-enabled immunomodulation by the protein corona

Abstract: New technologies with the capacity to tune immune system activity are highly desired in clinical practice and disease management. Here we demonstrate that nanoparticles with a protein corona enriched by...

Cited by 7 publications

References 57 publications

Optimizing Transfection Efficiency in CAR-T Cell Manufacturing through Multiple Administrations of Lipid-Based Nanoparticles

Optimizing Transfection Efficiency in CAR-T Cell Manufacturing through Multiple Administrations of Lipid-Based Nanoparticles

Nano-Immunomodulation: A New Strategy for Skeletal Muscle Diseases and Aging?

Looking Back, Moving Forward: Lipid Nanoparticles as a Promising Frontier in Gene Delivery

Contact Info

Product

Resources

About