Library Of Nanoparticles Research Articles

Metal Ion-Condensed DNA Nanoparticle Library: Phase Separation and Transition and Antisense Therapy Applications.

DNA condensation has long been investigated as a fundamental cellular activity and is known to be driven by the mediation of diverse condensing agents. The phase behaviors of DNA during condensation are particularly interesting because the complicated molecular structure of natural nucleotides fundamentally allows electrostatic, coordinate covalent, and various other secondary interactions with the condensing agents. Recently, metal ion (Mn+)-induced DNA condensation has emerged as a powerful approach to synthesizing nanoparticulate DNA structures suitable for therapeutic gene delivery. However, how the DNA phase changes during Mn+-induced DNA condensation has rarely been observed and is not understood yet. In this study, a library of Mn+-condensed DNA nanoparticles (Mn+-CDNPs) was established using 30 different types of Mn+s, and their phase behaviors during condensation were elucidated using spherical nucleic acids (SNAs) as electron microscopic labels. Importantly, the phase transition and separation of DNA were demonstrated to be driven by the Mn+s into either the growth of individual DNA particles or the fission of bulky DNA aggregates. Pt2+ and Eu3+ were chosen as model systems for the demonstration. The hard and soft acid nature of Mn+ is presumably the underlying driving force of these phase transitions. In addition, the Mn+-controlled anticancer therapeutic efficiency of the Mn+-CDNP library as a state-of-the-art gene delivery platform was demonstrated even for unmodified antisense oligonucleotides in association with the potential toxicity of the Mn+s released from the Mn+-CDNPs. This comprehensive study of the Mn+-dependent condensation of nucleic acids provides profound insights into the chemistry of the nucleic acid-Mn+ interactions and the reliable theragnostic applications of Mn+-CDNPs as functional nucleic acid nanostructures.

Hierarchical Self-Assembly of Water-Soluble Fullerene Derivatives into Supramolecular Hydrogels.

Controlling the self-assembly of nanoparticle building blocks into macroscale soft matter structures is an open question and of fundamental importance to fields as diverse as nanomedicine and next-generation energy storage. Within the vast library of nanoparticles, the fullerenes-a family of quasi-spherical carbon allotropes-are not explored beyond the most common, C60. Herein, a facile one-pot method is demonstrated for functionalizing fullerenes of different sizes (C60, C70, C84, and C90-92), yielding derivatives that self-assemble in aqueous solution into supramolecular hydrogels with distinct hierarchical structures. It is shown that the mechanical properties of these resultant structures vary drastically depending on the starting material. This work opens new avenues in the search for control of macroscale soft matter structures through tuning of nanoscale building blocks.

Integrating synthetic hydrogel nanoparticles with carbon dots for selective detection of hemoglobin

Synthetic hydrogel nanoparticles (NPs) display high affinity to biomacromolecules target and have significant potential as protein capture agents for medical and biotechnological applications. In this study, we proposed a strategy to modify Carbon dots (CDs) with engineered NPs to address their detection specificity in complex biological sensing matrices. A library of NPs tagged CDs (NPs@CDs) incorporating hydrophobic and carboxylate groups that bound with high affinity to hemoglobin (Hb) was prepared by precipitation polymerization and screened based on the fluorescence quenching effect. NPs and CDs were integrated through noncovalent interaction, which was confirmed by Transmission electron microscopy (TEM), Scanning electron microscopy (SEM), Dynamic Light Scattering (DLS) and Fourier transform infrared spectroscopy analyses. The specificity of NPs@CDs was prominently attributed to the unique affinity between the screened NPs and Hb, compared with the contrast fluorescence quenching study by corresponding CDs to structural analogues and coexistence substances of Hb. Moreover, the Hb-induced fluorescence quenching process was dominated by a synergistic effect of internal filter effect (IFE) and static quenching proved by UV–vis absorption spectrometry and time-resolved fluorescence spectrometry. The screened NPs@CDs showed strong recognition capability to Hb, and the fluorescence quenching rate was about 97 % which was 3.2 times that of CDs. A biosensing platform based on NPs@CDs was constructed, and the fluorescence signal of the nanoprobe decreased linearly as the concentration of Hb over the range of 0.2–80 μM L−1 with a detection limit of 7.5 nM L−1. The synergy between CDs and NPs resulted in high-speed real-time detection capability, high sensitivity, considerable specificity, low interference and good stability of this nanoprobe.

Differential Sensing Approach as a Pattern-based Discrimination for Biological Samples.

The differential sensing approach uses fingerprint patterning to distinguish uncharacterized biological samples. Inspired by natural sensory systems, an array of cross-reactive sensors generates unique response fingerprint depending on the samples. Until today, this array system has been developed using various materials, including the library of surface-charged nanoparticles and chemosensors. Many differential array systems have demonstrated accurate identification of bacterial species, viral subtypes, and cancer cells, as well as distinguishing disease states in blood or urine. This capability is particularly important for distinguishing between normal and abnormal states when specific marker molecules have not yet been identified, providing a powerful diagnostic tool. In this concept, we summarized representative outcomes of differential sensing applications for biological sample discrimination.

Disentangling Competitive and Synergistic Chemical Reactivities During the Seeded Growth of High-Entropy Alloys on High-Entropy Metal Sulfide Nanoparticles.

The seeded growth of one type of nanoparticle on the surface of another is foundational to synthesizing many multifunctional nanostructures. High-entropy nanoparticles that randomly incorporate five or more elements offer enhanced properties due to synergistic interactions. Incorporating high-entropy nanoparticles into seeded growth platforms is essential for merging their unique properties with the functional enhancements that arise from particle-particle interactions. However, the complex compositions of high-entropy materials complicate the seeded growth process due to competing particle growth and chemical reactivity pathways. Here, we design and synthesize a 36-member nanoparticle library to identify and disentangle these competitive interactions, ultimately defining chemical characteristics that underpin the seeded growth of high-entropy alloys on high-entropy metal sulfide nanoparticles. As a model system, we focus on (Cu,Zn,Co,In,Ga)S-SnPdPtRhIr, which combines a high-entropy metal sulfide semiconductor with a high-entropy alloy catalyst. We study the seeded growth of all possible pairwise combinations of Sn, Pd, Pt, Rh, Ir, and SnPdPtRhIr on the metal sulfides Cu1.8S, ZnS, Co9S8, CuInS2, CuGaS2, and (Cu,Zn,Co,In,Ga)S, which have comparable morphologies and sizes. Through these studies, we uncover unexpected chemical reactivities, including cation exchange, redox reactions, and diffusion. Reaction temperature, threshold reduction potentials, metal/sulfide chemical reactivity, and the relative strengths of the various bonds that could be formed during particle growth emerge as the primary factors that underpin seeded growth. Finally, we disentangle these competitive and synergistic chemical reactivities to generate a reactivity map that provides practical guidelines for achieving seeded growth in compositionally complex systems.

Synthetic Roadmap to a Large Library of Colloidal High-Entropy Rare Earth Oxyhalide Nanoparticles Containing up to Thirteen Metals.

Nanoparticles of high-entropy materials that incorporate five or more elements randomized on a crystalline lattice often exhibit synergistic properties that can be influenced by both the identity and number of elements combined. These considerations are especially important for structurally and compositionally complex materials such as multimetal multianion compounds, where cation and anion mixing can influence properties in competitive and contradictory ways. Here, we demonstrate the synthesis of a large library of colloidal high-entropy rare earth oxyhalide (REOX) nanoparticles. We begin with the synthesis of (LaCePrNdSmEuGdDyHoErYbScY)OCl, which homogeneously incorporates 13 distinct rare earth elements. Through time point studies, we find that (LaNdSmGdDy)OCl, a 5-metal analogue, forms through in situ generation of compositionally segregated core@shell@shell intermediates that convert to homogeneously mixed products through apparent core-shell interdiffusion. Assuming that all possible combinations of 5 through 13 rare earth metals are synthetically accessible, we propose the existence of a 7099-member REOCl nanoparticle library, of which we synthesize and characterize 40 distinct members. We experimentally validate the incorporation of a large number of rare earth elements using energy dispersive X-ray spectra, despite closely spaced and overlapping X-ray energy lines, using several fingerprint matching strategies to uniquely correlate experimental and simulated spectra. We confirm homogeneous mixing by analyzing elemental distributions in high-entropy nanoparticles versus physical mixtures of their constituent compounds. Finally, we characterize the band gaps of the 5- and 13-metal REOCl nanoparticles and find a significantly narrowed band gap, relative to the constituent REOCl phases, in (LaCePrNdSmEuGdDyHoErYbScY)OCl but not in (LaNdSmGdDy)OCl.

Laser Precise Synthesis of Oxidation-Free High-Entropy Alloy Nanoparticle Libraries.

High-entropy alloy nanoparticles (HEA-NPs) show exceptional properties and great potential as a new generation of functional materials, yet a universal and facile synthetic strategy in air toward nonoxidized and precisely controlled composition remains a huge challenge. Here we provide a laser scribing method to prepare single-phase solid solution HEA-NPs libraries in air with tunable composition at the atomic level, taking advantage of the laser-induced metastable thermodynamics and substrate-assisted confinement effect. The three-dimensional porous graphene substrate functions as a microreactor during the fast heating/cooling process, which is conductive to the generation of the pure alloy phase by effectively blocking the binding of oxygen and metals, but is also beneficial for realizing accurate composition control via microstructure confinement-endowed favorable vapor pressure. Furthermore, by combining an active learning approach based on an adaptive design strategy, we discover an optimal composition of quinary HEA-NP catalysts with an ultralow overpotential for Li-CO2 batteries. This method provides a simple, fast, and universal in-air route toward the controllable synthesis of HEA-NPs, potentially integrated with machine learning to accelerate the research on HEAs.

DIPG-29. PHENOTYPIC SCREENS IDENTIFY GENETIC REGULATORS OF NANOPARTICLE DELIVERY IN DIFFUSE MIDLINE GLIOMAS

Abstract BACKGROUND Diffuse midline gliomas (DMGs) are a universally fatal pediatric brain tumor with a median survival of 12 months. There is a dire need to develop novel treatment strategies for these patients. Nanomedicine harbors great potential to encapsulate cargoes across tissue barriers and selectively target cells of interest. To effectively utilize existing nanomedicines and develop design criteria for novel formulations, an improved understanding of nano-bio interactions within the context of disease and development is required. METHODS We harnessed a lentiviral all-in-one CRISPR/Cas9 knock-out library paired with fluorescence-activated cell sorting to probe the functional impact of 700 genes on the delivery of lipid-based nanoparticles to DMGs. Layer-by-layer electrostatic assembly was utilized to synthesize a library of liposomal nanoparticles with polymer surface coatings. RESULTS Two patient-derived neurosphere models of DMG were transduced with the pooled, barcoded all-in-one CRISPR/Cas9 library. Following selection, cells were incubated with nanoparticles for 1, 4, or 24 hours and dynamically sorted into populations based on nanoparticle affinity. Genomic DNA was isolated from sorted cells and subjected to next generation sequencing for barcode identification. The relative enrichment of each single guide RNA in each population was determined relative to matched controls. We found strong agreement between biologic replicates and both cell lines, and our phenotypic screens revealed strong genetic regulators of nanoparticle uptake. We identified formulation-dependent and independent regulators, as well as regulators that were time-dependent. The top target genes are currently being validated in DMG and human neural stem cell models. CONCLUSIONS In ongoing work, we are leveraging genetic and chemical inhibition to sensitize DMG cells to nanoparticle therapeutics. We plan to leverage our screening technique to develop novel combination therapy approaches that could be implemented across multiple administration routes to enhance the use of nanomedicine for DMG and other pediatric brain tumors.

Improvement of Hydrogen Peroxide and Glucose Detection in Serum using Persistent Luminescent Nanoparticles: Impact of Synthesis Parameters and Particle Size

AbstractPersistent Luminescent Nanoparticles (PLNPs) emit prolonged light after excitation, allowing getting images with high signal‐to‐noise ratio, making them particularly beneficial for in vitro biodetection. In this study, we report the preparation of a library of gallium and zinc‐based Persistent Luminescent Nanoparticles doped with chromium (ZnGa2O4:Cr3+) through two distinct syntheses conducted at 120 °C for 12 hours or 24 hours, followed by a calcination at 500 °C or 750 °C. Using centrifugation, and starting from polydisperse samples, we demonstrated the ability to achieve different sizes in a monodisperse manner. Furthermore, our research revealed that ZnGa2O4:Cr3+ (ZGO) nanoparticles react differently with hydrogen peroxide (H2O2) based on their size, synthesis duration, and calcination temperature. We observed the most significant amplification of persistent luminescence for ZGO particles prepared at 120 °C for 12 hours and calcinated at 500 °C, with a size of approximately 100 nm, in the presence of 100 mM H2O2 (later named ZGO‐2). This study enabled the detection of H2O2 in serum without autofluorescence using the developed PLNPs, with a detection limit of up to 2.1 μM and a recovery rate of around 99 %. Additionally, we designed a glucose test for based on the responsiveness of ZGO PLNPs to H2O2. Using glucose oxidase, the glucose detection limit in serum was established around 0.13 μM, with a detection range between 0–5 μM. These findings could open new avenues to further control the size, synthesis, calcination temperature, and persistent luminescence of PLNPs, thus enhancing their utility in the development of new in vitro biosensors.

A universal and ultrafast method for fabricating a library of nanocellulose-supported metal nanoparticles

Using nanocatalysts to catalyze water electrolysis for hydrogen production is an ideal solution to address the energy crisis. The most well-adopted fabrication methods for nanocatalysts are tube furnace annealing, Hydrothermal method, etc., hardly satisfying the trade-off among coarsening, dispersity, and particle size due to mutual restrictions. Herein, a universal, ultrafast and facile cellulose nanometer whiskers-high temperature shock (CNW-HTS) method was reported for fabricating a library of ultrafine metal nanoparticles with uniform dispersion and narrow size distribution. The metal-anchor functional groups in CNW (i.e., –OH and –COOH) and the characteristics of the HTS method for ultrafast heating and powerful quenching synergistically contribute to the successful synthesis of metal nanoparticles. As an initial demonstration, the as-prepared Pt nanocatalyst (η10 ​mA ​cm−2 ​= ​51.8 ​mV) shows more excellent catalytic hydrogen evolution reaction (HER) performance than the Pt catalyst prepared in the tubular furnace (η10 ​mA ​cm−2 ​= ​169.4 ​mV). This rapid and universal CNW-HTS method can pave the way for nanomanufacturing to produce high-quality metal nanoparticles, thereby expanding applications of energy conversion and electrocatalysis.

Abstract 492: Investigating solute carrier SLC46A3 to enhance nanotherapeutic delivery for cancer

Abstract BACKGROUND: Nanoparticle therapeutics have multifaceted potential to improve efficacy and decrease toxicity of cancer treatments. Nanotherapeutics can encapsulate a range of cargos and have tunable surface chemistry that can enhance drug delivery to tumors. Cellular features also play a role in successful drug delivery, and predictive biomarkers may improve patient selection for clinical trials. In previous work, we identified SLC46A3, a poorly characterized lysosomal solute carrier, as a strong negative regulator of lipid-based nanoparticle uptake across many cancer models. We hypothesize that the expression of SLC46A3 will act as a predictive biomarker for therapeutic nanoparticles. METHODS: To test this hypothesis, we used two engineered model systems to probe the relationship between SLC46A3 expression and therapeutic response: lentiviral overexpression in a natively low-expressing melanoma model and genetic deactivation in a natively high-expressing breast cancer model. A library of FDA-approved and experimental therapeutic nanoparticles were screened in both models under a range of conditions. Liposomal formulations of chemotherapeutics were included with high and low surface functionalization of polyethylene glycol to investigate the impact of this ubiquitous ‘stealth’ polymer on our biomarker of interest. Solid lipid nanoparticles with nucleic acid cargo were also included. Orthogonal methods were used to probe these interactions and the therapeutic efficacy of nanotherapeutics, including pulsed dosing viability assays and label-free live-cell imaging. RESULTS: We found that SLC46A3 expression is a class-specific biomarker of lipid-based nanoparticle uptake and is predictive of distinctive patterns of response to therapy-loaded nanoparticles in both formulation and time-dependent manners. Additionally, we present a new technology for long-term, live cell imaging of nanoparticle uptake and trafficking that can discriminate mechanisms of cytotoxicity, accelerating the study of nano-bio interactions. These findings highlight a promising approach for the functional validation of nanoparticle-specific biomarkers using clinically relevant formulations that have the potential for rapid repurposing. Citation Format: Julianna Kenny-Serrano, Elizabeth B. Nelson, Gil Covarrubias, Namita Nabar, Joelle Straehla, Aanchal Gupta, Natalie Boehnke, Paula Hammond. Investigating solute carrier SLC46A3 to enhance nanotherapeutic delivery for cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 492.

Glycocalyx-Mimicking Nanoparticles with Differential Organ Selectivity for Drug Delivery and Therapy.

Organ-selective drug delivery is expected to maximize the efficacy of various therapeutic modalities while minimizing their systemic toxicity. Lipid nanoparticles and polymersomes can direct the organ-selective delivery of mRNAs or gene editing machineries, but their delivery is limited to mostly liver, spleen, and lung. A platform that enables delivery to these and other target organs is urgently needed. Here, a library of glycocalyx-mimicking nanoparticles (GlyNPs) comprising five randomly combined sugar moieties is generated, and direct in vivo library screening is used to identify GlyNPs with preferential biodistribution in liver, spleen, lung, kidneys, heart, and brain. Each organ-targeting GlyNP hit show cellular tropism within the organ. Liver, kidney, and spleen-targeting GlyNP hits equipped with therapeutics effectively can alleviate the symptoms of acetaminophen-induced liver injury, cisplatin-induced kidney injury, and immune thrombocytopenia in mice, respectively. Furthermore, the differential organ targeting of GlyNP hits is influenced not by the protein corona but by the sugar moieties displayed on their surface. It is envisioned that the GlyNP-based platform may enable the organ- and cell-targeted delivery of therapeutic cargoes.

Varying the hydrophobic core composition of polymeric nanoparticles affects NLRP3 inflammasome activation.

Understanding the interactions of nanoparticle carriers with innate immune cells is crucial for informing the design and efficacy of future nano-immunotherapies. An intriguing aspect of their interaction with the immune system has recently emerged, i.e., their ability to activate the NLRP3 inflammasome, a key component of the innate immune response. While the effect of the surface properties of nanoparticles has been extensively investigated in the context of nanoparticle-immune cell interactions, the influence of core composition remains largely unexplored, particularly regarding its impact on inflammasome activation. To shed light on these interactions, we developed a library of supramolecular polymer nanoparticles (SNPs) with different core compositions, varying their hydrophobic quotient by virtue of the side chain length and the repeating units in the polymer construct. The impact of modulating SNP core hydrophobic properties was investigated in macrophages by evaluating their cellular internalization, cytokine release, lysosomal rupture-calcium signaling, calcium flux-mitochondrial ROS production and their ability to activate the NLRP3 inflammasome, providing mechanistic insights into inflammasome activation. We established a direct correlation between increasing the side chain length of the polymer construct, thereby increasing the core hydrophobicity of SNPs and enhanced NLRP3 complex formation, as indicated by ASC speck imaging analysis and the elevated 1L-1β expression. Furthermore, the results demonstrated that the inflammasome signaling cascades and kinetics varied based on the SNP's hydrophobic side chain length and repeating units. Specifically, the nanoparticle with the longest alkyl side chain effectuated NLRP3 activation preferentially through the mitochondrial damage pathway. In vivo evaluation of SNPs in C57BL/6 mice confirmed elevated proinflammatory cytokines, notably with the SNP having the longest C12-alkyl side chain. This confirms that the higher core hydrophobicity composition of the SNP results in inflammasome activation in vivo. In summary, this study established SNP core composition as a novel nanoparticle-associated molecular pattern (NAMP) responsible for NLRP3 inflammasome activation, shedding light on intricate cellular pathways for informed nanoparticle design in immunotherapy and vaccine applications.

A highly tuneable inverse emulsion polymerization for the synthesis of stimuli-responsive nanoparticles for biomedical applications.

Polymeric nanomaterials have seen widespread use in biomedical applications as they are highly tuneable to achieve the desired stimuli-responsiveness, targeting, biocompatibility, and degradation needed for fields such as drug delivery and biosensing. However, adjustments to composition and the introduction of new monomers often necessitate reoptimization of the polymer synthesis to achieve the target parameters. In this study, we explored the use of inverse emulsion polymerization to prepare a library of polymeric nanoparticles with variations in pH and temperature response and examined the impact of overall batch volume and the volume of the aqueous phase on nanoparticle size and composition. We were able to prepare copolymeric nanoparticles using three different nonionic and three different anionic comonomers. Varying the non-ionizable comonomers, acrylamide (AAm), 2-hydroxyethyl methacrylate, and N-isopropylacrylamide (NIPAM), was found to alter the mass percentage of methacrylic acid (MAA) incorporated (from 26.7 ± 3.5 to 45.8 ± 1.8 mass%), the critical swelling pH (from 5.687 ± 0.194 to 6.637 ± 0.318), and the volume swelling ratio (from 1.389 ± 0.064 to 2.148 ± 0.037). Additionally, the use of NIPAM was found to allow for temperature-responsive behavior. Varying the ionizable comonomers, MAA, itaconic acid, and 2-acrylamido-2-methylpropane sulfonic acid (AMPSA), was found to significantly alter the critical swelling pH and, in the case of AMPSA, remove the pH-responsive behavior entirely. Finally, we found that for the base P(AAm-co-MAA) formulation, the pH-responsive swelling behavior was independent of the scale of the reaction; however, variations in the aqueous volume relative to the volume of the continuous phase significantly affected both the nanoparticle size and the critical swelling pH.

Semi-supervised learning of images with strong rotational disorder: assembling nanoparticle libraries

ss-rVAE classification can generalize from a small labeled data subset with weak orientational disorder to a larger unlabeled dataset with stronger disorder. We apply it to nanoparticle datasets to train a robust classifier and understand physical factors of data variation.

Magnetic nano-Fe3O4-based oleophilic tracer for stability studies of nano-tracer in oilfields condition

Nano-tracers have demonstrated exceptional performance in the petroleum engineering. However, the challenges of oleophilic nano-tracers persist in the complex separation and detection of these tracers from products, and the stability of nano-tracers in oilfields conditions is not currently explored in detail. In this work, magnetic nanoparticle libraries modified with stearic acid (SA) were harnessed to create a novel oleophilic oilfields tracer (M-Fe3O4 @SA). The quantities of tracer were expanded by doping various metal elements, which have the potential to match the number of fracturing stages. Additionally, the tracers were modified with stearic acid, provides ability to selectively tracing the oil. M-Fe3O4 @SA was examined by X-ray diffraction, Fourier-transform infrared spectroscopy, inductively coupled plasma mass spectrometry, thermogravimetric analysis coupled with differential scanning calorimetry, and vibrating sample magnetometry. The results revealed that the M-Fe3O4 @SA can be approximated as stearic acid-modified Fe3O4 nanoparticles, and its properties are not affected by the doped elements. As an oleophilic tracer, the wettability of the tracer was evaluated according to the static water contact angle. The contact angle reaches its maximum value (151.94°) when modified with 21% stearic acid. The thermal stability evaluation demonstrated that the tracer maintains a high retention rate (93%−73%) at 30 ℃− 90 ℃. The stable radius of the tracer dispersed in dodecane is about 2000 nm, and in ethanol is 1000 nm, which consistent with the theoretical calculation of the extended DLVO theory.

Lipid Nanoparticle Library Towards Development of Next Generation Genomic Medicines

Ionizable amino lipids are a major constituent of the lipid nanoparticles for delivering nucleic acid therapeutics (e.g., DLin-MC3-DMA in ONPATTRO®, ALC-0315 in Comirnaty®, SM-102 in Spikevax®). Scarcity of lipids that are suitable for cell therapy, vaccination, and gene therapies continue to be a problem in advancing many potential diagnostic/therapeutic/vaccine candidates to the clinic. Herein, we describe the development of novel ionizable lipids to be used as functional excipients for designing vehicles for nucleic acid therapeutics/vaccines in vivo or ex vivo use in cell therapy applications. We first studied the transfection efficiency (TE) of LNP-based mRNA formulations of these ionizable lipid candidates in primary human T cells and established a workflow for engineering of primary immune T cells. We then adapted this workflow towards bioengineering of CAR constructs to T cells towards non-viral CAR T therapy. Lipids were also tested in rodents for vaccine applications using self-amplifying RNA (saRNA) encoding various antigens. We have then evaluated various ionizable lipid candidates and their biodistribution along with the mRNA/DNA translation exploration using various LNP compositions. Further, using ionizable lipids from the library, we have shown gene editing of various targets in rodents. We believe that these studies will pave the path to the advancement in nucleic acid based therapeutics and vaccines, or cell & gene therapy agents for early diagnosis and detection of cancer, and for targeted genomic medicines towards cancer treatment and diagnosis.

Surface Geometry of Cargo-less Gold Nanoparticles Is a Driving Force for Selective Targeting of Activated Neutrophils to Reduce Thrombosis in Antiphospholipid Syndrome.

Antiphospholipid syndrome (APS) is an autoimmune disease characterized by recurrent arterial, venous, and microvascular thrombosis where activated neutrophils play a determinant role. However, neutrophils are challenging to target given their short lifespan in circulation and spontaneous activation. Screening of a small library of gold nanoparticles (AuNPs) led to the discovery of a formulation capable of targeting activated neutrophil attachment and has demonstrated that star-shaped, anti-PSGL-1-antibody-coated AuNPs (aPSGL-1-AuNPs) were more efficacious compared with other shapes of AuNPs. Our findings further revealed an exciting and safe targeting mode toward activated neutrophils in the APS mouse model induced by human-patient-derived antiphospholipid IgGs. Our studies demonstrate that targeting is dependent on the specific topographical features of the highly segregated PSGL-1 on the activated neutrophil surface for which a high affinity shape-driven nanomedicine can be designed and implemented. As such, star-shaped aPSGL-1-AuNPs serve as a promising nanoimmunotherapy for immunothrombosis associated with neutrophil adhesion in APS.

Single-Cell Diagnosis of Cancer Drug Resistance through the Differential Endocytosis of Nanoparticles between Drug-Resistant and Drug-Sensitive Cancer Cells.

Single-cell diagnosis of cancer drug resistance is highly relevant for cancer treatment, as it can be used to identify the subpopulations of drug-resistant cancer cells, reveal the sensitivity of cancer cells to treatment, and monitor the progress of cancer drug resistance. However, simple and effective methods for cancer drug resistance detection at the single-cell level are still lacking in laboratory and clinical studies. Inspired by the fact that nanoparticles with diverse physicochemical properties would generate distinct and specific interactions with drug-resistant and drug-sensitive cancer cells, which have distinctive molecular signatures, here, we have synthesized a library of fluorescent nanoparticles with various sizes, surface charges, and compositions (SiO2 nanoparticles (SNPs), organic PS-co-PAA nanoparticles (ONPs), and ZIF-8 nanoparticles (ZNPs)), thus demonstrating that the composition has a critical influence on the interaction of nanoparticles with drug-resistant cancer cells. Furthermore, the clathrin/caveolae-independent endocytosis of ZNPs together with the P-glycoprotein-related decreased cell membrane fluidity resulted in a lower cellular accumulation of ZNPs in drug-resistant cancer cells, consequently causing the distinct cellular accumulation of ZNPs between the drug-resistant and drug-sensitive cancer cells. This difference was further quantified by detecting the fluorescence signals generated by the accumulation of nanoparticles at the single-cell level via flow cytometry. Our findings provide another insight into the nanoparticle-cell interactions and offer a promising platform for the diagnosis of cancer drug resistance of various cancer cells and clinical cancer samples at the single-cell level.

Designed nanoparticles elicit cross-reactive antibody responses to conserved influenza virus hemagglutinin stem epitopes.

Despite the availability of seasonal vaccines and antiviral medications, influenza virus continues to be a major health concern and pandemic threat due to the continually changing antigenic regions of the major surface glycoprotein, hemagglutinin (HA). One emerging strategy for the development of more efficacious seasonal and universal influenza vaccines is structure-guided design of nanoparticles that display conserved regions of HA, such as the stem. Using the H1 HA subtype to establish proof of concept, we found that tandem copies of an alpha-helical fragment from the conserved stem region (helix-A) can be displayed on the protruding spikes structures of a capsid scaffold. The stem region of HA on these designed chimeric nanoparticles is immunogenic and the nanoparticles are biochemically robust in that heat exposure did not destroy the particles and immunogenicity was retained. Furthermore, mice vaccinated with H1-nanoparticles were protected from lethal challenge with H1N1 influenza virus. By using a nanoparticle library approach with this helix-A nanoparticle design, we show that this vaccine nanoparticle construct design could be applicable to different influenza HA subtypes. Importantly, antibodies elicited by H1, H5, and H7 nanoparticles demonstrated homosubtypic and heterosubtypic cross-reactivity binding to different HA subtypes. Also, helix-A nanoparticle immunizations were used to isolate mouse monoclonal antibodies that demonstrated heterosubtypic cross-reactivity and provided protection to mice from viral challenge via passive-transfer. This tandem helix-A nanoparticle construct represents a novel design to display several hundred copies of non-trimeric conserved HA stem epitopes on vaccine nanoparticles. This design concept provides a new approach to universal influenza vaccine development strategies and opens opportunities for the development of nanoparticles with broad coverage over many antigenically diverse influenza HA subtypes.